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Monoamine oxidase inhibitors in Amazonian hallucinogenic plants : ethnobotanical, phytochemical, and… McKenna, Dennis Jon

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MONOAMINE OXIDASE INHIBITORS IN AMAZONIAN HALLUCINOGENIC PLANTS: ETHNOBOTANICAL, PHYTOCHEMICAL, AND PHARMACOLOGICAL INVESTIGATIONS By DENNIS JON MCKENNA B. A . , U n i v e r s i t y of C o l o r a d o , 1973 M . S c , U n i v e r s i t y of Hawa i i , 1979 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Department of Botany) We accept t h i s t h e s i s as conforming to the r e q u i r e d s tandard THE UNIVERSITY OF BRITISH COLUMBIA February 1984 © Dennis Jon McKenna, 1984 In p r e s e n t i n g t h i s t h e s i s i n p a r t i a l f u l f i l m e n t o f the requ i rements f o r an advanced degree a t the U n i v e r s i t y o f B r i t i s h Co lumb ia , I agree t h a t the L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r r e f e r e n c e and s tudy . I f u r t h e r agree t h a t p e r m i s s i o n f o r e x t e n s i v e copy ing o f t h i s t h e s i s f o r s c h o l a r l y purposes may be g ran ted by the head o f my department o r by h i s o r her r e p r e s e n t a t i v e s . I t i s unders tood tha t copy ing o r p u b l i c a t i o n o f t h i s t h e s i s f o r f i n a n c i a l ga i n s h a l l not be a l l owed w i thou t my w r i t t e n p e r m i s s i o n . Department o f Botany The U n i v e r s i t y o f B r i t i s h Columbia 1956 Main Mall Vancouver , Canada V6T 1Y3 26 Ap r i l , 1984 Date )E-6 (3/81) i i ABSTRACT E t h n o b o t a n i c a l , p h y t o c h e m i c a l , and pha rmaco log i c a l i n v e s t i g a t i o n s of two Amazonian h a l l u c i n o g e n s are p r e s e n t e d . The extant l i t e r a t u r e on the botany , c h e m i s t r y , and ethnopharmacology of the Ma lp i gh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s i s rev iewed. The h a l l u c i n o g e n i c beverage ayahuasca i s p repared from the woody l i a n a B a n i s t e r i o p s i s c aap i (Ma lp igh iaceae ) and v a r i o u s admixture p l a n t s which s t rengthen or modify the e f f e c t . The genus V i r o l a ( M y r i s t i c a c e a e ) i s the source of the o ther h a l l u c i n o g e n s i n v e s t i g a t e d ; the cambia l r e s i n of c e r t a i n V i r o l a spp . i s made i n t o h a l l u c i n o g e n i c s n u f f s by some Amazonian t r i b e s , wh i l e o the r s p repare an o r a l l y -i nges t ed drug from the r e s i n . A l though d e r i v e d from e n t i r e l y d i f f e r e n t b o t a n i c a l s o u r c e s , both ayahuasca and the V i r o l a drugs owe t h e i r h a l l u c i n o g e n i c a c t i v i t y to i ndo l e a l k a l o i d s , v i z . , t r yp tamines and /3-carbo l ines . The major t r yp tamines found in these p r e p a r a t i o n s are N ,N-d imethy l t r yp tamine and/or 5-methoxy-N,N-d imethy l t r yp tamine ; both are potent h a l l u c i n o g e n s but are i n a c t i v e when i nges ted o r a l l y , presumably due to o x i d a t i v e deaminat ion by v i s c e r a l monoamine ox idase (MAO). The /3-carboline a l k a l o i d s , a l t hough hav ing l i m i t e d a c t i v i t y as h a l l u c i n o g e n s , are potent r e v e r s i b l e i n h i b i t o r s of MAO; thus they may p r o t e c t the t r yp tamines from v i s c e r a l MAO and render them o r a l l y a c t i v e . T h i s mechanism may under l y the o r a l a c t i v i t y of ayahuasca and the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s d r u g s . Ayahuasca i s an i n t e g r a l pa r t of mes t i zo f o l k med ic ine among the lower s o c i o -economic c l a s s e s l i v i n g in semi-urban Amazonian c e n t e r s , but the u t i l i z a t i o n of V i r o l a spp . as the source of h a l l u c i n o g e n i c p r e p a r a t i o n s i s c o n f i n e d to a few ind igenous Amazonian t r i b e s , and even among some of these groups i s a r a p i d l y d i s a p p e a r i n g p r a c t i c e . The pharmacology and b i o c h e m i s t r y of t r yp tamine and 0 -c a r b o l i n e d e r i v a t i v e s i s reviewed (Chapter I I ) . T h e i r b i o s y n t h e s i s , d i s t r i b u t i o n , s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s , i n t e r a c t i o n s wi th MAO i n h i b i t o r s , endogenous s y n t h e s i s and deg rada t i v e metabol ism in mammals, h a l l u c i n o g e n i c p r o p e r t i e s , and other b i o l o g i c a l a c t i v i t i e s a re rev iewed. R e s u l t s of e thnograph i c and e t h n o b o t a n i c a l f i e l dwork in the Amazon Bas in are p resen ted (Chapter I I I ) . Herbar ium voucher c o l l e c t i o n s are documented (Appendix I I ) , and the methods used i n the c o l l e c t i o n of drug samples and p l an t m a t e r i a l s f o r chemica l a n a l y s i s are d e s c r i b e d . E thnograph ic o b s e r v a t i o n s on the use of ayahuasca and the o r a l l y - i n g e s t e d V i r o l a pas tes are p r e s e n t e d , and the methods used in t h e i r p r e p a r a t i o n are documented.. The f o l k - m e d i c a l use of ayahuasca by an ayahuasquero l i v i n g near P u c a l l p a , Pe ru , i s d e s c r i b e d . The procedures f o l l owed by Bora and Wi to to in fo rmants in the p r e p a r a t i o n of the o r a l l y - i n g e s t e d V i r o l a pas tes are a l s o d e s c r i b e d . Obse r va t i ons on the b i o l o g i c a l a c t i v i t y of ayahuasca and the V i r o l a pas t e s in s e l f - e x p e r i m e n t s are i n c l u d e d . The contemporary use of ayahuasca i n mes t i zo f o l k medic ine i s compared to the use of the o r a l M y r i s t i c a c e o u s d rugs , which has remained e t h n o l o g i c a l l y r e s t r i c t e d to the Bora and Wi to to t r i b e s . The p o s s i b i l i t y i s r a i s e d tha t the somewhat u n r e l i a b l e pha rmaco log i c a l a c t i v i t y of the M y r i s t i c a c e o u s d rugs , a r e f l e c t i o n of t h e i r chemica l v a r i a b i l i t y , may have c o n t r i b u t e d to a d e c l i n e in the use of the p a s t e s . The a l k a l o i d s of a number of ayahuasca brews, c u l t i v a r s of B. c a a p i , and admixture p l a n t s were q u a l i t a t i v e l y and q u a l i t a t i v e l y ana l yzed us ing t h i n - l a y e r chromatography (TLC) , h i gh-p res su re l i q u i d chromatography (HPLC) and gas chromatography/mass spect rometroy (GC/MS) The ayahuasca samples con t a i ned i n s u f f i c i e n t l e v e l s of 0- c a r b o l i n e s to account fo r t h e i r h a l l u c i n o g e n i c p r o p e r t i e s at the doses t y p i c a l l y used ; however i n most samples the c o n c e n t r a t i o n of DMT was we l l above the t h r e s h o l d l e v e l , assuming tha t i t i s o r a l l y a c t i v a t e d by the b lockade of v i s c e r a l MAO. D i f f e r e n t batches of ayahuasca had s i m i l a r a l k a l o i d c o m p o s i t i o n s , however the c o n c e n t r a t i o n s of t o t a l a l k a l o i d s and the p r o p o r t i o n s of i n d i v i d u a l c o n s t i t u e n t s v a r i e d c o n s i d e r a b l y in batches of ayahuasca p repared by d i f f e r e n t ayahuasqueros in v a r i o u s p a r t s of Pe ru . D i f f e r e n t batches prepared by the same p r a c t i t i o n e r were remarkably c o n s i s t e n t both in t o t a l a l k a l o i d c o n c e n t r a t i o n and in the p r o p o r t i o n s of c o n s t i t u e n t s . C o n s i d e r a b l e v a r i a t i o n in a l k a l o i d c o n c e n t r a t i o n in s e v e r a l r e cogn i zed c u l t i v a r s of B. caap i were found but may be due to env i ronmenta l f a c t o r s r a the r than g e n e t i c d i f f e r e n c e s . S u b s t a n t i a l c o n c e n t r a t i o n s of DMT were found in a l l Psychot r i a v i r i d i s samples a n a l y z e d , and in one c o l l e c t i o n of D i p l o p t e r y s cabre rana but DMT was not de t e c t ed in P s y c h o t r i a c a r t h a g e n e n s i s . DMT was the s i n g l e major base de t e c t ed in these admix tu res ; on l y t r a c e s of o ther a l k a l o i d s were d e t e c t e d . Seve ra l uncommon admixture p l a n t s were screened f o r a l k a l o i d s , but on ly one, Abuta g r a n d i f o l i a , (Menispermaceae) gave an unambiguously p o s i t i v e r e a c t i o n . V A l k a l o i d s in twenty-e ight M y r i s t i c a c e o u s bark and l e a f samples were q u a l i t a t i v e l y and q u a n t i t a t i v e l y determined us ing TLC , GC, p r e c i p i t a t i o n t e s t s , and GC/MS. S i x t e e n of the twenty-e i g h t samples con t a i ned d e t e c t a b l e a l k a l o i d s . DMT and 5-MeO-DMT were the major bases , w i th much sma l l e r amounts of NMT and/or t r yp tamine a l s o p resen t i n most samples . D e t e c t a b l e l e v e l s of 0 -c a r b o l i n e s were not found i n the bark and l e a f samples . Four teen of the e igh teen V i r o l a samples con t a i ned a l k a l o i d s ; none of the s i x I r yan the ra s p e c i e s c o n t a i n e d d e t e c t a b l e a l k a l o i d s . An i n d o l i c base , i d e n t i f i e d as N-methyl-tryptophan methyl e s t e r , was found in Osteophloem platyspermum. Seven samples of o r a l l y -inges ted drugs made from V i r o l a spp . were a n a l y z e d . A l l but one con t a i ned s u b s t a n t i a l amounts of t r yp t am ines , but the types and p r o p o r t i o n s v a r i e d g r e a t l y between samples . Samples of V i r o l a snu f f i n c l u d i n g v a r i o u s admixtures were ana l yzed and a l l but one con t a i ned t r y p t a m i n e s . Drug samples wi th the h i ghes t c o n c e n t r a t i o n s of a l k a l o i d s con t a i ned 15-20 mg/g d wt; the bark and l e a f samples had c o n c e n t r a t i o n s rang ing from 0.04 to 0.25 mg/g d wt. Only two V i r o l a pas te samples con t a i ned d e t e c t a b l e l e v e l s of 0 - c a r b o l i n e s , which were i d e n t i f i e d as MTH/3C and DMTH/3C. The 0 - c a r b o l i n e s were t r a ce c o n s t i t u e n t s . Methods were d e v i s e d f o r the i_n v i t r o assay of r a t - l i v e r monoamine ox idase (MAO) us i ng 1 * -C-se ro ton in as s u b s t r a t e . S t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of v a r i o u s t r yp tamine and 0 -c a r b o l i n e d e r i v a t i v e s as MAOI were de te rm ined . The MAOI a c t i v i t y of /3-carbol ine d e r i v a t i v e s was s e v e r a l o rde r s of magnitude g r ea t e r than the a c t i v i t y measured wi th t r yp tamine d e r i v a t i v e s . DMT was the most a c t i v e MAOI of the t r yp tamines t e s t e d wh i le v i harmine was the most a c t i v e of the 0 - c a r b o l i n e s . Mult i-component m ix tu res of 0 - c a r b o l i n e s were not s i g n i f i c a n t l y more e f f e c t i v e than the s i n g l e most a c t i v e component, i n d i c a t i n g an a d d i t i v e r a t he r than a s y n e r g i s t i c mechanism of a c t i o n . Samples of ayahuasca were h i g h l y a c t i v e as MAOI even when d i l u t e d by s e v e r a l o rde r s of magni tude. The a c t i v i t y was comparable to m ix tu res of p'-carbol ines hav ing s i m i l a r c o n c e n t r a t i o n s and p r o p o r t i o n s . Samples of o r a l l y - i n g e s t e d V i r o l a pas tes were l e s s e f f e c t i v e than ayahuasca as MAOI. The i n h i b i t i o n which they e l i c i t e d was c l o s e l y matched by m ix tu res of t r yp tamine s tandards hav ing comparable p r o p o r t i o n s and c o n c e n t r a t i o n s . An a l k a l o i d -f r ee pas te sample and a crude l i g n a n f r a c t i o n from V. e longa ta e l i c i t e d on l y a s l i g h t degree of n o n - s p e c i f i c i n h i b i t i o n at the h i ghes t c o n c e n t r a t i o n s . These o b s e r v a t i o n s i n d i c a t e tha t the l i m i t e d MAOI a c t i v i t y of the pas t e s i s due p r i m a r i l y to the t r y p t a m i n e s ; the t r a c e s of 0 - c a r b o l i n e s or non-n i t rogenous i n h i b i t o r s p resent p robab l y do not c o n t r i b u t e s i g n i f i c a n t l y to the t o t a l i n h i b i t i o n . These r e s u l t s suggest tha t the i n h i b i t i o n of p e r i p h e r a l MAO by 0 - c a r b o l i n e s may e x p l a i n the o r a l a c t i v i t y of ayahuasca , but i t i s u n l i k e l y that t h i s mechanism can account fo r the o r a l a c t i v i t y of the M y r i s t i c a c e o u s p a s t e s . Some a l t e r n a t i v e mechanism must t h e r e f o r e be c o n s i d e r e d . One p o s s i b l e a l t e r n a t i v e i s tha t V i r o l a spp . may c o n t a i n n o n - a l k a l o i d c o n s t i t u e n t s which are a c t i v e as i n h i b i t o r s of hepa t i c microsomal m ixed- func t i on ox idases (MFOs). Expe r imen ta l ev idence i s rev iewed which i n d i c a t e s tha t these enzymes, r a the r than h e p a t i c MAO, may a c t u a l l y be more important in the p e r i p h e r a l metabol i sm and i n a c t i v a t i o n of o r a l l y - a d m i n i s t e r e d DMT and r e l a t e d compounds. v i i i TABLE OF CONTENTS A b s t r a c t i i Tab l e of Contents v i i i L i s t of Tab l es x i v L i s t of F i g u r e s xv i Acknowledgements x v i i i A b b r e v i a t i o n s Used in Text x x i i i D e d i c a t i o n xxv Par t I: I n t r o d u c t i o n and L i t e r a t u r e Review 1 Chapter I: The Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s of South America 2 I. I n t r o d u c t i o n . . . . . 2 I I . Botany and Chemis t ry of the Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s 5 A. Botany and Chemist ry of Ma lp igh i a ceous H a l l u c i n o g e n s 5 B. Botany and Chemist ry of M y r i s t i c a c e o u s H a l l u c i n o g e n s 7 I I I . Ethnopharmacology of Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s 8 A. Source P l an t s and Methods of P r epa r a t i on 9 B. P o s t u l a t e d Mechanism of A c t i o n 11 C. Admixture P l an t s 14 IV. E thnograph i c Aspec ts of Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s 16 V. Scope and O b j e c t i v e s of the Present I n v e s t i g a t i o n 20 A. F i e ldwork 21 B. L abo ra to r y I n v e s t i g a t i o n s 22 ix V I . L i t e r a t u r e C i t e d 25 Chapter I I : Chemistry and Pharmacology of Tryptamines and | 3-carbo l ines 29 I. Occu r r ence , D i s t r i b u t i o n , and B i o s y n t h e s i s of Tryptamines and /3-carbol ines 29 I I . Pharmacology of H a l l u c i n o g e n i c Tryptamines 33 A. S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s 33 B. Metabo l i sm of H a l l u c i n o g e n i c Tryptamines 36 C. H a l l u c i n o g e n i c Tryptamines and MAO I n h i b i t i o n 39 I I I . B i o chemis t r y and Pharmacology of /3-carboline d e r i v a t i v e s 43 A. Psychopharmacology of /3-carboline d e r i v a t i v e s 44 1. /3-carbol ines as Monoamine Oxidase I n h i b i t o r s .44 2. /3-carbol ines as H a l l u c i n o g e n s 46 3. Endogenous s y n t h e s i s of /3-carbol ines 47 B. Other N e u r o l o g i c a l and B i o l o g i c a l A c t i v i t i e s of /?-carbol ines 48 IV. L i t e r a t u r e C i t e d 51 Part I I : E t h n o b i o l o g i c a l I n v e s t i g a t i o n s of Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s 58 Chapter I I I : E t h n o b o t a n i c a l C o l l e c t i o n s and E thnograph i c O b s e r v a t i o n s 59 I . E t h n o b o t a n i c a l C o l l e c t i o n s 59 A. C o l l e c t i o n and I d e n t i f i c a t i o n of Herbar ium Voucher Specimens 59 B. C o l l e c t i o n of P l an t M a t e r i a l and Drug Samples fo r Phytochemica l A n a l y s i s 60 I I . E thnograph i c Obse r va t i ons 60 A. E thnograph ic Aspec ts of Ayahuasca 61 B. E thnograph ic Aspec ts of O r a l l y - a c t i v e M y r i s t i c a c e o u s Pastes 69 I I I . L i t e r a t u r e C i t e d 83 Par t I I I : Phy tochemica l and Pha rmaco log i ca l I n v e s t i g a t i o n s • 84 Chapter IV: A l k a l o i d C o n s t i t u e n t s of Ayahuasca Brews, S o u r c e - p l a n t s , and Admixture P l a n t s 85 I. I n t r o d u c t i o n 85 I I . M a t e r i a l s and Methods 87 A. F i e l d C o l l e c t i o n s of Drug Samples and P l an t M a t e r i a l s 87 B. Two-dimensional T h i n - l a y e r Chromatography 88 C. H igh P ressure L i q u i d Chromatography 94 1. A n a l y t i c a l C o n d i t i o n s 94 2. Q u a n t i t a t i v e Methods 96 3. Sample P r e p a r a t i o n 97 D. GC/MS Ana lyses 99 E. A l k a l o i d Tes t s and TLC of Uncommon Admixture P l an t s 100 I I I . R e s u l t s and D i s c u s s i o n 101 A. Ayahuasca Brews 101 B. A l k a l o i d Content of B a n i s t e r i o p s i s caap i C u l t i v a r s 107 C. A l k a l o i d Content of Ayahuasca Admixture P l a n t s . . . . 1 1 0 1. DMT-conta in ing Admixtures 110 2. Uncommon Admixture P l a n t s 113 xi IV. Summary 116 V. L i t e r a t u r e C i t e d .* 118 Chapter V: A l k a l o i d C o n s t i t u e n t s of O r a l l y - a c t i v e M y r i s t i c a c e o u s H a l l u c i n o g e n s 121 I. I n t r o d u c t i o n 121 I I . M a t e r i a l s and Methods 122 A. D e t e c t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Samples . .122 B. Q u a n t i t a t i v e A n a l y s i s of Tryptamine S tandards 125 C. P r e p a r a t i o n of M y r i s t i c a c e o u s Samples fo r GC Q u a n t i t a t i o n 128 I I I . R e s u l t s and D i s c u s s i o n 130 A. D e t e c t i o n and I d e n t i f i c a t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Samples 130 1. Bark and l e a f samples 130 2. Snuff and pas te samples 140 B. Q u a n t i t a t i v e Ana l y ses of M y r i s t i c a c e o u s Samples . . . 1 4 8 IV. Summary 153 V. L i t e r a t u r e C i t e d 155 Chapter V I : O r a l l y - a c t i v e Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s as Monoamine Oxidase I n h i b i t o r s 158 I. I n t r o d u c t i o n 158 I I . M a t e r i a l s and Methods 161 A. P r e p a r a t i o n of R a t - l i v e r C y t o s o l 161 B. P r e p a r a t i o n of Reac t i on M ix tu re and A d d i t i o n of L a b e l l e d Subs t r a t e 162 x i i C . Assays Us ing Tryptamine and /3-carboline S tandards 163 D. P r epa r a t i on and Q u a n t i t a t i o n of Ayahuasca Samples f o r MAO Assay 165 E. P r epa r a t i on and Q u a n t i t a t i o n of M y r i s t i c a c e o u s Paste Samples fo r MAO Assay 166 I I I . R e s u l t s and D i s c u s s i o n 168 A. /3-carbolines and Tryptamines as MAOI : S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s 168 1. /3-carbol ines 168 2. Tryptamines 173 B. A c t i v i t y of Ayahuasca and Ayahuasca Ana logs as MAOI 177 C . A c t i v i t y of M y r i s t i c a c e o u s Pas tes and Paste Analogs as MAOI 184 IV. Summary 190 V. L i t e r a t u r e C i t e d 192 Chapter V I I : The Comparat ive Ethnopharmacology of Ma lp igh i a ceous and M y r i s t i c a c e o u s H a l l u c i n o g e n s : Summary and C o n c l u s i o n s 195 I. I n t r o d u c t i o n 195 I I . Ayahuasca 196 I I I . M y r i s t i c a c e o u s H a l l u c i n o g e n s 198 IV. C o n c l u s i o n 205 V. L i t e r a t u r e C i t e d 207 x i i i Appendix I: Biodynamic C o n s t i t u e n t s in Ayahuasca Admixture P l a n t s 208 L i t e r a t u r e C i t e d •. 214 Appendix I I : L i s t of Herbar ium Voucher C o l l e c t i o n s 223 x i v LIST OF TABLES Tab le I - O r a l l y and P a r e n t e r a l l y A c t i v e P s y c h o t r o p i c Tryptamine D e r i v a t i v e s 34 Tab le II - hRf Va lues of Tryptamine and /3-carboline Standards in So lvent System I & II 89 Tab le III - HPLC Q u a n t i t a t i o n of U n d i l u t e d Ayahuasca Samples 104 Tab le IV - HPLC Q u a n t i t a t i o n of L y o p h i l i z e d Ayahuasca Samples 105 Tab le V - HPLC Q u a n t i t a t i o n of B a n i s t e r i o p s i s caap i C u l t i v a r s 109 Tab le VI - DMT C o n t a i n i n g Admixture P l a n t s - A n a l y s i s by TLC, HPLC, and GC/MS 111 Tab le VII - D e t e c t i o n of A l k a l o i d s in Uncommon Admixture P l an t s .114 Tab le VII I - Tryptamine Standards - GC A n a l y t i c a l Data . . . . 1 2 6 Tab le IX - D e t e c t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Bark and Leaf Samples 132 Tab le X - D e t e c t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Paste and Snuff Samples 135 Tab l e XI - Q u a n t i t a t i v e A n a l y s i s of Tryptamines in M y r i s t i c a c e o u s Samples 149 Tab le XII - A l k a l o i d C o n c e n t r a t i o n i n M y r i s t i c a c e o u s Paste Samples Asayed f o r MAOI A c t i v i t y 167 Tab le XII I - S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s of 0 - c a r b o l i n e s as MAO I n h i b i t o r s 170 Tab le XIV - MAOI A c t i v i t y of Ayahuasca Samples and M ix tu r e s of /3-carbol ines 171 X V Tab le XV - MAOI A c t i v i t y of Tryptamine D e r i v a t i v e s and M y r i s t i c a c e o u s Pas tes 174 Tab le XVI - B i o l o g i c a l l y A c t i v e C o n s t i t u e n t s of Ayahuasca Admixture P l an t s 209 Tab le XVII - L i s t of Herbar ium Voucher C o l l e c t i o n s 225 xv i LIST OF FIGURES F i g u r e 1 - Some N a t u r a l l y - O c c u r r i n g Tryptamine D e r i v a t i v e s 30 F i g u r e 2 - Some N a t u r a l l y - O c c u r r i n g /3-carboline D e r i v a t i v e s 31 F i g u r e 3 - P o s i t i o n s of Tryptamine and /3-carboline S tandards in Two-dimensional T h i n - l a y e r Chromatography 91 F i g u r e 4 - One-dimens iona l TLC of Tryptamine Standards in So l ven t s I and II 92 F i g u r e 5 - HPLC E l u t i o n P r o f i l e of Ayahuasca Sample 95 F i g u r e 6 - TLC A n a l y s i s of Peruv ian Ayahuasca Samples 102 F i g u r e 7 - GC Q u a n t i t a t i o n of Tryptamine Standards 127 F i g u r e 8 - UV Spectrum of N-methyl-tryptophan Methy l E s t e r from Leaves of Osteophloem platyspermum 139 F i g u r e 9 - Mass Spectrum of N-methyl-tryptophan Methy l E s t e r from Leaves of Osteophloem platyspermum 139 F i gu re 10 - Mass Spectrum of MTH/3C from La Chor re ra Qo '-koey 142 F i g u r e 1 1 - Mass Spectrum of DMTH/3C from La Chor re ra Qo' -koey . . . 1 4 2 F i g u r e 12 - MAOI A c t i v i t y of Ayahuasca 179 F i g u r e 13 - MAOI A c t i v i t y of /3-carboline M ix tu res 181 F i g u r e 14 - MAOI A c t i v i t y of /3-carboline/DMT M ix tu res 182 F i g u r e 15 - MAO I n h i b i t i o n by M y r i s t i c a c e o u s Pas tes - La Chor re ra Qo '-koey 185 xv i i F i gu r e 16 - MAO I n h i b i t i o n by M y r i s t i c a c e o u s Pastes -A l f r e d o Moreno Sample #1 187 F i g u r e 17 - MAO I n h i b i t i o n by M y r i s t i c a c e o u s Pas tes -Marcos F l o r e s Sample #1 188 F i g u r e 18 - MAOI A c t i v i t y of N o n - a l k a l o i d C o n s t i t u e n t s of M y r i s t i c a c e o u s Pas tes 189 xv i i i ACKNOWLEDGEMENTS An i n t e r d i s c i p l i n a r y r e sea r ch p r o j e c t of the so r t r epor ted here owes i t s s u c c e s s f u l comp le t i on to the c o - o p e r a t i o n , encouragement, and a s s i s t a n c e of numerous i n d i v i d u a l s . The f i e l dwo rk formed an i n d i s p e n s a b l e p re lude to the l a b o r a t o r y i n v e s t i g a t i o n s ; i t was c a r r i e d out with the pe rm i s s i on of the Pe ruv ian government and but fo r t h e i r co-ope r a t i on c o u l d not have been completed at a l l . The f a c t that t h i s r e sea r ch has been accompl i shed i s testament to tha t s p i r i t of i n t e r n a t i o n a l g o o d w i l l that i s one of the most exemplary c h a r a c t e r i s t i c s of the s c i e n t i f i c endeavor in f r ee s o c i e t i e s . A c c o r d i n g l y , I am indebted to many p e o p l e , in Canada, the Un i t ed S t a t e s , and Peru , fo r h e l p i n g to make t h i s work p o s s i b l e . Only some can be mentioned by name but to a l l I g i v e my deepest thanks . In Lima we were a s s i s t e d by D r . Ramon F e r r e y r a , D i r e c t o r of the Museo H i s t o r i a Na tu r a l " J a v i e r P r a d o " . Use of the herbar ium f a c i l i t i e s of the Museo and Dr . F e r r e y r a ' s h e l p in o b t a i n i n g c o l l e c t i o n permi ts are g r e a t l y a p p r e c i a t e d . S r s . E l i a s Mucha, f o rmer l y of the F a c u l t y of F o r e s t r y at UBC, and Jose P u r i s a c a of the M i n i s t e r i o de A g r i c u l t u r a were a l s o very h e l p f u l i n g u i d i n g two i n e x p e r i e n c e d g r i ngos through the maze of Pe ruv ian bu reauc racy , and we thank them fo r t h e i r p a t i e n c e . In P u c a l l p a we were r e c e i v e d h o s p i t a b l y and he lped in numerous ways by the members of the I n s t i t u t o L i n g u i s t i c o de Verano , p a r t i c u l a r l y Ted Long and h i s f a m i l y , D ick R u t t e r , and Wes T h e i s o n . We were f i l l e d w i th good f o o d , good wine, and good s t o r i e s on s e v e r a l o c c a s i o n s by Conner and Mary N i x o n , owners of La B r i s a at Ya r ina Cocha . N i c o l e Maxwell became a va lued i n fo rman t , an en joyab l e t r a v e l i n g companion, and an e x c e l l e n t f r i e n d ; she was xix i n s t r u m e n t a l i n h e l p i n g us to accompl i sh many of our g o a l s , and no thanks can p o s s i b l y repay her adequa t e l y . On my second stay in P u c a l l p a , I en joyed the h o s p i t a l i t y of S r . F r a n c i s c o Montes Shuna and h i s b ro the r Pablo Shuna and I thank them fo r t h e i r k i n d n e s s . In I q u i t o s we were g r e a t l y a s s i s t e d by D r . F r a n k l i n Aya la F l o r e s , Dean of Q u a l i f i c a c i o n and d i r e c t o r of the He rba r i o Amazonense, U n i v e r s i d a d Nac i ona l Amazonia Peruana (UNAP). Dr . A y a l a ' s e x t e n s i v e knowledge of the Rio Ampiyacu area made i t p o s s i b l e to s u c c e s s f u l l y pursue our f i e l dwork t h e r e . At Dr . A y a l a ' s i n v i t a t i o n , we made f r ee use of the herbar ium f a c i l i t i e s at UNAP. The e x c e l l e n t taxonomic f i e l d a s s i s t a n c e of S r . Juan Ruiz and S r . Jose To r r e s c o n t r i b u t e d g r e a t l y to our c o l l e c t i n g e f f o r t s . We owe a l l of these men a s i n c e r e thanks ; t h e i r g e n e r o s i t y and e x p e r t i s e has c o n t r i b u t e d s i g n i f i c a n t l y to t h i s work. No l e s s a c o n t r i b u t i o n was made by S r a . Ad r i ana Loyaza , head of the I q u i t o s o f f i c e of the Amazon Na tu r a l Drug Company; she was i n s t rumen ta l in h e l p i n g us o b t a i n p h y t o s a n i t a r y pe rmi t s fo r the expor t of l i v e spec imens , and a l lowed us to ho ld our l i v e c o l l e c t i o n s in the garden of the ANDC u n t i l they cou ld be s a f e l y a i r l i f t e d to Canada. Ad r i ana a l s o p r o v i d e d our i n t r o d u c t i o n to A l f r e d o Moreno, one of our most v a l u a b l e in fo rmants at Puco U r q u i l l o . A d r i a n a ' s many f a v o r s , q u i e t s u p p o r t , and good humor were a l l i n v a l u a b l e and we are very g r a t e f u l f o r the time and e f f o r t she put in on our b e h a l f . Other peop le in I q u i t o s who he lped out in some way are D r . A l Gen t r y , a s s i s t a n t c u r a t o r of the M i s s o u r i B o t a n i c a l Gardens ; Mr. Wade D a v i s , of the Harvard B o t a n i c a l Museum, who was i n s t rumen ta l to the s u c c e s s f u l comp le t i on of our f i e l dwork in the R io Ampiyacu XX a r e a ; and the crew of the RV H e r a c l i t u s , who p r o v i d e d p a r t i a l l o g i s t i c a l support fo r tha t p o r t i o n of the e x p e d i t i o n ; the c o n t r i b u t i o n of a l l of these i n d i v i d u a l s or g roups , however g rea t or s m a l l , i s very much a p p r e c i a t e d . S eve ra l employees of the M i n i s t e r i o de A g r i c u l t u r a in I qu i t o s a l s o a s s i s t e d us wi th the export of l i v e p l a n t s and herbar ium vouche r s ; among them Ing . C a r l o s Sa l aza r Marchand, and Ing. E m i l i o V i l l a r C a s t r o . We are g r a t e f u l f o r t h e i r c o - o p e r a t i o n . Many people in North Amer ica a l s o a s s i s t e d , a d v i s e d , or he lped us in some way. Among them are Dr . Timothy Plowman of the Ch icago F i e l d Museum; not on ly has Dr . Plowman encouraged and suppor ted every aspect of t h i s p r o j e c t , from i t s i n i t i a l concep t i on to i t s c o m p l e t i o n , he has a l s o i d e n t i f i e d most of the herbar ium voucher c o l l e c t i o n s , a t ime-consuming and monumental task which i s f a r beyond the c a p a b i l i t i e s of t h i s r e s e a r c h e r . H i s e f f o r t s in t h i s r e spec t and every other deserve the warmest acknowledgement. Other peop le at the F i e l d Museum and e lsewhere c o n t r i b u t e d to the voucher d e t e r m i n a t i o n s , . i n c l u d i n g Penny M a t a k a i t i s , D r s . W. R. Anderson and Bronwen Gates of the U n i v e r s i t y of M i c h i g a n , A l Gentry at M i s s o u r i , and Dr . W. A. Rodr igues of the I n s t i t u t o Pesqu isas do Amazonense(INPA) in Manaus. We thank them a l l f o r t ak i ng time away from more p r e s s i n g and important mat te rs to devote to these d e t e r m i n a t i o n s . D r . Bo Holmstedt of the K a r o l i n s k a I n s t i t u t e , P r o f . R. E. S chu l t e s of the Harvard B o t a n i c a l Museum, and Dr . Laurent R i v i e r of the U n i v e r s i t e de Lausanne, a l l p rov ided support and encouragement f o r the p r o j e c t through co r respondence , u s e f u l s u g g e s t i o n s , and sage a d v i c e ; we s i n c e r e l y thank them fo r t h e i r in formed i n s i g h t s and fo r the xxi example they set f o r a s p i r i n g e thnopharmaco log i s t s everywhere. Comple t ion of v a r i o u s t e c h n i c a l phases of t h i s work depended on the co-ope ra t i on and goodw i l l of s e v e r a l p e o p l e ; among them Dr . C .-K. Wat and F e l i p e B a l z a , who p r o v i d e d e x c e l l e n t t e c h n i c a l a s s i s t a n c e in the GC/MS ana l y ses of v a r i ous samples ; D r . G. B e l lwa rd , who pe rm i t t ed the use of her r a d i o i s o t o p e l a b o r a t o r y f a c i l i t i e s fo r the development of the methods used in the MAO a s s a y s ; and Malcolm F i n l a y s o n , who gave u s e f u l adv i ce on the work-up of the r a t - l i v e r enzyme p r e p a r a t i o n . T h e i r c o n t r i b u t i o n s to these a spec t s of the work are g r a t e f u l l y acknowledged. I would a l s o l i k e to thank s e v e r a l o ther peop le or i n s t i t u t i o n s who have c o n t r i b u t e d to the success of t h i s p r o j e c t through some form of suppo r t , e i t h e r mora l , e m o t i o n a l , s p i r i t u a l , or f i n a n c i a l . H igh on t h i s l i s t i s Don MacRae, the o ther h a l f of the pronoun "we" used in these acknowlegements. He was a s t e a d f a s t t r a v e l i n g companion in Peru and always kept h i s head l e v e l and h i s sense of humor engaged whatever the p r e v a i l i n g p s y c h o l o g i c a l weather ; there were many t imes when i t was r e a s s u r i n g to have someone l i k e that a round . He i s a f i n e f r i e n d and an e x c e l l e n t c o l l e a g u e and I wish him luck in whatever he under takes in the f u t u r e , knowing he w i l l handle i t e a s i l y and w e l l . Other peop le I wish to thank f o r t h e i r many t a n g i b l e and i n t a n g i b l e c o n t r i b u t i o n s to t h i s p r o j e c t are S h e i l a Humphrey, L u i s Luna , Terence McKenna, and Joe McKenna; each of you knows how much you have g iven to make i t a l l worth i t . I would l i k e to express my a p p r e c i a t i o n to the F a c u l t y of Graduate S t ud i e s at UBC fo r f i n a n c i a l support in the form of UBC Graduate F e l l o w s h i p s ; and to the Department of Botany f o r support in the xx i i form of a Teach ing A s s i s t a n c e s h i p . I would a l s o l i k e to thank the members of my graduate r e sea r ch committee who have c o n t r i b u t e d c o n s i d e r a b l e time and e f f o r t to the review of t h i s t h e s i s . F i n a l l y , but c e r t a i n l y not l e a s t of a l l , I wish to thank my f r i e n d , who a l s o happens to be my s u p e r v i s o r , D r . N e i l Towers. H i s enthus iasm fo r t h i s p r o j e c t has been u n s t i n t i n g to the very end ; and h i s support has gone f a r beyond the mundane f i n a n c i a l v a r i e t y ( a l though he has f u r n i s h e d p l e n t y of t h a t , t o o ) . To each and a l l of these p e o p l e , and a l s o to anyone I have ove r l ooked i n a d v e r t e n t l y who deserved to be ment ioned , I g i ve my warmest and s i n c e r e s t thanks . ABBREVIATIONS USED IN TEXT TLC - t h i n - l a y e r chromatography GC - gas chromatography GC/MS - gas chromatography/mass spect rometry UV - u l t r a - v i o l e t HPLC - h igh p ressu re l i q u i d chromatography SAR - s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s MAO - monoamine ox idase MAOI - monoamine ox idase i n h i b i t o r MFO - mixed f u n c t i o n ox idase CNS - c e n t r a l nervous system MTHF - 5-me thy l t e t r ahyd ro fo l a t e SAM - S-adenosy l-meth ion ine INMT - indole-N-methy l t r a n s f e r a s e TA - t r yp tamine NMT - N-methyl t ryptamine DMT - N ,N-d imethy l t ryptamine DMT-NO - N,N-d imethy l t ryptamine-N-oxide 5-MeO-TA - 5-methoxy-tryptamine 5-MeO-DMT -5-methoxy-N,N-dimethyl t ryptamine 5HT - 5-hydroxy-tryptamine ( s e ro ton in ) 5HO-DMT - 5-hydroxy-N,N-dimethy l t ryptamine (bu fo ten ine ) IAA - i n d o l e a c e t i c a c i d DET - N ,N-d i e thy l t r yp t am ine DPT - N ,N-d ip ropy l t r yp tamine DIPT - N , N - d i i s o p r o p y l t r y p t a m i n e DAT - N , N - d i a l l y l t r y p t a m i n e DBT - N ,N-d ibu t y l t r yp t am ine THH - te t rahydroharmine xx iv 6-MeO-THH - 6~MeO-tetrahydroharman 6-MeO-MTH/3C - 6-methoxy-2-methyl-tetrahydro-p , -carbol ine 6-MeO-DMTH|3C - 6-methoxy- 1 , 2-dimethy l-tet rahydro-/3-carbol ine TH/JC - tetrahydro-/3-c 'arbol ine MTH/3C - 2-methyl-tetrahydro-/?-carbol ine DMTH/3C - 1 , 2 - d i m e t h y l - t e t r a h y d r o - 0 - c a r b o l i n e THH3C - te t rahydroharman-3-carboxy la te /3CC - /3-carbo l ine-3-carboxy la te /3CEE - /3-carbo l ine-3-carboxy la te e t h y l e s t e r T h i s t h e s i s i s r e s p e c t f u l l y d e d i c a t e d to the ind igenous f o l k h e a l e r s of the Amazon, whose knowledge of b o t a n i c a l medic ine c o n s t i t u t e s a l egacy of va lue fo r a l l of humanity . 1 PART I: INTRODUCTION AND LITERATURE REVIEW 2 CHAPTER I: THE MALPIGHIACEOUS AND MYRISTICACEOUS* HALLUCINOGENS OF SOUTH AMERICA I. I n t r o d u c t i o n Dur ing the l a s t th ree decades our s c i e n t i f i c knowledge of the botany and chemis t r y of p l a n t h a l l u c i n o g e n s has expanded enormous ly . T h i s has been accompl i shed through the co-ope ra t i v e e f f o r t s of e t h n o b o t a n i s t s , working in the f i e l d to i d e n t i f y and c o l l e c t the sou r ce-p l an t s u t i l i z e d by a b o r i g i n a l p e o p l e s , and phy tochemis ts working in the l a b o r a t o r y , who have in many cases succeeded in i s o l a t i n g and c h a r a c t e r i z i n g the biodynamic c o n s t i t u e n t s r e s p o n s i b l e fo r these p r o p e r t i e s [ 1 , 2 ] . The number of h ighe r p l a n t s p e c i e s i s e s t ima ted at between 400,000 and 800,000; of t h e s e , on ly an i n s i g n i f i c a n t number—somewhat fewer than 100—are known to ' be e x p l o i t e d as h a l l u c i n o g e n s throughout the w o r l d , and l e s s than twenty of these s p e c i e s may be d e s c r i b e d as major [2 ] . Most h a l l u c i n o g e n i c p l a n t s owe t h e i r p r o p e r t i e s to a l k a l o i d a l c o n s t i t u e n t s , and s i n c e at l e a s t 5000 s p e c i e s of h ighe r p l a n t s c o n t a i n a l k a l o i d s , there i s a s t rong l i k e l i h o o d tha t many more h a l l u c i n o g e n i c s p e c i e s remain und i s cove red [2 ] . S c i e n t i f i c i n t e r e s t in the b o t a n i c a l and chemica l sources * No te : Usage of the terms ' M a l p i g h i a c e o u s ' and ' M y r i s t i c a c e o u s ' in t h i s t h e s i s f o l l o w s tha t e s t a b l i s h e d by S c h u l t e s [ 7 , 8 ] ; used in r e f e r e n c e to p l an t specimens or p l a n t m a t e r i a l they r e f e r to members of the Ma lp igh i a ceae and M y r i s t i c a c e a e , r e s p e c t i v e l y ; used in r e f e r e n c e to drug samples or h a l l u c i n o g e n s , they r e f e r to drugs d e r i v e d p r i m a r i l y from sou r ce-p l an t s in the Ma lp igh i a ceae or M y r i s t i c a c e a e . 3 of h a l l u c i n o g e n s has p a r t l y f o l l owed upon,, and p a r t l y r e s u l t e d f rom, the i n c r e a s e d " r e c r e a t i o n a l " use of such compounds in our own s o c i e t y . Whi le the use of h a l l u c i n o g e n s , or " p s y c h e d e l i c s , " as they are sometimes c a l l e d , i s not as widespread a p r a c t i c e today as i t was ten or twelve years ago [3 ] , h a l l u c i n o g e n s n e v e r t h e l e s s con t inue to be used in contemporary s o c i e t y . T h i s has g iven r i s e to the m i sconcep t i on that they are something r e l a t i v e l y new. In f a c t , h a l l u c i n o g e n s have been known and va lued in human c u l t u r e s at l e a s t s i n ce the l a t e P a l e o l i t h i c , and p robab l y even e a r l i e r . A b o r i g i n a l p e o p l e s , f o r c ed to r e l y on nature fo r f o o d , m e d i c i n e s , and p a l l i a t i v e s of a l l s o r t s , have l i k e w i s e a c t i v e l y sought and used p s y choac t i v e p l a n t s fo r v a r i o u s m a g i c a l , r e l i g i o u s , m e d i c i n a l , or r e c r e a t i o n a l pu rposes . Perhaps nowhere in the wor ld has the knowledge and use of endemic h a l l u c i n o g e n i c p l a n t s deve loped to the extent tha t i t has in the upper Amazon Bas in of South Amer i ca . Of the hundred or so s p e c i e s of p l a n t s employed as h a l l u c i n o g e n s throughout the w o r l d , f u l l y 60% of them are n a t i v e to the New Wor ld ; t h i s f a c t i s s i g n i f i c a n t in i t s e l f and has been r epea ted l y remarked upon in the e t h n o b o t a n i c a l l i t e r a t u r e [ 4 , 5 ] . Even more c u r i o u s i s tha t of the ten or so New World h a l l u c i n o g e n s that have major e t h n o b o t a n i c a l s i g n i f i c a n c e , f u l l y two- th i rds have t h e i r geograph ic and e t h n o l o g i c a l c e n t e r s of d i s t r i b u t i o n in the Northwest Amazon—the no tab le excep t i ons be ing the peyote cac tus (Lophophora w i11 i ams i i ) of the American Southwest and the P s i l o c y b e mushrooms of Oaxaca, Mex i co . T h i s odd ly asymmet r i ca l d i s t r i b u t i o n of the New World h a l l u c i n o g e n s may be p a r t l y due to e c o l o g i c a l f a c t o r s and p a r t l y to e t h n o l o g i c a l ones . E c o l o g i c a l 4 f a c t o r s may c o n t r i b u t e in tha t few other p l a c e s on e a r t h can boast of the sheer s p e c i e s d i v e r s i t y which abounds in the Amazon B a s i n , hence i t i s not s u r p r i s i n g that a t y p i c a l c r o s s - s e c t i o n of r e p r e s e n t a t i v e s p e c i e s has a g r ea t e r p r o p o r t i o n of p l a n t s c o n t a i n i n g p s y c h o a c t i v e c o n s t i t u e n t s . E t h n o l o g i c f a c t o r s may p l a y a r o l e in that no o ther ind igenous peop les in the wor ld depend as t o t a l l y on the v e g e t a l environment as do those i n h a b i t i n g the Amazon B a s i n , nor does any o ther group e x p l o i t the v e g e t a l environment as tho rough l y as do the Amazonian t r i b e s . Given t h i s j u x t a p o s i t i o n of s p e c i e s d i v e r s i t y and abundance, and in tense e x p l o i t a t i o n of the v e g e t a l env i ronment , we would expect to f i n d tha t t h i s knowledge of the p l a n t kingdom would extend as we l l ( i f not e s p e c i a l l y ) to the l o c a l l y a v a i l a b l e p s y c h o a c t i v e p l a n t s . L a r g e l y because of i n t e r d i s c i p l i n a r y e t h n o b o t a n i c a l , e t h n o l o g i c a l , and phy tochemica l i n v e s t i g a t i o n s c a r r i e d out w i t h i n the l a s t two decades , the b o t a n i c a l sources and most of the a c t i v e c o n s t i t u e n t s of the major h a l l u c i n o g e n i c p l a n t s used by Northwest Amazon t r i b e s have now been i d e n t i f i e d [ 6 ] , W i th in the lowland a reas of' the upper Amazon Bas in p r o p e r , i . e . , w i t h i n the r eg ion bounded on the West by the Andes, on the nor th by the R io O r i n o c o , on the south by the R io U c a y a l i d r a i n a g e , and on the eas t by the c o n j u n c t i o n of the borders of P e ru , Co lomb ia , and B r a z i l , the ind igenous complex of h a l l u c i n o g e n i c p l a n t s i s t o t a l l y dominated by two gene ra : B a n i s t e r i o p s i s i n the Ma lp igh i a ceae and V i r o l a in the M y r i s t i c a c e a e . The former i s the b a s i s fo r the h a l l u c i n o g e n i c brew known v a r i o u s l y as ayahuasca , c a a p i , or yage, wh i le c e r t a i n V i r o l a spp . a re most we l l known 5 as the source of p o t e n t l y h a l l u c i n o g e n i c s n u f f s whose cen te r of u t i l i z a t i o n extends from the Puinave Ind ians of the Rio Apapor i s r eg ion of the Colombian Vaupes , no r th to the Waika Ind ian groups-of the Or inoco headwaters in southern Venezue la [ 7 ] , One might suppose that s i n ce the main source p l a n t s and chemica l c o n s t i t u e n t s of the pr imary Northwest Amazon h a l l u c i n o g e n s have been i d e n t i f i e d , there remains very l i t t l e to i n t e r e s t the b o t a n i s t or phy tochemis t . T h i s assumption i s unwarranted, however; a c a r e f u l study of the l i t e r a t u r e p e r t a i n i n g to these ind igenous drugs r e vea l s tha t many of the most i n t e r e s t i n g prob lems , from the po in t of view of the b i o c h e m i s t , p h a r m a c o l o g i s t , and t o x i c o l o g i s t , have been a l l but i g n o r e d . Some of the more i n t e r e s t i n g of these problems, are the sub jec t of the i n v e s t i g a t i o n s r e p o r t e d in t h i s t h e s i s . 11 . Botany and Chemist ry of Ma lp igh i a ceous and Myr i s t i caceous  H a l l u c inogens A. Botany and Chemis t ry of the Ma lp igh i a ceous Ha l l u c i nogens The woody l i a n a B a n i s t e r i o p s i s caap i (Spruce ex G r i s e b . ) M o r t o n (Ma lp igh iaceae ) forms the b a s i s of the h a l l u c i n o g e n i c beverage commonly known as ayahuasca (Quechua fo r " v i n e of the s o u l s " ) a l t hough in d i f f e r e n t r eg ions of the Amazon i t i s known by other v e r n a c u l a r names, i n c l u d i n g yage, c a a p i , natema, and p inde [8 ] . A l though B. c aap i i s u s u a l l y the spec i e s used , B. i n e b r i a n s Mor ton , B. q u i t e n s i s (Ndz.) Mor ton , and T e t r a p t e r y s me thys t i c a S c h u l t e s have a l l been r e p o r t e d as sources of the d r i nk [ 8 ] . On o c ca s i on ayahuasca i s p repared from 6 the b o i l e d bark or stems of one of these spec i e s without the a d d i t i o n of any other b o t a n i c a l i n g r e d i e n t s ; more commonly, however, the l eaves or bark of v a r i o u s admixture p l a n t s are added i n t o the brew in o rder to s t reng then or modify the e f f e c t [ 9 ] . The l eaves of one Ma lp igh i a ceous spec i e s have been repor ted [10] as an admix tu re , D i p l o p t e r y s cabrerana (Cuat recasas ) Ga tes . Former ly known as B a n i s t e r i o p s i s rusbyana , the name of t h i s s p e c i e s has been changed in a recent taxonomic r e v i s i o n [11, Plowman, T . , p e r s . Comm., 1980]. A l though sun.dry spec i e s from numerous f a m i l i e s are u t i l i z e d as admixtures to ayahuasca ( c f . Appendix I, a l s o [ 9 , 1 2 , 1 3 , 1 4 , 1 5 ] ) , the admixtures used most commonly are D i p l o p t e r y s cabre rana and P s y c h o t r i a v i r i d i s R.&P. and P s y c h o t r i a c a r t h a q e n e h s i s J a c q . (Rub iaceae ) . Admixtures in the Solanaceae are a l s o common, i n c l u d i n g tobacco ( N i c o t i a n a s p p . ) , Brugmansia s p p . , and B r u n f e l s i a spp. The most d e t a i l e d chemica l study to date of ayahuasca and i t s b o t a n i c a l i n g r e d i e n t s i s tha t of R i v i e r and L indgren [15 ] . Us ing GC/MS a n a l y s i s , these i n v e s t i g a t o r s found that the major a c t i v e c o n s t i t u e n t s of ayahuasca are the 0-ca rbo l i ne a l k a l o i d s harmine, ha rma l i ne , and t e t r ahyd roha rm ine , and N,N-d ime thy l t r yp t am ine ( c f . F i g 1 & 2, Chapter I I ) . The /3-carbol ines are c o n s t i t u e n t s of B. c aap i [16] wh i le DMT has been i s o l a t e d as a c o n s t i t u e n t of D. cabre rana [17] and has a l s o been r epo r t ed [15] in both P s y c h o t r i a c a r t h a g e n e n s i s and P. v i r i d i s . Hashimoto et a l . [18,19] r epo r t ed the i s o l a t i o n of 6 /3-carboline bases , harmic amide, a c e t y l norharmine , ke to te t r ahyd ronorha rm ine , harmine N-oxide, harmic a c i d methyl e s t e r , and h a r m a l i n i c a c i d ) from the l eaves of B. c aap i in a d d i t i o n to the three main 7 c o n s t i t u e n t s . These workers a l s o r epo r t ed the i s o l a t i o n of the p y r r o l i d i n e bases sh ihun ine and d i h y d r o s h i h u h i n e [20] from l eaves of B. c a a p i . These compounds are t r a c e c o n s t i t u e n t s in the p l a n t , however, and t h e i r ext remely low c o n c e n t r a t i o n s (0.007 - 0.0001 %) make i t u n l i k e l y tha t they c o n t r i b u t e s i g n i f i c a n t l y to the pha rmaco log i c a l a c t i v i t y of ayahuasca . B. Botany and Chemis t ry of the M y r i s t i c a c e o u s H a l l u c i n o g e n s The genus V i r o l a ( M y r i s t i c a c e a e ) i n c l u d e s some 45 to 60 s p e c i e s of t r o p i c a l t r e e s , n a t i v e to the f o r e s t s of C e n t r a l and South Amer i ca , and forming an e s p e c i a l l y abundant component of the Amazonian f l o r a . At p resen t about s i x V i r o l a spp . , v i z . , V. c a l o p h y l l a , V. c a l o p h y l l o i d e a , V . e l o n g a t a , V . c u s p i d a t a , V i r o l a t h e i o d o r a , and p o s s i b l y V. p e r u v i a n a , a re known to be u t i l i z e d in the p r e p a r a t i o n of h a l l u c i n o g e n s . Spec ies concepts w i t h i n the genus are p o o r l y e l u c i d a t e d , and t h i s has l ed to c o n s i d e r a b l e c o n f u s i o n in the e t h n o b o t a n i c a l and phy tochemica l l i t e r a t u r e . For i n s t a n c e , the most recent s y s t ema t i c r e v i s i o n of the genus [21] s t a t e s tha t V i r o l a t h e i o d o r a and V. c u s p i d a t a are e q u i v a l e n t to V. e l o n g a t a , wh i le V. c a l o p h y l l o i d e a i s equated wi th V. c a l o p h y l l a . Thus , depending on which s p e c i e s concepts are a c c e p t e d , one may say that as many as s i x or as few as th ree V i r o l a spp . have been i m p l i c a t e d as h a l l u c i n o g e n s ! The phy tochemica l b a s i s f o r the use of V i r o l a spp . as h a l l u c i n o g e n s i s we l l e s t a b l i s h e d , s i n c e the ba rk , sap , l e a v e s , and roo t s of a number of V i r o l a s p e c i e s have been shown to c o n t a i n s u b s t a n t i a l amounts of h a l l u c i n o g e n i c t r yp tamines such as N , N - d i m e t h y l t r y p t a m i n e , 5-methoxy-N,N-d imethy l t r yp tamine , and 8 other t r yp tamine d e r i v a t i v e s , i n c l u d i n g N-methy l t ryptamine , 5-methoxy-N-methy l t ryptamine, 5-methoxy-tryptamine, and t ryptamine ( c f . F i g . 1) . /3-carboline a l k a l o i d s ( F i g . 2) have a l s o been i s o l a t e d from some V i r o l a spp . A g u r e l l et a l . [22] r epo r t ed the i s o l a t i o n of 6-MeO-2-methyl-tetrahydro-/3-carbol ine from V i r o l a  t h e i o d o r a and V. r u f u l a , wh i le a s i m i l a r compound, 6-MeO-l,2-dimethyl-TH/3C, was i s o l a t e d from Anadenanthera p e r e g r i n a L. (Leguminoseae) which a l s o c o n t a i n s t ryptamine d e r i v a t i v e s and i s the source of a h a l l u c i n o g e n i c snu f f in the Car ibbean and B r a z i l [ 2 ] . In a l a t e r p u b l i c a t i o n [23] these workers de tec ted both compounds in V. pe ruv i ana and an inde te rmina te V i r o l a s p e c i e s ; the r e l a t e d compound 2-methyl-TH/3C was de t e c t ed in V. c a l o p h y l l a , V. e l o n g a t a , and V i r o l a t h e i o d o r a . In a l l i n s t a n c e s the |3-carbol ines were t r a ce components compared to the t r y p t a m i n e s ; V. c u s p i d a t a i s the on ly spec i e s so f a r i n v e s t i g a t e d in which the major a l k a l o i d s are /3-carbol i nes . Cassady and co-workers [24] i s o l a t e d 6-MeO-harman, 6-MeO-harmalan, and 6-MeO-tetrahydroharman from the l eaves and stems of t h i s s p e c i e s , but no t r yp tamines were d e t e c t e d . T h i s r a the r d i s t i n c t i v e b i o chemi ca l c h a r a c t e r i n d i c a t e s that V. c u s p i d a t a i s p robab l y d i s t i n c t from V. e l onga ta and the o ther V i r o l a spp . which have been c l a imed to be e q u i v a l e n t to V. e longa ta by Rodr igues [21 ] . I I I . Ethnopharmacoloqy of Ma lp igh i a ceous and M y r i s t i c a c e o u s  H a l l u c inogens A l though the most important sou r ce-p l an t s and the presumed p s y c h o a c t i v e c o n s t i t u e n t s of the Ma lp igh i a ceous and 9 M y r i s t i c a c e o u s h a l l u c i n o g e n s have been i d e n t i f i e d , the ethnopharmacology of both groups of h a l l u c i n o g e n s i s s t i l l i n c o m p l e t e l y unde rs tood . A. Source P l a n t s and Methods of P r e p a r a t i o n The h a l l u c i n o g e n ayahuasca i s prepared by b o i l i n g the bark or c rushed stems of B. caap i t oge the r wi th one or more of the sundry admixture p l a n t s mentioned above or l i s t e d in Appendix I. The exact mode of p r e p a r a t i o n v a r i e s from reg ion to r e g i o n ; in some areas the stems and admixtures are b o i l e d fo r many hou r s , and the brew i s concen t r a t ed over a low f i r e to a f r a c t i o n of i t s o r i g i n a l volume; in other r e g i o n s , the brew i s b o i l e d fo r a r e l a t i v e l y sho r t time (1-2 hours) and i s not c o n c e n t r a t e d , whi le in s t i l l o the r c a s e s , a co ld-water i n f u s i o n i s p r e p a r e d . O b v i o u s l y the method of p r e p a r a t i o n would be expected to a f f e c t the c o n c e n t r a t i o n , p r o p o r t i o n s , and perhaps the k inds of a c t i v e a l k a l o i d s p resen t in the m i x t u r e . 0 - c a r b o l i n e s can be r e a d i l y formed from t r yp tamines by a ldehyde condensa t ion f o l l owed by c y c l i z a t i o n ( P i c t e t - Speng l e r r e a c t i o n ) [25 ] , and thus might we l l be formed as a r t i f a c t s du r i ng the p ro longed b o i l i n g and c o n c e n t r a t i o n of the brew. In any case the p r e p a r a t i o n as f i n a l l y i n g e s t e d would c o n t a i n a r e l a t i v e l y h igh p r o p o r t i o n of harmine , h a r m a l i n e , and t e t r ahyd roha rm ine , and p o s s i b l y other 0 -c a r b o l i n e s , and a low to moderate amount of DMT, depending on how much admixture was added. The use of M y r i s t i c a c e o u s s p e c i e s fo r h a l l u c i n o g e n s c e n t e r s around the i n g e s t i o n of p r e p a r a t i o n s d e r i v e d from the " r e s i n " (sap) of the t r e e , even though the l e a v e s , ba rk , or r oo t s of the 10 t r ee may a l s o con t a i n the a c t i v e a l k a l o i d s . The mode of i n g e s t i o n of the r e s i n appears to be d i v i d e d g e o g r a p h i c a l l y and e t h n o l o g i c a l l y i n t o two major c a t e g o r i e s . In the r eg i on cen te red around the R io Or inoco-R io Vaupes dra ingage of Co lomb ia , Venezue l a , and B r a z i l , the ind igenous people prepare a snu f f from the d r i e d , powdered r e s i n of v a r i o u s V i r o l a s p e c i e s . These t r i b e s i n c l ude the Pu inave , Kubeo, Tukano, and v a r i o u s l i n g u i s t i c subgroups of the w ide ly d i s p e r s e d Waika (Yanomamo) t r i b e . The snuf f may be p repared from the V i r o l a r e s i n a lone but more o f t en i s mixed wi th the ashes or powdered l eaves of other p l a n t s [26, see be low] . The second mode of u t i l i z a t i o n of V i r o l a r e s i n as an h a l l u c i n o g e n i s by o r a l i n g e s t i o n , e i t h e r of the r e s i n d i r e c t l y or of p e l l e t s p repared from the r e s i n and r o l l e d in the ashes of other p l a n t s , whose i d e n t i t y may v a r y . The p r a c t i c e of o r a l i n g e s t i o n of V i r o l a r e s i n has come to the a t t e n t i o n of e t h n o b o t a n i s t s on ly r e c e n t l y and seems to be g e o g r a p h i c a l l y and e t h n o l o g i c a l l y separa ted from the use of V i r o l a r e s i n as a s n u f f . The o r a l use of V i r o l a i s found p r i m a r i l y among the Bo ra , Muinane and Wi to to t r i b e s i n h a b i t i n g the Caraparana- Igaraparana l o c a l i t y of the Colombian Putamayo. F i e ld-work c a r r i e d out by S chu l t e s and Swain [7 ,27 ,28 ] near the v i l l a g e of E l Encanto on the R io Caraparana i n d i c a t e d that the best " k i n d " of oo-koo '-na (as the drug i s c a l l e d in Wi toto ) was d e r i v e d from V i r o l a t h e i o d o r a . Subsequent f i e l d-work was c a r r i e d out by S c h u l t e s , Swain, and Plowman [29] i n the Bora and Wi toto s e t t l emen t s of T i e r r a F i r m a , B r i l l o Nuevo, and Puco U r q u i l l o on the R io Ampiyacu near the Peruv ian se t t l ement of Pebas. The best sources i n d i c a t e d by the Bora fo r the p r e p a r a t i o n of the o r a l 11 drug (known in Bora as ku'-ru-k_u) was V. e l o n g a t a ; V. pavon i s, V. l o r e t e n s i s , and V. su r inamens i s were a l s o i n d i c a t e d . The Wi to to in formants at Puco U r q u i l l o i n d i c a t e d tha t V . e l o n g a t a , V . s u r i n a m e n s i s , and s p e c i e s in the r e l a t e d genus I r y a n t h e r a , v i z . , I_. m a c r o p h y l l a , I. u l e i , and I. tessmanni i , were a l l s u i t a b l e f o r the p r e p a r a t i o n of the d r u g . The genus I r yan the ra had not been i m p l i c a t e d p r e v i o u s l y as an h a l l u c i n o g e n ; subsequent a n a l y s i s of I_. u l e i de t e c t ed a t r a c e (.00013 mg/g) of 5-MeO-DMT in the bark . Both the M y r i s t i c a c e o u s s n u f f s and the o r a l l y i nges t ed M y r i s t i c a c e o u s pas tes would be expected to c o n t a i n s u b s t a n t i a l c o n c e n t r a t i o n s of DMT and/or 5-MeO-DMT, as we l l as o ther t r yp t am ines ; l e s s e r c o n c e n t r a t i o n s of /3-carbolines might a l s o be p r e s e n t , e i t h e r because they are o r i g i n a l l y p resen t in the r e s i n of some V i r o l a s p e c i e s , ' or as a r e s u l t of fo rmat ion from t ryp tamines du r i ng the cook ing p r o c e s s . The a l k a l o i d c o n s t i t u e n t s of some V i r o l a s n u f f s have been c h a r a c t e r i z e d [30] but the compos i t i on of the o r a l l y - i n g e s t e d pas tes has not been p r e v i o u s l y i n v e s t i g a t e d . B. P o s t u l a t e d Mechanism of A c t i o n A l though the sou r ce-p l an t s and admixture p l a n t s u t i l i z e d in the p r e p a r a t i o n of the Ma lp igh i a ceous h a l l u c i n o g e n s are d i f f e r e n t from those used f o r the M y r i s t i c a c e o u s s n u f f s and p a s t e s , s i m i l a r a c t i v e a l k a l o i d s are r e s p o n s i b l e f o r the p h a r m a c o l o g i c a l a c t i v i t y of both of these Amazonian h a l l u c i n o g e n s . The ayahuasca brews c o n t a i n /3-carbol ines as the major a l k a l o i d s wi th DMT d e r i v e d from the admixture p l a n t s p resen t i n l e s s e r c o n c e n t r a t i o n s ; the M y r i s t i c a c e o u s d r u g s , on 12 the o ther hand, c o n t a i n h a l l u c i n o g e n i c t r yp tamine d e r i v a t i v e s as the pr imary a c t i v e c o n s t i t u e n t s , but 0-ca rbo l i ne s may be present as minor components. A l though DMT and 5-MeO-DMT are ext remely po t en t-psycho tom ime t i c s , a c t i v e in the 5 to 100 mg range, a p e c u l i a r i t y of t h e i r pharmacology i s tha t they are not o r a l l y a c t i v e [31] appa r en t l y because they are deaminated by p e r i p h e r a l monoamine ox idase (MAO), the m i t o c h o n d r i a l l y l o c a l i z e d enzyme tha t i s r e s p o n s i b l e fo r the o x i d a t i v e deaminat ion of b i ogen i c amines . The r e fo r e these t r yp tamines e i t h e r must be adm in i s t e r ed p a r e n t e r a l l y in order to man i fes t t h e i r a c t i v i t y , or they can be rendered o r a l l y a c t i v e i f taken toge ther wi th a MAO i n h i b i t o r . On the o ther hand, the 0 - c a r b o l i n e s , a l t hough hav ing l i m i t e d a c t i v i t y as h a l l u c i n o g e n s (some d e r i v a t i v e s are a c t i v e in the 500-1000 mg range ; c f . Chapter I I ) , a re ext remely potent sho r t-a c t i n g i n h i b i t o r s of monoamine o x i d a s e , e f f e c t i v e _in. v i t r o at c o n c e n t r a t i o n s on the order of 1 0 " 6 - 10~ 8 M [ 3 2 , 3 3 , 3 4 ] . The s i g n i f i c a n c e of t h i s to the ethnopharmacology of these drugs may thus be unde r s tood : a l though d i f f e r e n t s p e c i e s and d i v e r s e admixtures are i n v o l v e d in the p r e p a r a t i o n of the h a l l u c i n o g e n i c B a n i s t e r i o p s i s brews, the M y r i s t i c a c e o u s s n u f f s used no r th of the R io Negro and the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s h a l l u c i n o g e n s used in Peru and Co lomb ia , in each c a s e , i t i s the presence of me thy l a t ed , psychotomimet ic t r yp tamines tha t i s ' t h e s i ne qua non fo r the h a l l u c i n o g e n i c p r o p e r t i e s of these na t i v e p r e p a r a t i o n s . The pha rmaco log i c a l r a t i o n a l e f o r the i n g e s t i o n of V i r o l a r e s i n in the form of a snu f f i s now c l e a r ; the a c t i v e t r yp tamines are not o r a l l y a c t i v e , so t h i s p a r e n t e r a l route of a d m i n i s t r a t i o n n e a t l y s i d e s t e p s t h e i r i n a c t i v a t i o n by p e r i p h e r a l 13 MAO. Presumably the t r yp tamines a lone are s u f f i c i e n t to account fo r the a c t i v i t y of the s n u f f s . In the case of the Ma lp igh i a ceous ayahuasca brews and the o r a l l y i nges t ed M y r i s t i c a c e o u s p a s t e s , the M A O - i n h i b i t i n g /3-carbol ines (or some o ther mechanism) i s r e q u i r e d in order to o r a l l y a c t i v a t e the t r y p t a m i n e s . The o r a l a c t i v a t i o n of the t r yp tamines by the /3-c a r b o l i n e s has been proposed [ 9 , 1 0 , 1 3 , 2 7 , 2 8 , 2 9 ] as the u n d e r l y i n g mechanism, but t h i s has not been e x p e r i m e n t a l l y c o n f i r m e d . Seve ra l q u e s t i o n s need to be i n v e s t i g a t e d e x p e r i m e n t a l l y be fo re t h i s p o s t u l a t e d mechanism can be a c c e p t e d . It must be shown t h a t : 1.) The t r yp tamines in q u e s t i o n are not o r a l l y a c t i v e by themse l ves ; enough data on the human pharmacology of these drugs has been accumulated in recent years ( c f . Chapter II and [31]) that t h i s po in t i s f a i r l y we l l e s t a b l i s h e d . 2.) The methy la ted t r yp tamines can be rendered o r a l l y a c t i v e by MAOIs, s p e c i f i c a l l y /3-carbol ines ; t r yp tamines have been shown to be p o t e n t i a t e d j_n v i v o by some MAOIs, but t h i s has not been s p e c i f i c a l l y demonstrated fo r /3-carbol ines ( c f . Chapter II and [35 ] ) . 3.) H a l l u c i n o g e n i c t r yp tamines are p resen t in these n a t i v e drug p r e p a r a t i o n s in s u f f i c i e n t c o n c e n t r a t i o n s that the i n g e s t i o n of an average dose would exceed the known t h r e s h o l d dose fo r these compounds, e . g . , i f DMT i s the a c t i v e t r yp tamine then at l e a s t 15 mg and p r e f e r a b l y 60-75 mg would have to be i nges ted f o r the drug to man i fes t i t s h a l l u c i n o g e n i c e f f e c t s . Under c o n d i t i o n s of p e r i p h e r a l MAO i n h i b i t i o n , s u b s t a n t i a l l y sma l l e r doses of DMT may e l i c i t h a l l u c i n o g e n i c e f f e c t s , s i n ce a g r ea t e r p r o p o r t i o n c o u l d escape deg rada t i v e metabol ism under these c o n d i t i o n s . 4.) /3-carbol ines , 1 4 or some o ther type of MAOI, are p resen t in the n a t i v e p r e p a r a t i o n in s u f f i c i e n t c o n c e n t r a t i o n to e f f e c t i v e l y i n h i b i t MAO and thus o r a l l y p o t e n t i a t e the t r y p t a m i n e s . Expe r imen ta l i n v e s t i g a t i o n s of some of these q u e s t i o n s were among the pr imary o b j e c t i v e s of the work d e s c r i b e d in t h i s t h e s i s . C. Admixture P l an t s Another aspect of these na t i v e h a l l u c i n o g e n s which deserves f u r t h e r i n v e s t i g a t i o n i s the i n f l u e n c e of admixture p l a n t s as de te rminan ts of t h e i r pha rmaco log i c a l a c t i v i t y . On oc cas i on the drugs may be prepared wi thout the a d d i t i o n of any admix tu res , but c u s t o m a r i l y , admixtures are employed. P a r t i c u l a r l y in the case of ayahuasca i t seems that the u t i l i z a t i o n of admixture p l a n t s has reached a r a the r h igh degree of b o t a n i c a l and p h a r m a c o l o g i c a l s o p h i s t i c a t i o n . Bes ides the t r yp tamine-c o n t a i n i n g admixtures which are a lmost always i n c l uded in ayahuasca , the re i s a v i r t u a l pharmacopoeia of admixtures which are o c c a s i o n a l l y used , depending on the m a g i c a l , r i t u a l , or med ica l purposes fo r which the drug i s be ing made and consumed [ 9 , 1 0 , 1 3 , 1 4 , 1 5 ] . Many of these ayahuasca admixtures have not been b o t a n i c a l l y i d e n t i f i e d , much l e s s c h e m i c a l l y c h a r a c t e r i z e d ; however the b o t a n i c a l i d e n t i t y of some admixtures has been e s t a b l i s h e d , and in many cases they are spec i e s which c o n t a i n b iodynamic c o n s t i t u e n t s of p o t e n t i a l med ica l v a l u e . The c o n t r i b u t i o n that the admixtures may make to the pha rmaco log i c a l a c t i v i t y of ayahuasca i s , a t t h i s t ime , a complete myste ry , and an area w e l l d e se r v i ng of f u r t h e r r e s e a r c h by e thnopha rmaco log i s t s and phy tochemi s t s . The phy tochemica l 1 5 i n f o r m a t i o n tha t i s a v a i l a b l e on approx imate l y 70 of the spec i e s known to be used as admixtures has been summarized in Appendix I . The admixture p l a n t s used in the p r e p a r a t i o n of the M y r i s t i c a c e o u s pas tes and s n u f f s have not r e c e i v e d a great dea l of a t t e n t i o n in the e thnopha rmaco log i ca l l i t e r a t u r e . The two admixtures o f t e n used in the p r e p a r a t i o n of V i r o l a snuf f are E l i z a b e t h a p r i n c e p s (Shomb. ex Bentham) Hooker (Leguminosae) and J u s t i c i a p e c t o r a l i s J a c q . v a r . s t e n o p h y l l a Leonard (Acanthaceae) [26 ] . Ashes made from the bark of E. pr inceps are o f t e n added to the s n u f f s , and f r e q u e n t l y the powdered l eaves of J u s t i c i a p e c t o r a l i s v a r . s t e n o p h y l l a are added to "make the snu f f sme l l b e t t e r . " [ 26 ] . There are a l s o r e p o r t s [36, P rance , G. T . , p e r s . comm., 1983] that o c c a s i o n a l l y a snu f f i s p repared from J . p e c t o r a l i s by i t s e l f . Whether t h i s snu f f i s p s y c h o a c t i v e or whether t h i s p l an t c o n t a i n s b iodynamic c o n s t i t u e n t s i s not known as no phy tochemica l data i s a v a i l a b l e on t h i s s p e c i e s . The on ly use of admixtures in the manufacture of the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s pas tes that has been noted [27 ,28 ,29 ] i s tha t o c a s s i o n a l l y the p e l l e t s of V i r o l a r e s i n are r o l l e d in the " s a l t s " of c e r t a i n p l a n t s , the s a l t s be ing the l e a c h i n g s of ashes made from the bark or l eaves of v a r i o u s s p e c i e s . S c h u l t e s and Swain [28] have d e s c r i b e d the p rocess of p r e p a r i n g t h i s " s a l t " . Among the p l a n t s used as the source of the s a l t a re E s chwe i l e r a i t a y e n s i s Kunth ( Lecy th idaceae ) and Gus t a v i a poepp ig i ana Berg ex . M a r t i u s ( L e c y t h i d a c e a e ) , Theobroma  subinacum Mar t . ( S t e r c u l i a c e a e ) and s e v e r a l palm s p e c i e s i n c l u d i n g Geonoma juruana Dammer. S ince on l y the ashes of these 16 p l a n t s are used in the p r e p a r a t i o n of the V i r o l a p e l l e t s i t i s very u n l i k e l y that any biodynamic c o n s t i t u e n t s are d e r i v e d from these admix tu res . One p o s s i b i l i t y , which seems to have been ove r l ooked in the l i t e r a t u r e , i s that the ashes are added to make the mixture a l k a l i n e and thus conve r t the a c t i v e a l k a l o i d s to the f r ee base form. T h i s may f a c i l i t a t e a b s o r p t i o n and uptake of the a l k a l o i d s . Ashes made from l eaves of Cec rop i a spp . (Moraceae) are s i m i l a r l y added to the coca powder which i s w ide ly used by the same groups that use the o r a l V i r o l a p r e p a r a t i o n s . I t i s c o n s i d e r e d common knowledge that ashes must be added to the coca in o rder to make i t s t r o n g . The a d d i t i o n of ashes to the M y r i s t i c a c e o u s pas tes may be an ex t ens i on of t h i s ' g ene ra l r u l e , and may a c t u a l l y i n f l u e n c e t h e i r b i o l o g i c a l a c t i v i t y . IV. E thnograph i c Aspec ts of Ma lp igh i a ceous and M y r i s t i c a c e o u s  H a l l u c i n o g e n s The focus of the r e s e a r c h r epo r t ed in t h i s t h e s i s i s on the b o t a n i c a l , c h e m i c a l , and pha rmaco log i c a l a spec t s of the Ma lp i gh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s ; however, a f u l l unde rs t and ing of these p l a n t drugs must a l s o c o n s i d e r the c u l t u r a l con tex t of t h e i r use . The e f f e c t s of h a l l u c i n o g e n s , u n l i k e most o ther pha rmaco log i c a l agen t s , are i n f l u e n c e d to a l a r g e degree by the e x p e c t a t i o n s of the u s e r . These e x p e c t a t i o n s , and the nature of the i n d i v i d u a l ' s drug e x p e r i e n c e , w i l l , in t u r n , p a r t i a l l y be determined by p s y c h o l o g i c a l and n e u r o p h y s i o l o g i c a l v a r i a b l e s , and p a r t i a l l y by the p r e v a i l i n g c u l t u r a l myths and b e l i e f s su r round ing the drug 17 in q u e s t i o n . T h i s i s t rue in our own c u l t u r e as we l l as in " t r a d i t i o n a l " non-Western c u l t u r e s . U n l i k e our own c u l t u r e , in most t r a d i t i o n a l s o c i e t i e s in which h a l l u c i n o g e n s are used , that use takes p l a ce in a m a g i c o - r e l i g i o u s c o n t e x t . Use of the h a l l u c i n o g e n i s surrounded by r i t u a l and ceremony, the set and s e t t i n g of the drug expe r i ence i s c a r e f u l l y chosen and man ipu la ted by the shaman or med ic ine man, and there i s u s u a l l y a s p e c i f i c purpose fo r consuming the drug--for d i v i n a t i o n , for i n s t a n c e , or to d i s c o v e r the cause of an i l l n e s s , or to communicate wi th the s p i r i t - w o r l d . In t r a d i t i o n a l c u l t u r e s the boundar ies between r e l i g i o n , magic , and medic ine are not c l e a r l y d e l i n e a t e d ; the f u n c t i o n of the shaman or t r a d i t i o n a l hea l e r l i e s somewhere between the Western concepts of p r i e s t , d o c t o r , and p s y c h o t h e r a p i s t ; i l l n e s s may be p r e c i p i t a t e d by p h y s i c a l , p s y c h o l o g i c a l , or s u p e r n a t u r a l c auses , or a combina t ion of t h e s e , and a l l a re amenable to treatment by the methods a v a i l a b l e to the shaman. In t h i s sense the " h o l i s t i c " t h e r a p i e s which have c h a r a c t e r i z e d recent t r ends in modern med ic ine are not tha t d i f f e r e n t from the t h e r a p e u t i c methods p r a c t i c e d by the t r a d i t i o n a l h e a l e r . Both proceed from the r e c o g n i t i o n that mind and body are an i n t e g r a t e d u n i t and the most e f f e c t i v e t h e r a p i e s are those tha t are d i r e c t e d at improv ing both p h y s i c a l and mental h e a l t h . Thus i t i s not s u r p r i s i n g that p l a n t h a l l u c i n o g e n s , which p ro found l y a f f e c t both the mind and the body, as we l l as a f f o r d i n g access to and a c e r t a i n degree of m a n i p u l a t i o n of ( r e a l or imagined) mag ica l or s u p e r n a t u r a l d imens ions , shou ld occupy such a prominent p o s i t i o n in the t r a d i t i o n a l h e a l e r ' s armamentarium. 1.8 I t i s not w i t h i n the scope of t h i s t h e s i s to review the voluminous e thnograph ic and a n t h r o p o l o g i c a l l i t e r a t u r e r e l a t e d to the use of p l a n t h a l l u c i n o g e n s in t r a d i t i o n a l h e a l i n g ; r e f e r e n c e s [37,38] p rov ide a gene ra l overv iew of t h i s s u b j e c t . Re i che l -Do lma to f f [39] has p rov ided a f a s c i n a t i n g and d e t a i l e d account of the c e n t r a l p o s i t i o n of ayahuasca and the M y r i s t i c a c e o u s s n u f f s in the r e l i g i o n and cosmology of the Tukano I nd i ans , whi le Chagnon [40] has d e s c r i b e d the use of V i r o l a snuf f among the Yanomamo. V i r t u a l l y no th ing i s known of the m e d i c a l , m a g i c a l , or r e l i g i o u s use of the o r a l l y i nges t ed M y r i s t i c a c e o u s pas tes beyond the f ragmentary i n f o rma t i on recorded by S chu l t e s [27] and the e q u a l l y f ragmentary o b s e r v a t i o n s made du r i ng the present i n v e s t i g a t i o n . Both are d i s c u s s e d more f u l l y in Chapter I I I . Other o b s e r v a t i o n s r e l a t e d to the methods of p r e p a r a t i o n , use , and s u b j e c t i v e e f f e c t s of ayahuasca and the o r a l l y i nges t ed pas tes are a l s o to be found in Chapter I I I . E thnograph ic a spec t s of the use of ayahuasca are more a c c e s s i b l e to the ou t s i de observe r than the M y r i s t i c a c e o u s pas tes c h i e f l y because ayahuasca i s used among urban mes t i zo p o p u l a t i o n s r e s i d i n g on the o u t s k i r t s of l a r g e r i v e r i n e i n d u s t r i a l and t rade c e n t e r s such as I q u i t o s and P u c a l l p a . Hence many of the c u l t u r a l and l i n g u i s t i c b a r r i e r s which are encountered in the e thnograph i c study of the M y r i s t i c a c e o u s pas tes are not a f a c t o r in the study of ayahuasca . Most members of the mes t i zo p o p u l a t i o n in these urban or semi-urban se t t l emen t s speak a d i a l e c t i c a l form of Span ish (though not n e c e s s a r i l y as the f i r s t l anguage ) . Face- to- face i n t e r v i e w s can 19 thus be conducted w i th i n f o r m a n t s , and t h i s i s g e n e r a l l y a more f r u i t f u l procedure than i n t e r v i ews conducted through a Span ish-speak ing i n t e r p r e t e r . A more open a t t i t u d e toward the ou t s i de i n v e s t i g a t o r i s o f t en found among mes t i zos who use ayahuasca and t h i s a l s o f a c i l i t a t e s g a t h e r i n g e thnograph i c i n f o r m a t i o n . Contemporary use of ayahuasca in Amazonian mes t i zo p o p u l a t i o n s appears to be an amalgam of d i v e r s e t r i b a l t r a d i t i o n s . The l a rge urban se t t l emen t s have become me l t i ng p o t s ; peop le of many d i f f e r e n t c u l t u r a l backgrounds have migra ted to these c en t e r s in sea rch of employment in the lumber, pe t ro l eum, and s i m i l a r r esource-based i n d u s t r i e s , and have brought wi th them t h e i r own t r i b a l t r a d i t i o n s and b e l i e f systems ( u s u a l l y s y n c r e t i c a l l y fused wi th C h r i s t i a n i t y due to p r i o r con tac t w i th m i s s i o n a r i e s ) . The c u l t u r a l background of these migrant l a b o r e r s o f t en extends to a knowledge of the m e d i c i n a l p l a n t s va lued in t h e i r own c u l t u r e . Over the years t h i s d rug-p lan t l o r e d e r i v e d from d i v e r s e sources has g r a d u a l l y d i f f u s e d through the l a r g e r mes t i zo s o c i e t y and become melded i n t o a coherent system of t r a d i t i o n a l med i c i ne . T h i s t r a d i t i o n , though i t i n c o r p o r a t e s e lements of i t s d i v e r s e t r i b a l o r i g i n s , i s at the same time unique to the mes t i zo s o c i a l c l a s s . T h i s p rocess of c u l t u r a l amalgamation has occu r r ed over the same p e r i o d of time that most of the t r i b a l s o c i e t i e s in which r e s i d e the an tecedents of mes t i zo f o l k ' m e d i c i n e have d i s i n t e g r a t e d and/or d i s a p p e a r e d . Hence mes t i zo f o l k-med i c i ne as i t i s p r a c t i c e d today in the urban c e n t e r s of the Amazon i s a l i v i n g system of t r a d i t i o n a l med ic ine based on the e thnomedic ine of many c u l t u r e s ; in many cases i t i s the on ly p l a ce where such knowledge has been 20 p r e s e r v e d . Hence i t i s impor tan t , even u rgen t , t ha t mes t i zo f o l k - m e d i c i n e and the p l a n t s that form i t s b a s i s be s t u d i e d by i n v e s t i g a t o r s w i th s c i e n t i f i c backgrounds in m e d i c i n e , pharmacology , phy tochemi s t r y , and botany whi le the o p p o r t u n i t y s t i l l e x i s t s . The ayahuasca admixtures l i s t e d in Appendix I p ro v i de an example. Some of the s p e c i e s known to be used as admixtures to ayahuasca c o n t a i n h i g h l y biodynamic and p o t e n t i a l l y m e d i c a l l y u s e f u l c o n s t i t u e n t s ; how many o the rs may have va lue tha t have so f a r escaped i n v e s t i g a t i o n i s not known. The i n s i s t e n c e , nea r l y u n i v e r s a l among ayahuasqueros , that ayahuasca teaches med i c i ne , i . e . , tha t one l e a r n s about the p r o p e r t i e s of p l a n t s and how to use them by t a k i n g them in c o n j u n c t i o n wi th ayahuasca [14 ] , cannot be d i s m i s s e d out of hand. Fu r the r i n s i g h t s i n t o the use of ayahuasca in contemporary mes t i zo f o l k med ic ine may be found in [ 1 4 , 3 8 , 4 1 , 4 2 , 4 3 ] . V. Scope and O b j e c t i v e s of the Present I n v e s t i g a t i o n It may be seen from the p r e ced ing d i s c u s s i o n that s e v e r a l a spec t s of the c h e m i s t r y , pharmacology, botany , and ethnography of the Ma lp igh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s remain p o o r l y unde r s tood . The o b j e c t i v e s of the r e sea r ch d e s c r i b e d in t h i s t h e s i s were to i n v e s t i g a t e some of these un reso l ved p rob lems . An i n t e r d i s c i p l i n a r y approach was adop ted , i n .wh i ch e thnograph i c and e t h n o b o t a n i c a l f i e l d i n v e s t i g a t i o n s were combined w i th l a b o r a t o r y i n v e s t i g a t i o n s of the chemis t r y and pharmacology of the Ma lp igh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s and the p l a n t s used in t h e i r manufac tu re . The s c i e n t i f i c r a t i o n a l e f o r s p e c i f i c a spec t s of the i n v e s t i g a t i o n 21 i s g iven in the a p p r o p r i a t e c h a p t e r s ; the o v e r - a l l o b j e c t i v e s of the r e sea r ch are enumerated below. A. F i e l d Work T h i s had s e v e r a l complementary o b j e c t i v e s : 1. C o l l e c t i o n of e t h n o b o t a n i c a l and e thnograph i c in format i o n . Th i s i n c l u d e d i n fo rma t i on r e l a t e d to the sou r ce-p l an t s used in the manufacture of the d rugs , the methods of p r e p a r a t i o n , the methods of i n g e s t i o n , the purposes fo r which the drugs are used , and the o c c a s i o n s when they are consumed, and the c u l t u r a l contex t of t h e i r use . Na t i ve v e r n a c u l a r names of the p l a n t s and i n fo rma t i on on t h e i r f o l k taxonomy was a l s o c o l l e c t e d . 2. C o l l e c t i o n of p l a n t s and drug samples . C o l l e c t i o n s of Ma lp igh i aceous and M y r i s t i c a c e o u s drug p r e p a r a t i o n s were made and, when p o s s i b l e , the source p l a n t s and admixtures used in t h e i r p r e p a r a t i o n were a l s o c o l l e c t e d . P l an t c o l l e c t i o n s i n c l u d e d both herbar ium voucher specimens and samples fo r phy tochemica l a n a l y s i s . As many c o l l e c t i o n s of Ma lp igh i a ceous and M y r i s t i c a c e o u s s p e c i e s as p o s s i b l e were made, even though they were not n e c e s a r i l y used to make drug p r e p a r a t i o n s or i n d i c a t e d as be ing so u s a b l e . S p e c i a l e f f o r t s were made to c o l l e c t any admixture p l a n t s i n d i c a t e d by i n fo rman t s . Any o ther p l a n t s which were i n d i c a t e d by in fo rmants to have m e d i c i n a l a p p l i c a t i o n s were a l s o c o l l e c t e d even though they were not c o n s t i t u e n t s of h a l l u c i n o g e n i c p r e p a r a t i o n s . 22 3. Assessment of the b i o l o g i c a l a c t i v i t y of o r a l l y -a c t i v e Ma lp igh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s . A l though there i s c o n s i d e r a b l e d i s c u s s i o n of the mechanism of a c t i o n of the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s and Ma lp igh i a ceous h a l l u c i n o g e n s in the e t h n o b o t a n i c a l l i t e r a t u r e , r e p o r t s of t h e i r e f f e c t s on o u t s i d e obse rve rs are rare or non-e x i s t e n t . Thus i t i s on l y assumed that these drugs are o r a l l y -a c t i v e as h a l l u c i n o g e n s ; s u b s t a n t i a t i n g ev idence i s r e q u i r e d from an o u t s i d e observer who i s not immersed in the c u l t u r a l b e l i e f system su r round ing these d r u g s . Thus one of the o b j e c t i v e s of the f i e ld-work was to determine whether these p r e p a r a t i o n s a r e , in f a c t , o r a l l y e f f e c t i v e . T h i s was p a r t i c u l a r l y important in the case of the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s p a s t e s , s i n ce t h e i r p h y s i o l o g i c a l and p s y c h o l o g i c a l e f f e c t s have not p r e v i o u s l y been expe r i enced by a non- Ind ian . B. L abo ra to r y I n v e s t i g a t i o n s These were des igned to complement the f i e l d i n v e s t i g a t i o n s and to r e s o l v e s e v e r a l q u e s t i o n s which c o u l d be approached e x p e r i m e n t a l l y : 1. Q u a n t i t a t i v e i n v e s t i g a t i o n s of the a c t i v e c o n s t i t u e n t s of the Ma lp igh i a ceous and M y r i s t i c a c e o u s h a l l u c i n o g e n s . A l though p r e v i o u s i n v e s t i g a t o r s have determined that the a c t i v e c o n s t i t u e n t s of these drugs are in a l l l i k e l y h o o d 23 psychotomimet ic t r yp tamines and /3-carbol ines , q u a n t i t a t i v e data on the l e v e l s of a l k a l o i d s in these p r e p a r a t i o n s i s r e l a t i v e l y s p a r s e . I t i s not known, fo r i n s t a n c e , whether the amount of DMT in a " t y p i c a l " dose of ayahuasca or V i r o l a snu f f exceeds the t h r e s h o l d dose r e q u i r e d to e l i c i t the psychotomimet ic e f f e c t s . S i m i l a r l y , a l though i t has been proposed that the DMT in ayahuasca and the o r a l l y i nges ted M y r i s t i c a c e o u s pas tes i s rendered o r a l l y a c t i v e by the M A O - i n h i b i t i n g /3-carbol ines , i t has not been shown that /3-carbol ines are p resen t in s u f f i c i e n t c o n c e n t r a t i o n s to e f f e c t i v e l y i n h i b i t MAO. The r e fo r e q u a n t i t a t i v e methods were dev i sed to determine the k inds and r e l a t i v e p r o p o r t i o n s of t r yp tamines and /3-carbol ines in the drug samples and in the p l a n t samples . 2. Comparison of the a l k a l o i d c o n s t i t u e n t s of the drug p r e p a r a t i o n s wi th the a l k a l o i d c o n s t i t u e n t s of the s o u r c e - p l a n t s . Q u a n t i t a t i v e and q u a l i t a t i v e methods were a p p l i e d in order to determine to what extent the a l k a l o i d compos i t i on of the drug samples d i f f e r e d from that of the s o u r c e - p l a n t s . Admixture p l a n t s were a l s o sc reened fo r a l k a l o i d s and in some cases q u a n t i t a t i v e l y a n a l y z e d . 3. I n v e s t i g a t i o n of the e f f e c t of drug samples and t h e i r a l k a l o i d c o n s t i t u e n t s on MAO in v i t r o . The mechanism of a c t i o n p o s t u l a t e d fo r the o r a l a c t i v i t y of these drugs i s tha t the /3-carbol ines p r o t e c t the p s y c h o a c t i v e t r yp tamines from deaminat ion by p e r i p h e r a l monoamine o x i d a s e , 24 thus p e r m i t t i n g t h e i r uptake i n t o the CNS. A l though r e a s o n a b l e , t h i s mechanism had not been e x p e r i m e n t a l l y demonst ra ted . T h e r e f o r e methods were deve loped fo r a s s e s s i n g the a c t i v i t y of v a r i o u s drug p r e p a r a t i o n s and t h e i r c o n s t i t u e n t s ' fo r a c t i v i t y as MAO i n h i b i t o r s . The MAOI a c t i v i t y of s y n t h e t i c t r yp tamine and 0-c a r b o l i n e d e r i v a t i v e s was a l s o assessed in an e f f o r t to e s t a b l i s h s t r u c t u r e / a c t i v i t y c o r r e l a t i o n s . M ix tu r e s of s y n t h e t i c compounds in c o n c e n t r a t i o n s approx imat ing those found in the drug p r e p a r a t i o n s were a l s o assayed in order to determine whether the compounds might ac t s y n e r g i s t i c a l l y in comb ina t i on . The MAOI a c t i v i t y observed w i th the m ix tu res of s y n t h e t i c s tandards was compared wi th that observed wi th the crude drug p r e p a r a t i o n s in order to determine whether . the a c t i v i t y was p a r t l y due to n o n - a l k a l o i d a l c o n s t i t u e n t s . I_n v i v o i n v e s t i g a t i o n s of the MAOI o c t i v i t y of the drug p r e p a r a t i o n s us i ng an ima ls were o r i g i n a l l y p lanned but were not c a r r i e d out due to the unacceptab le l eng th of t ime r e q u i r e d to deve lop and p e r f e c t the necessary t e c h n i q u e s . 25 V I . L i t e r a t u r e C i t e d 1. S c h u l t e s , R. E. (1970) The B o t a n i c a l and Chemica l D i s t r i b u t i o n of H a l l u c i n o g e n s . Annual Review of P l an t  P h y s i o l o g y 21:571-98 2. S c h u l t e s , R. E. & A. Hofmann (1980) The Botany and Chemist ry  of H a l l u c i n o g e n s . 2nd E d i t i o n • Cha r l e s C. Thomas, P u b l i s h e r s , S p r i n g f i e l d , 111. 3. G r i n s p o o n , L. & L. B. B a k a l a r . (1979) P s y c h e d e l i c Drugs  R e c o n s i d e r e d . Bas i c Books, Inc . New York . 4. S c h u l t e s , R. E. (1963) The B o t a n i c a l Sources of the New World N a r c o t i c s . The P s y c h e d e l i c Review 1:145-66 5. S c h u l t e s , R. E. (1970) The New World Ind ians and T h e i r H a l l u c i n o g e n i c P l a n t s . M o r r i s Arboretum B u l l e t in 21:3-14 6. E f r o n , D. H . , B. Ho lmstedt , & N. S. K l i n e (eds . ) (1967) E thnopharmaco log ic Search fo r P sychoac t i ve Drugs . U. S. P u b l i c Hea l t h S e r v i c e P u b l i c a t i o n # 1645 7. R. E. S chu l t e s (1979) E v o l u t i o n of the I d e n t i f i c a t i o n of the M y r i s t i c a c e o u s H a l l u c i n o g e n s of South Amer i c a . J ou rna l of  Ethnopharmacoloqy 1:211-239 1979 8. S c h u l t e s , R. E. (1957) The I d e n t i t y of the M a i i p i g h i a c e o u s N a r c o t i c s of South Amer i ca . Harvard B o t a n i c a l Museum L e a f l e t s 18:1-56 9. P i n k l e y , H. V. (1969) P l an t Admixtures to Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k . L l o y d i a 32:305-14 10. Der M a r d e r o s i a n , A. H . , H. V. P i n k l e y , & M. F. Dobbins IV. (1968) Na t i ve Use and Occur rence of N ,N-d imethy l t ryptamine in the Leaves of B a n i s t e r i o p s i s rusbyana . American J ou rna l  of Pharmacy 140:137-147 11. Ga t e s , B. (1979) New Names in B a n i s t e r i o p s i s and D i p l o p t e r y s (Ma lp igh iaceae ) of the Guayana H i g h l a n d . B r i t t o n i a 31:108-9 26 12. S c h u l t e s , R. E . , (1979) New Data on the Ma lp igh i a ceous N a r c o t i c s of South Amer i ca . Harvard B o t a n i c a l Museum L e a f l e t s 23:137-47 13. S c h u l t e s , R. E. (1972) E t h n o t o x i c o l o g i c a l S i g n i f i c a n c e of A d d i t i v e s to New World H a l l u c i n o g e n s . P l an t Sc i ence B u l l e t i n 18: 34-41 14. Luna , L. E. (1983) The Concept of P l an t s as Teachers Among Four Mes t i zo Shamans of I q u i t o s , Nor theas t Pe ru . Paper p resen ted at the Symposium on Shamanism, X l t h I n t e r n a t i o n a l Congress of A n t h r o p o l o g i c a l and E t h n o l o g i c a l S c i e n c e s , Phase 2. Vancouver , B. C , August 20-23 1983. 15. R i v i e r , L. & J . L i ndgren (1972) Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k : E t h n o b o t a n i c a l and Chemica l I n v e s t i g a t i o n s . Economic Botany 29:101-129 16. D e u l o f e u , V. (1967) Chemica l Compounds I s o l a t e d from B a n i s t e r i o p s i s and Re l a t ed S p e c i e s , pp. 393-402 in D. H. E f r o n , B. Ho lmstedt , & N. S. K l i n e (eds . ) E thnopharmaco loq ic Search fo r P s ychoac t i v e Drugs . U. S. P u b l i c Hea l t h S e r v i c e P u b l i c a t i o n .#1645 17. A g u r e l l , S . , B. Ho lms ted t , & J . E. L i ndgren (1968) A l k a l o i d Content of B a n i s t e r i o p s i s rusbyana . American J o u r n a l of  Pharmacy 140:148-151 18. Hashimoto, Y. & K. Kawanishi (1975) New Organ ic Bases from Amazonian B a n i s t e r i o p s i s c a a p i . Phy tochemis t ry 1 4:1633-35 19. Hashimoto, Y. & K. Kawanishi (1976) New A l k a l o i d s from B a n i s t e r i o p s i s c a a p i . Phytochemi s t r y 15:1559-60 20. Kawan i sh i , Y . , Y. Uhara & Y. Hashimoto (1982) Sh ihun ine and D ihyd rosh ihun ine from B a n i s t e r i o p s i s c a a p i . J ou rna l of Na tu r a l P roduc ts 45:637-8 21. W. A. Rod r i gues . (1980) Rev isao Taxonomica das E s p e c i e s de V i r o l a Aub le t ( M y r i s t i c a c e a e ) do B r a s i l . Ac ta Amazonica 1:1-123 22. A g u r e l l , S . , B. Ho lms ted t , J . E. L i n d g r e n , & R. E. S c h u l t e s . (1968) I d e n t i f i c a t i o n of Two New /J-carboline A l k a l o i d s in South American H a l l u c i n o g e n i c P l a n t s . B i o chemica l  Pharmacology 17:2487-88 27 23. B. Ho lmsted t , J . E. L i n d g r e n , T . Plowman, L. R i v i e r , R. E. S c h u l t e s & 0 . T o v a r . (1980) Indole A l k a l o i d s i n Amazonian M y r i s t i c a c e a e : F i e l d and Labora to r y Resea rch . Harvard B o t a n i c a l Museum L e a f l e t s 28:215-234 24. J . M. Cassady , G. E. B l a i r , R. F. Ra f fau f & V. E. T y l e r . (1971) The I s o l a t i o n of 6-methoxyharmalan and 6-methoxyharman from V i r o l a c u s p i d a t a . L l o y d i a 34:161-162 25. A l l e n , J . R. F . , & B. Holmstedt (1980) The Simple 0-c a r b o l i n e A l k a l o i d s . Phy tochemis t ry 19:1573-1582 26. S c h u l t e s , R. E. & B. Holmstedt (1968) The V e g e t a l I ng r ed i en t s of the M y r i s t i c a c e o u s Snu f f s of the Northwest Amazon. Rhodora 70:113-160 27. S c h u l t e s , R. E. (1969) V i r o l a as an O r a l l y - a d m i n i s t e r e d H a l l u c i n o g e n . Harvard B o t a n i c a l Museum Leaf l e t s 22:229-40 28. S c h u l t e s , R. E. & T . Swain (1976) Fu r the r Notes on V i r o l a as an O r a l H a l l u c i n o g e n . J ou rna l of P s y c h e d e l i c Drugs 8:317-324 29. S c h u l t e s , R. E . , T . Swain, & T . Plowman (1977) V i r o l a as an O r a l H a l l u c i n o g e n Among the Boras of Pe ru . Harvard B o t a n i c a l Museum L e a f l e t s 25:259-272 30. A g u r e l l , S . , B. Ho lmsted t , & J . E. L i ndgren (1969) A l k a l o i d s i n C e r t a i n Spec ies of V i r o l a and Other South American P l a n t s of E thnopharmaco log ic I n t e r e s t . Ac ta Chemica S cand inav i c a 23:903-916 31. S h u l g i n , A. T . (1976) Psychotomimet ic Agen t s . c h . 4 in Maxwell Gordon (ed. ) P sychopharmaco log i ca l Agents V o l . IV. Academic Press 32. U d e n f r i e n d , S. , B. Wi tkop, B. G. R e d f i e l d , & H. Weissbach (1958) S tud i e s wi th R e v e r s i b l e I n h i b i t o r s of Monoamine O x i d a s e : Harmal ine and Re l a t ed Compounds. B iochemica l  Pharmacology 1:160-165 33. M c l s a a c , W. M. & V. E s t evez (1966) S t r u c t u r e - a c t i o n R e l a t i o n s h i p s of /3-carbol ines as Mono-amine Oxidase I n h i b i t o r s . B i ochemica l Pharmacology 26:1625-27 28 34. B u c k h o l t z , N. S. & W. 0 . Boggan (1977) Monoamine Oxidase I n h i b i t i o n in B ra in and L i v e r Produced by 0 - c a r b o l i n e s : S t r u c t u r e - a c t i v i t y R e l a t i o n s h i p s and Subs t r a t e S p e c i f i c i t y . B i o chemica l Pharmacology 26:1991-96 35. S q u i r e s , R. F. (1978) Monoamine Oxidase I n h i b i t o r s : Animal Pharmacology. in L. L. I v e r son , S. D. I v e r s o n , & S. H. Snyder (eds . ) Handbook of Psychopharmacology V o l . 14. Plenum P r e s s , New York . 36. Chagnon, N . , P. LeQuesne, & J . M. Cook (1971) Yanomamo H a l l u c i n o g e n s : A n t h r o p o l o g i c a l , B o t a n i c a l , and Chemical F i n d i n g s . Cur rent Anthropo logy 12:72-74 37. Harner , M. (ed. ) (1973) H a l l u c i n o g e n s and Shamanism. Oxford U n i v e r s i t y P r e s s , London. 38. F u r s t , P. T . (ed.) (1976) Ha l l u c i nogens and C u l t u r e . Chand le r and Sharp, San F r a n c i s c o . 39. Reichel-Dol-matof f , G. (1975) The Shaman and the J aguar : A Study of N a r c o t i c Drugs Among the Ind ians of Co lomb ia . Temple U n i v e r s i t y P r e s s , P h i l a d e l p h i a . 40. Chagnon, N. (1968) Yanomamo: The F i e r c e People• H o l t , R i n e h a r t , and Winston, New York . 41. De R i o s , M. Dobkin (1972) V i s i o n a r y V i n e : P s y c h e d e l i c H e a l i n g in the Peruv ian Amazon. Chand le r , San F r a n c i s c o . 42. De R i o s , M. Dobkin (1971) Ayahuasca : The H e a l i n g V i n e . I n t e rna t i o n a l J ou rna l of Soc i a l P s y c h i a t r y 17:256-69 43. De R i o s , M. Dobkin (1970) B a n i s t e r i o p s i s i n W i t c h c r a f t and H e a l i n g in I q u i t o s , Pe ru . Economic Botany 24:296-300 29 CHAPTER 11: CHEMISTRY AND PHARMACOLOGY OF TRYPTAMINES AND 0-CARBOLINES I. Occu r r ence , D i s t r i b u t i o n , and B i o s y n t h e s i s of Tryptamines and fl-carbolines The s imple d e r i v a t i v e s of t r yp tamine and 0 - c a r b o l i n e ( F i g . 1 & 2) are among the most w ide ly d i s t r i b u t e d a l k a l o i d s in the . p l a n t , a n i m a l , and funga l kingdoms [ 1 , 2 ] . As of 1977, some 19 s imple d e r i v a t i v e s of t ryptamine had been i d e n t i f i e d in na tu re , d i s t r i b u t e d among 26 h igher p l an t f a m i l i e s and the A g a r i c a l e s [ 1 ] ; new s p e c i e s are c o n t i n u a l l y be ing added to the l i s t ( c f . , e . g . , [3]) and undoubtedly the t a b u l a t i o n p resen ted in [1] s tands in need of r e v i s i o n . At the t ime of p u b l i c a t i o n of [2] in 1980, 64 s imp le /3-carboline d e r i v a t i v e s d i s t r i b u t e d among 25 h ighe r p l a n t f a m i l i e s and 3 s p e c i e s of fung i had been i d e n t i f i e d . Comparison of the s p e c i e s c i t e d in [1] and [2] shows tha t in many i n s t a n c e s , p l a n t s c o n t a i n i n g t ryptamine d e r i v a t i v e s a l s o c o n t a i n s t r u c t u r a l l y r e l a t e d /3-carbol ines ; t h i s obse r v a t i on i s not s u r p r i s i n g s i n ce /3-carbol ines are b i o s y n t h e s i z e d from t ryp tophan and/or t ryptamine v i a condensa t ion wi th 1 or 2 carbon m o i e t i e s [4 ,5 ] or v i a N-ace t y l a t i on of t r yp tamine fo l l owed by c y c l o d e h y d r a t ion to 3 , 4-dihydro-/3-carbol ines [ 6 ] . The b i o s y n t h e s i s of t ryptamine d e r i v a t i v e s has been rev iewed by Smith [ 7 ] , Both t ryptamine d e r i v a t i v e s and /3-carbolines have been d e t e c t e d as endogenous m e t a b o l i t e s in mammals, i n c l u d i n g man. Bu fo ten ine and v a r i o u s r e l a t e d 5-hydroxy- indo le thy lamines are c o n s t i t u e n t s of f r og and toad venoms [ 8 ] , be ing common in the genera H y l a , L e p t o d a c t y l u s , Rana, and Bu fo . A wider spectrum of Figure 1 - Some Naturally-Occurring Tryptamine Derivatives* Name of Abbreviation Substitution Pattern: Compound R i R> R , R a R> R t tryptophan H H COOH H H H 5-hydroxy-tryptophan H H COOH H OH H tryptamine TA H H H H H H 5-hydroxy-tryptamine 5HT H H H H OH H N-methy1 -tryptamine NMT H CH, H H H H 5-methoxy-tryptamine 5-MeO-T H H H H OCH, H 6-methoxy-tryptamine 6-MeO-T H H H H H OCH, 5-methoxy-N-methy1 -tryptamine 5-MeO-NMT H CH, H H OCH, H 5-methoxy-N-acety1 -tryptamine H 0=CCHJ H H OCH, H (melatonin) N.N-dlmethyl tryptamine DMT C H i CH, H H H H 5-hydroxy-N,N-dimethyl-tryptamine 5H0-DMT CH, CH, H H OH H (bufotenine) 5-methoxy-N,N-d1methyl-tryptamine 5MeO-DMT C H i CH, H H OCH, H 4-hydroxy-N,N-dimethyl-tryptamine 4H0-DMT C H i CH, H OH H H ( p s i l o c l n ) 4-phosphoryl-N.N-dlmethyl -tryptamine C H i CH, H OPO,H H H (psilocybin) 4-phosphoryl-N-methy 1 -tryptamine H CH, H 0PO,H H H (baeocystIne) 4-phosphoryl-tryptamine H H . H OPO,H H H (norbaeocys 11ne) N-methyl-tryptophan-methyl ester H CH, COOCH, H H H (abrlne methyl ester) N.N-dlmethyl-tryptophan - CH, CH, COOH H H H *c f . Table 1 for structures of some psychoactive synthetic tryptamine derivatives. Figure 2 - Some Naturally-Occurring 0-carboline Derivatives Additional Substitutents T r i v i a l Name Ring Type R = & Their Location Abbreviat ion norharman A H - -norharma1 an B H - -tetrahydro-norharman C H - -harman A C H , - -harma1 an B C H , - -tetrahydro-harman C C H , - -harmine A C H , c -0CH, -harma1ine B C H , c -0CH, -tetrahydro-harmine C C H , c -0CH, -harmic amide A CONH. c -0CH, -acetyl norharmine A C=OCH, c -0CH, -harmine N-oxide A C H , c -0CH,. Ni-->0 -ketotetrahydro-norharm i ne C =0 c -OCH, -harmic acid methyl ester A C00CH, c -OCH, -harma1inic acid B C O O H c -OCH. -harmol A C H , c - O H -harmalol B C H , c - O H -6-methoxy-harman A C H , c . - O C H i -6-methoxy-harma1 an B C H , C i -OCH, -6-methoxy-tetrahydro-harman C C H , C I -OCH, -6-methoxy-2-methy1-tetrahydro-0-carbol1ne C H N, - C H , , C.-0CH, 6-MeO-MTHjC 6-methoxy-1,2-dlmethyl-tetrahydro 0-carboline c C H , N i - C H , , Ct-OCH, 6-Me0-DMTHflC 2-methyl-tetrahydro-0-carboline c H N i - C H , MTHflC 1,2-dimethy 1-tetrahydro-0-carboline c C H , N i - C H , DMTHjC 0-carboline-3-carboxylate A H c , -COOH flCC 0-carboline-3-carboxy1 ate ethyl ester A H c , -C00CH, 0CCEE tetrahydroharman-3-carboxylate C C H , c , i— -COOH ~ b r e v i c o l i n e A C H , c u> 32 bu fo t en ine d e r i v a t i v e s i s found in the l a s t genus, i n c l u d i n g b u f o v i r i d i n e , the s u l f a t e e s t e r of b u f o t e n i n e , dehyd robu fo t en ine , and i t s s u l f a t e e s t e r , b u f o t h i o n i n e . A l though i t i s c l o s e l y r e l a t e d to the h a l l u c i n o g e n i c t r yp tamine d e r i v a t i v e s , bu fo ten ine i t s e l f i s not h a l l u c i n o g e n i c (see below, and [9 ] ) , a c t i n g as a p r e s so r r a the r than a h a l l u c i n o g e n in man. The sk in of Bufo a l v a r i u s c o n t a i n s 5-MeO-DMT at the r a the r s t a g g e r i n g c o n c e n t r a t i o n of 50-160 mg/g of s k i n [ 8 ] ; t h i s i s the on l y Bufo spec i e s known to c o n t a i n a h a l l u c i n o g e n i c t r yp t am ine . Methy l t r a n s f e r a s e s which c a t a l y z e the s y n t h e s i s of t r yp tamines i n c l u d i n g DMT, 5-MeO-DMT, and bu fo t en ine have been c h a r a c t e r i z e d in human l u n g , b r a i n , b l o o d , c e r e b r o s p i n a l f l u i d , l i v e r , and h e a r t , and a l s o in r a b b i t l u n g , t o a d , mouse, s t e e r , gu inea p i g , and baboon b r a i n s , as we l l as in o ther t i s s u e s in these s p e c i e s [10 ] . The _i_n v i vo fo rmat ion of DMT from l a b e l l e d p r e c u r s o r s has been demonstrated in r a b b i t lung and in ra t b r a i n [10 ] . The s t a t e of knowledge of DMT b i o s y n t h e s i s in mammals as of 1981 has been summarized in [10 ] . Endogenous /3-carboline d e r i v a t i v e s have a l s o been de t e c t ed in human and ra t t i s s u e s ( c f . s e c t i o n I I I .A , be low) . Both t e t r ahydro - / 3~ca rbo l ines and the f u l l y a romat i c d e r i v a t i v e s have been de t e c t ed [11 ] . An aromat ic /3-carboline has been i s o l a t e d from ag ing human l ens p r o t e i n [12] which i s i n t e r e s t i n g in l i g h t of the r e c e n t l y d i s c o v e r e d photo-c y t o t o x i c i t y of a romat i c /3-carbol ines [ 13 ] . Endogenous /3-c a r b o l i n e d e r i v a t i v e s and t e t r a h y d r o i s o q u i n o l i n e s have been de t e c t ed in human u r i ne f o l l o w i n g e thano l l o a d i n g . Re fe rences [11,14] p rov ide comprehensive recent r ev i ews . 33 11. Pharmacology of H a l l u c i n o g e n i c Tryptamines A. S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s Most c l i n i c a l and pha rmaco log i c a l s t u d i e s w i th h a l l u c i n o g e n i c t r yp tamines have focused on t h e i r _i_n v i t r o metabol i sm and/or the p o s s i b l e endogenous s y n t h e s i s of methy la ted t r yp tamine d e r i v a t i v e s in p a t h o l o g i c a l s t a t e s such as s c h i z o p h r e n i a . There i s a p a u c i t y of s c i e n t i f i c l i t e r a t u r e on the sys temat i c i n v e s t i g a t i o n of s t r u c t u r a l i n f l u e n c e s on the h a l l u c i n o g e n i c a c t i v i t y of t r yp tamine d e r i v a t i v e s in humans. I n v e s t i g a t i o n s of s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of h a l l u c i n o g e n s are comp l i c a t ed by the f a c t that no adequate an imal m o d e l . e x i s t s fo r t h i s pu rpose ; the drug d i s c r i m i n a t i o n t e s t deve loped by Appel and co-workers [15] r ep r e sen t s a s tep in the r i g h t d i r e c t i o n , however. As a r e s u l t , much of the c u r r e n t l y a v a i l a b l e i n f o rma t i on i s d e r i v e d from "underground" sources of dub ious c r e d i b i l i t y . Shu lg in [16,17] has summarized the a v a i l a b l e i n f o rma t i on in two recent r ev i ews . The s u b s t i t u t i o n s i t e s on the t r yp tamine nuc l eus tha t are important de te rminants of h a l l u c i n o g e n i c a c t i v i t y a re the i n d o l e r i n g , the s i de-cha in c a r b o n s , or the s i d e - c h a i n n i t r o g e n ( c f . Tab le I ) . The N-a lky l homologs of DMT in which the N,N-dimethyl s u b s t i t u e n t s are r e p l a c e d wi th more a l i p h a t i c m o i e t i e s i n c l u d e N,N-d i e t h y l t r y p t a m i n e (DET), N ,N-d ip ropy l t r yp tamine (DPT), N,N-d i i s o p r o p y l t r y p t a m i n e (DIPT) , N , N - d i a l l y l t r y p t a m i n e (DAT), and N ,N-d ibu t y l t r yp t am ine (DBT). A l l of these homologs are p s y c h o a c t i v e i n man except fo r N , N - d i b u t y l t r y p t a m i n e , and a l l a re a p p a r e n t l y o r a l l y a c t i v e except f o r DMT i t s e l f , which i s Table I - Orally and Parenterally Active Psychotropic Tryptamine Derivatives* Name of Compound R, R i Substitution R, Pattern R« R. Dosage (mg) Route: Oral/Parenteral tryptamine H H H H H 100t par/ora1? DMT CH, CH, H H H 60 par DET C H , CH, H H H 60 par/ora1 DPT n-prop n-prop H H H 60 par/ora1 OAT C,H» CH, H H H 60 par/ora1 DIPT i-prop 1-prop H H H 30 ora 1 5Me0-DIPT i-prop i-prop H H OCH, 12 ora 1 5MeO-DMT CH, CH, H H OCH i 6 par p s i l o c i n CH, CH, H OH H 12* ora 1 CZ-74 CiH% C H» H OH H 15* ora 1 serotonin H H H H OH 100* ora 1 bufotenine CH, CH, H H OH 16b par IT-290 H H CH, H H 30 oral 4-hydroxy-a-methy1 -tryptamine H H CH, OH H 20# ora 1 MP-809 H H CH, H CH, 606 ora 1 5-f1uoro-a-methy1 -tryptam i ne H H CH, H F 25* ora 1 5-methoxy-a-methy 1 -tryptamlne H H CH, H OCH, 3 ora 1 4-hydroxy-d i i sopropy1 -tryptamine 1 -prop 1-prop H OH H 12* ora 1 4-hydroxy-N-1sopropyl,N-methy1 -tryptamine i-prop CH, H OH H 6* ora 1 N-t-butyl-. tryptamine H t-butyl H H H ?* ora 1 3-[2-(2,5-dimethylpyrrolyl)ethyl ]-indole H H H ? ? ( sIde-cha1n=1-ethyl(2,5-dimethylIpyrrole) * Data compiled from [9,16,17.18]. t Autonomic symptoms; l i t t l e central a c t i v i t y . * The phosphate esters are psilocybin and CEY-19. respectively; both are stoichiometr1ca11y equivalent to the 4-hydroxy isomers. * Cardiovascular and autonomic symptoms; l i t t l e central a c t i v i t y , b A pressor amine rather than a hallucinogen in man. ^ An antidepressant rather than a hallucinogen In man. * Based on anonymous reports in the lay press. No c l i n i c a l studies have been published. 35 o r a l l y i n a c t i v e in doses exceed ing 1000 mg ( c f . Tab l e I ) . Presumably the o r a l i n a c t i v i t y of DMT i s due to i t s deaminat ion by monoamine ox idase ( c f . d i s c u s s i o n below) and those d e r i v a t i v e s hav ing b u l k i e r N-a lky l s u b s t i t u e n t s are o r a l l y a c t i v e due to s t e r i c h indrance of the enzyme. Potency of a l l of the N , N - d i a l k y l d e r i v a t i v e s ment ioned above i s c o n s i d e r a b l y enhanced by hyd roxy l s u b s t i t u t i o n at the i n d o l e 4 - p o s i t i o n ; t h i s s u b s t i t u t i o n a l s o c o n f e r s o r a l a c t i v i t y on the parent compound, DMT ( p s i l o c i n = 4-hydroxy-DMT). The mechanism u n d e r l y i n g the o r a l a c t i v i t y of p s i l o c i n i s u n e l u c i d a t e d , but i t p robab l y i s due to the fo rmat ion of an i n t r a m o l e c u l a r i o n i c bond between the a n i o n i c r i n g hyd roxy l and the charged s i d e - c h a i n n i t r o g e n ; t h i s c o n f i g u r a t i o n c o u l d form a "pseudo" C r i n g and thus p r o t e c t the s i d e - c h a i n from deaminat ion [ 9 3 . I n t e r e s t i n g l y the 5-hydroxyl isomer of p s i l o c i n , b u f o t e n i n e , i s i n a c t i v e as an h a l l u c i n o g e n and a c t s p r i m a r i l y on the p e r i p h e r a l autonomic sys tems, c aus i ng severe c a r d i o v a s c u l a r s t i m u l a t i o n , s a l i v a t i o n , h y p e r t e n s i o n , l a c h r y m a t i o n , and h y p e r v e n t i l a t i o n , but no c e n t r a l e f f e c t s [17 ] . Appa ren t l y the 5-hydroxyl s u b s t i t u t i o n does not permi t the fo rmat ion of the i n t r a m o l e c u l a r z w i t t e r i o n i c bond and the a n i o n i c c h a r a c t e r of the oxygen s u b s t i t u e n t i n t e r f e r e s w i th the uptake of the compound i n t o the c e n t r a l nervous sys tem. The 5-0-methyl ana logue of bu fo t en ine (5-MeO-DMT) i s p a r e n t e r a l l y a c t i v e as an h a l l u c i n o g e n in man but i s not o r a l l y a c t i v e . 5-MeO-DMT e x h i b i t s s i m i l a r p s y c h o l o g i c a l and somat ic symptoms in man as DMT, but i s approx imate l y an o rder of magnitude more potent on a m i l l i g r a m b a s i s ( c f . Tab le I ) . Both 5-MeO-DMT and b u f o t e n i n e are c l o s e l y r e l a t e d s t r u c t u r a l l y to the CNS n e u r o t r a n s m i t t e r 36 s e r o t o n i n . The N , N - d i i s o p r o p y l ana logues of DMT and 5-MeO-DMT are both o r a l l y a c t i v e as h a l l u c i n o g e n s in man [18 ] . L i t t l e i s known of the h a l l u c i n o g e n i c a c t i v i t y of the N-monosubst i tu ted, N - d e a l k y l , or N - c y c l o a l k y l t r y p t a m i n e s ; the mono-te r t-buty l d e r i v a t i v e i s " r e p u t e d " to be o r a l l y a c t i v e in the underground 1 i t e r a t u r e [17 ] . In a d d i t i o n to i n d o l e r i n g s u b s t i t u e n t s and a l i p h a t i c N-a l k y l s u b s t i t u e n t s , the s ide cha in a-ca'rbon r ep r e sen t s a t h i r d s u b s t i t u t i o n s i t e a f f e c t i n g h a l l u c i n o g e n i c a c t i v i t y . Methy l s u b s t i t u t i o n of the a-carbon c o n f e r s o r a l psychotomimet ic a c t i v i t y on the compounds a-methy l t ryptamine and 5-MeO-a-methy l t r yp tamine [ 9 , 1 7 ] . The mechanism of o r a l a c t i v i t y in the case of these analogues i s undoubtedly r e l a t e d to s t e r i c h indrance of enzymatic deaminat ion by the a - s u b s t i t u e n t . a-methy l t r yp tamine and a-e thy l t r yp tamine have been shown to ac t as c o m p e t i t i v e i n h i b i t o r s of MAO [19 ] . No i n fo rma t i on i s a v a i l a b l e on the a c t i v i t y of a - s u b s t i t u t e d N , N - d i a l k y l t r y p t a m i n e s . B. Metabo l i sm of H a l l u c i n o g e n i c Tryptamines The s y n t h e t i c and deg rada t i v e metabol ism of DMT in mammals has been r e c e n t l y reviewed [10 ] . Indo le N- and O-methyl t r a n s f e r a s e s which c a t a l y z e the s y n t h e s i s of DMT, 5-MeO-DMT, and bu fo t en ine have been c h a r a c t e r i z e d in human l u n g , b r a i n , b l o o d , and c e r e b r o s p i n a l f l u i d 1 2 0 ] . T r yp tamine , 5-hyd roxy t r yp tamine , and N-methyl t ryptamine have been i d e n t i f i e d as s u b s t r a t e s fo r i ndo l e-N-me thy l t r ans f e r a ses but the re i s c o n s i d e r a b l e v a r i a t i o n in s u b s t r a t e s p e c i f i c i t y in d i f f e r e n t organisms and t i s s u e s . Two INMTs have been c h a r a c t e r i z e d in the A u s t r a l i a n grass P h a l a r i s 37 tube rosa which have d i f f e r e n t a f f i n i t i e s fo r the pr imary amine ( t r yp tamine ) and the secondary amine (NMT), i n d i c a t i n g that both are r e q u i r e d in the b i o s y n t h e s i s of the t e r t i a r y amine [21 ] . Presence of two or more types of INMT in mammals has not been proven but would e x p l a i n the v a r y i n g subs t r a t e a f f i n i t i e s in d i f f e r e n t t i s s u e s of the same s p e c i e s . S-adenosyl meth ion ine (SAM) f u n c t i o n s as the methyl donors in t h i s t r a n s m e t h y l a t i o n r e a c t i o n ; however, both SAM and 5 - m e t h y l t e t r a h y d r o f o l i c a c i d (MTHF) have been found to p a r t i c i p a t e in the s y n t h e s i s of 2-m e t h y l - t e t r a h y d r o - 0 - c a r b o l i n e (MTH/3C) and tet rahydro-/3-carbol ine (TH0C) when incubated i_n v i t r o wi th NMT and t r yp t am ine , r e s p e c t i v e l y [22 ] . The te t rahydro-/3-carbo l ine fo rmat ion probab ly occu rs v i a the enzymatic fo rmat ion of HCHO from the methyl donors f o l l o w e d by non-enzymatic condensa t ion w i th the i n d o l e s u b s t r a t e s v i a a P i c t e t - S p e n g l e r r e a c t i o n . Both DMT and SAH, the de-methyl d e r i v a t i v e of SAM, are potent i n h i b i t o r s of the INMT a c t i v i t y [22 ] . P o s s i b l e mechanisms i n v o l v e d in the r e g u l a t i o n of the INMT a c t i v i t y have been reviewed by Ba rke r , et a l . [10 ] . Degrada t i ve metabol ism of DMT has a l s o been r e c e n t l y i n v e s t i g a t e d [22] and i s rev iewed i n [10 ] , A q u a n t i t a t i v e study of DMT metabol ism in ra t whole b r a i n homogenates us ing deu t e r a t ed DMT [22] found I AA, NMT, MTH/JC, and DMT-N-ox ide as m e t a b o l i t e s . IAA was the major m e t a b o l i t e when DMT was incubated at 6 x 1 0 ~ 8 M, but at 2 x I O ' 5 M, DMT-NO was the major m e t a b o l i t e . I ncuba t ion of 6 x 1 0 " 8 M DMT in homogenates obta ined from r a t s p r e t r e a t e d w i th the MAO i n h i b i t o r i p r o n i a z i d r e s u l t e d in the i n h i b i t i o n of IAA fo rmat ion by 83%; however NMT and DMT-NO fo rma t i on , was i n h i b i t e d by 90%, and no MTH/3C was formed. 38 Based on these o b s e r v a t i o n s , the au thors [22] s p e c u l a t e d that a h igh p r o p o r t i o n of IAA probab ly a rose as a secondary m e t a b o l i t e r e s u l t i n g from the ox idat ive- deaminat ion of NMT. DMT i t s e l f i s a poor s u b s t r a t e fo r MAO [ 2 3 , 2 4 ] ; the r e l a t i v e r a te of o x i d a t i o n of NMT i s some 9 t imes f a s t e r than DMT and 280 t imes f a s t e r than DMT-NO. The t e t r a h y d r o - 0 - c a r b o l i n e s de t e c t ed as t r a c e m e t a b o l i t e s may be formed from the nonenzymatic condensa t ion of t r yp tamine and/or NMT wi th the HCHO formed as an i n t e rmed i a t e in the N-demethylat ion of DMT. Barker et a l . [22] have made the i n t e r e s t i n g o b s e r v a t i o n that d i r e c t C-hyd roxy l a t i on of t e r t i a r y amines and t e r t i a r y amine N-oxide rearrangement r e s u l t s in the fo rmat ion of i d e n t i c a l i n t e rmed i a t e s ( c a r b i n o l a m i n e s , i m i n i u m ions ) which can undergo i n t r a m o l e c u l a r c y c l i z a t i o n to t e t r ahydro-/3-ca rbo l i nes . E a r l i e r s t u d i e s of DMT metabol i sm in ra t l i v e r microsomes ob ta ined from ao ima ls p r e t r e a t e d w i th i p r o n i a z i d de t e c t ed 6-hydroxy-DMT and 6-hydroxy-DMT-NO in a d d i t i o n to t r yp t am ine , NMT, and DMT-NO, g i v i n g r i s e to s p e c u l a t i o n tha t 6-hyd roxy l a t i on might be an important me tabo l i c pathway fo r DMT [25 ] . More recent s t u d i e s have shown t h a t , wh i le 6-hyd roxy l a t i on i s c h a r a c t e r i s t i c of DMT metabol ism in p e r i p h e r a l t i s s u e s , i t appa r en t l y does not occur in b r a i n [22 ] . Other i n v e s t i g a t o r s have focused on the metabo l i sm of 4-hydroxy-DMT ( p s i l o c i n ) [ 2 6 , 2 7 ] . These s t u d i e s i n d i c a t e that o x i d a t i v e deaminat ion of the s i d e - c h a i n may be a r e l a t i v e l y minor route of deg rada t i on fo r t h i s compound. A me tabo l i c study of the f a t e of 1 " C - p s i l o c i n in the ra t [26] i n d i c a t e d tha t unchanged p s i l o c i n and 4-hydroxy-IAA accounted fo r on l y 40% of t o t a l u r i n a r y m e t a b o l i t e s , the remainder be ing p resen t as 39 ext remely h y d r o p h i l i c m e t a b o l i t e s , which cou ld not be p o s i t i v e l y i d e n t i f i e d as g l u c u r o n i d e s . H o r i t a and Weber [27] s t u d i e d the i n c u b a t i o n of p s i l o c y b i n in ra t k idney homogenates. They found tha t p s i l o c y b i n was r e a d i l y dephosphory l a t ed to p s i l o c i n by a l k a l i n e phosphatase ; p s i l o c i n was then r a p i d l y me tabo l i zed to a b l u e - c o l o r e d product which they specu l a t ed was the o-quinone of p s i l o c i n . The fo rmat ion of the b lue product was u n a f f e c t e d by the presence of MAO i n h i b i t o r s , but c o u l d be i n h i b i t e d by KCN. Subsequent l y , p s i l o c i n and other h yd roxy- indo l e s i n c l u d i n g s e r o t o n i n and bu fo t en ine were shown to be o x i d i z e d to c o l o r e d p roduc t s in the presence of mammalian cytochrome ox idase [28 ] . The s i g n i f i c a n c e of these metabo l i c i n v e s t i g a t i o n s of DMT, p s i l o c i n , and r e l a t e d t r yp tamines l i e s in the r e c o g n i t i o n that o x i d a t i v e deaminat ion by MAO i s not n e c e s s a r i l y the o n l y , or even the major , pathway a v a i l a b l e fo r the deg rada t i v e metabol ism of these compounds. C. H a l l u c i n o g e n i c Tryptamines and MAO I n h i b i t i o n The a c t i v i t y of some t ryp tamine d e r i v a t i v e s as MAO i n h i b i t o r s has been i n v e s t i g a t e d [ 2 3 , 2 4 , 2 9 ] . U n l i k e the /3-c a r b o l i n e s , however, ex t ens i v e s t u d i e s of the s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of t r yp tamines wi th respec t to MAOI a c t i v i t y have not been c a r r i e d o u t . L e s s i n and co-workers [29] examined the SAR of a s e r i e s of s u b s t i t u t e d t r yp tamines and /3-carbol ines on a c t i v i t y as MAOI and i n h i b i t o r s of 5HT uptake . Among the t r yp tamines the most potent ana log was 6-MeO-a,a-d ime thy l t r yp t am ine which had 1.5% of the i n h i b i t o r y potency of h a r m a l i n e . The a c t i v i t y of N ,N-d imethy l t r yp tamine , 2-methyl-DMT 40 and 5-benzyloxy-DMT as MAOI in the guinea p i g l i v e r was i n v e s t i g a t e d [23] at v a r i o u s i n h i b i t o r and s u b s t r a t e c o n c e n t r a t i o n s . In a l l cases DMT had s i g n i f i c a n t l y more a c t i v i t y than the o ther d e r i v a t i v e s . The a c t i v i t y of DMT as MAOI was a l s o i n f l u e n c e d by the s u b s t r a t e ; g r e a t e s t i n h i b i t i o n was observed wi th 5HT as s u b s t r a t e , i n t e rmed i a t e i n h i b i t i o n wi th tyramine as s u b s t r a t e , wh i le the lowest a c t i v i t y was found wi th t ryptamine as s u b s t r a t e . These r e s u l t s are c o n s i s t e n t wi th the p o s t u l a t e tha t DMT may be a s p e c i f i c i n h i b i t o r of MAO-A ( c f . Chapter V I ) . Ho et a l . [24] i n v e s t i g a t e d the MAOI a c t i v i t y of a s e r i e s of 5-s u b s t i t u t e d gramines , a-methy l t r yp tamines , and d ime thy l t r yp t am ines us ing t r yp tamine as s u b s t r a t e . The order of MAOI a c t i v i t y of the t ryptamine d e r i v a t i v e s t e s t e d was DMT>5-methyl-DMT>5 1MeO-DMT>5-hydroxyl-DMT. The DMT d e r i v a t i v e s were g e n e r a l l y more potent than the gramine d e r i v a t i v e s except fo r 5-bromogramine which gave comparable i n h i b i t i o n at 72% of the c o n c e n t r a t i o n of DMT. The p o t e n t i a t i o n of the b e h a v i o u r a l and pha rmaco log i c a l e f f e c t s of t r yp tamine d e r i v a t i v e s by MAO i n h i b i t o r s has been i n v e s t i g a t e d , a l though the s p e c i f i c q u e s t i o n of the o r a l p o t e n t i a t i o n of DMT and other p a r e n t e r a l l y - a c t i v e d e r i v a t i v e s has a p p a r e n t l y not been i n v e s t i g a t e d . The e f f e c t of DMT in human v o l u n t e e r s was assessed be fo re and 3 days a f t e r t reatment wi th the MAOI i p r o n i a z i d [30 ] . P a t i e n t s r e c e i v i n g DMT at a reduced dose f o l l o w i n g the i p r o n i a z i d t reatment expe r i enced none of the v i s u a l h a l l u c i n a t i o n s or d i s t u r b a n c e s of t ime and space p e r c e p t i o n which t y p i f y the symptoms of the d r u g ; they r epo r t ed on l y a f e e l i n g of " s t r a n g e n e s s " . P a t i e n t s r e c e i v i n g a dose 41 e q u i v a l e n t to tha t g iven p r i o r to i p r o n i a z i d had a two-phase r e sponse . The f i r s t stage was s i m i l a r to the usua l DMT p s y c h o s i s , but l e s s pronounced; i l l u s i o n s and h a l l u c i n a t i o n s were p r e s e n t , but l e s s c o l o r f u l , and on ly man i f e s t ed w i th the eyes c l o s e d ; the second phase was c h a r a c t e r i z e d by a p e r s i s t e n t f e e l i n g of " s t r a n g e n e s s " to which the p a t i e n t s o f t en r eac t ed n e g a t i v e l y , or i n d i f f e r e n t l y . Based on these t r i a l s the au thors [30] specu l a t ed that the reduced e f f e c t s may have been due to the h ighe r 5HT c o n c e n t r a t i o n in the b r a i n due to MAO i n h i b i t i o n , thus m i t i g a t i n g the 5HT b l o c k i n g e f f e c t s of DMT. T h i s o b s e r v a t i o n was a l s o suppor ted by the o b s e r v a t i o n that p r i o r a d m i n i s t r a t i o n of 1-methy l-d- lyserg ic a c i d bu tano lamide , a power fu l s e r o t o n i n a n t a g o n i s t , g r e a t l y exacerba ted the psychotomimet ic e f f e c t s of DMT [31 ] . Moore and co-workers [32] s t u d i e d the e f f e c t s of i p r o n i a z i d , ch l op romaz ine , and meth io thep in on DMT-induced changes in body tempera tu re , p u p i l l a r y d i l a t a t i o n , b lood p r e s su re and EEG in r a b b i t s . Ch lo rpromaz ine a t t e n t u a t e d or b locked the e f f e c t s of DMT on these parameters but i p r o n i a z i d p ro longed the e l e v a t e d r e c t a l temperatures and m y d r i a s i s induced by DMT. A r t e r i a l b lood p r e s s u r e and EEG were not markedly a l t e r e d by pre-t reatment w i th i p r o n i a z i d . Wang-Lu and Domino [33] i n v e s t i g a t e d the e f f e c t of the MAOI i p r o n i a z i d and t r any l c yp romine on DMT h a l f - l i f e in r a t l i v e r and b r a i n and found a g r e a t l y i n c r e a s e d h a l f - l i f e in both t i s s u e s a f t e r t r ea tment . T rany l cyp romine p ro longed DMT h a l f - l i f e more than i p r o n i a z i d . I n t e r e s t i n g l y , t reatment w i th a l a r g e r dose of DMT in the absence of MAOI p ro longed the h a l f - l i f e in b r a i n but not in l i v e r . The au tho rs s p e c u l a t e d tha t t h i s may be 42 due to the f a c t that DMT i t s e l f i s a weak MAOI and that i t s c l e a r a n c e from b r a i n i s ma in ly dependent on MAO. L i v e r may have other enzymes capab le of m e t a b o l i z i n g DMT even though MAO i s i n h i b i t e d . H a l f - l i f e in b r a i n and l i v e r was s i m i l a r in the presence of MAOI i n d i c a t i n g tha t s i m i l a r enzymes p a r t i c i p a t e in the metabol ism of DMT f o l l o w i n g MAO i n h i b i t i o n . Shah and Hedden [34] s t u d i e d behav i ou r a l e f f e c t s and metabol ism of DMT in mice p r e t r e a t e d w i th SKF-525A, i p r o n i a z i d , and ch lo rp romaz ine and found tha t i p r o n i a z i d p ro longed the b e h a v i o u r a l e f f e c t s of DMT and a l s o s i g n i f i c a n t l y e l e v a t e d DMT l e v e l s in p lasma, b r a i n , and l i v e r w i th r espec t to c o n t r o l s . SKF-525A, an i n h i b i t o r of a wide v a r i e t y of hepa t i c microsomal enzymes, d i d not p ro l ong the b e h a v i o u r a l e f f e c t s of DMT or r e s u l t in i n c r e a s e d t i s s u e or plasma l e v e l s , thus p r o v i d i n g f u r t h e r ev idence tha t MAO i s the pr imary enzyme i n v o l v e d in the metabol ism of DMT i_n v i v o . It a l s o i n d i c a t e d that the c e n t r a l e f f e c t s of DMT are due to the parent compound r a the r than the 6-hydroxy-metabol i te de t e c t ed in some s t u d i e s [25 ] . 6-hyd roxy-de r i v a t i v e s of DMT and r e l a t e d compounds are i n a c t i v e as h a l l u c i n o g e n s [16 ] . A l though the a v a i l a b l e l i t e r a t u r e i n d i c a t e s tha t MAOI do have s i g n i f i c a n t i n f l u e n c e s on both the metabol ism and behav i ou r a l e f f e c t s of DMT, a p p a r e n t l y the s p e c i f i c i n t e r a c t i o n s of DMT wi th /3-c a r b o l i n e s have not been i n v e s t i g a t e d . T h i s apparent o v e r s i g h t i s e s p e c i a l l y remarkable in view of the c l o s e s t r u c t u r a l r e l a t i o n s h i p s of t r yp tamine d e r i v a t i v e s and 0 - c a r b o l i n e s [ 1 , 2 ] , the p robab le metabo l i c i n t e r c o n v e r s i o n of DMT, o ther t r y p t a m i n e s , and /3-carbol ines [ 2 2 , 5 7 ] , the involvement of both c l a s s e s of compounds in important neu ro- r egu l a to r y f u n c t i o n s 43 such as MAO a c t i v i t y and amine uptake [11 ,14 ,29] and the p robab l e r o l e of t ryptamine//3-carbol ine combina t ions in the mechanism of o r a l a c t i v i t y of the h a l l u c i n o g e n s ayahuasca and the o r a l l y - i n g e s t e d V i r o l a pas tes ( c f . Chapter I ) . The a v a i l a b l e ev idence s t r o n g l y suggests that t r yp tamines and /3-carbol ines are c l o s e l y r e l a t e d , not on ly s t r u c t u r a l l y but a l s o p h a r m a c o l o g i c a l l y , yet very l i t t l e i s known about t h e i r i n t e r a c t i o n s in mammalian sys tems. I I I . B i o chemis t r y and Pharmacology of g-ca rbo l ine D e r i v a t i v e s /3-carbol ines have been known to s c i ence s i n c e the i s o l a t i o n of harmal ine from Peganum harmala by Goebel in 1847, f o l l owed a. few yea rs l a t e r by the i s o l a t i o n of harmine from the same s p e c i e s by J . F r i t s c h e [35 ] . The s t r u c t u r e e l u c i d a t i o n of both compounds was accompl i shed by Manske in 1927, and t h e i r t o t a l s y n t h e s i s was p u b l i s h e d by Spath and Lederer in 1930 [35 ] . In s p i t e of t h e i r long h i s t o r y , many a spec t s of the b i o c h e m i s t r y and pharmacology of /3-carboline d e r i v a t i v e s remain p o o r l y unde r s tood . S ince the d i s c o v e r y in 1959 that harma l ine and ' r e l a t e d d e r i v a t i v e s are c o m p e t i t i v e r e v e r s i b l e i n h i b i t o r s of monoamine ox idase [36 ] , the re has been a resurgence of i n t e r e s t i n /3-carbo l ines . The p resen t s e c t i o n focuses p r i m a r i l y on the psychopharmacology of /3-carbol ines ; o ther a spec t s of t h e i r pharmacology have been d i s c u s s e d in [37] and w i l l on l y be ment ioned he re . 44 A. Psychopharmacology of /3-carboline D e r i v a t i v e s 1. /3-carbol ines as Monoamine Oxidase I n h i b i t o r s The a c t i v i t y of j3-carbol ine d e r i v a t i v e s as c o m p e t i t i v e r e v e r s i b l e i n h i b i t o r s of MAO was f i r s t demonstrated by Uden f r i end and co-workers [36 ] . Subsequent l y , s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s were i n v e s t i g a t e d by Mc lsaac & Es tevez [38 ] , Ho et a l . [39 ] , and Buckho l tz & Boggan [ 4 0 , 4 1 ] , D i r e c t comparison of the r e s u l t s of these s t u d i e s are comp l i c a t ed by the use of d i f f e r e n t an ima ls and t i s s u e s as the source of MAO, and a l s o by the use of d i f f e r e n t s u b s t r a t e s . Other i n v e s t i g a t i o n s [42 ,43 ,44 ,45 ] have p r o v i d e d ev idence that at l e a s t two s p e c i e s of MAO e x i s t ; these forms of MAO, des igna ted MAO-A and MAO-B, have d i f f e r e n t s u b s t r a t e s p e c i f i c i t i e s , a re • d i f f e r e n t i a l l y s e n s i t i v e to v a r i o u s MAO i n h i b i t o r s , and have d i f f e r e n t k i n e t i c p r o p e r t i e s . F u l l e r [42] demonstrated that harmal ine (and presumably o ther /3-carbol ines) i s a s e l e c t i v e i n h i b i t o r of MAO-A. Some SAR s t u d i e s of /3-carboline MAOI a c t i v i t y have used tyramine as s u b s t r a t e [38 ] , o the r s have used t ryptamine [39 ,40 ,41 ] and s t i l l o the r s have used t r yp t am ine , 5HT, and /3-phenylethylamine as s u b s t r a t e [41 ] . 5HT i s a s p e c i f i c s u b s t r a t e of MAO-A whi le t ryptamine and tyramine are s u b s t r a t e s of both MAO A and B [46 ] ; /3-phenylethylamine i s p robab ly me tabo l i zed ma in ly by MAO-B in v i v o [46 ] . In view of the d i f f e r e n t s u b s t r a t e s and enzyme s p e c i e s used in d i f f e r e n t s t u d i e s i t i s not s u r p r i s i n g that there are c o n s i d e r a b l e d i f f e r e n c e s in the I 5 0 v a lues and o ther s t r u c t u r e / a c t i v i t y pa ramete rs . Mc lsaac and Es tevez [38] u s i ng tyramine as s u b s t r a t e 45 found tha t f u l l y a romat ic (3-carbol ines were most a c t i v e as MAOI wh i l e te t rahydro- (3-carbo l ines had the l e a s t a c t i v i t y ; d i h yd ro-d e r i v a t i v e s were i n t e rmed ia t e in po tency , a r e s u l t that agrees w i th other s t u d i e s . L i t t l e d i f f e r e n c e in potency was found between 6- or 7- methoxy l a ted , or u n s u b s t i t u t e d /3-carbolines but h y d r o x y l s u b s t i t u t i o n reduced the a c t i v i t y . Buckho l tz and Boggan [41] u s i ng t ryptamine as s u b s t r a t e r epo r t ed r e s u l t s in genera l agreement w i th those of [38] except that 7-methoxy-/3-carbolines were more potent than 6-methoxy- or u n s u b s t i t u t e d d e r i v a t i v e s ; methyl s u b s t i t u t i o n at C, decreased a c t i v i t y f o r l i v e r MAO but i n c r e a s e d i t fo r b r a i n MAO. Other d i f f e r e n c e s in potency between l i v e r and b r a i n MAO were a l s o noted fo r v a r i o u s d e r i v a t i v e s . G e n e r a l l y most $-carbo l ines gave lower I 5 0 v a l ues wi th 5HT as s u b s t r a t e than wi th t ryptamine as s u b s t r a t e . See Chapter IV for a compar ison of these r e s u l t s w i th a study c a r r i e d out in the p resen t work. Ho and co-workers [39] s t u d i e d the i n f l u e n c e of v a r i o u s s u b s t i t u e n t s on C , , N 2 , and N 9 of the /3-carboline nuc leus on MAOI a c t i v i t y . M e t h y l , e t h y l , or c a r b o x y l s u b s t i t u t i o n of C, r e s u l t e d in p r o g r e s s i v e decrease in a c t i v i t y , the c a r b o x y l s u b s t i t u t e d compounds be ing some e igh teen t imes l e s s a c t i v e than the u n s u b s t i t u t e d d e r i v a t i v e s . E t h y l or N-propyl s u b s t i t u t i o n of N 2 d i d not s i g n i f i c a n t l y reduce a c t i v i t y , a l t hough N 2 _ a c e t y l s u b s t i t u t i o n e s s e n t i a l l y a b o l i s h e d the a c t i v i t y . Methy l s u b s t i t u t i o n of the i n d o l i c n i t r o g e n (N 9 ) s i g n i f i c a n t l y enhanced a c t i v i t y of the te t rahydro-/3-carbo l ines but on l y s l i g h t l y enhanced the aromat ic d e r i v a t i v e s . Buckho l tz and Boggan [40] s t u d i e d te t rahydro-/3-carbo l ines in mouse b r a i n and l i v e r and found tha t they were more potent i n h i b i t o r s i n b r a i n than in 46 l i v e r . 6-MeO-TH/3C was wi thout i n h i b i t o r y a c t i v i t y in l i v e r but had about the same a c t i v i t y in b r a i n as TH/3C. 2. /3-carbol ines as H a l l u c i n o g e n s The l i t t l e that i s known about the h a l l u c i n o g e n i c a c t i v i t y of /3-carbol ines stems p r i m a r i l y from the s t u d i e s of Pennes and Hoch [47] and Naranjo [48 ] . No sys t emat i c human s t u d i e s of the h a l l u c i n o g e n i c a c t i v i t y of /3-carboline d e r i v a t i v e s have been conducted in over 25 y e a r s ; many of the r e s u l t s r epo r t ed by Naran jo [48] were ob ta ined from a s i n g l e human t r i a l . What does seem c l e a r , however, i s that the h a l l u c i n o g e n i c a c t i o n of /3-c a r b o l i n e s on l y man i f e s t s at t h r e s h o l d dosage l e v e l s which are s e v e r a l o r d e r s of magnitude g rea t e r than the doses r e q u i r e d to man i f es t t h e i r a c t i v i t y as MAOI or as c o m p e t i t i v e i n h i b i t o r s of 5HT and e p i n e p h r i n e uptake [ c f . 4 0 , 4 1 ] . It i s u n l i k e l y , t h e r e f o r e , tha t these a c t i v i t i e s can be invoked as the mechanism r e s p o n s i b l e f o r the h a l l u c i n o g e n i c a c t i o n of some /3-carbol ines . Hoch [47] r epo r t ed that harmine was o r a l l y i n a c t i v e at doses i n excess of one gram, but observed t h r e s h o l d h a l l u c i n o g e n i c e f f e c t s at i . v . doses between 200-250 mg. S l o t k i n [49] observed comparable e f f e c t s at 50 mg i . v . Naranjo [48] r e p o r t e d that harma l ine was o r a l l y a c t i v e at 4 mg/kg; t e t rahydroharmine was a p p a r e n t l y l e s s a c t i v e , however the equa t ion 300 mg tet rahydroharmine=100 mg harma l ine r e s u l t e d from a s i n g l e t r a i l . Naran jo [50] r epo r t ed that 6-MeO-harmalan, the 6 - s u b s t i t u t e d ana log of ha rma l i ne , gave t h r e s h o l d o r a l a c t i v i t y at 1.5 mg/kg; comparable l e v e l s of 6-MeO-tetrahydroharmine gave " m i l d e r e f f e c t s " . 6-MeO-harman, the ana log of harmine, has a p p a r e n t l y 47 not been i n v e s t i g a t e d . 3. Endogenous S yn thes i s of /3-carbol ines in Mammals The hypo thes i s that endogenously s y n t h e s i z e d h a l l u c i n o g e n s might p l ay a r o l e in the e t i o l o g y of s c h i z o p h r e n i a or o ther mental d i s o r d e r s has f a l l e n in and out of favor among r e sea r che r s ever s i n ce Osmund and Smythies f i r s t proposed the idea in 1952 [51 ] . The i s sue has s t i l l not been r e s o l v e d , even though unequ ivoca l ev idence [ c f . 20] has been ob ta ined i n d i c a t i n g that h a l l u c i n o g e n s can a r i s e in mammalian systems under c e r t a i n c o n d i t i o n s . Mc lsaac [52] was the f i r s t to suggest that an endogenous /3-carbol ine, formed from the c y c l o d e h y d r a t i o n of the p i n e a l hormone me la ton in (5-methoxy-N-acety l t ryptamine ) , c o u l d be an e t i o l o g i c a l f a c t o r in mental i l l n e s s . Subsequent ly Mc lsaac [53] demonstrated that 1-methyl-6-MeO-TH/3C c o u l d be formed from 6-MeO-tryptamine and ace ta ldehyde under m i l d p h y s i o l o g i c a l c o n d i t i o n s i_n v i t r o and a l s o ir\ v i v o in r a t s p r e t r e a t e d wi th i p r o n i a z i d and d i s u l f i r a m (Antabuse ) . The hypo thes i s appeared to be con f i rmed when F a r r e l and Mclsaac [54] c l a imed to have i s o l a t e d a p i n e a l hormone, a d r e n o g l o m e r u l o t r o p i n , which was i d e n t i c a l to 1-methyl-6-MeO TH/3C. T h i s i n i t i a l t e n t a t i v e i d e n t i f i c a t i o n cou ld not be con f i rmed and e v e n t u a l l y the c l a i m was withdrawn [55 ] . A l though M c l s a a c ' s o r i g i n a l h ypo thes i s tha t the p i n e a l g land i s a l i k e l y s i t e of endogenous /3-carboline fo rmat ion has been con f i rmed on ly fo r b i r d s [5'6], the ev idence that /3-carbol ines are s y n t h e s i z e d in a v a r i e t y of t i s s u e s i s now overwhelming. /3-carbol ines have been u n e q u i v o c a l l y i d e n t i f i e d in human plasma and p l a t e l e t s , and 48 in the ra t whole b r a i n , f o r e b r a i n , a r cua te n u c l e u s , and ad rena l g l a n d . Most of the endogenous /3-carbol ines so fa r c h a r a c t e r i z e d are 6-methoxy-, 6-hydroxy-, or u n s u b s t i t u t e d tetrahydro-/3-c a r b o l i n e s , however the f u l l y a romat ic d e r i v a t i v e harman has been found in the ra t a r cua te nuc leus [11 ] . It has been shown [57,58] that in some i n s t a n c e s endogenous /3-carbol ines are formed as a r e s u l t of condensa t ion of indo lamines wi th formaldehyde r e l e a s e d non-enzymat i ca l l y from 5-methyl-t e t r a h y d r o f o l i c a c i d (MTHF). Recent i n t e r e s t has c en te r ed on the p o s s i b l e involvement of endogenous /3-carbol ines , t e t r a h y d r o i s o q u i n o l i n e s , and other amine/aldehyde condensa t ion p roduc t s in the e t i o l o g y of a l c o h o l i s m [ 5 9 , 6 0 ] , 2-methyl-TH/3C and harman were i d e n t i f i e d in human u r i ne a f t e r e thano l l o a d i n g [61 ] , Presumably these c o n s t i t u e n t s are formed v i a the condensa t ion of b i o g e n i c amines such as s e r o t o n i n or dopamine w i th a c e t a l d e h y d e , the pr imary m e t a b o l i t e of e t h a n o l . See r e f e r e n c e s [11,14] fo r comprehensive recent rev iews of the b i o c h e m i s t r y , pharmacology, and pa tho logy of endogenous mammalian a l k a l o i d s . B. Other N e u r o l o g i c a l and B i o l o g i c a l A c t i v i t i e s of /3-c a r b o l i n e s /3-carbol ines e x h i b i t a wide spectrum of n e u r o p h y s i o l o g i c a l and b i o l o g i c a l a c t i v i t i e s in a d d i t i o n to those d i s c u s s e d above. I t i s not w i t h i n the scope of t h i s review to d i s c u s s each of them in d e t a i l ; they are ment ioned here fo r the sake of comp le teness . The a p p r o p r i a t e r e f e r e n c e s shou ld be c o n s u l t e d fo r 49 f u r t h e r i n f o r m a t i o n . A number of /3-carbol ines have been shown to i n h i b i t the uptake of 5HT, dopamine, n o r - e p i n e p h r i n e , and ep ineph r i ne i n t o synaptosomal suspens ions [ 3 7 , 4 0 ] , Other /3-c a r b o l i n e d e r i v a t i v e s are i n h i b i t o r s of membrane ATPases in human e r y t h r o c y t e s , ra t b r a i n , and squ id r e t i n a l axon [ 3 7 , 6 4 ] . I n t e r f e r e n c e wi th s y n t h e s i s of b i o g e n i c amines by some /3-c a r b o l i n e s has a l s o been r e p o r t e d [37 ] , /3-carbol ine-3-c a r b o x y l a t e e t h y l e s t e r (BCCE) and o ther /3-carboline d e r i v a t i v e s have been i m p l i c a t e d [62,63] as p o s s i b l e endogenous l i g a n d s fo r the benzod iazep ine (Val ium) r e c e p t o r s , a l though other compounds, i n c l u d i n g p u r i n e s and kynuramines, have a l s o been proposed [ c f . 63 fo r a comprehensive r e v i e w ] . Harmal ine and r e l a t e d d e r i v a t i v e s exe r t a potent " v a s o p r e s s i n " - l i k e e f f e c t on Na and water t r a n s p o r t in i s o l a t e d toad s k i n , s t i m u l a t i n g hydrosmot ic f low a c ros s the membrane [ 6 4 , 6 5 ] , F a i l u r e of harma l ine to e l i c i t any e f f e c t i n p r e p a r a t i o n s p r e t r e a t e d w i th v a s o p r e s s i n and/or no r-ep ineph r ine suggested a c o m p e t i t i v e mechanism of a c t i o n . Harmine, harmal ine and r e l a t e d compounds have t remorogen ic e f f e c t s , c a r d i o v a s c u l a r e f f e c t s , and a l s o i n f l u e n c e homeothermic mechanisms, c aus ing hypothermia in some an ima ls ( r a t s , mice) and hyper thermia in o thers ( r a b b i t s ) [37 ] , 0- ca rbo l i ne s a l s o have b i o l o g i c a l a c t i v i t i e s o ther than t h e i r e f f e c t s on n e u r o p h y s i o l o g i c a l sys tems. For i n s t ance Hopp and co-workers [66] found that harmine e x h i b i t e d s i g n i f i c a n t an t i - t r ypanosoma l a c t i v i t y aga in s t Trypanosoma l e w i s i . The mutagenic or co-mutagenic e f f e c t of c e r t a i n /3-carbol ines has been noted [67] and the mechanism r e s p o n s i b l e may be r e l a t e d to the i n t e r a c t i o n of /3-carbol ines w i th n u c l e i c a c i d s [ 6 8 , 6 9 ] . More 50 r e c e n t l y , the u l t r a - v i o l e t - m e d i a t e d p h o t o c y t o t o x i c and photogenotox i c a c t i v i t y of some 0 - c a r b o l i n e d e r i v a t i v e s has been d e s c r i b e d [ 1 3 , 7 0 ] , 51 IV. L i t e r a t u r e C i t e d 1. Smi th , T . A. (1977) Tryptamine and Re la ted Compounds in P l a n t s . Phy tochemis t ry 16:171-75 2. A l l e n , J . R. F . , & B. Holmstedt (1980) The Simple 0 - c a r b o l i n e A l k a l o i d s . Phy tochemis t ry 19 :1573-1582. 1980 3. S k a l t s o u n i s , A. L . , F. T i l l e q u i n , & M. Koch . (1983) P l an tes de Nouve l l e -Ca l edon i e LXXXI I I . A l c a l o i d e s des T i g e s F e u i l l e e s de Me l i cope l e p t o c o c c a . J ou rna l of N a t u r a l P roducts 46:732-35 .4. O 'Donovan, D . G . , & M. F. Kennea l l y (1967) The B i o s y n t h e s i s of E l e a g n i n e . J ou rna l of the Chemica l S o c i e t y S e c t . C:1109-10 5. O 'Donovan, D. G. (1976) P roceed ings of the Royal I r i s h  Academy Se c t i on B. 76:187 6. S l a t o r , M. , & I. J . McFa r l ane . (1968) The B i o s y n t h e s i s and Metabo l i sm of Harman in P a s s i f l o r a e d u l i s . Phy tochemis t ry 7:605-11 7. Smi th , T . A. (1977) Recent Advances in the B i ochemis t r y of P l an t Amines. Chapter 2 in P rogress in Phy tochemis t ry V. 4. L. R e i n h o l d , J . B. Harbourne, and T . Swain ( e d s . ) . Academic P r e s s . 8. Da ly & B. Wi tkop. (1971) Chemist ry and Pharmacology of F rog Venoms. Chapter 39 in Venomous Animals and T h e i r Venoms V. I I , W. B i i cher l & E. E. E. Buckley ( e d s . ) . Academic P ress 9. Kan to r , R. E . , S. D. D u d l e t t e s , & A. T . S h u l g i n . (1980) 5-Methoxy-a-Methy l-t ryptamine ( a , 0 - d i m e t h y l s e r o t o n i n ) , a H a l l u c i n o g e n i c Homolog of S e r o t o n i n . B i o l o g i c a l P s y c h i a t r y 15:349-52 10. Ba rke r , S. A . , J . A. M o n t i , & S. T . C h r i s t i a n . (1981) N,N-d i m e t h y l t r y p t a m i n e : An Endogenous H a l l u c i n o g e n . I n t e r n a t i o n a l Review of Neurob io logy 22:83-110 11. M e l c h i o r , C , and M. A. C o l l i n s . (1982) The Route and S i g n i f i c a n c e of Endogenous S yn thes i s of A l k a l o i d s in Mammals. CRC C r i t i c a l Reviews in T o x i c o l o g y 9:313-356 52 12. D i l l o n , J . , A. Spec to r , & K. N a k a n i s h i . (1976) I d e n t i f i c a t i o n of /3-carbol ines I s o l a t e d from F l u o r e s c e n t Human Lens P r o t e i n s . Nature 259:422-3 13. McKenna, D. & G. H. N. Towers. (1981) U l t r a - v i o l e t Media ted C y t o t o x i c A c t i v i t y of /3-carboline A l k a l o i d s . Phytochemi s t r y 20: 1001-1004 14. Bloom, F . , J . Barchas , M. S a n d l e r , & E. Usd in (eds . ) (1982) g - c a r b o l i n e s and T e t r a h y d r o i s o q u i n o l i n e s . p roceed ings of a symposium h e l d at the Sa lk I n s t i t u t e , La J o l l a , Ca , Dec. 12-13, 1981. A lan R. L i s s , p u b l i s h e r , New York 15. A p p e l , J . B. f F. J . Whi te , & A. M. Holohean (1982) A n a l y z i n g Mechanism(s) of H a l l u c i n o g e n i c Drug A c t i o n w i th Drug D i s c r i m i n a t i o n P rocedures . Neurosc ience and B i o b e h a v i o u r a l Reviews 6:529-36 16. S h u l g i n , A. T . (1976) Psychotomimet ic Agen t s , chapte r 4 in Maxwell Gordon (ed. ) P sychopharmaco log i ca l Agents V. IV. Academic P ress 17. S h u l g i n , A. T . (1982) Chemis t ry of Psycho tomimet i c s . Chapter 1 in F. H o f f m e i s t e r & G. S t i l l e (eds . ) Handbook of  Expe r imenta l Pharmacology V o l . 55 P t . I l l : P s y c h o t r o p i c  Agents P t . I l l : A l c o h o l & Psychotomimet i cs , P s y c h o t r o p i c  Ef f e c t s of C e n t r a l - A c t ing Drugs . S p r i n g e r - V e r l a g 18. S h u l g i n , A. T . & M. F. C a r t e r (1980) N,N-d i i s o p r o p y l t r y p t a m i n e (DIPT) and 5-Methoxy-N,N-d i i s o p r o p y l t r y p t a m i n e (5-MeO-DIPT). Two O r a l l y A c t i v e Tryptamine Analogs wi th CNS A c t i v i t y . Communicat ions in  Psychopharmacology 4:363-69 19. G r e i g , M. K., R. A. Walk, & A. J . Gibbon (1959) The E f f e c t of Three Tryptamine D e r i v a t i v e s on Se ro ton in Metabo l i sm i_n v i t r o and in v i v o . B r i t i s h J o u r n a l of Pharmacology 127:110-20. Rosengar ten , H. & A. J . F r i e d h o f f (1976) A Review of Recent S tud i e s of the B i o s y n t h e s i s and E x c r e t i o n of H a l l u c i n o g e n s Formed by Me thy l a t i on of N e u r o t r a n s m i t t e r s or Re l a t ed Subs tances . S c h i z o p h r e n i a B u l l e t i n 2:90-105 21. Mack, J . P. G . , & M. S l a y t o r (1979) Indo le thy l amine N-methyl T r a n s f e r a s e s of P h a l a r i s t u b e r o s a : P u r i f i c a t i o n and P r o p e r t i e s . Phy tochemis t ry 18:1921-5 53 22. Ba rke r , S. A . , J . A. M o n t i , and S. T . C h r i s t i a n (1980) Metabo l i sm of the H a l l u c i n o g e n N,N-d imethy l t ryptamine in Rat B ra in Homogenates. B i ochemica l Pharmacology 29:1049-57 23. R. B. Bar low. (1961) E f f e c t s on Amine Oxidase of Substances Which Antagon ize 5-hydroxytryptamine More Than Tryptamine on the Rat Fundus S t r i p . B r i t i s h J ou rna l of Pharmacology 16:153-162. 24. B. T . Ho, W. M. Mc l s aa c , R. An , R. T . H a r r i s , K. E. Walker , & P. M. K r a l i k . (1970) B i o l o g i c a l A c t i v i t i e s of Some 5-S u b s t i t u t e d N ,N-d imethy l t r yp tamines , a-methy l t r yp tamines , and Gramines . Psychopharmacolog ia 16:385-394. 25. S z a r a , S. & J . A x e l r o d . (1959) H y d r o x y l a t i o n and N-demethy l a t ion of N ,N-d imethy l t r yp tamine . E x p e r i e n t i a 15:216-17 26. K a l b e r e r , F . , W. K r e i s & J . Rutschmann (1962) Fate of P s i l o c i n in the Rat . B i o chemica l Pharmacology 11:261-69 27. H o r i t a , A. and L. J . Weber (1961) The Enzymic Dephosphory l a t i on and O x i d a t i o n of P s i l o c y b i n and P s i l o c i n by Mammalian T i s sue Homogenates. B iochemica l Pharmacology 7 : 47-54 28. Weber, L. J . & A. H o r i t a (1963) O x i d a t i o n of 4- and 5-hyd roxy indo l e D e r i v a t i v e s by Mammalian Cytochrome O x i d a s e . L i f e S c i ences 1: 44-49 29. L e s s i n , A. W., R. F. Long , & M. W. Parkes (1967) The C e n t r a l S t imu lan t P r o p e r t i e s of some S u b s t i t u t e d I n d o l y l a l k y l a m i n e s ' and /3-carbol ines and T h e i r A c t i v i t i e s as I n h i b i t o r s of Monoamine Oxidase and the Uptake of 5-hydroxyt ryptamine . B r i t i s h J ou rna l of Pharmacology and Chemotherapy 29:70-79 30. S a i - H a l a s z , A. (1963) The E f f e c t of MAO I n h i b i t i o n on the Expe r imen ta l P s ychos i s Induced by D ime thy l t r yp t am ine . Psychopharmacolog ia 4:385-88 31. S a i - H a l a s z , A. (1962) The E f f e c t of A n t i s e r o t o n i n on the Expe r imen ta l P sychos i s Induced by D ime thy l t r yp t am ine . E x p e r i e n t i a 18:137-138 54 32. Moore, R. H. S. K. Demetr iou & E. F. Domino (1975) E f f e c t s of I p r o n i a z i d , Ch loropromaz ine and Me th io thep in on DMT-induced Changes in Body Temperature , P u p i l l a r y D i l a t a t i o n , B lood P ressure and EEG in the R a b b i t . A r c h i v e s  I n t e r n a t i o n e l s de pharmacodynamic et de t h e r a p i e 213:64-72 33. Wang Lu , L. J . , & E. F. Domino (1976) E f f e c t s of I p r o n i a z i d and Trany l cypromine on the H a l f - L i f e of N,N-d ime thy l t r yp t am ine in Rat B ra in and L i v e r . B i o chemica l  Pharmacology 25:1521-27 34. Shah, N. S . , & M. P. Hedden (1977) Behav i ou ra l E f f e c t s and M e t a b o l i c Fate of N ,N-d imethy l t ryptamine in Mice P r e t r e a t e d wi th /3-d i e thy l am inoe thy l -d ipheny lp ropy l a ce t a t e (SKF-525-A) , I p r o n i a z i d , and Ch lo rp romaz ine . Pharmacology, B i ochemis t r y and Behaviour 8:351-56 35. Ho lms ted t , B. (1982) /3-carbol ines and T e t r a h y d r o i s o q u i n o l i n e s : H i s t o r i c a l and E thnopha rmaco log i ca l Background, in F. Bloom, et a l . , ( eds . ) op . c i t . 36. U d e n f r i e n d , S . , B. W i tkop, B. G. R e d f i e l d , & H. Weissbach (1958) S tud i e s wi th R e v e r s i b l e I n h i b i t o r s of Monoamine O x i d a s e : Harmal ine and Re la ted Compounds. B i o chemica l  Pharmacology 1:160-165 37. Ho, B. T . (1977) Pha rmaco log i ca l and B iochemica l S tud i e s wi th /?-carboline A n a l o g s , chapte r VI in W. B. Essman & L. V a z e l l i , ( eds . ) Cur ren t Developments in Psychopharmacoloqy V. 4. Spectrum P u b l i c a t i o n s , Inc . 38. M c l s a a c , W. M. & V. E s t evez ( 1 9 6 6 ) S t r u c t u r e - a c t i o n R e l a t i o n s h i p s of /3-carbol ines as Mono-amine Oxidase I n h i b i t o r s . B i ochemica l Pharmacology 26:1625-27 39. Ho, B. T . , W. M. M c l s a a c , K. E. Walker , & V. E s t evez (1968) I n h i b i t i o n of Monoamine Ox idase : I n f l uence of Methy l S u b s t i t u t i o n on the I n h i b i t o r y A c t i v i t y of /3-carbol ines . J o u r n a l of Pha rmaceut i ca l S c i ences 57:269-74 40. B u c k h o l t z , N. S . , and W. O. Boggan (1976) E f f e c t s of Te t rahydro - /3-ca rbo l ines on Monoamine Oxidase and Se ro ton in Uptake in Mouse B r a i n . B i o chemica l Pharmacology 25:2319-2321 55 41. B u c k h o l t z , N. S. £ W. 0. Boggan (1977) Monoamine Oxidase I n h i b i t i o n i n B r a i n and L i v e r Produced by /3-carbo l ines : S t r u c t u r e - a c t i v i t y R e l a t i o n s h i p s and Subs t r a t e S p e c i f i c i t y . B iochemica l Pharmacology 26:1991-96 42. F u l l e r , R. W., B. J . Warren, & B. B. Mo l loy (1970) S e l e c t i v e I n h i b i t i o n of Monoamine Oxidase in Rat B r a i n M i t o c h o n d r i a . B iochemica l Pharmacology 19:2934-2936 43. D o n n e l l y , C. H . , E. R i che l son & D. L. Murphy (1976) P r o p e r t i e s of Monoamine Oxidase in Mouse Neuroblastoma NIE-115 C e l l s . B i ochemica l Pharmacology 25:1629-1643 44. D o n n e l l y , C. H . , & D. L. murphy (1977) S u b s t r a t e - and I n h i b i t o r - r e l a t e d C h a r a c t e r i s t i c s of Human P l a t e l e t Monoamine Ox idase . B iochemica l Pharmacology 26:853-858 45. Hous l ay , M. D. £ K. F. T i p t o n (1974) A K i n e t i c E v a l u a t i o n of Monoamine Oxidase A c t i v i t y in Rat L i v e r M i t o c h o n d r i a l Outer Membranes. B i ochemica l J ou rna l 139:645-652 46. N e f f , N. H. and H.-Y. T . Yang (1974) Another Look at the Monoamine Ox idases and the Monoamine Ox idase I n h i b i t o r Drugs . L i f e S c i ences 14:2061-2074 47. Pennes, H. H. & P. H. Hoch (1957) Psycho tomimet i c s , C l i n i c a l and T h e o r e t i c a l C o n s i d e r a t i o n s : Harmine, WIN-299 and N a l l i n e . American J ou rna l of P s y c h i a t r y 113:887-92 48. Na ran jo , C. (1967) P s y c h o t r o p i c P r o p e r t i e s of the Harmala A l k a l o i d s , pp . 385-391 in D. H. Efr .on, B. Ho lms ted t , & N. S. K l i n e (eds . ) E thnopharmaco log ic Search f o r P sychoac t i ve  Drugs . U. S. P u b l i c Hea l th S e r v i c e P u b l i c a t i o n #1645 49. S l o t k i n , T . A . , . V . D i S t e f ano & W. Y. W. Yu (1970) B lood L e v e l s and U r i n a r y E x c r e t i o n of Harmine and I t s M e t a b o l i t e s in Man and Ra t s . The J ou rna l of Pharmacology and  Exper imenta l T h e r a p e u t i c s 173:26-30 50. Na ran jo , C. (1969) P s y cho the rapeu t i c P o s s i b i l i t i e s of New Fantasy-enhanc ing Drugs . C I i n i c a l T o x i c o l o g y 2:209-224 51. Osmund, H . , and J . R. Smythies (1952) S c h i z o p h r e n i a : A New Approach . J ou rna l of Menta l S c i ences 98:309-315 56 52. Mc l s aa c , W. M. (1961) A B iochemica l Concept of Mental D i s e a s e . Pos tgraduate Med ic ine 30:111-118 53. Mc l s aa c , W. M. (1961) Format ion of 1-methyl-6-methoxy-1 , 2 , 3 , 4 - t e t r a - h y d r o - 0 - c a r b o l i n e under P h y s i o l o g i c a l C o n d i t i o n s . B i o ch im i ca et B i o p h y s i c a Acta 52:604-606 54. F a r r e l l , G. & W. M. Mclsaac (1961) A d r e n o g l o m e r u l o t r o p i n . A r c h i v e s of B i ochemis t r y & B i o p h y s i c s 94:543-44 55. F a r r e l l , G . , W. M. M c l s a a c , & A. N. T a y l o r (1964) Neurohumoral F a c t o r s from the Bra ins tem E p i p h y s i s . pp. 141-47 in L. M a r t i n i & A. P e c i l e (eds . ) Hormonal S t e r i o d s ^ B i o c h e m i s t r y , Pharmacology, and The r apeu t i c s V. I. Academic P r e s s . 56. K a r i , I. (1 98 1 ) 6-methoxy-1 , 2 , 3 , 4-tetrahydro-/3-carbol ine in P i n e a l G land of Ch icken and Cock. FEBS L e t t e r s 127:277-280 57. Hsu, L. L . , & A. J . Mande l l (1975) Enzymatic Format ion of T e t r a h y d r o - 0 - c a r b o l i n e s from Tryptamine and 5-M e t h y l t e t r a h y d r o f o l i c A c i d in Rat B ra in F r a c t i o n s : Reg iona l and S u b c e l l u l a r D i s t r i b u t i o n . J ou rna l of Neurochemis t ry 24:631-36 58. Rommelspacher, H . , H. Caper & S. S t rauss (1976) On the Mode of Format ion of Te t rahydro-/3-carbo l ines . L i f e S c i ences 18:81-88 59. Rahwan, R. G. (1975) Tox i c E f f e c t s of E thano l - P o s s i b l e Role of Ace t a l dehyde , T e t r a h y d r o i s o q u i n o l i n e s , & Te t rahydro-/3-carbo l ines . T o x i c o l o g y and A p p l i e d  Pharmacology 34:3-27 60. D a v i s , V. E. and M. A. Walsh (1970) A l c o h o l , Amines, and A l k a l o i d s : A P o s s i b l e B iochemica l Bas i s fo r A l c o h o l A d d i c t i o n . Sc ience 167:1005-7 61. Rommelspacher, H . , S. S t rauss & J . Lindemann (1980) E x c r e t i o n of Tetrahydroharmane and Harmane i n t o the U r ine of Man and Rat A f t e r a Load w i th E t h a n o l . FEBS L e t t e r s 109:209-12 57 62. M e n n i n i , T . , S. C o t e c c h i a , S. G a r a t t i n i ( 1982) Does Ethyl-/3-c a r b o l i n e - 3 - c a r b o x y l a t e I n t e r a c t w i th Mouse B r a in Benzod iazep ine Receptors in V i vo? J ou rna l of Pharmacy and  Pharmacology 34:394-5 63. L a p i n , I. P. (1983) S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s in Kynuren ine , Diazepam and Some P u t a t i v e Endogenous L igands of the Benzod iazep ine Recep to r s . Neurosc i ences and B i o b e h a v i o u r a l Reviews 7:107-118 64. Canessa , M. , E. J a i m o v i c h , & M. de l a Fuente (1972) The E f f e c t of Harmane D e r i v a t i v e s on the Na-dependent Pho spho r y l a t i on from the Membrane ATPase System, p. 28ff V th I n t e r n a t i o n a l Congress of Nephro logy . Mexico C i t y 65. de Sousa , R. C , & A. Grosso (1978) V a s o p r e s s i n - l i k e E f f e c t s of a H a l l u c i n o g e n i c Drug - Harmal ine - on Sodium and Water T r a n s p o r t . J ou rna l of Membrane B i o l ogy 40:77-94 66. Hopp, K. H. , L. V . Cunningham, M. C . B romel , L. J . S che rme is te r and S. K. W. K a h l i l (1976) In V i t r o An t i t r ypanosoma l A c t i v i t y of C e r t a i n A l k a l o i d s Aga ins t Trypanosoma l e w i s i . L l o y d i a 39:375-77 67. Umezawa, K., A. S h i r a i , T . Matsushima, & T . Sugimura (1978) Comutagenic E f f e c t of Norharman and Harman wi th 2-A c e t y l a m i n o f l u o r e n e D e r i v a t i v e s . P roceed ings of the  N a t i o n a l Academy of S c i ences 75:928-30 68. Remson, J . F. & P. A. C e r u t t i (1979) I n h i b i t i o n of DNA Repa i r and DNA S yn thes i s by Harman in Human A l v e o l a r Tumor C e l l s . B i o chemica l and B i o p h y s i c a l Research Communications 86:124-129 69. H a y a s h i , K., M. Nagao, & T . Sugimura (1977) I n t e r a c t i o n s of Harman and Norharman wi th DNA. N u c l e i c Ac i ds Research 4:3679-85 70. Towers, G. H. N . , & Z. Abramowsky. (1983) UV-mediated G e n o t o x i c i t y of F u r a n o q u i n o l i n e s and of C e r t a i n Tryptophan-De r i v ed A l k a l o i d s . J ou rna l of N a t u r a l P roduc ts 46:576-81 58 PART I I : ETHNOBIOLOGICAL INVESTIGATIONS OF MALPIGHIACEOUS AND MYRISTICACEOUS HALLUCINOGENS 59 CHAPTER I I I : ETHNOBOTANICAL COLLECTIONS AND ETHNOGRAPHIC OBSERVATIONS I. E t h n o b o t a n i c a l C o l l e c t i o n s A. C o l l e c t i o n and I d e n t i f i c a t i o n of Herbar ium Voucher Spec imens Dur ing the f i e ld-work conducted in c o n n e c t i o n wi th t h i s m r e s e a r c h , herbar ium voucher specimens were c o l l e c t e d for a l l p l an t s p e c i e s i n d i c a t e d by in formants to have m e d i c i n a l p r o p e r t i e s or other e t h n o b o t a n i c a l l y s i g n i f i c a n t u se s . P a r t i c u l a r e f f o r t s were made to secure voucher specimens fo r a l l s ou r ce-p l an t s and admixture p l a n t s used in the p r e p a r a t i o n of the Ma lp igh i a ceous and M y r i s t i c a c e o u s drug samples which have been ana l yzed in t h i s s tudy . These e f f o r t s were not always t o t a l l y s u c c e s s f u l , i . e . in some i n s t ances i t was p o s s i b l e to c o l l e c t on l y l i m i t e d amounts of voucher m a t e r i a l (as , fo r example, when c o l l e c t i n g from the c u l t i v a t e d p l o t of an ayahuasquero ) , or the a c t u a l s ou r ce-p l an t s used in the manufacture of a p a r t i c u l a r sample were not a v a i l a b l e . In these i n s t a n c e s p l a n t s of the same s p e c i e s from the same gene ra l l o c a l i t y were c o l l e c t e d fo r comparat i ve pu rposes . C o l l e c t i o n numbers c i t e d throughout r e f e r to the p e r s o n a l c o l l e c t i o n numbers of D. McKenna and are p r e f a ced wi th "DMK-". Excep t i ons are Plowman 6040 ( D i p l o p t e r y s c a b r e r a n a ) , Plowman 6041 (B. caap i v a r . " c i e l o " ) and Plowman 12,218 ( V i r o l a spp . ). These samples were k i n d l y s u p p l i e d by D r . Timothy Plowman of the F i e l d Museum of N a t u r a l H i s t o r y in C h i c a g o . Yanomamo snu f f samples were a g i f t to the author by D r . E rnes to M i g l i a z z a , f o rme r l y of the 60 Department of An th ropo logy , U n i v e r s i t y of Mary l and , C o l l e g e Park ; no accompanying voucher specimens are a v a i l a b l e f o r the snuf f samples . A u t h e n t i c a t e d herbar ium vouchers f o r a l l "DMK-" c o l l e c t i o n numbers have been d e p o s i t e d in the Herbar ium of the Department of Botany, U n i v e r s i t y of B r i t i s h Co lumb ia . D u p l i c a t e vouchers f o r most c o l l e c t i o n s are a l s o on d e p o s i t at the F i e l d Museum; o ther h e r b a r i a in North and South America have d u p l i c a t e s of some but not a l l c o l l e c t i o n s . A l l voucher c o l l e c t i o n s of e t h n o b o t a n i c a l s i g n i f i c a n c e which were made in connec t i on w i th t h i s work are l i s t e d in Appendix I I . B. C o l l e c t i o n of P l an t M a t e r i a l and Drug Samples fo r Phy tochemica l A n a l y s i s Bark and l e a f samples used in the phy tochemica l ana l y ses c o n s i s t e d e i t h e r of a i r - d r i e d p l an t m a t e r i a l or of m a t e r i a l p rese r ved in 100% methanol at the t ime of c o l l e c t i o n . Most of the drug samples were a l s o p rese r ved in methanol at the t ime of c o l l e c t i o n ; e x c e p t i o n s were two ayahuasca samples , the o r a l l y -inges ted pas te sample from La C h o r r e r a , Colombia ( c o l l e c t e d in 1971—no voucher a v a i l a b l e ) and the Yanomamo snu f f samples . D e t a i l s of the p r e p a r a t i o n of the p l a n t m a t e r i a l s and drug samples fo r phy tochemica l a n a l y s i s a re c i t e d in the " M a t e r i a l s and Methods" s e c t i o n s of the a p p r o p r i a t e c h a p t e r s . 11. E thnograph i c Observat ions The e thnograph i c a spec t s of the Ma lp igh i a ceous and 61 M y r i s t i c a c e o u s h a l l u c i n o g e n s have been the sub jec t of study by a n t h r o p o l o g i s t s , e t h n o b o t a n i s t s , s c h o l a r s and e x p l o r e r s s i n ce be fo re the tu rn of the c e n t u r y . In some r e s p e c t s the ethnography of these n a t i v e h a l l u c i n o g e n s i s much more thorough ly documented than the b o t a n i c a l , p h y t o c h e m i c a l , or pha rmaco log i c a l f a c e t s , as these have on l y begun to r e c e i v e c l a r i f i c a t i o n w i t h i n the l a s t t h i r t y y e a r s . The r e l e v a n t e t h n o b o t a n i c a l l i t e r a t u r e on the sub j e c t was reviewed in chapte r I; t h i s chapte r documents pe r sona l o b s e r v a t i o n s made, d u r i n g the course of f i e l d w o r k . A. E thnograph i c Aspec ts of ayahuasca Dur ing the f i e l dwo rk c a r r i e d out fo r t h i s s tudy , v a r i o u s ayahuasqueros (persons who s p e c i a l i z e in the p r e p a r a t i o n and use of ayahuasca in f o l k med ic ine ) were con t a c t ed in the v i c i n i t i e s of I q u i t o s , P u c a l l p a , and Ta r apo to . An e f f o r t was made to i n t e r v i ew and to c o l l e c t ayahuasca samples from as many d i f f e r e n t p r a c t i t i o n e r s as p o s s i b l e . T h i s was done in an attempt to l e a r n a.) whether there were s i g n i f i c a n t d i f f e r e n c e s in the a l k a l o i d compos i t i on of ayahuasca samples p repared by d i f f e r e n t p r a c t i t i o n e r s , e i t h e r in terms of types of a l k a l o i d s found , or i n c o n c e n t r a t i o n , b.) whether e thnograph i c e lements of ayahuasca use are d i f f e r e n t from one r eg ion to ano the r , or from one p r a c t i t i o n e r to ano the r . In a l l , ayahuasca samples prepared by s i x d i f f e r e n t p r a c t i t i o n e r s were c o l l e c t e d , and in some c a s e s , samples of d i f f e r e n t batches p repared by the same person were c o l l e c t e d . The phy tochemica l r e s u l t s are d i s c u s s e d in Chapter IV. In some i n s t a n c e s , con t a c t w i th the ayahuasquero was s u p e r f i c i a l , and c o n s i s t e d e i t h e r of a t t e n d i n g the weekly 62 ayahuasca s e s s i o n as a pay ing p a r t i c i p a n t or of v i s i t i n g h i s home to purchase samples of ayahuasca ; * in o ther i n s t a n c e s , repeated v i s i t s were made and i t was p o s s i b l e to conduct more d e t a i l e d i n t e r v i e w s . Of the s e v e r a l ayahuasqueros c o n t a c t e d , I became best a cqua in ted wi th Don F i d e l Mosambite, who l i v e d in the sma l l v i l l a g e of Jose O l a y a , not f a r from P u c a l l p a . Most of the remarks which f o l l ow have been d e r i v e d from our c o n v e r s a t i o n s . Don F i d e l was q u i t e w i l l i n g to t a l k about the use of ayahuasca and other m e d i c i n a l p l a n t s . He had some unders tand ing of Western medic ine and s c i ence and seemed impressed by the f a c t tha t we were b o t a n i c o s . Don F i d e l seemed to look upon Western medic ine as complementary to h i s own e thnomedica l t r a d i t i o n ; he was q u i t e aware tha t some d i s e a s e s are caused by microbes or d i e t a r y d e f i c i e n c i e s and c o u l d be t r e a t e d wi th a n t i b i o t i c s or v i t a m i n s . He would o f t en send h i s p a t i e n t s to seek t reatment from Western med ica l p r a c t i t i o n e r s , i f a n t i b i o t i c s were c a l l e d f o r , and o c c a s i o n a l l y he would supp ly them with v i t am ins h i m s e l f . In most i n s t a n c e s , however, he f e l t tha t the causes of i l l n e s s were s u p e r n a t u r a l in o r i g i n or were amenable to t reatment wi th any of s e v e r a l hundred m e d i c i n a l p l a n t s wi th which he was f a m i l i a r . In these cases he would undertake the treatment h i m s e l f . Ayahuasca was u s u a l l y i n c o r p o r a t e d i n t o the t r ea tment , a l though i t was not always g i ven to the p a t i e n t ; at t imes on l y Don F i d e l consumed ayahuasca , in order to d iagnose the i l l n e s s , and at o ther t imes the p a t i e n t would r e c e i v e i t as w e l l . In o ther c a s e s , the •Al though the mascu l ine pronoun i s used in t h i s d e s c r i p t i o n , ayahuasqueros can be (and f r e q u e n t l y are ) women. 63 t reatment i n v o l v e d m e d i c i n a l p l a n t s o ther than ayahuasca and n e i t h e r Don F i d e l or the p a t i e n t would take ayahuasca . In a l l cases tha t I obse r ved , however, the a d m i n i s t r a t i o n of the b o t a n i c a l remedy, which might take the form of a t e a , p o t i o n , p o u l t i c e , or t i n c t u r e , was accompanied by mag ica l songs , c h a n t s , and w h i s t l i n g . The p a t i e n t , the remedy, and the gene ra l v i c i n i t y were l i b e r a l l y censed wi th tobacco smoke which Don F i d e l burned in a s p e c i a l p ipe which he had made h i m s e l f . Some ayahuasqueros make passes over the p a t i e n t ' s body wi th a r a t t l e made of d r i e d palm f ronds in o r d e r , presumably , to d r i v e out e v i l s p i r i t s or purge the p a t i e n t of ma levo lent i n f l u e n c e s . I never saw Don F i d e l use such a r a t t l e , however, he would o f t en massage the a f f e c t e d pa r t of the p a t i e n t ' s body, and accompany t h i s wi th a v i go rous and no i s y suck ing of the a f f e c t e d a r e a . T h i s i s a common element of shamanic p r a c t i c e everywhere, the purpose of the suck ing be ing to draw out a mag i ca l ob jec t which has been p l a ced in the body of the p a t i e n t by a s o r c e r e r or ma levo lent shaman. Luna [1] has d e s c r i b e d in d e t a i l the mag ica l symbols , o b j e c t s , and powers which are i n t r i n s i c to the shamanic p r a c t i c e s of the mes t izo ayahuasqueros . A l though Luna ' s f i e l d i n v e s t i g a t i o n s were conducted among ayahuasqueros in I q u i t o s , Don F i d e l ' s mag ica l cosmology c o n t a i n e d e s s e n t i a l l y the same e l ements , even though he l i v e d s e v e r a l hundred m i l e s to the sou th . Ev idence based on two samples i s h a rd l y c o n c l u s i v e , but i n d i c a t e s that the mag ica l cosmology shared among ayahuasqueros i s remarkably c o n s i s t e n t from reg ion to r eg ion and between i n d i v i d u a l s ; t h i s may mean tha t i t i s d e r i v e d from some much o l d e r , g e o g r a p h i c a l l y and c u l t u r a l l y r e s t r i c t e d , t r i b a l 64 t r a d i t i o n that has g r a d u a l l y been a s s i m i l a t e d i n t o mes t i zo f o l k m e d i c i n e . A l though I v i s i t e d the house of Don F i d e l f r e q u e n t l y du r i ng my f i e l d s t u d i e s in the P u c a l l p a a r e a , I was unable to w i tness the a c t u a l p r e p a r a t i o n of ayahuasca . The ayahuasca which Don F i d e l used was u s u a l l y p repared by h i s u n c l e , Juan Sa las Coumar i , a much o l de r man of c o n s i d e r a b l e repute as an ayahuasquero , who l i v e d twenty km from P u c a l l p a . On one occas ion I v i s i t e d the house of Don Juan accompanied by Don F i d e l and was a b l e to see the ayahuasca , a l r eady cut up and set to b o i l , a long w i th the l eaves of the " chac runa " admixture ( P s y c h o t r i a  v i r i d i s ) , in a l a r g e aluminum pot set over the f i r e in Don J uan ' s cook ing shed . P r e p a r a t i o n of ayahuasca i s a m u l t i s t a g e p r o c e s s , i n v o l v i n g p ro longed b o i l i n g of the p l a n t m a t e r i a l w i th s e v e r a l changes of f r e s h water , f o l l owed f i n a l l y by combining the f r a c t i o n s and c o n c e n t r a t i n g them over a low f i r e to a f r a c t i o n of the o r i g i n a l volume. Luna [2] has p rov ided e x c e l l e n t pho tog raph i c documentat ion of the s teps in the p r e p a r a t i o n of ayahuasca . The l eng th of b o i l i n g , the amount of p l an t m a t e r i a l i n i t i a l l y e x t r a c t e d , and the degree of f i n a l c o n c e n t r a t i o n , must a l l vary s i g n i f i c a n t l y , s i n ce c o n s i d e r a b l e v a r i a t i o n in a l k a l o i d con ten t i s found in brews prepared by d i f f e r e n t ayahuasqueros ( c f . Chapter IV ) . An e t h n o b o t a n i c a l l y i n t e r e s t i n g aspect of f o l k taxonomy r e l a t e d to the i d e n t i f i c a t i o n of the " p r o p e r " P s y c h o t r i a spp . f o r use as admixtures came to l i g h t d u r i n g the course of f i e l d w o r k . Some P s y c h o t r i a s p e c i e s , i n c l u d i n g P. v i r i d i s , possess t i n y s p i n e - l i k e ex t ens i ons of the m i d - r i b on the a b a x i a l 65 s u r f a c e of the l e a f . These appear to be s l i g h t l y swo l len g l a n d u l a r s t r u c t u r e s which may be e q u i v a l e n t to the "domat i a " found in some P s y c h o t r i a spp . (Gentry , A . , p e r s . comm., 1981). A domatium i s a pa r t of a l e a f , p e t i o l e , stem or o ther p l an t par t tha t i s i n h a b i t e d by a n t s , m i t e s , or other i n s e c t s . The domatia-l i k e s t r u c t u r e s of the Psychot r i a v i r i d i s specimens I examined appeared to be much too sma l l to accomodate ants of any v i s i b l e s i z e , but may be i n h a b i t e d by mi tes (Prance, G. T . , p e r s . comm., 1983). These t i n y doma t i a- l i k e s t r u c t u r e s were p o i n t e d out by the ayahuasqueros I i n t e r v i ewed as the key f e a tu r e used to i d e n t i f y the chacrunas s u i t a b l e fo r use as adm ix tu r e s . "Chac runa " i s the v e rnacu l a r term fo r Psychot r i a spp . A l l of the ayahuasqueros save one i n s i s t e d tha t these s t r u c t u r e s — t e r m e d by them esp inas ( sp ines )--had to be p r e s e n t ; p l a n t s l a c k i n g esp inas were regarded as f a l s e chacrunas and were c o n s i d e r e d to have no va lue as admix tu res . Indeed, a l l of the P. v i r i d i s c o l l e c t i o n s which were ana l yzed ( c f . Tab l e VI) possessed these s t r u c t u r e s , and a l l c o n t a i n e d DMT; the s i n g l e specimen which l a cked these s t r u c t u r e s a l s o con ta ined no t r yp tamines or o ther a l k a l o i d s . T h i s specimen (DMCK #109) may co r respond to P s y c h o t r i a  c a r t h a g e n e n s i s J a c q . , a l t hough the c o l l e c t i o n i s s t e r i l e and the i d e n t i f i c a t i o n t h e r e f o r e t e n t a t i v e ; f u r t h e r doubt i s cas t on the i d e n t i f i c a t i o n by the f a c t tha t R i v i e r & L i ndg ren [3] r epo r t ed DMT in P s y c h o t r i a c a r t h a g e n e n s i s , wh i le none was de t e c t ed in .DMCK #109. I a t t ended the weekly ayahuasca s e s s i o n s h e l d at Don F i d e l ' s house on s e v e r a l o c c a s i o n s . They were always h e l d at n i g h t , and would commence soon a f t e r da rk . V a r i o u s people from 66 the ne ighborhood a t t ended , t r i c k l i n g by ones and twos i n t o Don F i d e l ' s d o o r y a r d , murmuring g r e e t i n g s to t h e i r f r i e n d s as they took up p o s i t i o n s in h i s " l i v i n g room"--ac tua l l y a tha tched veranda open on one s ide to the c o o l even ing a i r . Everyone sat a round , smoking and t a l k i n g q u i e t l y among themselves whi le they wa i ted fo r the s t r a g g l e r s to a r r i v e . Some p a t i e n t s d e s i r i n g t reatment from Don F i d e l might come from c o n s i d e r a b l e d i s t a n c e s to a t t end th'e. s e s s i o n s ; people were o f t en l a t e . Those a t t e n d i n g appeared to be a t y p i c a l c r o s s - s e c t i o n of l o c a l mes t i zo s o c i e t y ; some were r e g u l a r p a r t i c i p a n t s who showed up every week; o the r s a t t ended on ly a s i n g l e s e s s i on fo r some s p e c i f i c t rea tment . Some of those a t t e n d i n g were p a t i e n t s seek ing t rea tment , wh i le one or two men were appa ren t l y a p p r e n t i c e s of Don F i d e l ' s , there to l e a r n the ayahuasca songs and the p l a n t m e d i c i n e s ; s t i l l o ther p a r t i c i p a n t s used ayahuasca " r e c r e a t i o n a l l y " , or were merely c u r i o u s about i t . Not everyone who a t tended would n e c e s s a r i l y take ayahuasca ; some r e c e i v e d some other form of t reatment from Don F i d e l , o the r s merely watched. U s u a l l y everyone had a r r i v e d by about 9 :30 . Don F i d e l would uncork the wine b o t t l e of ayahuasca , which by t h i s time had been a p p r o p r i a t e l y b l e s s e d w i th songs and w h i s t l i n g and thorough ly fumigated wi th t obacco , and pour about 70 ml of the dark brown, i n t e n s e l y b i t t e r l i q u i d i n t o a sma l l gourd cup which he passed to each of the p a r t i c i p a n t s i n t u r n . Everyone d r a i n e d the cup at one g u l p , g r imac ing as they handed i t back so Don F i d e l c o u l d pour the next l i b a t i o n . Don F i d e l took the l a s t draught h i m s e l f , o f t en t a k i n g two c u p f u l s to everyone e l s e ' s one. A f t e r the ayahuasca had been consumed, c o n v e r s a t i o n f e l l to a minimum. Everyone sat 67 q u i e t l y , wa i t i ng fo r the i n i t i a l e f f e c t s of the d r u g . P r e s e n t l y Don F i d e l would e x t i n g u i s h the o i l lamp, p l ung ing the room in to b l a c k n e s s . Not long a f t e r , the f i r s t of the many ayahuasca songs and chants would b e g i n , and con t inue o f f and on throughout the n i g h t . A few of these were in Span i sh , most were in Quechua, and some were a combina t ion of the two l anguages . T h e i r tenor and tempo was low and s t a t e l y , reminding me of some med ieva l Russ ian Orthodox l i t u r g i c a l chants which I had heard years b e f o r e . The v i s i o n s and h a l l u c i n a t i o n s began at about the same time as the s i n g i n g , and seemed to be keyed i n t o and s u s t a i n e d by the s i n g i n g . The v i s i o n s faded or became l e s s i n t e r e s t i n g whenever the chan t i ng and s i n g i n g s topped ; then a new song would t r i g g e r a renewed e x f o l i a t i o n of hypnagogic imagery. The nature of the v i s i o n s i s d i f f i c u l t to c h a r a c t e r i z e . They d i d not have the i n t e n s e , hard-edge, g e o m e t r i c a l q u a l i t y t y p i c a l of " p s y c h e d e l i c " imagery ; r a the r t h e i r c o l o r s were muted, r i c h in s o f t browns, g r eens , and r e d s , the s o r t s of c o l o r s one might encounter on the f o r e s t f l o o r , or in anc i en t t a p e s t r i e s . Jung le p l a n t s w i th enormous l e a v e s , tw in ing l i a n a s , and underwater scenes of l a rge bony-p la ted f i s h e s were common m o t i f s . The drug d i d not always e l i c i t p ro found mental changes in me; on l y on two o c c a s i o n s , when I persuaded Don F i d e l to double my usua l a l l o t m e n t , d i d I expe r i ence more than the very m i l d e s t e f f e c t s . How much of the content , of the v i s i o n s , and the degree of response to the d r u g , were f u n c t i o n s of my own p h y s i c a l and p s y c h o l o g i c a l s t a t e , i s hard to de te rm ine . I suspect that o n e ' s d i e t , as we l l as body we igh t , may i n f l u e n c e the amount of ayahuasca r e q u i r e d to a ch i eve t h r e s h o l d e f f e c t s . C e r t a i n l y the ayahuasqueros 68 themselves a l l i n s i s t that d i e t i s a major f a c t o r [ 1 ] . S ince I was heav i e r than most p a r t i c i p a n t s , and unaccustomed to the t y p i c a l mes t i zo d i e t of yuco , p l a n t a i n s , and smoked f i s h , my a b s o r p t i o n and metabol ism of the drug may have been d i f f e r e n t and thus I r e q u i r e d a h ighe r dose . I never expe r i enced the nausea or vomi t i ng that u s u a l l y forms an i n t r i n s i c par t of the ayahuasca exper i ence and which g i v e s i t a r e p u t a t i o n as an e x c e l l e n t pu rge . Most of the o ther p a r t i c i p a n t s excused themselves to go have t h e i r purge in the backyard du r i ng the s e s s i o n s ; some went s e v e r a l t imes . Everyone kept an outward demeanor of calm du r i ng the s e s s i o n s , and I d i d not observe any nega t i ve or f e a r f u l r e a c t i o n s . L i k e the host in C a t h o l i c i s m , ayahuasca i s the c e n t r a l mystery but the expe r i ence i s g iven coherence and meaning by the s o c i a l and p s y c h o l o g i c a l set and s e t t i n g . ^This was very c a r e f u l l y o r c h e s t r a t e d and c o n t r o l l e d by Don F i d e l through h i s use of songs , c h a n t s , t obacco-b low ing , and o ther r i t u a l m a n i p u l a t i o n s . By t h i s means, he was ab l e to i n su re tha t everyone had a p o s i t i v e expe r i ence and came away from the s e s s i o n wi th a sense of p e r s o n a l and s p i r i t u a l renewa l . The s e s s i o n s would c o n t i n u e , punc tua ted by chants and s i n g i n g , a lmost u n t i l dawn. No one ever took more than the i n i t i a l dose of ayahuasca . O c c a s i o n a l l y Don F i d e l and one of h i s a s s i s t a n t s would take time to m i n i s t e r to one of the p a t i e n t s ( i f any were p resen t fo r t r e a tmen t ) . T h i s m i n i s t r a t i o n i n v a r i a b l y i n v o l v e d more s i n g i n g , t obacco-b low ing , and u s u a l l y no i s y and v i g o r o u s s u c k i n g , des igned to draw out the mag ica l po isonous ob j ec t d i s c u s s e d above and in [ 1 ] . E v e n t u a l l y , a f t e r every p a t i e n t had been a t tended to and the e f f e c t s of the drug had begun to wane, 69 the s e s s i o n would draw to a c l o s e , and the p a r t i c i p a n t s would f i l e out in the c o o l predawn hours to r e tu rn to t h e i r own homes and d a i l y l i v e s . B. E thnograph ic Aspec ts of O r a l l y A c t i v e M y r i s t i c a c e o u s Pastes A l though mes t izo f o l k medic ine has changed as d i v e r s e t r i b a l e lements have been s y n c r e t i c a l l y a s s i m i l a t e d i n t o i t , n e v e r t h e l e s s i t remains a v i t a l and l i v i n g t r a d i t i o n . Ayahuasca i s an i n t r i n s i c par t of t h i s t r a d i t i o n and the re i s l i t t l e danger that i t s use w i l l d i sappear in the f o r e seeab l e f u t u r e . The s i t u a t i o n i s much d i f f e r e n t wi th respec t to the o r a l l y -i nges t ed M y r i s t i c a c e o u s p a s t e s . Knowledge of the p r e p a r a t i o n and uses of the o r a l l y a c t i v e V i r o l a pas tes r ep resen t s a body of t r i b a l l o r e which i s r a p i d l y d i s a p p e a r i n g as the f r a g i l e t r i b a l c u l t u r e s become i n c r e a s i n g l y fragmented under the impact of Western c u l t u r e . U n l i k e ayahuasca , whose contemporary use among mes t i zos i s an amalgam of many t r a d i t i o n s , the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s drugs were never used ou t s i de the Bora , W i t o t o , and p o s s i b l y the Muinane t r i b e s [ 4 , 5 , 6 ] ; even w i t h i n these groups t h e i r use was a j e a l o u s l y guarded s e c r e t known on ly to the medic ine men ( C a l l e , H . , p e r s . comm., 1971). The most d e t a i l e d knowledge of the M y r i s t i c a c e o u s drug i s the pos ses s i on of the o l d e r men.* It i s they who know which of the many V i r o l a * U n l i k e ayahuasca , in which both men and women may p a r t i c i p a t e , the use of the o r a l l y - a c t i v e M y r i s t i c a c e o u s drug i s appa ren t l y an a c t i v i t y from which women are e x c l u d e d . 70 s p e c i e s are s u i t a b l e fo r the p r e p a r a t i o n of the d r u g ; i t i s they who know the r i g h t admixture p l a n t s , and the proper way to e x t r a c t and concen t r a t e the t r yp tamine- laden V i r o l a r e s i n . Most of these men have long s i n ce d i e d , and wi th them have d i e d the e thnomedica l t r a d i t i o n s of t h e i r p e o p l e . In the meantime, o u t s i d e i n f l u e n c e s brought by the white m e n — a l c o h o l i s m , C h r i s t i a n i t y , f o r e i g n d i s e a s e s fo r which the ind igenous people have l i t t l e r e s i s t a n c e , c u l t u r a l and g e o g r a p h i c a l d i s l o c a t i o n occas ioned by the r u t h l e s s e x p l o i t a t i o n of these t r i b e s in the rubber i ndus t r y in the e a r l y pa r t of t h i s c e n t u r y — a l l of these c i r cums tances have had a s h a t t e r i n g impact on these once proud and s e l f - s u f f i c i e n t j ung l e p e o p l e s . T h e i r e thnomedica l knowledge has been swept away by the ons laught of " p r o g r e s s , " a long wi th t h e i r r e l i g i o n , cosmo logy , .mag i c , a r t , poe t r y and mus i c . They are a people wa i t i ng to d i e ; they are regarded as l i t t l e more than an embarass ing reminder of the b r u t a l past and an impediment to the f u t u r e . As a r e s u l t , the Bora and Wi toto have become the v i c t i m s of a not-so-ben ign n e g l e c t . The sooner contemporary h i s t o r i c a l , s o c i a l , and economic f o r c e s have completed the dec imat ion of these p e o p l e , the sooner the way w i l l be c l e a r to implement the grand p lan of Amazonian "deve lopment " . T h i s a t t i t u d e , which u n f o r t u n a t e l y p r e v a i l s among many L a t i n Americans today , i s t y p i c a l of that which has too o f t e n c h a r a c t e r i z e d the a t t i t u d e s of Western European man toward a b o r i g i n a l peop les in o ther t imes and p l a c e s . A l though the p r e ced ing remarks r e f l e c t my own o p i n i o n s , formed as a r e s u l t of n e c e s s a r i l y incomplete pe r sona l o b s e r v a t i o n s , o the r s [7 ,8] have documented the a t r o c i t i e s committed in the name of the rubber 71 i n d u s t r y and the cu r r en t l e s s than o p t i m i s t i c ou t l ook fo r the con t i nued s u r v i v a l of the ind igenous Amazonian p e o p l e s . The o r a l l y - i n g e s t e d M y r i s t i c a c e o u s p a s t e s , l i k e so much of Bora and W i to to c u l t u r e , are on l y a f ad ing memory. They are remembered, by some of the middle-aged and o l d e r men, as something that was once used by t h e i r f a t h e r s and g r a n d f a t h e r s . The purposes for which the drug was used seem to have been l a r g e l y f o r g o t t e n , the i d e n t i f i c a t i o n of the proper source-p l a n t s has become t e n t a t i v e , the method of p r e p a r a t i o n i s now u n c e r t a i n , a sub jec t of debate among themse lves . T h i s i s e s s e n t i a l l y the s i t u a t i o n I encountered when our pa r t y a r r i v e d at the l i t t l e Wi to to/Bora v i l l a g e of Puco U r q u i l l o an the R io Ampiyacu, j u s t a shor t r i d e by motorboat from the Peruv ian r i v e r town of Pebas, at the con f l uence of the Ampiyacu and the Amazon. My o b s e r v a t i o n s are not un ique ; o the r s [ 5 , 6 ] have remarked on the f a i r l y l oose grasp in formants in t h i s r eg i on had on the b o t a n i c a l and e thnopharmaceut i ca l f i n e p o i n t s a s s o c i a t e d wi th the p r e p a r a t i o n of the M y r i s t i c a c e o u s p a s t e s . The North American s c i e n t i s t comes in h i s motorboat , b r i n g i n g money and sugar and s a l t , shotgun s h e l l s and c i g a r e t t e l i g h t e r s and m a l a r i a p i l l s , and many o ther coveted i t ems . He asks the in fo rmants to teach him the s e c r e t s of t h e i r f a t h e r s and t h e i r f a t h e r s ' f a t h e r s , a s s u r i n g them that these mat ters are of i n t e r e s t to h i s white-coa ted c o l l e a g u e s in t h e i r s h i n i n g h o s p i t a l s in Amer i ca , Canada, Europe . W e l l , i t seems a sma l l enough favor to a sk ; they are impressed by the f r i e n d l y v i s i t o r s , .and are eager to a c q u i r e the a t t r a c t i v e i tems they have brought to t r a d e ; s o , they are happy to h e l p o u t , they are anx ious to show t h e i r g o o d w i l l . Perhaps i f 72 they co-opera te they w i l l r e c e i v e a few e x t r a shot gun s h e l l s , or a p r e t t y p i e ce of c l o t h fo r t h e i r w i fe or s i s t e r . So they set ou t , w i th the best w i l l in the w o r l d , to l ead us to the p l a n t s , to show us how the paste i s made, to take us through the procedure s t ep by s t e p , as best they can r e c o n s t r u c t i t a c c o r d i n g to the formula of the long dead medic ine men. What does i t matter i f a few d e t a i l s have been f o r g o t t e n , i f one or two of the p l a n t s which were fo rmer l y used , which grew, perhaps , in t h e i r o r i g i n a l home but not in t h i s new v i l l a g e spawned by the m i g r a t i o n , what does i t matter i f these minor d e t a i l s have been l o s t or f o r g o t t e n or d e l i b e r a t e l y omit ted? The American s c i e n t i s t w i l l go away s a t i s f i e d . How i s he to know i f the drug has been prepared in e x a c t l y the manner p r e s c r i b e d by the a n c i e n t med ic ine men? How, fo r that mat te r , a re the Bora and Wi to to themselves to know? I encountered th ree in fo rmants who were w i l l i n g to p rov ide samples of the o r a l V i r o l a pas tes du r i ng our two-week s tay in the R io Ampiyacu r e g i o n . Two of these i n f o rman t s , Jorge Churay and Marcos Vega F l o r e s , were Bora I n d i a n s ; Jorge Churay l i v e d in Puco U r q u i l l o wh i l e Marcos F l o r e s l i v e d in B r i l l o Nuevo, a sma l l v i l l a g e about h a l f a day by motorboat u p r i v e r from Puco U r q u i l l o , on the banks of the Yaguasyacu, a t r i b u t a r y of the Ampiyacu. The p o p u l a t i o n of B r i l l o Nuevo was Bora whi le that of Puco U r q u i l l o was rough ly h a l f Bora and h a l f W i t o to ; each group i n h a b i t e d a d i f f e r e n t pa r t of the v i l l a g e separa ted by about h a l f a k i l o m e t e r . The Bora and Wi to to in Puco U r q u i l l o d i d not get a l ong very w e l l ; the re was an uneasy t ruce between them that reminded me of the s i t u a t i o n tha t might e x i s t between 73 P r o t e s t a n t s and C a t h o l i c s sha r i ng the same Boston g h e t t o . The t h i r d i n fo rman t , A l f r e d o Moreno, in a d d i t i o n to be ing the l o c a l shaman, was the headman of the Wi toto pa r t of Puco U r q u i l l o . Don A l f r e d o was by f a r the o l d e s t of these i n f o rman t s , and a l s o was the most s e c r e t i v e about the method of making the p a s t e . He agreed to p rov ide me wi th a sample of " o o ' - k o e y " as he c a l l e d i t , but would not a l low me to accompany him to f i n d the t r ee or to observe or photograph the procedure f o l l owed in i t s manufac ture . Only a f t e r he had d e l i v e r e d h i s f i r s t sample, s e v e r a l days a f t e r we had s t ru ck our b a r g a i n , d i d he take me to v i s i t the cumaia that he had cut down (cumala i s a gene r i c v e r n a c u l a r term for M y r i s t i c a c e o u s s p e c i e s in Peru) so that I c o u l d c o l l e c t bark and voucher spec imens . The f i r s t sample, d e r i v e d from V. s e b i f e r a , (DMK-40) turned out to have h igh l e v e l s of t r yp tamines ( c f . Chapter V) and d e f i n i t e l y e x h i b i t e d some o r a l a c t i v i t y in a subsequent b ioassay (see be low) . Before d e p a r t i n g f o r B r i l l o Nuevo on the second l eg of our f i e l d w o r k , I a r ranged w i th Don A l f r e d o to purchase three more paste samples ; I was to p i ck them up on our way back downr iver in about ten days t ime . I p a i d fo r the samples in advance , the enormous sum of nea r l y t h i r t y d o l l a r s , which fo r Don A l f r e d o was p robab l y a sma l l f o r t u n e . It was not enough, a p p a r e n t l y , to make an honest man out of h im. The samples were wa i t i ng fo r us on our way back th rough , each smal l banana-leaf packet of pas te accompanied by i t s own bundle of V i r o l a l eaves c o l l e c t e d as vouchers (DMK-67 ,68 , and 69 ) ; however, ana l y ses (Chapter V) showed very much lower l e v e l s of t r yp tamines in these pas tes than in the f i r s t sample. A l s o , the s i m i l a r i t i e s of the voucher specimens and the 74 almost i d e n t i c a l chemica l p r o f i l e s of these samples l e d me to conc lude that I had been g iven th ree samples which had a l l been d e r i v e d from the same t r e e . T h i s was c o n t r a r y to our agreement but of course d i d not become c l e a r u n t i l months l a t e r when the samples were ana l yzed in Canada. The task of l o c a t i n g three d i f f e r e n t s u i t a b l e specimens to prepare three d i f f e r e n t pas te samples i s c o n s i d e r a b l e , and I do not f a u l t Don A l f r e d o fo r t a k i n g the easy way ou t ; a f t e r a l l , he was j u s t t r y i n g to make some easy money. I do not know to t h i s day whether the e x c e p t i o n a l l y low t ryptamine l e v e l s found in these th ree pas te samples were a r e s u l t of d e l i b e r a t e decep t i on on the pa r t of Don A l f r e d o , or whether he j u s t happened to s e l e c t a specimen tha t was poor in the a c t i v e compounds. Subsequent ana l y ses (Chapter -V) have shown that there i s c o n s i d e r a b l e v a r i a t i o n from p l an t to p l a n t . Desp i t e the f a c t that the l a s t three samples r e c e i v e d « from Don A l f r e d o were d i s a p p o i n t i n g (the q u e s t i o n of t h e i r o r a l a c t i v i t y remains u n r e s o l v e d , s i n ce these three samples were immediate ly p r e se r ved in methanol and were not b ioassayed ) I s t i l l f e e l tha t Don A l f r e d o ' s knowledge of the pas tes was the most ex t ens i v e and the c l o s e s t to the " r e a l " t r a d i t i o n . T h i s f e e l i n g i s p a r t l y based on h i s f i r s t sample, which had h igh l e v e l s of t r yp tamines and d i d e x h i b i t o r a l a c t i v i t y , and p a r t l y on the f a c t tha t he p o i n t e d out s e v e r a l o ther " s u i t a b l e " V i r o l a specimens which proved to c o n t a i n s u b s t a n t i a l amounts of the r i g h t k inds of t r yp t am ines ; he a l s o seemed to have a more e x t e n s i v e knowledge of admixtures than the o ther in fo rmants I c o n t a c t e d . In f a c t , Don A l f r e d o u t i l i z e d two unusua l admixtures in making the pas te which have not been r e p o r t e d in the 75 l i t e r a t u r e . One of these was a f e r n , Anemia s p . , (DMK-39), the l eaves of which are made i n t o a t e a ; the water from t h i s i n f u s i o n i s then used to cook the V i r o l a r e s i n . The other admixture was a green c r u s t o s e l i c h e n of inde te rminant i d e n t i t y , which was growing on the bark of a sma l l t r e e , R inora racemosa (Mart . & Zucc . )Kuntze (DMK-38) ( V i o l a c e a e ) ; s c r a p i n g s of t h i s l i c h e n were added to the V i r o l a r e s i n whi le i t was s immer ing. Jorge Churay and h i s b ro the r- in- l aw Rey were our Bora in fo rmants in Puco U r q u i l l o . They f r e e l y admi t ted that they had never used the p a s t e s , but c l a imed that t h e i r f a t h e r s had done so and that they had watched i t s manufacture many t imes . They agreed to take us to c o l l e c t bark samples to make the " k u ' - r u -k u " , the Bora name fo r the M y r i s t i c a c e o u s p a s t e s , and a l s o to p o i n t out some of the d i f f e r e n t k inds of cumalas which are r e c o g n i z e d by the Bora . The Bora d i s t i n g u i s h 15 k inds of cumala and have d i f f e r e n t v e r n a c u l a r d e s i g n a t i o n s f o r each . Not a l l of them would be c o n s i d e r e d d i f f e r e n t by a t axonomis t . ( c f . Appendix I I ) . One morning we set ou t , in the company of Jorge Churay , Rey, and two o ther Bora to f i n d a s u i t a b l e cumala to make " k u ' - r u - k u " . The Boras p o i n t e d out s e v e r a l cumalas to us a l ong the way, but these were passed over as "not s t r o n g " . F i n a l l y we came to a l a r g e t r e e , about 70 fee t h igh wi th a DBH of about 25 cm (DMK-34, V. p a v o n i s ) . They removed the bark in long s t r i p s from the lower pa r t of the t r u n k . The inner cambia l l a y e r exuded a cop ious r e s i n wi th a very a romat i c s p i c y o d o r ; t h i s r e s i n was c l e a r but immediate ly o x i d i z e d to a r e d d i s h -orange c o l o r . A f t e r we had brought the bark back to the main Bora malocca in Puco U r q u i l l o , the Bora proceeded to c h i p away 76 the oute r l a y e r s of the bark us ing a machete. The inner phloem l a y e r wi th the cambium was then pounded i n t o long f i b r o u s s t r i p s wi th an ax-head. These were s t u f f e d i n t o a l a r g e aluminum pot and tamped down i n to the bottom of the v e s s e l . A min imal amount of wate r--bare ly enough to cover the s t r i p s of b a r k — was added and then the mixture was b o i l e d over a low f i r e fo r about 3 hours . A f t e r c o o l i n g , the bark s t r i p s were removed and tho rough l y rung out to remove excess water ; the r e s u l t i n g reddish-brown l i q u i d was concen t r a t ed f u r t h e r to a t h i c k p a s t e ; t h i s was mixed with an approx imate l y equ i v a l en t amount of s i f t e d ashes of C e c r o p i a sp . Ashes of the same s p e c i e s are a l s o added to the powdered coca which most of the Bora men chew f r e q u e n t l y . No other admixtures were added to t h i s paste sample . The completed " k u ' - r u - k u " was a dark reddisH-brown, f l e c k e d wi th b l a c k , and was e s s e n t i a l l y o d o r l e s s and t a s t e l e s s ; i t had l o s t most of the s p i c y , sharp odor which had c h a r a c t e r i z e d the "raw" V i r o l a r e s i n . The f o l l o w i n g evening I b i oassayed t h i s sample in an attempt to determine whether i t was in f a c t an o r a l l y a c t i v e h a l l u c i n o g e n . It produced no e f f e c t at a l l , d e s p i t e the f a c t tha t I had f a s t e d p r e v i o u s l y and over the course of the evening i nges t ed s e v e r a l t imes the recommended dose . Subsequent a n a l y s i s r e v e a l e d tha t t h i s p a r t i c u l a r pas te sample as we l l as the sou r ce-p l an t from which i t was d e r i v e d (DMK-34) d i d not c o n t a i n a l k a l o i d s of any so r t (Chapter V ) . A second sample made by Jorge Churay ( fo r which vouchers are u n a v a i l a b l e ) was a l s o comp le te l y wi thout a c t i v i t y . I t h ink that t h e i r f a i l u r e to s e l e c t a s u i t a b l e s p e c i e s on two o c ca s i ons i s an i n d i c a t i o n of j u s t how 77 much the t r i b a l knowledge of t h i s o r a l h a l l u c i n o g e n has d e t e r i o r a t e d over the l a s t few decades . S chu l t e s [5 ,6 ] has a l s o po in t ed out that sometimes s p e c i e s i n d i c a t e d by n a t i v e in formants as s u i t a b l e fo r making the pas tes were found not to c o n t a i n any a c t i v e t r yp t am ines . A l though some in formants remember how the pas tes were once p r epa r ed , s i n c e they themselves no longer u t i l i z e the d r u g , t h e i r b o t a n i c a l grasp of j u s t which s p e c i e s are s u i t a b l e has become somewhat l o o s e . Another paste sample was made by Marcos ( "e l n i n o " ) Vega F l o r e s , a Bora from the v i l l a g e of B r i l l o Nuevo, u p r i v e r from Puco U r q u i l l o . Don Marcos , a l though not a p a r t i c u l a r l y o l d man (I p l a c ed him at about 45 years o ld ) was h i g h l y regarded in B r i l l o Nuevo as an exper t on d r u g s , p o i s o n s , and med ic ines of a l l s o r t s . He had l ea rned most of. i t from h i s f a t h e r , Eugenio F l o r e s , now in h i s mid-70s, from whom he had i n h e r i t e d the o f f i c e of v i l l a g e medic ine man. Don Marcos p repared the pas te q u i t e d i f f e r e n t l y than the Bora at Puco U r q u i l l o had done. For one t h i n g , he took a, l e s s e r amount of bark from the t r e e , and was c a r e f u l to take i t from one s i d e o n l y . He e x p l a i n e d tha t t h i s was so tha t the t r ee would not d i e . The t r ee was a f l o w e r i n g specimen of V. e longa ta (DMK-59). Once back at h i s hu t , i n s t e a d of c h i p p i n g o f f the oute r bark wi th a machete, he c a r e f u l l y removed on ly the innermost cambia l l a y e r . T h i s he s t r i p p e d o f f in long f l e x i b l e f i b r o u s s t r i p s , on l y a few m i l l i m e t e r s t h i c k . The remainder of the bark was d i s c a r d e d . He beat the cambia l s t r i p s wi th a wooden m a l l e t , then d ipped them i n t o a v e s s e l of water and rung the j u i c e i n t o a sma l l enamel bowl . He repeated t h i s wr ing ing two or three t imes fo r each 78 s t r i p , then d i s c a r d e d i t . The bowl c o n t a i n i n g the cambia l w r ing ings was simmered u n t i l the l i q u i d was c o n c e n t r a t e d to a t h i c k pas te about the c o l o r and c o n s i s t e n c y of c h o c o l a t e s y rup . T h i s was then mixed w i th ashes made from the d r i e d f r u i t husk of Theobroma b i c o l o r (DMK-64) u n t i l i t was the c o n s i s t e n c y of t h i c k dough, and c o u l d be r o l l e d between the f i n g e r s i n t o a l i t t l e b a l l . P r i o r to a d d i t i o n of the a shes , I d ipped my f i n g e r i n t o the syrupy pas te and p l a ced a dab on the t i p of my tongue, as Don Marcos i n d i c a t e d I shou ld do . I immediate ly f e l t a sha rp , bu rn ing s e n s a t i o n , which turned my tongue numb fo r a few m inu tes ; at such a low dose , however, I d i d not expect any h a l l u c i n o g e n i c a c t i v i t y , nor was there any. Four of the seven pas te samples which were c o l l e c t e d in the R io Ampiyacu area were t e s t e d fo r o r a l a c t i v i t y , e i t h e r by myse l f or by o ther persons in our p a r t y ; in a d d i t i o n , the La Cho r r e r a " o o ' - k o e y " , c o l l e c t e d at La C h o r r e r a , Co lomb ia , in 1971, was a l s o t e s t e d . The two pas te samples-made by Jorge Churay at Puco U r q u i l l o were comp le t e l y i n a c t i v e as h a l l u c i n o g e n s and appeared to be p h y s i o l o g i c a l l y i n e r t , even though an amount s e v e r a l t imes tha t supposed ly r e q u i r e d was i n g e s t e d . The f i r s t sample made by A l f r e d o Moreno at Puco U r q u i l l o (made from V. s e b i f e r a , DMK-40) appeared to e x h i b i t some degree of o r a l a c t i v i t y at the dosage a s s a y e d . No p h y s i o l o g i c a l symptoms were n o t e d . The a c t i v i t y d e t e c t e d c o n s i s t e d of the e l i c i t a t i o n of hypnagogic imagery wi th the eyes c l o s e d . The sub jec t who assayed the sample f e l t tha t o r a l a c t i v i t y was d e f i n i t e l y p r e s e n t , and that a l a r g e r dose might have t r i g g e r e d the f u l l spectrum of h a l l u c i n o g e n i c e f f e c t s . On 79 a n a l y s i s , t h i s sample proved to c o n t a i n s u b s t a n t i a l c o n c e n t r a t i o n s of DMT and 5-MeO-DMT, a l though no 0-ca rbo l i ne s were d e t e c t e d . The f o u r t h sample assayed was d e r i v e d from V. e l onga t a (DMK-59) and was made by Marcos F l o r e s at B r i l l o Nuevo. T h i s sample proved to be the most p h y s i o l o g i c a l l y a c t i v e of any of the four a s s a y e d . O r a l i n g e s t i o n of approx imate l y a gram and a h a l f of the pas te e l i c i t e d a r a p i d and pro found r esponse , c h a r a c t e r i z e d by c o n s i d e r a b l e p h y s i o l o g i c a l d i s t r e s s r a t he r than by the p e r c e p t u a l and p s y c h o l o g i c a l d i s t u r b a n c e s u s u a l l y t y p i c a l of h a l l u c i n o g e n s . The i n i t i a l e f f e c t s , which man i f e s t ed w i t h i n ten minutes f o l l o w i n g i n g e s t i o n , c o n s i s t e d of a s t rong burn ing s ensa t i on in the mouth and t h r o a t , q u i c k l y deve l op ing i n t o a f e e l i n g of numbness in the l i p s , tongue, and t h r o a t . Swal lowing became d i f f i c u l t and b r e a t h i n g was l a b o r e d . The numbness g r a d u a l l y spread over the r e s t of my body, and my l imbs f e l t a t i n g l i n g s e n s a t i o n in t h e i r e x t r e m i t i e s . My body f e l t heavy and i n e r t , and I became c h i l l e d , My mind was r a c i n g and f e l t d i s s o c i a t e d from my body. I became somewhat a larmed at the r a p i d i t y of onset of the symptoms and f e l t p h y s i c a l l y most uncomfo r t ab l e . I focused on my own b r e a t h i n g , which was i r r e g u l a r and sha l l ow , and concen t r a t ed on t r y i n g to ma in ta in a s t e a d i e r , deeper rhythm of b r e a t h i n g . I found myse l f r e f l e c t i n g on the use of V i r o l a as an arrow p o i s o n , and f e l t t ha t I now unders tood how i t might be e f f e c t i v e in t h i s way; I f e l t i n capab l e of any p h y s i c a l e x e r t i o n , and thought that any f u r t h e r s t r e s s p l a c e d upon my r e s p i r a t o r y . s y s t e m - might be dangerous i f not l e t h a l . There were no hypnagogic images, p e r c e p t u a l changes , or h a l l u c i n a t i o n s , but I had the f e e l i n g that they might have 80 man i f e s t ed had I taken a s l i g h t l y h ighe r dose . A c u i t y of hea r ing proved to be g r e a t l y he igh t ened , and I was ext remely s e n s i t i v e to the cons tan t background hum of i n s e c t no i ses from the f o r e s t . The symptoms of p h y s i c a l d i s t r e s s p e r s i s t e d fo r about one h a l f -hour to 45 minutes , than g r a d u a l l y f aded . B rea th ing became e a s i e r , and I became drowsy and f e l l i n t o a s t a t e of " t w i l i g h t " s l e e p which may have l a s t e d ten or f i f t e e n minutes . G r a d u a l l y most of the symptoms d i s a p p e a r e d , but I con t inued to f e e l c h i l l e d fo r most of the n i g h t . The o v e r - a l l e f f e c t of the paste sample was more ak in to the a c t i o n s of a p resso r amine or gene ra l a n a e s t h e t i c than a h a l l u c i n o g e n , but the b i oa s s y l e f t no doubt that the sample d i d d e f i n i t e l y possess o r a l a c t i v i t y . It was not p a r t i c u l a r l y p l easan t or e n j o y a b l e , however. On l a t e r a n a l y s i s , t h i s sample was found to c o n t a i n a h igh c o n c e n t r a t i o n of 5-MeO-DMT and a t r a c e of NMT; s e v e r a l b i o l o g i c a l l y a c t i v e l i g n a n s were i s o l a t e d from DMK-59 by Don MacRae (MacRae, unpub l i shed d a t a , 1982), and these may we l l have c o n t r i b u t e d to the pha rmaco log i c a l a c t i v i t y . No /3-carbolines were de t e c t ed in t h i s pas te sample. The f i f t h pas te sample which has been sub j e c t ed to human b ioassay i s the La Chor re ra " o o ' - k o e y " , c o l l e c t e d in 1971 in Co lomb ia . No h a l l u c i n o g e n i c or o ther p h y s i o l o g i c a l a c t i v i t y was ever d e t e c t e d with t h i s sample in s e v e r a l t r i a l s , even though e x c e s s i v e doses were i nges t ed on some o c c a s i o n s . I n t e r e s t i n g l y , t h i s sample con t a i ned s u b s t a n t i a l amounts of t r yp tamines and d e t e c t a b l e amounts of tetrahydro-/3-c a r b o l i n e s ( c f . Chapter V ) . A l though the l i m i t e d numbers of human b ioassays r epo r t ed here are not c o n c l u s i v e , they do i n d i c a t e tha t the o r a l a c t i v i t y 81 of these M y r i s t i c a c e o u s pas tes may be determined by v a r i a b l e s o ther than the f o r t u i t o u s combinat ion of t r yp tamine d e r i v a t i v e s and /3-carbol ines . The pas te sample showing the g r e a t e s t a c t i v i t y c o n t a i n e d on l y a s i n g l e a l k a l o i d , 5-MeO-DMT, which i s supposed ly not o r a l l y a c t i v e ; on the o ther hand, the La Chor re ra sample, which d i d c o n t a i n the a c t i v e t r yp tamines and /3-carbol ines in combina t ion was comp le te l y wi thout o r a l p h y s i o l o g i c a l a c t i v i t y . Samples l a c k i n g a l k a l o i d s a l s o had no a c t i v i t y , and t h i s r e s u l t i s at l e a s t c o n s i s t e n t wi th e x p e c t a t i o n s . The f a c t o r s de te rm in ing presence or absence of o r a l a c t i v i t y in those samples which do c o n t a i n a l k a l o i d s s t i l l remain u n e l u c i d a t e d . The r e s u l t s of these human t r i a l s , however, c o n s i d e r e d together w i th those ob ta ined from the ir\ v i t r o MAOI assays of the pas te samples ( c f . Chapter VI) i n d i c a t e tha t the o r a l a c t i v i t y i s not due s imply (or s o l e l y ) to the o r a l p o t e n t i a t i o n of the t r yp tamines by the /3-carbol ines . In f a c t , the a l k a l o i d a l p r o f i l e s of the M y r i s t i c a c e o u s pas te samples has been shown to be h i g h l y v a r i a b l e , r e f l e c t i n g the chemica l v a r i a b i l i t y in the s o u r c e - p l a n t s used to prepare the drug ( c f . Chapter V ) . T h i s , in t u r n , would p robab ly r e s u l t in c o n s i d e r a b l e v a r i a t i o n in p h a r m a c o l o g i c a l a c t i v i t y from sample to sample. In f a c t , the d i f f i c u l t i e s encountered in e x e r c i s i n g some degree of pha rmaceu t i c a l q u a l i t y - c o n t r o l over the compos i t i on of d i f f e r e n t batches of the drug may e x p l a i n some of the e thnopha rmaco log i ca l p u z z l e s about these p a s t e s , e . g . , the f a c t tha t they are made and used s e c r e t l y by the medic ine men, the f a c t that they have never been used o u t s i d e the Bora , W i t o to , and Muinane groups a l t hough V i r o l a s n u f f s are w ide ly used in the Amazon B a s i n ; and 82 the f a c t tha t t h e i r use even in these t r i b e s has d i m i n i s h e d or d i s appea red as con t a c t s wi th o u t s i d e c u l t u r e s have i n c r e a s e d . 83 I I I . L i t e r a t u r e C i t e d 1. Luna , L. E. (1983) The Concept of P l an t s as Teachers Among Four M e s t i z o Shamans of I q u i t o s , Nor theas t Pe ru . Paper p resen ted at the Symposium on Shamanism, X l t h I n t e r n a t i o n a l Congress of A n t h r o p o l o g i c a l and E t h n o l o g i c a l S c i e n c e s , Phase 2. Vancouver , B. C . , August 20-23 1983. 2. Luna , L. E. ( 1983) Don E m i l i o y sus D o c t o r i c i t o s . F i l m p resen ted at X l t h I n t e r n a t i o n a l Congress of A n t h r o p o l o g i c a l and E t h n o l o g i c a l S c i e n c e s , Phase 2. Vancouver , B. C , August 20-23 1983. 3. R i v i e r , L. & J . L i ndgren (1972) Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k : E t h n o b o t a n i c a l and Chemica l I n v e s t i g a t i o n s . Economic Botany 29:101-129 4. S c h u l t e s , R. E. (1969) V i r o l a as an O r a l l y - a d m i n i s t e r e d H a l l u c i n o g e n . Harvard B o t a n i c a l Museum L e a f l e t s 22:229-40 5. S c h u l t e s , R. E . , T . Swain, & T . Plowman (1977) V i r o l a as an O r a l H a l l u c i n o g e n Among the Boras of Pe ru . Harvard B o t a n i c a l Museum L e a f l e t s 25:259-272 6. S c h u l t e s , R. E. & T . Swain (1976) Fu r the r Notes on V i r o l a as an O r a l H a l l u c i n o g e n . J ou rna l of P s y chede l i c  Drugs 8:317-324 7. C o l l i e r , R i cha rd (1968) The R i ve r tha t God F o r g o t : The S to ry  of the Amazon Rubber Boom. E. P. Dutton & C o . , New York . 8. Shoumatof f , A lex (1978) The R i v e r s Amazon. S i e r r a C lub Books, San F r a n c i s c o . 84 PART I I I : PHYTOCHEMICAL AND PHARMACOLOGICAL INVESTIGATIONS 85 CHAPTER IV: ALKALOID CONSTITUENTS OF AYAHUASCA BREWS, SOURCE-PLANTS, AND ADMIXTURE PLANTS I. I n t r o d u c t i o n The h a l l u c i n o g e n i c beverage ayahuasca (Quechua fo r " v i ne of the s o u l s " ) i s w ide ly used fo r m e d i c i n a l , r i t u a l , and r e c r e a t i o n a l purposes by the a b o r i g i n a l and mes t i zo p o p u l a t i o n s i n h a b i t i n g the Amazon B a s i n . While ayahuasca i s the most common term fo r the drug in the Peruv ian Amazon, in d i f f e r e n t r eg ions i t i s known by v a r i ous v e r n a c u l a r names, i n c l u d i n g yage, c a a p i , natema, and p inde [1 ] , The b i t t e r , c o f f e e - c o l o r e d beverage i s p repared by b o i l i n g the bark of the jung le l i a n a B a n i s t e r i o p s i s  c aap i (Spruce ex G r i s e b . ) Morton (Ma lp igh iaceae ) toge ther wi th the l eaves of v a r i o u s admixture p l a n t s , the a d d i t i o n of which i s b e l i e v e d to i n t e n s i f y or modify the e f f e c t [ 2 ] . Many of the admixture p l a n t s used remain u n c h a r a c t e r i z e d e i t h e r b o t a n i c a l l y or c h e m i c a l l y , however those used most commonly are P s y c h o t r i a  v i r i d i s Ruiz & Pavon, P s y c h o t r i a c a r t hagenens i s J a c q . , and D i p l o p t e r y s cabrerana (Cua t r e casas )Ga tes ; the l a t t e r i s a l s o a l i a n a in the Ma lp igh i aceae fo rmer l y known as B a n i s t e r i o p s i s  rusbyana [3 , Plowman, T . , p e r s . comm., 1980]. The most complete study to date of the chemis t r y of the ayahuasca beverage , and of the sou r ce-p l an t s and admixture p l a n t s used in i t s manufac ture , i s tha t of R i v i e r and L indgren [ 4 ] ; these i n v e s t i g a t o r s s t u d i e d ayahuasca samples and vouchered b o t a n i c a l m a t e r i a l c o l l e c t e d among the Sharanahua and C u l i n a t r i b e s of the upper R io Purus i n Pe ru . Us ing GC/MS as the p r imary a n a l y t i c a l method, R i v i e r and L indgren found that the 0-c a r b o l i n e a l k a l o i d s harmine, ha rma l i ne , and t e t r ahyd roha rm ine , 86 and DMT were the major a c t i v e c o n s t i t u e n t s of ayahuasca ( c f . F i g . 1 & 2 ) . The /3-carbolines are c o n s t i t u e n t s of B a n i s t e r i o p s i s  caap i [5] wh i le N ,N-d imethy l t ryptamine has been r epo r t ed in the two P s y c h o t r i a spp. [4] as we l l as in D. cabrerana [6] Hashimoto et a l . [7 ,8 ] have r epo r t ed the i s o l a t i o n of 6 /3-c a r b o l i n e bases from the l eaves of B a n i s t e r i o p s i s caap i in a d d i t i o n to the three main c o n s t i t u e n t s ( v i z . : harmic amide, a c e t y l norharmine , ke to t e t r ahyd rono rha rm ine , harmine N-oxide, harmic a c i d methyl e s t e r , and h a r m a l i n i c a c i d ) . However the ext remely low c o n c e n t r a t i o n s of these compounds in the p l an t (.007-.0001 %) make i t u n l i k e l y that they c o n t r i b u t e s i g n i f i c a n t l y to the pha rmaco log i c a l a c t i v i t y of ayahuasca . A l though the a c t i v e a l k a l o i d s of ayahuasca are now.known, c e r t a i n a spec t s of the pharmacology of the drug remain to be c l a r i f i e d . DMT i s known to be a potent h a l l u c i n o g e n but i s a l s o known to be i n a c t i v e when i nges ted o r a l l y [9 ] , p robab l y due to deaminat ion by i n t e s t i n a l and hepa t i c monoamine ox idase (MAO). The /3-carbol ines p resent in B. caap i are known to be h i g h l y a c t i v e r e v e r s i b l e i n h i b i t o r s of MAO [10 ,11 ,12] and are p robab ly a l s o h a l l u c i n o g e n i c . The psychotomimet ic a c t i v i t y of the /3-c a r b o l i n e s i s not we l l unders tood because there are r e l a t i v e l y few c l i n i c a l i n v e s t i g a t i o n s of the e f f e c t s of /3-carbol ines on human s u b j e c t s . Naranjo [13] r epo r t ed harmal ine to be h a l l u c i n o g e n i c at o r a l doses of 4 mg/kg but cou ld not r epor t s i m i l a r l y unequ i voca l r e s u l t s f o r harmine or t e t r ahyd roha rm ine . Pennes & Hoch [14] r epo r t ed harmine was o r a l l y i n a c t i v e at l e v e l s approach ing 12 mg/kg. I t has been suggested [ 2 , 6 , 9 , 1 5 , 1 6 ] tha t the h a l l u c i n o g e n i c p r o p e r t i e s of the crude ayahuasca brew 87 r e s u l t from a s y n e r g i s t i c i n t e r a c t i o n among the v a r i o u s c o n s t i t u e n t s ; s p e c i f i c a l l y , tha t i t r e s u l t s from an o r a l a c t i v a t i o n of the DMT through the i n h i b i t i o n of MAO by the 0-c a r b o l i n e s . T h i s mechanism would render the DMT o r a l l y a c t i v e by b l o c k i n g i t s deg rada t i on by v i s c e r a l MAO. A l though t h i s mechanism i s reasonab le and has long been accepted in the e thnopha rmaco log i ca l l i t e r a t u r e , the e f f e c t of ayahuasca on monoamine ox idase has not been e x p e r i m e n t a l l y de te rmined . T h i s chap te r p r e sen t s the r e s u l t s of phy tochemica l i n v e s t i g a t i o n s of the a l k a l o i d c o n s t i t u e n t s of ayahuasca and the sou r ce-p l an t s used in i t s manufac ture . An e v a l u a t i o n of the e f f e c t of ayahuasca on MAO i_n v i t r o i s r epo r t ed in Chapter V I . 11. M a t e r i a l s and Methods A. F i e l d C o l l e c t i o n of Drug Samples and P lan t M a t e r i a l s C o l l e c t i o n numbers c i t e d throughout t h i s paper r e f e r to the p e r s o n a l c o l l e c t i o n numbers of D. McKenna, w i th the excep t i on of Plowman 6040 ( D i p l o p t e r y s cabrerana ) and Plowman 6041 (B. c aap i v a r . " c i e l o " ) . P l an t m a t e r i a l f o r Plowman 6040 and 6041 was k i n d l y s u p p l i e d by Dr . Timothy Plowman of the F i e l d Museum in C h i c a g o . A u t h e n t i c a t e d herbar ium vouchers fo r a l l c o l l e c t i o n numbers c i t e d have been d e p o s i t e d in the Herbar ium of the Dept . of Botany, U n i v e r s i t y of B r i t i s h Co lumbia . D u p l i c a t e vouchers of most c o l l e c t i o n s are a l s o on d e p o s i t at the Ch icago F i e l d Museum. Dur ing e t h n o b o t a n i c a l f i e l dwo rk i n the s p r i n g of 1981, e i g h t samples of ayahuasca p r e p a r a t i o n s were ob t a i ned from 88 ayahuasqueros l i v i n g on the o u t s k i r t s of the Pe ruv ian towns of I q u i t o s , P u c a l l p a , and Ta r apo to . These samples were q u a l i t a t i v e l y ana l yzed us i ng 2-d imens iona l TLC and q u a n t i f i e d us ing HPLC. I d e n t i f i c a t i o n of a l k a l o i d s was based on compar ison wi th a u t h e n t i c s t anda rds . Ayahuasca samples used f o r a n a l y s i s are i d e n t i f i e d in t h i s chapte r by the name of the ayahuasquero from whom they were o b t a i n e d , and a l s o by a number. In some cases more than one sample was ob ta ined from the same p e r s o n . See Chapter III f o r d e t a i l s of methods used in the c o l l e c t i o n of drug samples and source p l a n t s fo r phy tochemica l a n a l y s i s ; i n f o r m a t i o n on herbar ium voucher c o l l e c t i o n s i s t a b u l a t e d in Appendix 1 1 . B. Two-dimensional T h i n - l a y e r Chromatography The f l u o r e s c e n c e c h a r a c t e r i s t i c s , E h r l i c h ' s c o l o r r e a c t i o n s , and Rf va lues in two so l v en t systems were determined fo r the t ryp tamine and /3-carboline s tandards used in t h i s study (Table I I ) . Two-dimensional TLC of the t r yp tamine and /3-c a r b o l i n e s tandards i s shown g r a p h i c a l l y in F i g . 3; F i g . 4 i l l u s t r a t e s the Rf va lues of s e l e c t e d mix tu res of t r yp tamine s tandards f o l l o w i n g 1-dimensional development i n so l ven t 1 and so l v en t 2. Two-dimensional t h i n - l a y e r chromatography was c a r r i e d out u s i ng 10 x 10 cm Polygram S i l i c a Ge l G U V 2 5 " p r ecoa ted p l a t e s (Brinkmann I ns t rumen ts ) . The o r i g i n was marked wi th p e n c i l in the lower l e f t hand co rne r of the p l a t e , 1.0 cm from the bottom and l e f t - h a n d edge. F i v e ul a l i q u o t s of the m a t e r i a l to be ana l yzed ( c o n s i s t i n g e i t h e r of the crude ayahuasca samples or in TABLE I I : HRF* VALUES OF TRYPTAMINE AND p-CARBOLINE STANDARDSt Compound UV E h r l i c h ' s Name S o l v e n t 1* S o l v e n t 2tt f 1 u o r e s c e n c e r e a c t i onb 0 - c a r b o l i n e s : norharman 62 . 7 84 .8 b l ue -harman 67 .9 84 .8 dark b l u e -harma1 an 49 .0 67 . 7 g r e y - g r e e n nt£ t e t r a h y d r o -harman 30 .0 40 . 3 nf - UVA* nt t e t r a h y d r o -3 - c a r b o x y - h a r m a n 0 .0 55 .9 nf - UVA nt harm 1ne 58 . 2 82 .9 dark b l u e -harma1i ne 33 . 2 45 . 7 aqua -t e t r a h y d r o -harmi ne 24 .0 35 . 7 nf - UVA I t . b l u e * harmol 35 . 7 8 1 .3 dark b l u e -harma1ol 17 . 3 35 . 7 aqua -6-MeO-harman 66 .0 82 .7 I t . b l u e -6-MeO-harma1 an 44 . 3 56 .8 tan -6 - M e O - t e t r a h y d r o -harman 26 .0 34 . 1 nf - UVA nt b r e v 1 c o 1 i ne 56 . 5 79 . 5 dark b l u e nt mean % s t a n d a r d d e v i a t i o n o = 6 . 24 5 . 5 T ryptami nes : 1 - t r y p t o p h a n 0. .0 51 . 5 nf - UVA b 1 u e - g r a y 5 - h y d r o x y -1 - t r y p t o p h a n 0. .0 46 . 2 " b l u e - g r a y t r y p t o p h o l 75 . 1 85 . 6 g rey t ryptam i ne 35 . 0 29 . 5 . " b 1 u e - g r a y 5 - h y d r o x y -t ryptam i ne 13. 5 18 . 9 " g r a y 5-MeO-tryptami ne 29 . 4 25. 0 " I t . b l u e 6 - M e O - t r y p t a m i n e 29. 3 24 . 0 " dk, b l u e gram 1ne 28 . 8 25. 9 " I t . v i o l e t N-methy1 -t ryptam i ne 10. 7 17 . 8 II b l u e - g r a y DMT 4 1 . 3 4 1 . 9 " b 1 u e - g r a y 5-hydroxy-DMT 16 . 4 28 . 7 " I t . b l u e 5-MeO-DMT 35 . 5 37. 1 " I t b l u e ps i1oc i n 42. 9 41 . 6 " dk. v i o l e t ps i1ocyb i n 0 . 0 3 . 2 " I t . v i o l e t me la ton i n 46 . 3 83 . 7 " I t . b l u e 5-MeO-di i s o p r o p y l -t ryptam i ne 93 . 7 54 . 1 " I t . b l u e mean % s t a n d a r d d e v i a t i o n o = 1 . 7 8 . 0 9 0 d i s t a n c e migra ted by compound * hRf = X 100 d i s t a n c e migra ted by so l v en t t A l l s t andards used in TLC and in the MAO assays r epo r t ed in Chapter VI were purchased from Sigma Chemica l C o . , S t . L o u i s , Mo, or A l d r i c h Chemica l C o . , Mi lwaukee, W i s . , w i th the f o l l o w i n g e x c e p t i o n s : P s i l o c y b i n and p s i l o c i n s tandards (Sandoz: l o t numbers 8001 & 5001, r e s p e c t i v e l y ) were g i f t s of the Ch i e f of S c i e n t i f i c S e r v i c e s , Hea l t h and Wel fare Canada, Ottawa, O n t a r i o ; 5-MeO-d i i sopropy l t r yp tamine and 3-[2-(2,5 d i m e t h y l ) p y r r o l y l e t h y l ] - i n d o l e were g i f t s of D r . B. Abeysekara , R a d i o p h a r m a c e u t i c a l s , I n c . , Vancouver , B. C ; 6-MeO-MTH/3B was a g i f t of D r . Bo Ho lmstedt , K a r o l i n s k a I n s t i t u t e , S tockho lm; b r e v i c o l i n e was a g i f t of D r . E. L e e t , Dept . Of Chemi s t r y , U n i v e r s i t y of M inneso ta , M i n n e a p o l i s , M i n n . ; t e t rahydroharmine was s y n t h e s i z e d from 6-MeO-tryptamine (Sigma) and a ce t a l dehyde , a c c o r d i n g to the method of Akabor i and S a i t o [29 ] . $ So l ven t 1: e the r/2-butanone/conc . NH«OH 5:4:1 (upper phase) # So l ven t 2: n-propano l/1 .5% NH«OH 9:2 b r e a c t i o n to E h r l i c h ' s reagent ( c f . [20]) $ nt= not t e s t e d 4> nf - UVA = not f l u o r e s c e n t ; v i s i b l e as UV-absorb ing spot under shor t wave UV ii Slow c o l o r - r e a c t i o n v i s i b l e a f t e r 24 hours mean s tandard d e v i a t i o n CJ mean % s tandard d e v i a t i o n = X 100 mean hRf o o a: Ld > _ J o 9Qf BQ-7Q. 5Q; 4Qf 3Qf iQ; 30X 14+ 17X 2+ 11 + 1+ 6+ 3+ 12+ 18X 26X 29X 9+ 20 21*X22 7+ 4+ 25X 13+ 8 + 10+ 19X 23X QMt-10- so. 30- 40< 16 15 -K r K — 50- G0> 70- BO. S O L V E N T 2 RF X 1 0 0 91 90 < 100-F i g u r e 3 - P o s i t i o n s of Tryptamine and 0 - c a r b o l i n e S tandards in Two D imens iona l Th in Layer Chromatography + = 0 - c a r b o l i n e s X = 1. Norharman 15. 2. Harman 16. 3. Harmalan 17. 4. Tetrahydroharman 18. 5. Tetrahydroharman- 19. 3-ca rboxy la te 20. 6. Harmine 21 . 7. Harmal ine 22. 8. Tet rahydroharmine 23. 9. Harmol 24. 10 . Harma lo l 25. N,N 1 1 . 6-Me0-harman 26. 12 . 6-Me0-harmalan 27. 13 . 6-Me0-tetrahydroharman 28. 14 . B r e v i c o l i n e 29. 30. T ryptamines L-t ryptophan 5-hydroxy- l- t ryptophan T r y p t o p h o l Tryptamine ' 5-hydroxy-tryptamine 5- MeO-tryptamine 6- MeO-tryptamine Gramine N-methyl-tryptamine N ,N-d imethy l t r yp tamine 5-hydroxy--d imethy l t r yp tamine 5-MeO-N,N-dimethyltryptamine P s i l o c i n P s i l o c y b i n Me l a ton in 5-MeO-di i s o p r o p y l t r y p t a m i n e 9Qr o o •c-l BQ. X 7Q; L _ Ol BQ. •<H 5Q> r -1 • 1 4Q. 1 ' 1 > _ J 3 Q r D 2 Q f i Q -Q-1 0 0 -9Q; O O r-i BQ. X 7Q: L _ C £ £ Q -5 Q r r -1 1 1 4Q.. L.l i > _ l 3Q? D L D E Q . l Q r Q-30+ 17+ 24+ 24+ IJt 18+ 26+ + 20+21 23+ 23+ 25+ 28. 18+ 20+ 19 + 29+ 24+ 24+ 24+ 22+ 19+ -15. I V 92 3 ' 4 . 5- B« 7» B« 9« 1 0 -5TANDARD COMBINATIONS 17+ 29+ 15+ 24+ 24 + 26+ 18+ 23+ 23+ 27+24 25+ 16+ » * 20 + 18+ 19+ 30+ 24+ 24+ 24 + 22+ 19 + 28+ H-0* 1' 2- 3' 4 . 5 . G • 7. 8- 9-5TANDARD COMBINATIONS F i g u r e 4 - One D imens iona l TLC of M ix tu res of Tryptamine Standards Compos i t ion of s tandard m i x t u r e s : * 1. Tryptamine + NMT + DMT 2. DMT + 5-MeO-DMT + NMT 3. DMT + 5-HO-DMT + p s i l o c i n + p s i l o c y b i n 4. 5-MeO-tryptamine + 6-MeO-tryptamine 5. Tryptamine + 5-MeO-tryptamine + 5HT 6. L-t ryptophan + t r y p t o p h o l 7. 5-hydroxy- l- t ryp tophan + 5HT 8. DMT + gramine 9 DMT + me la ton in l O - l l -10. DMT + 5-MeO-d i i sopropy l t r yp tamine * Numbering of i n d i v i d u a l c o n s t i t u e n t s F i g . 3. i s a c c o r d i n g to key in 93 the case of the admixture p l a n t s of a methanol s o l u t i o n of the p u r i f i e d a l k a l o i d f r a c t i o n ) were a p p l i e d to the o r i g i n us ing a M i c rocap a p p l i c a t o r . The a p p l i e d sample was d r i e d under a gen t l e stream of a i r , and then deve loped in the f i r s t d i r e c t i o n us ing e ther/2-butanone/conc . NH aOH 5:4:1 (So lvent 1) . So lvent 1 was f r e s h l y p repared in a sepa ra to r y funne l and the upper phase was c o l l e c t e d f o r TLC . F o l l o w i n g development in So lvent 1, the p l a t e s were removed and a l l owed to a i r dry in a fume hood. Development in the second dimension was commenced when the p l a t e s were comple te l y f r e e of So lvent 1, i n d i c a t e d by the absence of any so l ven t odo r . P l a t e s were then r o t a t e d 90° to the l e f t w i th r e spec t to t h e i r p o s i t i o n in So lvent 1, and deve loped in So l ven t 2, c o n s i s t i n g of n-propanol/1.5% NH„OH 9 :2 ; t h i s so l v en t was s t a b l e for 2-3 days at room temperature i f kept s e a l e d in a g round-g lass s toppered f l a s k . Development in both So l ven t 1 & 2 was c a r r i e d out at ambient temperature in an u n l i n e d 10 x 30 x 26 cm g l a s s chromatographic tank c o n t a i n i n g 50 ml ± 5 ml of s o l v e n t . F o l l o w i n g development in So lvent 2, p l a t e s were removed and a i r - d r i e d in a fume hood fo r 30-60 min or o ve rn igh t on the l a b o r a t o r y bench. P l a t e s were examined under sho r t - and long-wave UV l i g h t to v i s u a l i z e the a l k a l o i d s . DMT and te t rahydro-/3-carbo l ines are v i s i b l e as dark spots under short-wave UV wh i le the a romat i c and d ihydro-/3-carbo l ines g i ve c h a r a c t e r i s t i c s t rong f l u o r e s c e n t c o l o r s under long-wave UV. D u p l i c a t e p l a t e s were sprayed wi th E h r l i c h ' s reagent [17] which g i v e s b lue to v i o l e t c o l o r s wi th DMT and other t r yp tamine d e r i v a t i v e s f o l l o w i n g exposure to HCI v apo r s . Aromat ic and d ihydro-/3-carbo l ines do not reac t w i th E h r l i c h ' s reagent however 94 te t rahydroharmine g i v e s a c h a r a c t e r i s t i c r o b i n ' s egg b lue c o l o r which deve lops over 24 h r . T h i s slow r e a c t i o n can be used to d i s t i n g u i s h te t rahydroharmine from i t s more aromat ic ana logues and a l s o from the t r yp tamines which g i ve darker b lue r e a c t i o n s tha t appear w i t h i n 30 min of exposure to HCI v a p o r s . A TLC p l a t e c o n t a i n i n g a l i q u o t s of known /3-carboline and t ryptamine s tandards was deve loped s imu l t aneous l y w i th the sample p l a t e s ; c o n s t i t u e n t s in the samples co r r e spond ing to known s tandards c o u l d thus be r e a d i l y i d e n t i f i e d by compar ison of the sample p l a t e s wi th the " s t a n d a r d " p l a t e ( c f . "ayahuasca a n a l o g u e " , F i g . 6 ) . C. High P ressure L i q u i d Chromatography (HPLC) 1. A n a l y t i c a l c o n d i t i o n s A V a r i a n model 5000 HPLC i n t e r f a c e d w i th a S p e c t r a - p h y s i c s model SP4100 computing i n t e g r a t o r was used f o r the q u a n t i t a t i v e a n a l y s i s of the ayahuasca samples , the B a n i s t e r i o p s i s caap i c u l t i v a r s , and the DMT-conta in ing admixture p l a n t s ( F i g . 5 ) . C o n s t i t u e n t s were d e t e c t e d by UV a b s o r p t i o n at 260 nm wi th a V a r i a n model 634 v a r i a b l e wavelength U V / v i s i b l e spec t ropho tomete r . Column c o n s i s t e d of a V a r i a n Micropak MCH-10 r eve r se phase co lumn, 30 cm x 4 mm i . d . So l v en t s were methanol/water c o n t a i n i n g 0.05% t r i e t h y l a m i n e . A g r ad i en t e l u t i o n program was used fo r the a n a l y s i s , from 60-90 % methanol a t a r a te of 1%/min. So l ven t f low ra te was 2 ml/min . Samples were a p p l i e d to the column v i a a Rheodyne model 7125 s y r i nge l o a d i n g sample i n j e c t o r f i t t e d w i th a 20 ul sample l o o p . Samples 95 B F i g u r e 5 - HPLC E l u t i o n P r o f i l e of Peruvian Ayahuasca A.) A l k a l o i d standards. B.) Ayahuasca sample. A n a l y t i c a l C o n d i t i o n s : S o l v e n t : methanol/H 20 c o n t a i n i n g 0.05% t r i e t h y l a m i n e Column: V a r i a n Micropak MCH-10, 30 cm x 4 mm i . d. Program: 60 - 90 % methanol, 1%/min; flow r a t e = 2 ml/min D e t e c t i o n : 260 nm, V a r i a n Model 634 U V / v i s i b l e spectrophotometer 96 were loaded onto the sample i n j e c t o r us ing a 25 ul Hami l ton #702 m i c r o l i t e r s y r i nge f i t t e d w i th a #22 gauge 90° beve led need l e . 2. Q u a n t i t a t i v e methods Q u a n t i t a t i v e ana l y ses were c a r r i e d out u s i ng the e x t e r n a l s tandard program s u p p l i e d wi th the SP4100 computing i n t e g r a t o r . In t h i s method, the column i s c a l i b r a t e d wi th a mixture c o n t a i n i n g known c o n c e n t r a t i o n s of s tandard compounds. The i n t e g r a t o r program c a l c u l a t e s the peak area of each s tandard in the c a l i b r a t i o n mixture and no rma l i zes the response f a c t o r . C a l i b r a t i o n at two or more c o n c e n t r a t i o n s enab les the i n t e g r a t o r to generate the c o e f f i c i e n t s of a l i n e a r equa t ion r e l a t i n g sample c o n c e n t r a t i o n to peak a r e a . T h i s c a l i b r a t i o n data i s s t o r ed in the memory c i r c u i t s of the i n t e g r a t o r and i s used to c a l c u l a t e the c o n c e n t r a t i o n of components of i n t e r e s t in subsequent sample runs by compar ison of the peak areas of sample components wi th the peak areas of s tandards in the c a l i b r a t i o n mix. Stock s o l u t i o n s of harmine, ha rma l i ne , t e t r ahydroharmine and DMT s tandards were made up to a p r e c i s e c o n c e n t r a t i o n of 1 mg/ml. Equa l a l i q u o t s from the s tock s o l u t i o n s were combined, g i v i n g a c a l i b r a t i o n mix ture of 0.25 mg/ml of each component; 1:1 d i l u t i o n o f . a p o r t i o n of t h i s mixture wi th methanol gave a second c a l i b r a t i o n mixture in which the c o n c e n t r a t i o n of each component was 0.125 mg/ml. The SP4100 i n t e g r a t o r was c a l i b r a t e d by making s i n g l e 20 ul i n j e c t i o n s of the c a l i b r a t i o n mix ture at each c o n c e n t r a t i o n l e v e l . The i n t e g r a t o r was r e c a l i b r a t e d f o l l o w i n g every f i v e sample i n j e c t i o n s . 97 3. Sample p r e p a r a t i o n a . Ayahuasca samples . The ayahuasca samples which had not been d i l u t e d w i th methanol on c o l l e c t i o n were d i l u t e d fo r a n a l y s i s so tha t the a l k a l o i d s p resen t were w i t h i n the c o n c e n t r a t i o n range of the c a l i b r a t i o n s t a n d a r d s . A 1.5 ml a l i q u o t of the crude p r e p a r a t i o n was d i l u t e d to 15 ml wi th c h i l l e d methanol and a wh i te , f l o c c u l e n t , p ro t e i na ceous p r e c i p i t a t e which separa ted from s o l u t i o n was removed by f i l t r a t i o n . T h i s d i l u t e d p r e p a r a t i o n , a f t e r f i l t r a t i o n through a Pas teur p i p e t t e p lugged wi th g l a s s wool , was i n j e c t e d d i r e c t l y i n t o the HPLC. Two r e p l i c a t e s of each ayahuasca sample were p r e p a r e d , and each r e p l i c a t e sample was i n j e c t e d twice du r i ng separa te runs f o l l o w i n g c a l i b r a t i o n of the i n t e g r a t o r us ing the s tandard m i x t u r e s . Va lues r epo r t ed in Tab le III are the means of these four r e p l i c a t e i n j e c t i o n s . Va lues g iven are 10x the a c t u a l va lue measured s i n c e the samples measured were 1/10 the c o n c e n t r a t i o n of the u n d i l u t e d brew. The ayahuasca samples which had been d i l u t e d wi th methanol in the f i e l d were q u a n t i f i e d f o r a l k a l o i d s in terms of mg/g dry weight of the l y o p h i l i z e d sample . F i f t e e n ml of the methanol-d i l u t e d sample was evaporated on a steam b a t h , f r o z e n , then l y o p h i l i z e d . A p o r t i o n of the f r e e z e - d r i e d r e s i due was ground to a f i n e powder and 100 mg was t r a n s f e r r e d to a 100 ml round bottom f l a s k . Ten ml 100% methanol was added and the s o l u t i o n was e x t r a c t e d over a steam bath fo r 5 min . The methanol was removed w i th a Pasteur p i p e t t e and f i l t e r e d through g l a s s woo l . The e x t r a c t i o n was repeated u s i n g a second ten ml a l i q u o t of methano l . The f i l t e r e d e x t r a c t s were combined, and the f i n a l 9 8 volume was a d j u s t e d to 10 m l . Twenty M1 a l i q u o t s of t h i s s o l u t i o n were i n j e c t e d onto the HPLC. As w i th the p r e v i o u s samples , two r e p l i c a t e s of each sample were p r e p a r e d , and each r e p l i c a t e was i n j e c t e d tw i ce . F i g u r e s g iven in Tab le IV are the average of these four r e p l i c a t e i n j e c t i o n s . b. B a n i s t e r i o p s i s caap i c u l t i v a r s . Q u a n t i t a t i o n of the a l k a l o i d content of the B a n i s t e r i o p s i s caap i c u l t i v a r s was c a r r i e d out on stem samples which had been d r i e d under low heat (<60 ° ) in a p l an t d r y e r . The a i r - d r i e d stems were ground to powder in a Wi ley m i l l and 5 g was e x t r a c t e d w i th 2x 100 ml methanol f o r 24 hrs on a r o t a r y shaker . The combined e x t r a c t s were concen t r a t ed under vacuum, f i l t e r e d through g l a s s wool and the vo lume•adjusted to 25 ml wi th methano l . Ten y l a l i q u o t s were i n j e c t e d d i r e c t l y onto the HPLC. Four r e p l i c a t e i n j e c t i o n s of each sample were made, and f i g u r e s g iven in Tab l e V r e f l e c t the mean of these four r e p l i c a t e i n j e c t i o n s . c . DMT-conta in ing admixture p l a n t s . A l l of the Psychot r i a  v i r i d i s samples were ana l yzed us ing methano l-preserved l e a f m a t e r i a l c o l l e c t e d in the f i e l d ; the D i p l o p t e r y s cabre rana sample c o n s i s t e d of f r e e z e - d r i e d l eaves d e r i v e d from a greenhouse propagated c l one of Plowman 6040. The methanol was decanted from the methano l-preserved m a t e r i a l and the remain ing s o l i d matter was f r o z e n , then l y o p h i l i z e d . The f r e e z e - d r i e d l e a f m a t e r i a l was powdered and e x t r a c t e d ove rn igh t on a r o t a r y shaker w i th methanol (10-20 ml/g d . w t . ) . The D. cab re rana l eaves were e x t r a c t e d d i r e c t l y wi th methanol and o therw ise t r e a t e d in the same manner as the P s y c h o t r i a samples . The methanol e x t r a c t s were f i l t e r e d , combined wi th the o r i g i n a l methanol used to 99 p rese rve the samples, and concen t r a t ed in a r o t a r y evapora tor to a known volume. The crude methanol e x t r a c t s were sea l ed and s t o r ed a t . 4 ° C. For purposes of a l k a l o i d q u a n t i t a t i o n , an a l i q u o t of the methano l i c e x t r a c t e q u i v a l e n t to 2.0 g dry wt of the f r e e z e - d r i e d l e a f m a t e r i a l was t r a n s f e r r e d to a 50 ml round-bottom f l a s k and evaporated to dryness on a r o t a r y e v a p o r a t o r . The r e s i due was shaken w i th 5 ml of 1 N HCI and f i l t e r e d . The a c i d i c f i l t r a t e was washed w i th 1 x 5 ml C H 2 C 1 2 , and the o rgan i c l a y e r d i s c a r d e d . The aqueous l a y e r was b a s i f i e d to pH 8-9 wi th s a t u r a t e d NaHC0 3 and e x t r a c t e d wi th 3 x 5 ml C H 2 C 1 2 . It was then f u r t h e r b a s i f i e d to pH 11-12 wi th 2 N NaOH and e x t r a c t e d wi th 2 x 5 ml C H 2 C 1 2 . The o rgan i c l a y e r s were combined, d r i e d over anhydrous N a 2 S O „ , evapora ted to d r y n e s s , r e d i s s o l v e d in methano l , and f i l t e r e d through g l a s s wool . Twenty ul a l i q u o t s of t h i s p u r i f i e d a l k a l o i d f r a c t i o n were i n j e c t e d i n t o the HPLC. C o n c e n t r a t i o n of the DMT p resen t was determined by compar ison with a s tandard curve c o n s t r u c t e d by i n j e c t i n g known amounts of DMT s t a n d a r d . F i gu r e s g i ven in Tab le VI are means ± s . e . of 2 to 5 r e p l i c a t e i n j e c t i o n s . D. GC/MS DMT-conta in ing admixtures were screened by GC/MS to c o n f i r m tha t the major i ndo l e base d e t e c t e d was N ,N-d imethy I t ryptamine . The ins t rument used was a F i n n i g a n model 1020 automated GC/MS i n t e r f a c e d w i th a Perk in-E lmer Sigma 3B gas chromatograph. The chromatograph was equipped wi th a 30 m x .25mm SE-54 fused s i l i c a c a p i l l a r y column (J&W S c i e n t i f i c ) . The chromatograph was temperature-programmed from an i n i t i a l temperature of 180° to a 100 f i n a l temperature of 2 5 0 ° . The ramp ra te was 3 ° /min i n i t i a t e d 3 min a f t e r i n j e c t i o n of the sample. I n j e c t o r b lock temperature was 2 5 0 ° , d e t e c t o r temperature 260° C . The c a r r i e r gas was h e l i u m . One M1 a l i q u o t s of DMT s t a n d a r d , or of the p u r i f i e d a l k a l o i d e x t r a c t s of the l e a f samples were i n j e c t e d . E l u t e d compounds were de t e c t ed as peaks in the r e c o n s t r u c t e d ion chromatogram (RIC) genera ted by the mass spect rometer data sys tem. Under these c o n d i t i o n s , DMT had a r e t e n t i o n time of 9.5-10 min , base peak 58, M* 188. The i n d o l e base p resen t in the D i p l o p t e r y s cabre rana and Psychot r i a l e a f e x t r a c t s had a mass spectrum and r e t e n t i o n time i d e n t i c a l w i th the s t a n d a r d . E. A l k a l o i d Tes t s & TLC of Uncommon Admixture P l a n t s The uncommon admixture p l a n t s were screened fo r a l k a l o i d s f o l l o w i n g the method of Farnsworth & Eu l e r [18 ] . M a t e r i a l used in the a n a l y s i s was p rese r ved in methano l , and t h i s was worked up in a manner i d e n t i c a l to that d e s c r i b e d above fo r the P s y c h o t r i a samples . A l i q u o t s of the a c i d i c f i l t r a t e were t e s t e d w i th 1-2 drops of e i t h e r M e y e r ' s , V a l s e r ' s , or D r a g e n d o r f f ' s r e agen t . Appearance of e i t h e r a marked t u r b i d i t y or a heavy p r e c i p i t a t e on a d d i t i o n of the reagent was i n t e r p r e t e d as a p o s i t i v e a l k a l o i d r e a c t i o n ; s l i g h t t u r b i d i t y i n d i c a t e d p o s s i b l e t r a c e s of a l k a l o i d . Compos i t ion of the reagents used i s g iven in M a r t e l l o & Farnsworth [ 19 ] . A f t e r t e s t i n g wi th the p r e c i p i t a t i o n r e a g e n t s , the a c i d i c aqueous s o l u t i o n s were b a s i f i e d to pH 8-9 w i th s a t u r a t e d NaHC0 3 and and e x t r a c t e d wi th 3 x 5 ml C H 2 C 1 2 ; i t was then b a s i f i e d to pH 11-12 and e x t r a c t e d w i th 2 x 5ml C H 2 C 1 2 . The combined o rgan i c l a y e r s were evapora ted to d ryness under 101 vacuum, and the res idue taken up in 2.0 ml methano l . F i ve M1 a l i q u o t s of t h i s f i n a l methanol f r a c t i o n were a p p l i e d to p recoa ted Polygram s i l i c a ge l p l a t e s U V 2 5 " , and the p l a t e was deve loped in one d i r e c t i o n in b u t a n o l / a c e t i c ac id/water 4 : 1 : 1 . F o l l o w i n g development, the p l a t e s were a i r - d r i e d , examined under sho r t - and long-wave UV l i g h t , and sprayed wi th D r a g e n d o r f f ' s m o d i f i e d reagent [20 ] . I I I . R e s u l t s and D i s c u s s i o n A. Ayahuasca Brews Comparison of the TLC p r o f i l e s of the e i g h t samples ( F i g . 6) shows that the major c o n s t i t u e n t s vary l i t t l e from sample to sample ; d i f f e r e n t batches made by the same ayahuasquero ( c f . Don F i d e l # 1 & 2, and Don Juan , # 1 & 2) are g e n e r a l l y s i m i l a r , and there i s a l s o l i t t l e v a r i a t i o n in the c o n s t i t u e n t s of brews made by d i f f e r e n t ayahuasqueros . Harmine, harmol , ha rma l i ne , and te t rahydroharmine were found to be the major /3-carbol ines p resen t in a l l of the samples , wh i le ha rma lo l was not de t ec t ed in any samples save one (Don M i l t o n # 1) . D imethy l t r yp tamine was found in a l l samples except that from T a r a p o t o . No o ther s i g n i f i c a n t E h r l i c h - p o s i t i v e spots were d e t e c t e d . Known c o n s t i t u e n t s were i d e n t i f i e d by compar ison w i th a mixture of a u t h e n t i c s tandards (ayahuasca " a n a l o g u e " , upper l e f t in F i g . 6) T races of o ther f l u o r e s c e n t compounds were a l s o d e t e c t e d in most samples ; i t i s assumed that these r ep resen t /3-carbol ines of undetermined s t r u c t u r e . Absence of DMT in the sample from Tarapoto i s s i g n i f i c a n t , s i n c e t h i s i s the on l y sample in which 102 AYAHUASCA "ANALOGUE" © DON FIDEL - SAMPLE ;? 1 PUCALLPA e ® DON FIDEL - SAMPLE 2 PUCALLPA (9 e DON JUAN - SAMPLE- if 1 PUCALLPA 0 * Cs) DON JUAN - SAMPLE ? 2 PUCALLPA 0 0 DON WILFREDO - SAMPLE # 1 TARAPOTO 9 ® DONA RIOS - SAMPLE // 1 IQUITOS ® ® © 3 • DOM MILTON - SAMPLE // 1 IQUITOS A (§0 DOM SOLON -- SAMPLE » 1 IQUITOS 9 $ <E+) CP <9 F i g u r e 6 - TLC A n a l y s i s of Peruvian Ayahuasca Samples A Dimethyltryptamine(DMT) S o l v e n t s : • Harmine I. ether/2-butanor.e/ • Harmaline 28% NH0OH 5:4:1 (upper phase) D Harmol I I . n-propanol/1.5% NH„OH 9:2 O Tetrahydroharmine P l a t e s : T Harmalol • Unknown Polygram s i l i c a g e l GF 2 5" (Brinkmann Instruments) (E+) = p o s i t i v e to E h r l i c h ' s Reagent 1 03 P s y c h o t r i a c a r thaqenens i s was employed as the admixture r a the r than the more commonly used P. v i r i d i s . The q u a n t i t a t i v e HPLC a n a l y s i s of f i v e u n d i l u t e d samples p repared by two ayahuasqueros in P u c a l l p a shows l i t t l e v a r i a t i o n from ba tch to b a t c h , e i t h e r in t o t a l a l k a l o i d conten t or in the p r o p o r t i o n s of c o n s t i t u e n t s (Table I I I ) . The me thano l -d i l u t ed samples were l y o p h i l i z e d and t h e i r a l k a l o i d con ten t s compared on a dry weight b a s i s (Table IV ) . These samples showed c o n s i d e r a b l e d i f f e r e n c e s in a l k a l o i d content (expressed as mg a l k a l o i d / g dry wt ) . The sample from P u c a l l p a had the h ighes t t o t a l a l k a l o i d content (75.7 mg/g d wt.) of which 76% was harmine , 10.6% was t e t r ahyd roha rm ine , and 7.6% was DMT. The samples from I q u i t o s and Tarapoto g e n e r a l l y had lower t o t a l a l k a l o i d l e v e l s , and a l s o d i f f e r e d in the p r o p o r t i o n s of d i f f e r e n t c o n s t i t u e n t s . The reason f o r the d i f f e r e n c e in r e l a t i v e p r o p o r t i o n of harmine and te t rahydroharmine in the P u c a l l p a samples and those from other r eg ions of Peru cou ld be r e l a t e d to the type of B. caap i c u l t i v a r employed to prepare the d r i n k , or to the method of p r e p a r a t i o n . Env i ronmenta l v a r i a b l e s , such as the type of s o i l in which the c u l t i v a r i s grown, or d i f f e r e n t c o n d i t i o n s of exposure to s u n l i g h t , may a l s o be i n v o l v e d . Tab le III shows, however, tha t a l l of the ayahuasca samples from P u c a l l p a c o n s i s t e n t l y show approx imate l y the same r e l a t i v e p r o p o r t i o n s of harmine to t e t rahydroharmine to ha rma l i ne . T h i s may i n d i c a t e tha t a l l of these samples were p repared e i t h e r from the same B. c aap i c u l t i v a r or c l o n e s of the same c u l t i v a r . T h i s would not be s u r p r i s i n g s i n ce Don Juan i s the unc le of Don F i d e l and they o f t e n c o l l a b o r a t e in the p r e p a r a t i o n of t h e i r brews. TABLE I I I HPLC QUANTITATION OF UNDILUTED AYAHUASCA SAMPLES Name of Sample A1ka1o i d Conc e n t r a t i on (mg/ml)* harmol harm 1ne % THH % harma11ne % DMT t o t a l Don Fide101 t r t r 3 ( .85 .01 ) 66 1 ( .06 .05) 18 0 ( .3 . 1 ) 5 0 ( .61 .01) 10 5 . 85 ( . 19) Don Fidel#2 t r 4 ( .64 • .3) 62 1 ( . 77 . 1) 24 0 (. . 45 1 ) 6 0 ( .6 . 1) 8 7 . 48 ( • 16) Don F1del*3 t r 3 ( . 4 2) 53 1 ( .94 .06) 30 0. ( 34 2) 5 0. ( . . 7 2) 1 1 6 . 38 ( . 1 ) Don Juan#1 t r 5 (-. . 3 36) 66 1 ( . 73 . 1 ) 21 0. (. 51 16) 6 0. (. 51 16) 6 8.05 ( .36) Don Juan#2 t r 5 . ( 51 .26) 67 1 ( .67 .07) 20 0. (. 4 1 14) 5 0. (. 6 2) 7 8 . 19 ( . 28) Average A l k a l o i d c o n t e n t t 4 . ( . 67 .2) 65 1 ( .60 .08) 22 0. (. 4 1 06) 6 0. (. 6 06 ) 8 7 . 28 ( .23) * f i g u r e s g i v e n a r e mg a l k a l o i d / m l of u n d i l u t e d sample, ± s. e.. shown i n pa r e n t h e s e s : p e r c e n t a g e s a r e % t o t a l a l k a l o i d , t average based on n=18 r e p l i c a t e i n j e c t i o n s . TABLE IV HPLC QUANTITATION OF LYOPHILIZED AYAHUASCA SAMPLES Sample & O r i g i n A l k a l o i d I C o n c e n t r a t i o n (mg/g d.wt. ).* harmine % THH ha r m a l i n e % DMT % t o t a l Don Fld e l / M P u c a l l p a 57 .6 ( .33) 76 8.0 ( .53) 1 1 4 . 2 ( .22) 5 5.8 ( .5) 8 75 . 6 (3.3) Don Sol on Iqu i t os 28 . 3 (1.3) 42 25.5 (1.4) 38 5.8 ( .9) 9 7 . 2 (1.7) 1 1 66.8 (3.4) Don Wi 1fredo T a r a p o t o 14 . 4 ( .44) 50 10.5 (1.2) 36 4 . 2 ( .3) 14 n.d. t - 29 . 1 ( 1 Don Ml 1 ton Iqu1tos 10. 2 ( .32) 33 10. 2 (1.7) 33 5.2 ( 1.7) 16 5 . 7 ( 1 .6) 18 31.3 (5.6) Dona R l o s Iqui tos 8.6 ( .3) 27 9.6 (1.8) 31 6.3 ( 1.8) 20 7 .O (2.1) 22 31.5 (5.6) Average A l k a l o i d 23 .8 (4.2) 51 11.1 (1.9) 24 5 . 1 ( .5) 1 1 6.4 ( .74) 14 46 . 9 (4.9) Content* * f i g u r e s g i v e n a re mg a l k a l o i d / g d. wt. of l y o p h i l i z e d sample: s t a n d a r d e r r o r s (n=4) are g i v e n i n par e n t h e s e s . Percentages g i v e n a r e % t o t a l a l k a l o i d . t n. d. = not d e t e c t e d * based on n=16 r e p l i c a t e i n j e c t i o n s 1 06 Summarizing the r e s u l t s of t h e i r q u a n t i t a t i v e s t u d i e s , R i v i e r and L indgren [4] s t a t e tha t a t y p i c a l 200 ml dose of ayahuasca c o n t a i n s a t o t a l of 65 mg a l k a l o i d , of which 30 mg i s harmine, 10 mg i s t e t r ahyd roha rm ine , and 25 mg, DMT. T h i s i s some ten to t h i r t y t imes l e s s than the dosage at which the (3-c a r b o l i n e s are h a l l u c i n o g e n i c a l l y a c t i v e in the pure form, ( c f . Chapter II) a l t hough i t i s w e l l w i t h i n the range at which they are e f f e c t i v e as MAO i n h i b i t o r s . Twenty f i v e mg i s j u s t above the t h r e s h o l d dose fo r DMT when t h i s compound i s i n j e c t e d i n t r a m u s c u l a r l y [21] but i t i s p o s s i b l e that the t h r e s h o l d may be lower under c o n d i t i o n s of MAO i n h i b i t i o n . Commenting on t h e i r f i n d i n g s , R i v i e r & L indgren [4] c o n c l u d e : "In view of these r e s u l t s , new pha rmaco log i c a l exper iments fo r a b e t t e r unde rs tand ing of the h a l l u c i n o g e n i c a c t i o n of ayahuasca seem n e c e s s a r y . " The a l k a l o i d l e v e l s found in the f i v e samples from P u c a l l p a (Table I I I ) exceed the l e v e l s r epo r t ed by R i v i e r and L indgren [4] in samples c o l l e c t e d on the upper R io Purus by at l e a s t an order of magni tude. Thus , (based on the average of the f i v e samples) a 100 ml dose of the P u c a l l p a ayahuasca ( c f . Tab le I I I ) c o n t a i n s 728 mg t o t a l a l k a l o i d , of which 467 mg i s harmine , 160 mg i s t e t r ahyd roha rm ine , 41 mg i s ha rma l i ne , and 60 mg i s DMT. T h i s i s we l l above the t h r e s h o l d dose f o r DMT but i s s t i l l c o n s i d e r a b l y below the h a l l u c i n o g e n i c dose l e v e l f o r the /?-c a r b o l i n e s . In p r a c t i c e the t y p i c a l dose i nges t ed in the P u c a l l p a ceremonies r a r e l y exceeds 75 ml and i s u s u a l l y c l o s e r to 55-60 m l . The r e l a t i v e l y l a r g e d i f f e r e n c e s in the a l k a l o i d content of the upper Purus ayahuasca ana l yzed by R i v i e r and 107 L indg ren and the P u c a l l p a ayahuasca ana l yzed in the p resen t study may be r e a d i l y e x p l a i n e d by the d i f f e r e n c e s in the method of p r e p a r a t i o n in the two r e g i o n s . In the upper Purus method, stems of B a n i s t e r i o p s i s caap i t o t a l i n g about 900 cm in l eng th and 1-4 cm diameter are cut i n t o shor t s e c t i o n s , .crushed, and packed in a 15 l i t e r meta l v e s s e l toge the r wi th a l t e r n a t i n g l a y e r s of l eaves of P s y c h o t r i a spp . Ten l i t e r s of water are added and the mixture i s b o i l e d fo r one hour , s t r a i n e d , and c o o l e d . The mixture i s then consumed without f u r t h e r p r o c e s s i n g . The method employed in P u c a l l p a s t a r t s out s i m i l a r l y but the mix ture i s b o i l e d fo r a much longer t ime , approx imate l y 10-15 hou r s . The water may be d r a i n e d o f f and r e p l a c e d wi th f r e s h water s e v e r a l t imes d u r i n g t h i s b o i l i n g p r o c e s s . The separa te ba tches are combined, a l l owed to c o o l , and f i l t e r e d through a s t r a i n e r or c h e e s e c l o t h . The p l an t m a t e r i a l i s removed from the cook ing pot and d i s c a r d e d , and then the s t r a i n e d ayahuasca i s poured back i n t o the pot and simmered over a low f i r e u n t i l i t has been concen t r a t ed to about h a l f i t s o r i g i n a l volume. F i v e or s i x l i t e r s of ayahuasca are ob ta ined from t h i s p r o c e s s ; these may be kept fo r up to s i x months wi thout r e f r i g e r a t i o n in wine or beer b o t t l e s s toppered wi th c o r k s . B. A l k a l o i d Content of B a n i s t e r i o p s i s caap i C u l t i v a r s A l l of the ayahuasqueros that we i n t e r v i ewed d u r i n g our f i e l d s t u d i e s in Peru r e c o g n i z e d s e v e r a l d i f f e r e n t " k i n d s " of ayahuasca which were c l a imed to vary in t h e i r p s y c h o l o g i c a l e f f e c t . The d i f f e r e n t i a t i o n of these v a r i e t i e s of ayahuasca was based in pa r t on the types of admixture p l a n t s which were added, 1 08 and in pa r t on the type of B. c aap i which was u t i l i z e d . Seve ra l types of B a n i s t e r i o p s i s caap i were g e n e r a l l y r e cogn i zed by these p r a c t i t i o n e r s and were d i s t i n g u i s h e d by d i f f e r e n t a d j e c t i v e s , e . g . , " c i e l o " ayahuasca , " l u c e r o " ayahuasca , " r u m i " ayahuasca . Some c l a imed to d i s t i n g u i s h as many as ten k inds of B a n i s t e r i o p s i s v ine (the term ayahuasca i s i n d i s c r i m i n a t e l y a p p l i e d e i t h e r to the B. caap i v i ne or to the beverage made from i t ) but most were f a m i l i a r wi th on l y two or th ree k i n d s . Presumably these " k i n d s " of B. c aap i are r e f e r a b l e to d i f f e r e n t c u l t i v a r s , r a c e s , or chemica l or mo rpho log i c a l v a r i e t i e s of B a n i s t e r i o p s i s c a a p i . There were no ou t s t and ing morpho log i c a l d i f f e r e n c e s between the three or four k inds of B. caap i which we c o l l e c t e d , and the r e l e van t voucher specimens have' a l l been determined as B a n i s t e r i o p s i s c aap i by taxonomic s p e c i a l i s t s in the Ma lp igh i a ceae (W. R. Anderson & B. Ga tes , U n i v e r s i t y of M i c h i g a n ) . HPLC a n a l y s i s of the a l k a l o i d l e v e l s in the d r i e d stems of th ree of the r e cogn i zed v a r i e t i e s p l u s one specimen (DMK#125) fo r which the v e r n a c u l a r name i s unknown has shown tha t the re i s c o n s i d e r a b l e v a r i a t i o n between samples (Table V ) . The lowest l e v e l was found i n DMK#126 which c o n t a i n e d 1.7 mg/g t o t a l a l k a l o i d , wh i le DMK#125 c o n t a i n e d the h i ghes t l e v e l , 13.6 mg/g. There appears to be no c o n s i s t e n t c o r r e l a t i o n of a l k a l o i d content w i th p a r t i c u l a r r e c o g n i z e d c u l t i v a r s , however. The v a r i a t i o n observed probab ly has more to do w i th the age of the p l a n t , and the s o i l , l i g h t , water , and other env i ronmenta l c o n d i t i o n s a f f e c t i n g the growth of the p a r t i c u l a r spec imen. The amounts of a l k a l o i d s are in the same gene ra l range as those de t e c t ed in the B. caap i samples ana l yzed by R i v i e r & L indgren TABLE V HPLC QUANTITATION OF BANISTERIOPSIS CAAPI CULTIVARS Col l e c t i o n #. Name. & O r i g i n ' DMK #110 "c 1 e 1 o" Tarapoto DMK #124 "Pucahuasca" Tarapoto DMK .#125 Iqu1tos DMK #126 " c i e 1 o " Iqu i tos DMK #128 " rum i " Iqu i t os Plowman 604 1 " c i e 1 o" Tarapoto-1976 (UBC-1982) A1ka1o1ds D e t e c t e d (mg/g d wt) ' . Harm i ne 5.3 72% 5.9 47% 6.35 47% 0.57 34% 4 . 4 51% 1 .0 35% THH 0.95 13% 3.3 26% 1.95 14% 0.25 15% 1 . 45 17% 1.3 47% harma11ne 1.1 15% 3.2 26% 3.8 28% 0.75 44% 2 .07 24% .5 18% harmo1 0.05 .7% 0.06 .5% 1 . 2 9% 0 . 1 6% 0.65 8% .01 .3% harma1ol n . d . + t r a c e - . 35 2% n.d. n.d. - n.d. t o t a l a 1ka1o1ds 7.4 12.5 13.6 1 .7 8.6 2 . 8 * n.d. = not d e t e c t e d 1 10 [ 4 ] , Fu r the r c l a r i f i c a t i o n of t h i s q u e s t i o n of p o s s i b l e chemica l or m o r p h o l o g i c a l d i f f e r e n c e s between r e c o g n i z e d types of B. caap i c u l t i v a r s would r e q u i r e a s ys temat i c sampl ing of as many d i f f e r e n t i n d i v i d u a l s of each type as p o s s i b l e ; c l i m a t i c , edaph i c , and other env i ronmenta l f a c t o r s shou ld a l s o be c o n s i d e r e d . In most of the B. caap i c u l t i v a r s examined, harma l ine c o n s t i t u t e d a g r ea t e r p r o p o r t i o n of the t o t a l a l k a l o i d content than in the ayahuasca brews. In the B. caap i c u l t i v a r s , harma l ine c o n s i s t e n t l y r ep resen ted 25-50% of the t o t a l a l k a l o i d s , wh i le in the ayahuasca samples , i t was approx imate l y 5-15% of the t o t a l a l k a l o i d s in most samples . T h i s i n d i c a t e s that the p rocess of b o i l i n g and c o n c e n t r a t i o n of the ayahuasca brews may r e s u l t in the o x i d a t i o n of a s i g n i f i c a n t f r a c t i o n of the harmal ine to the more aromat ic d e r i v a t i v e , harmine . C . A l k a l o i d Content of Ayahuasca Admixture P l a n t s 1. DMT-conta in ing Admixtures In Pe ru , the admixture p l an t employed most f r e q u e n t l y in the p r e p a r a t i o n of ayahuasca appears to be P s y c h o t r i a v i r i d i s R. & P. We encountered on ly one ayahuasquero du r i ng our f i e l dwo rk who p r e f e r r e d to use another s p e c i e s of P s y c h o t r i a , t e n t a t i v e l y i d e n t i f i e d as P s y c h o t r i a c a r t h a q e n e n s i s J a c q . I n t e r e s t i n g l y , no a l k a l o i d s of any k ind were de t e c t ed in t h i s c o l l e c t i o n (DMK#109, Tarapoto ) however, a l l of the P. v i r i d i s c o l l e c t i o n s con t a i ned N ,N-d imethy l t ryptamine as the s i n g l e major base . I d e n t i t y of the compound was con f i rmed by GC/MS and compar ison of i t s HPLC TABLE VI OMT CONTAINING ADMIXTURE PLANTS: ANALYSIS BY TLC. HPLC. 8. GC/MS Col 1 e c t i o n <* . Name. & O r i g i n DMK#21 Psy c h o t r i a v i r i d i s "chacruna" Iqu i tos DMK#108 Psyc h o t r i a v i r i d i s 1 "su i j a" Tarapoto DMK#109 Ps y c h o t r i a carthagenens i s "yage-chacruna" Tarapoto DMK#139 Psychot r i a v i r i d i s "chacruna" P u c a l l p a Plowman 6040 D i p i o p t e r y s c a b r e r a n a "chagro-panga" Tarapoto DMT S ta n d a r d TLC: s o l v e n t 1 hRf* 42 s o l v e n t 2 hRff 23 r e a c t i o n to E h r l i c h ' s reagent +(blue) 42 24 -M b 1 ue) n.d.t 38 27 +(blue) 42 25 + ( v i o l e t ) 4 1.3 25 ••(blue) HPLC : Ret. t i me (min) mg/g d wt . 18.4( . 1)b 1.58(.3) 18.3(.03) 1 .02( .04 ) 18.7(.05) 1 . 2 ( . 1 7 ) 1 8 . 4 ( 1 5 ) 1 .74( .4 ) 19 . 1 GC/MS: t r a c e n.d. n.d. n.d. 2-Me- 5-H0- M' = 188 c o n s t i t u e n t s THpC DMT m/z 58=1007, * S o l v e n t 1: ether/2-butanone/conc NH»0H 5:4:1 (upper phase): hRf= Rf x 100 t S o l v e n t 2: n-propano1/1.5% NH«OH 9:2 $ n.d.= not d e t e c t e d b F i g u r e s i n p a r e n t h e s e s are s t a n d a r d e r r o r s 1 12 r e t e n t i o n t ime , TLC hRf and E h r l i c h ' s c o l o r r e a c t i o n wi th that of an a u t h e n t i c s tandard (Table V I ) . The P. v i r i d i s samples ana l yzed con t a i ned f a i r l y s u b s t a n t i a l amounts of DMT, between 1 and 1.6 mg/g dry wt. in the l e a v e s . No a l k a l o i d was de tec t ed in f r u i t s or stems of P. v i r i d i s . No o ther a l k a l o i d s were de tec ted in any of the P s y c h o t r i a samples wi th the e x c e p t i o n of DMK #139, in which a t r a ce c o n s t i t u e n t wi th a mass spectrum co r r e spond ing to that r epo r t ed [4] f o r 2-methy l-tet rahydro-/3-carbol ine was d e t e c t e d . A s i n g l e sample of D i p l o p t e r y s cabre rana (Plowman 6040) , the Ma lp igh i a ceous admix tu re , was a v a i l a b l e fo r a n a l y s i s and t h i s a l s o con ta ined N ,N-d imethy l t ryptamine toge the r wi th a t r a c e amount of 5-hydroxy-DMT. The a l k a l o i d e x t r a c t of d r i e d l eaves of the o r i g i n a l (1976) c o l l e c t i o n of Plowman 6040 had an ion chromatogram that was e s s e n t i a l l y i d e n t i c a l to l e a f e x t r a c t s of greenhouse propagated c l o n e s of t h i s spec imen. Plowman 6040 con t a i ned s l i g h t l y h ighe r l e v e l s of DMT (1.74, mg/g d wt.) than the P. v i r i d i s samples , but o therw ise was i n d i s t i n g u i s h a b l e in-terms of a l k a l o i d c o n t e n t . A l though Plowman 6040 was c o l l e c t e d in Tarapoto where i t was be ing u t i l i z e d as an ayahuasca admix tu re , t h i s use of D i p l o p t e r y s cabre rana in Peru i s uncommon; t h i s s p e c i e s i s the usua l admixture in Southern Colombia and Ecuador [22] and in f a c t Plowman 6040 was o r i g i n a l l y brought to Tarapoto as a l i v e c u t t i n g from the Rio Pas taza in Ecuador (Plowman, T . , p e r s . comm., 1980). P s y c h o t r i a  v i r i d i s , or l e s s f r e q u e n t l y , P s y c h o t r i a c a r t h a g e n e n s i s are the admixtures of cho i c e in Peru and few of my in fo rmants in Peru were f a m i l i a r wi th D. cabre rana under i t s common names, chagro- panga or oco-yage. 1 1 3 2. Uncommon Admixture P l a n t s Ayahuasca i s u s u a l l y p repared us ing one of the DMT— c o n t a i n i n g admixture p l a n t s ment ioned above, e i t h e r D i p l o p t e r y s  cabre rana or a Psychot r i a s p . ; l e s s commonly, however, o ther admixtures are u t i l i z e d , e i t h e r i n c o n j u n c t i o n wi th the t r y p t a m i n e - c o n t a i n i n g admix tu res , or in p l a ce of them. Many of these admixtures have been i d e n t i f i e d in the e t h n o b o t a n i c a l l i t e r a t u r e [ 2 , 4 , 2 2 , 2 3 , ] a l though l i t t l e * i s known of t h e i r chemica l or biodynamic p r o p e r t i e s , ( c f . Appendix I ) . T h i s would appear to be a p romis ing area fo r f u r t h e r r e s e a r c h . Three c o l l e c t i o n s were made of p l a n t s which were s t a t e d by in formants to be used as admixtures to ayahuasca (Table V I I ) . One of these , T e l i o s t a c h y a l a n c e o l a t a , has been d i s c u s s e d by S chu l t e s [23] as an admix tu re , but the o ther two, Abuta q r a n d i f o l i a (Menispermaceae) and C o r n u t i a odora ta (Verbenaceae) have not p r e v i o u s l y been r epo r t ed as adm ix tu r e s . P l an t m a t e r i a l from these c o l l e c t i o n s , p rese r ved in methano l , were screened fo r a l k a l o i d s u s i ng a l k a l o i d p r e c i p i t a t i o n t e s t s and TLC (Table V I I ) . I n s u f f i c i e n t m a t e r i a l was a v a i l a b l e to permit f u r t h e r chemica l c h a r a c t e r i z a t i o n . The on l y c o l l e c t i o n g i v i n g an unambiguously p o s i t i v e t e s t was Abuta g r a n d i f o l i a (DMK #74). T h i s s p e c i e s has r e c e n t l y been r e p o r t e d [24] to c o n t a i n p a l m a t i n e , a t y p i c a l qua te rna ry base of the b i s -b e n z y l i s o q u i n o l i n e f am i l y which c h a r a c t e r i z e s the Menispermaceae. A l though pa lmat ine i s one of the commonest a l k a l o i d s in n a t u r e , i n v e s t i g a t i o n s of i t s pharmacology are s u r p r i s i n g l y s p a r s e . One study [25] found that pa lmat ine i n h i b i t e d the e f f e c t of ep i neph r i ne on b lood p ressu re of TABLE VII TEST FOR ALKALOIDS IN UNCOMMON ADMIXTURE PLANTS Col 1 .H DMK*74 DMK#22 DMK0 119 DMK# 1 Genus: Abuta T e l i o s t a c h y a Cornut i a Jus t i d a s p e c i e s : grand 1 f o l i a 1 anceolata o d o r a t a p e c t o r a 1 i s fam i1y: Men 1spermaceae Acanthaceae Verbenaceae Acanthaceae Par t e x t ' d ( g ) bark(2 ) 1 eaves(2.7) 1 eaves(5 . 7 ) 1eaves(1.2) React i on to: Meyer's r g t . + + - + -Va1ser's r g t + + - + -D r a g e n d o r f f ' s ++ TLC: D r a g e n d o r f f ' s * ++ F1uorescent Spots d e t e c t e d : Long wave UV + Short wave UV + * m o d i f i e d f o r TLC a c c o r d i n g to St a h l [21] 1 1 c r a b b i t s , on the i s o l a t e d r a t semina l v e s i c l e and on the toad h i n d - l e g ; i t s d e r i v a t i v e d l - t e t r a h y d r o p a l m a t i n e i n h i b i t e d the e f f e c t of 5-hydroxytryptamine on i s o l a t e d r a t u t e r i , c o l o n , and stomach. Pa lmat ine a l s o e x h i b i t e d a n t i c h o l i n e s t e r a s e a c t i v i t y . Both a l k a l o i d s had ACTH and b a c t e r i c i d a l a c t i v i t y . Some of these p r o p e r t i e s may be a n t a g o n i s t i c to the e f f e c t s of the 0-c a r b o l i n e s whi le o thers may be s y n e r g i s t i c . For ins tance ' harmal ine causes an i n c r ease in 5HT c o n c e n t r a t i o n in the whole b r a i n , wh i le harmine causes a s i g n i f i c a n t decrease in a c e t y l c h o l i n e in b r a i n ; on the o ther hand, harmine s t r o n g l y i n h i b i t s the ATP-Mg + + dependent uptake of no rep ineph r i ne i n t o i s o l a t e d ad r ena l medul la ry v e s i c l e s . [ 26 ] . Whether the o v e r a l l e f f e c t of p a l m i t i n e i s a g o n i s t i c or a n t a g o n i s t i c to the a c t i o n of the 0 - c a r b o l i n e s , there seems l i t t l e doubt that a d d i t i o n of the bark of Abuta q r a n d i f o l i a to ayahuasca c o u l d modify i t s e f f e c t . F u r t h e r i n v e s t i g a t i o n s of the pharmacology of t h i s and many other admixture p l a n t s are needed in order to c l a r i f y t h e i r c o n t r i b u t i o n to the e f f e c t s of ayahuasca . Just i c i a p e c t o r a l i s (Acanthaceae) i s a l s o i n c l u d e d in Tab le V I I . J u s t i c i a p e c t o r a l i s v a r . s t e n o p h y l l a i s not used as an ayahuasca admixture but has been r epo r t ed as an admixture to the V i r o l a s n u f f s , ( c f . Chapter V I , and [27]) and these au thors have suggested that i t may be used by i t s e l f as an h a l l u c i n o g e n i c s n u f f . No t r yp tamines or a l k a l o i d s of any o ther type were d e t e c t e d in our c o l l e c t i o n of t h i s s p e c i e s . GC/MS a n a l y s i s of e x t r a c t s of J us t i c i a p e c t o r a l i s i n d i c a t e tha t i t c o n t a i n s the coumarin d e r i v a t i v e u m b e l l i f e r o n e and the qua te rna ry n i t r o g e n d e r i v a t i v e be ta ine ( c f . Chapter V I , & [ 28 ] ) . 1 1 6 IV. Summary The a l k a l o i d a l c o n s t i t u e n t s of a number of ayahuasca brews, c u l t i v a r s of B. caap i and a v a r i e t y of admixture p l a n t s were q u a l i t a t i v e l y and q u a n t i t a t i v e l y i n v e s t i g a t e d us i ng 2-d imens iona l t h i n - l a y e r chromatography (TLC) and h igh-pressu re l i q u i d chromatography (HPLC) as the a n a l y t i c a l methods. Admixture samples were a l s o ana l yzed us ing gas chromatography/ mass spec t romet ry (GC/MS). Some admixture p l a n t s were screened f o r a l k a l o i d s us ing p r e c i p i t a t i o n t e s t s and TLC . The l e v e l s of /3-carbol ines found in most ayahuasca samples were, i n s u f f i c i e n t to account f o r the h a l l u c i n o g e n i c p r o p e r t i e s of ayahuasca at the doses t y p i c a l l y used , however the c o n c e n t r a t i o n of DMT would be we l l above the t h r e s h o l d l e v e l in most samples ; a p p a r e n t l y DMT i s r e s p o n s i b l e fo r the h a l l u c i n o g e n i c a c t i o n of ayahuasca , assuming that i t can be o r a l l y - a c t i v a t e d by the b lockade of v i s c e r a l MAO. D i f f e r e n t batches of ayahuasca p repared by the same person were remarkably c o n s i s t e n t , both i n terms of amount of t o t a l a l k a l o i d s and p r o p o r t i o n s of i n d i v i d u a l c o n s t i t u e n t s . C o n s i d e r a b l e v a r i a t i o n was found in samples p repa red by d i f f e r e n t p r a c t i t i o n e r s . V a r i a t i o n in a l k a l o i d content of B. c aap i c u l t i v a r s was a l s o found but may be due to env i ronmenta l f a c t o r s r a the r than a c t u a l g e n e t i c d i f f e r e n c e s between c l o n e s . S u b s t a n t i a l c o n c e n t r a t i o n s of DMT were found in s e v e r a l c o l l e c t i o n s of Psychot r i a v i r i d i s , and in one c o l l e c t i o n of D i p l o p t e r y s c a b r e r a n a , but was not de t ec t ed in P s y c h o t r i a  c a r t h a g e n e n s i s . DMT was the s i n g l e major base d e t e c t e d in these a d m i x t u r e s ; on l y t r a c e s of o ther a l k a l o i d s were p r e s e n t . Of 1 1 7 s e v e r a l uncommon admixture p l a n t s a l k a l o i d s , on ly Abuta g rand i f o l i a r e a c t i o n . which were screened f o r gave an unambiguously p o s i t i v e 1 18 V. L i t e r a t u r e C i t e d 1. S c h u l t e s , R. E. (1957) The I d e n t i t y of the M a i i p i g h i a c e o u s N a r c o t i c s of South Amer i ca . Harvard B o t a n i c a l Museum L e a f l e t s 18:1-56 2. S c h u l t e s , R. E. (1972) E t h n o t o x i c o l o g i c a l S i g n i f i c a n c e of A d d i t i v e s to New World H a l l u c i n o g e n s . P l an t Sc ience B u l l e t i n 18:34-41 3. Ga tes , B. (1979) New Names in B a n i s t e r i o p s i s and D i p l o p t e r y s (Ma lp igh iaceae ) of the Guayana H i g h l a n d . B r i t t o n i a 31:108-9 4. R i v i e r , L. & J . L i ndgren (1972) Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k : E t h n o b o t a n i c a l and Chemica l I n v e s t i g a t i o n s . Economic Botany 29:101-129 5. D e u l o f e u , V. (1967) Chemica l Compounds I s o l a t e d from B a n i s t e r i o p s i s and Re l a t ed Spec ies pp . 393-402 in D. H. E f r o n , B. Ho lmstedt , & N. S. K l i n e (eds . ) E thnopharmacolog ic Search fo r P sychoac t i ve Drugs U. S. P u b l i c Hea l th S e r v i c e P u b l i c a t i o n #1645 6. A g u r e l l , S . , B. Ho lmsted t , & J . E. L indgren (1968) A l k a l o i d Content of B a n i s t e r i o p s i s rusbyana . American J ou rna l of  Pharmacy 140:148-151 7. Hashimoto, Y. & K. Kawanishi (1975) New Organ ic Bases from Amazonian B a n i s t e r i o p s i s c a a p i . Phytochemi s t r y 14:1633-35 8. Hashimoto, Y. & K. Kawanishi (1976) New A l k a l o i d s from B a n i s t e r i o p s i s c a a p i . Phytochemi s t r y 15:1559-60 9. S h u l g i n , A. T . (1976) Psychotomimet ic Agen t s . Ch . 4 in Maxwell Gordon (ed. ) P sychopharmaco loq i ca l Agents V o l . IV. Academic Press 1 19 10. U d e n f r i e n d , S . , B. Wi tkop, B. G. R e d f i e l d , & H. Weissbach (1958) S t ud i e s w i th R e v e r s i b l e I n h i b i t o r s of Monoamine Ox idase : Harmal ine and Re l a t ed Compounds. B i o chemica l  Pharmacology 1:160-165 11. B u c k h o l t z , N. S. & W. 0. Boggan (1977) Monoamine Oxidase I n h i b i t i o n in B r a in and L i v e r Produced by /3-carbol ines : S t r u c t u r e - a c t i v i t y R e l a t i o n s h i p s and Subs t r a t e S p e c i f i c i t y B i o chemica l Pharmacology 26:1991-96 12. M c l s a a c , W. M. & V. Es tevez (1966) S t r u c t u r e - a c t i o n R e l a t i o n s h i p s of /3-carbol ines as Mono-amine Oxidase I n h i b i t o r s B iochemica l Pharmacology 26:1625-27 13. Na ran jo , C . (1967) P s y c h o t r o p i c P r o p e r t i e s of the Harmala A l k a l o i d s pp. 385-391 in D. H. E f r o n , et al., ( eds . ) op . c i t . 14. Pennes, H. H. & P. H. Hoch (1957) Psychotomimet i cs , C l i n i c a l and T h e o r e t i c a l C o n s i d e r a t i o n s : Harmine, WIN-299 and N a l l i n e . American J ou rna l of P s y c h i a t r y 113:887-92 15. Der M a r d e r o s i a n , A. H . , H. V. P i n k l e y , & M. F. Dobbins IV. (1968) Na t i ve Use and Occur rence of N ,N-d imethy l t ryptamine in the Leaves of B a n i s t e r i o p s i s rusbyana . American J ou rna l  of Pharmacy 140:137^147 16. A g u r e l l , S . , B. Ho lmsted t , J . E. L i n d g r e n , & R. E. S c h u l t e s . (1968) I d e n t i f i c a t i o n of Two New /3-carboline A l k a l o i d s in South American H a l l u c i n o g e n i c P l a n t s . B i ochemica l  Pharmacology 17:2487-88 17. Repke, D. B., D. T . L e s l i e & G. Guzman (1977) Baeocys t i n in P s i l o c y b e , Conocybe, and Panaeo lus . L l o y d i a 40:566-78 18. Fa rnswor th , N. R. & K. L. E u l e r (1962) An A l k a l o i d - s c r e e n i n g Procedure U t i l i z i n g T h i n - l a y e r Chromatography L l o y d i a 25: 186-195 19. M a r t e l l o , R. & N. R. Farnsworth (1962) Obse r va t i ons on the S e n s i t i v i t y of S eve ra l Common A l k a l o i d P r e c i p i t a t i n g Reagents . L l o y d i a 25:176-185 20. S t a h l , E . , (1969) Handbook of T h i n - l a y e r Chromatography 2nd. e d . S p r i n g e r - V e r l a g , New York . 120 21. S za r a , S. (1957) The Comparison of the P s y cho t i c E f f e c t of Tryptamine D e r i v a t i v e s w i th the E f f e c t s of Mesca l i ne and LSD-25 in S e l f - e x p e r i m e n t s . pp . 460-467 in S. G a r a t t i n i & V. G h e t t i ( eds . ) p s y c h o t r o p i c d r u g s . E l s e v i e r , amsterdam. 22. P i n k l e y , H. V. (1969) P l an t Admixtures to Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k . L l o y d i a 32:305 23. S c h u l t e s , R. E . , (1979) New Data on the Ma lp igh i a ceous N a r c o t i c s of South Amer i ca . Harvard B o t a n i c a l Museum L e a f l e t s 23 :137f f 24. Setor de F i t o q u i m i c a , INPA', Manaus, Amazonas, B r a z i l . (1971) Chemica l Compos i t ion of Amazonian P l a n t s . Ac ta Amazonica 1:83-86 25. C h ' e n , Mu-Ch'un & Chen-Yu C h ' i (1965) Comparat ive Pharmacology of Pa lmat ine and d l - t e t r a h y d r o p a l m a t i n e . Ya l  Hseuh Hseuh Pao 12:185-92. C i t e d in Chemica l A b s t r a c t s 63:3492g (19657 26. Ho, B. T . (1977) Pha rmaco log i ca l and B iochemica l S tud i e s wi th 0-ca rbo l i ne A n a l o g s . Cur ren t Developments in  Psychopharmacoloqy V o l . 4:153-177 27. Chagnon, N . , P. LeQuesne, & J . M. Cook (1971) Yanomamo H a l l u c i n o g e n s : A n t h r o p o l o g i c a l , B o t a n i c a l , and Chemica l F i n d i n g s . Cur rent Anthropo logy 1: 2-74 28. MacRae, D. & G. H. N. Towers (1984) J u s t i c i a p e c t o r a l i s - A Study of the Bas i s fo r i t s Use as a V i r o l a Snuff Admix ture . J ou rna l of E thnopharmacology. Submi t t ed . 29. A k a b o r i , S. & K. S a i t o (1930) S yn the t i s che Versuche in der Indo l-Gruppe , V I I I . : Synthese von Harman und Harmin. Chemische B e r i c h t e 63:2245-2248 121 CHAPTER V: ALKALOID CONSTITUENTS OF ORALLY-ACTIVE MYRISTICACEOUS HALLUCINOGENS I. I n t r o d u c t i o n V a r i o u s spec i e s of the genus V i r o l a ( M y r i s t i c a c e a e ) have been employed by Amazon Ind ian t r i b e s as the b a s i s of h a l l u c i n o g e n i c p r e p a r a t i o n s [ 1 ] . The drug ob ta ined from the V i r o l a t r e e s i s in a l l cases d e r i v e d from the " r e s i n " , the c o l o r l e s s to r edd i sh exudate of the cambia l l a y e r of the ba rk . Most commonly, the r e s i n i s evaporated to d ryness over a low f i r e and ground i n t o a powder, which i s i nges t ed in the form of a s n u f f . The exact method of p r e p a r a t i o n v a r i e s from reg ion to r e g i o n and t r i b e to t r i b e ; in some i n s t a n c e s the snu f f i s p repared wi thout the a d d i t i o n of other i n g r e d i e n t s , wh i le in o t h e r s , the powdered l eaves or ashes of o ther p l a n t s are added [ 2 ] , A l though the use of V i r o l a r e s i n as a snu f f has the widest g e o g r a p h i c a l and e t h n o l o g i c a l d i s t r i b u t i o n in the Amazon B a s i n , a few t r i b e s u t i l i z e o r a l l y - i n g e s t e d V i r o l a p r e p a r a t i o n s [ l ] , and the re are a l s o r e p o r t s [3] that the bark of c e r t a i n V i r o l a spp . may be smoked fo r h a l l u c i n o g e n i c e f f e c t s . Ex t ens i v e chemica l i n v e s t i g a t i o n s have been c a r r i e d out on many V i r o l a spp . and r e l a t e d genera [4] and the a c t i v e h a l l u c i n o g e n i c c o n s t i t u e n t s of the V i r o l a s n u f f s have been determined to be i n d o l e bases r e l a t e d to t r yp t am ine , e . g . , N,N-d i m e t h y l t r y p t a m i n e , 5-methoxy-N,N-dimethy l t ryptamine, and r e l a t e d d e r i v a t i v e s [ 5 , 6 ] . Trace amounts of /3-carbol ines have been r e p o r t e d from some s p e c i e s [ 6 ] , and in one s p e c i e s , not known to be used as an h a l l u c i n o g e n , /3-carbol ines are the major 1 22 a l k a l o i d s p resen t [7 ] . The a l k a l o i d a l c o n s t i t u e n t s of the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s p r e p a r a t i o n s have not p r e v i o u s l y been i n v e s t i g a t e d . A more d e t a i l e d d i s c u s s i o n of the e t h n o b o t a n i c a l , c h e m i c a l , and pha rmaco log i c a l a spec t s of the M y r i s t i c a c e o u s h a l l u c i n o g e n s may be found in Chapter I. In the present study a number of M y r i s t i c a c e o u s bark and l e a f samples c o l l e c t e d in the R io Ampiyacu r eg ion of Peru were screened fo r a l k a l o i d s us ing a l k a l o i d p r e c i p i t a t i o n t e s t s , TLC, GC, HPLC, and GC/MS. In a d d i t i o n , the a l k a l o i d c o n s t i t u e n t s of s e v e r a l n a t i v e drug samples were i n v e s t i g a t e d ; these c o n s i s t e d of s i x samples of o r a l l y - i n g e s t e d V i r o l a pas te c o l l e c t e d in the Rio Ampiyacu r eg ion in s p r i n g , 1981; one o r a l l y - i n g e s t e d V i r o l a pas te sample c o l l e c t e d at La C h o r r e r a , Co lomb ia , in 1971; and four samples of Yanomama snu f f c o l l e c t e d in Venezue la in 1972. I I . M a t e r i a l s and Methods A. D e t e c t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Samples C o l l e c t i o n numbers of M y r i s t i c a c e o u s bark and l e a f samples c i t e d in t h i s work r e f e r to the p e r s o n a l c o l l e c t i o n numbers of D. McKenna ( c f . Appendix II f o r a complete l i s t of herbar ium voucher c o l l e c t i o n s r e l e van t to t h i s work) . Herbar ium vouchers are a v a i l a b l e fo r a l l of the o r a l l y - i n g e s t e d pas tes c o l l e c t e d on the R io Ampiyacu, however no vouchers are a v a i l a b l e fo r the La Cho r r e r a pas te sample or f o r the Yanomama snu f f samples . A l l of the M y r i s t i c a c e o u s bark and l e a f samples were p r e se r ved in 100% methanol a t the time of c o l l e c t i o n . The methanol was decanted from the p r e se r ved m a t e r i a l and the remainder was f r o z e n , then 123 l y o p h i l i z e d . The f r e e z e - d r i e d m a t e r i a l was powdered and e x t r a c t e d ove rn igh t on a r o t a r y shaker (10-20 ml/g dry wt ) . The e x t r a c t s were f i l t e r e d , combined wi th the o r i g i n a l methanol used to p rese rve the sample, and concen t r a t ed under vacuum to a known volume. The crude e x t r a c t s were sea l ed and s t o r e d at 4 ° C. For a l k a l o i d s c r e e n i n g , a l i q u o t s of the e x t r a c t s e q u i v a l e n t to 3 g dry wt were evaporated to dryness in a 50 ml round bottom f l a s k . The r e s idue was shaken wi th 5 ml 1 N HCI, and f i l t e r e d . A sma l l a l i q u o t of the a c i d i c f i l t r a t e ( ca . 0.5 ml) was removed and used fo r the a l k a l o i d p r e c i p i t a t i o n t e s t s . The remainder was washed w i th 1 x 5 ml C H 2 C 1 2 and the o rgan i c l a y e r d i s c a r d e d . The aqueous l a ye r was b a s i f i e d to pH 8-9 wi th s a t u r a t e d NaHC0 3 and e x t r a c t e d wi th 15 ml p o r t i o n s of C H 2 C 1 2 ; i t was f u r t h e r b a s i f i e d to pH 10-12 w i th 2 N NaOH and e x t r a c t e d u n t i l a c i d i f i e d a l i q u o t s of the aqueous f r a c t i o n no longer gave a p o s i t i v e r e a c t i o n to Meye r ' s r eagen t . The o rgan i c f r a c t i o n s were combined, d r i e d over anhydrous N a 2 S O „ , evaporated to d ryness and r e d i s s o l v e d in 3 ml methano l . The o r a l l y - i n g e s t e d pas te samples c o l l e c t e d on the R io Ampiyacu were a l s o p rese r ved in methanol and were t r e a t e d in the same manner as the bark and l e a f samples . The La Cho r r e r a pas te sample was f r o z e n , l y o p h i l i z e d , and then e x t r a c t e d w i th methanol (20 ml/g dry wt ) . The Yanomama snuf f samples were d r i e d , powdered p l an t m a t e r i a l s ; these were e x t r a c t e d d i r e c t l y w i th methanol and prepared in the same manner as the o ther samples . A l k a l o i d p r e c i p i t a t i o n t e s t s were c a r r i e d out on sma l l a l i q u o t s of the a c i d i c f i l t r a t e du r i ng the a l k a l o i d i s o l a t i o n p rocedu re . T e s t s were made w i th V a l s e r ' s , M e y e r ' s , and D r a g e n d o r f f ' s reagent [ 8 ] ; a l l of these reagents form 124 p r e c i p i t a t e s or t u r b i d s o l u t i o n s in the presence of a l k a l o i d s , wh i l e s l i g h t t u r b i d i t y i n d i c a t e s p o s s i b l e t r a c e s of a l k a l o i d . V a l s e r ' s reagent i s approx imate l y an order of magnitude more s e n s i t i v e than the o ther two. TLC of the samples was conducted w i th the p u r i f i e d base e x t r a c t s which were r e d i s s o l v e d in methano l . Three /il a l i q u o t s were a p p l i e d to Polygram S i l i c a Ge l UV 2 5 " p recoa ted TLC p l a t e s (Brinkmann Intruments) at a po in t 1.0 cm from the bottom edge. F o l l o w i n g a p p l i c a t i o n the samples were d r i e d under a stream of a i r , and deve loped to a d i s t a n c e of 9 cm in e ther/2-butanone/conc . NH aOH 5 : 4 : 1 . The so l v en t was f r e s h l y p repared in a sepa ra to r y funne l and the upper phase c o l l e c t e d f o r TLC . Development was c a r r i e d out at ambient temperature in an u n l i n e d 10x23x26 cm g l a s s chromatographic tank c o n t a i n i n g 50 ± 5 ml of s o l v e n t . F o l l o w i n g development , p l a t e s were a i r - d r i e d in a fume hood fo r 30-60 min , then examined under l ong- and short-wave UV l i g h t . Tryptamine bases and tetrahydro-/3-c a r b o l i n e s appear as dark UV-absorbing spots under short-wave UV wh i le a romat i c and d ihydro-/3~carbo l ines have c h a r a c t e r i s t i c f l u o r e s c e n c e s under long-wave UV. F o l l o w i n g examinat ion under UV, p l a t e s were sprayed w i th E h r l i c h ' s reagent [9] which g i v e s b lue to v i o l e t c o l o r s wi th t ryptamine bases upon exposure to HCI v a p o r s . Tryptamines d e t e c t e d in the samples c o u l d be t e n t a t i v e l y i d e n t i f i e d from TLC based on compar ison of the Rfs and c o l o r r e a c t i o n s w i th au then t i c s tandards ( c f . Tab le II and F i g s . 3 and 4, Chapter IV fo r complete TLC data of the t r yp tamine and 0 -c a r b o l i n e s t a n d a r d s ) . For some samples , i d e n t i t i e s of c o n s t i t u e n t s de t e c t ed us i ng TLC were c o n f i r m e d by GC/MS. C o n d i t i o n s f o r the GC/MS a n a l y s i s were i d e n t i c a l to those 125 d e s c r i b e d in Chapter IV. For some samples , GC was used to con f i rm the i d e n t i t i e s of the t ryp tamine bases d e t e c t e d , based on compar ison wi th the r e t e n t i o n t imes of a u t h e n t i c s t anda rds . C o n d i t i o n s of the GC ana l y ses are d e s c r i b e d below. F o l l o w i n g TLC those samples which were p o s i t i v e fo r a l k a l o i d s were evaporated to d ryness under n i t r o g e n and s t o r e d at -20 ° C . B. Q u a n t i t a t i v e A n a l y s i s of Tryptamine S tandards Those M y r i s t i c a c e o u s samples which were a l k a l o i d - p o s i t i v e were q u a n t i f i e d us ing gas chromatography (GC). The inst rument used was a Sigma 3B Gas Chromatograph (Pe rk in Elmer) equipped wi th a hydrogen flame i o n i z a t i o n d e t e c t o r . The column was a 15 M x 0.25mm SE-30 fused ' s i l i c a c a p i l l a r y column (J & W S c i e n t i f i c ) . C a r r i e r gas was he l i um ; i n l e t p r e s su re fo r the c a r r i e r gas was 18 p s i ; i n l e t p r e s su re fo r both the hydrogen and a i r was 30 p s i ; s p l i t r a t i o was 0 .67 . A t t e n u a t i o n of the GC d e t e c t o r was 1x4 mv and the a t t e n u a t i o n of the cha r t r e co rde r was 1x5 mv. The chromatograph was temperature programmed, from an i n i t i a l temperature of 120° C to a f i n a l temperature of 200° C. I n i t i a l temperature was he l d fo r 3 min f o l l o w i n g i n j e c t i o n and then i n c r e a s e d at a r a te of l 0 ° / m i n . Re t en t i on t imes , peak h e i g h t s , and peak he igh t r a t i o s of t r yp tamine d e r i v a t i v e s were determined us ing s t anda rds (Table V I I I ) . Re t en t i on t imes were c a l c u l a t e d as the mean ± s tandard e r r o r of a minimum of 10 i n j e c t i o n s . M ix tu res of t r yp tamine s tandards were used to c o n s t r u c t c a l i b r a t i o n cu rves used in the q u a n t i t a t i v e ana l y ses of the M y r i s t i c a c e o u s samples . C o n c e n t r a t i o n of the t r yp tamine c a l i b r a t i o n m ix tu res ranged from TABLE V I I I - TRYPTAMINE STANDARDS: GC ANALYTICAL DATA tryp t a m i n e r e t e n t i o n c o n c e n t r a t i o n (mg/ml) s t a n d a r d time (min) .0625 .125 .25 .5 1.0 tryptamine 6.6+.15 peak h e i g h t (cm) n.d.* n.d. 1.6 7.5 phrt - - 6 2.8 5-MeO-T 9.3±.15 peak h e i g h t n.d. n.d n.d 2.1 9.3 phr - - - .8 3.5 NMT peak h e i g h t phr 7. 1+.21 n.d . 42 . 1 1 1 . 7 . 43 3 . 2 .82 DMT 7.3+.13 peak h e i g h t .94 2.9 6.7 17.7 phr .24 .74 1.7 4.5 5-MeO-DMT 9.7±. 13 peak h e i g h t .65 1.9 4.3 10.3 phr .16 .48 1.1 2.6 * n.d.= not d e t e c t e d t phr - peak h e i g h t r a t i o . h e i g h t of s t a n d a r d peak at c o n c e n t r a t i o n X phr= h e i g h t of gramine peak at 0.25 mg/ml value of denominator was 3.91 f o r n=42 i n j e c t i o n s . 1 27 F i g u r e 7 - GC Q u a n t i t a t i o n of Tryptamine S tandards 1 28 0.0625 to 0.5 mg/ml fo r DMT, 5-MeO-DMT, and NMT, and from 0.25 to 1.0 mg/ml fo r t ryptamine and 5-methoxy-tryptamine. S tandards were d i s s o l v e d in me thano l /py r id ine 9:1 and a l i q u o t s of 1 n l were i n j e c t e d . Tryptamine and 5-MeO-tryptamine, be ing p o l a r and r e a d i l y i o n i z e d compounds, cou ld not be r e l i a b l y de t ec t ed i f l e s s than 250 ng was i n j e c t e d ; we l l below 50 ng of the l e s s p o l a r methy la ted t ryp tamines cou ld e a s i l y be d e t e c t e d , however. Ten r e p l i c a t e i n j e c t i o n s of each c a l i b r a t i o n mix ture were made at each c o n c e n t r a t i o n l e v e l . Gramine was i n c l u d e d as an i n t e r n a l s tandard in a l l of the c a l i b r a t i o n m i x t u r e s , at a cons tan t c o n c e n t r a t i o n of 0.25 mg/ml. The mean peak he igh t r a t i o fo r each s tandard at each c o n c e n t r a t i o n , w i th respec t to the i n t e r n a l s t a n d a r d , was determined ( c f . Tab le V I I I ) . A l i n e a r r e l a t i o n s h i p was observed over the c o n c e n t r a t i o n ranges s t a t e d , when the peak he igh t r a t i o was p l o t t e d aga in s t concentrat ion* ( F i g . 7 ) . C. P r e p a r a t i o n of M y r i s t i c a c e o u s Samples f o r GC Q u a n t i t a t i o n The base f r a c t i o n s of the M y r i s t i c a c e o u s bark and l e a f samples had been p r e v i o u s l y evaporated to d ryness and s t o r e d at -20° C. Each sample r ep resen ted the a l k a l o i d s e x t r a c t e d from a known amount of dry weight of p l a n t m a t e r i a l , u s u a l l y 3.0 g . One ml of me thano l /py r i d i ne 9:1 was added to the samples , which were heated b r i e f l y over a steam bath and f i l t e r e d through a c o t t o n p lugged Pas teur p i p e t t e . The f i l t r a t e was c o l l e c t e d in a sma l l graduated c y l i n d e r , the p i p e t t e was washed wi th a d d i t i o n a l methanol and the sample was ad ju s t ed to a f i n a l volume of 1.5 m l . An a l i q u o t e q u i v a l e n t to 1.0 g dry wt was removed and 129 a d j u s t e d to 0.5 ml wi th a d d i t i o n a l s o l v e n t ; t h i s sample was f u r t h e r d i l u t e d to 1.0 ml by adding 0.5 ml of so l v en t c o n t a i n i n g gramine at a c o n c e n t r a t i o n of 0.5 mg/ml. Thus the f i n a l c o n c e n t r a t i o n of the sample was 1.0 g dry wt/ml, and the f i n a l c o n c e n t r a t i o n of the gramine i n t e r n a l s tandard was 0.25 mg/ml. One ul a l i q u o t s were i n j e c t e d i n t o the GC. F i v e r e p l i c a t e i n j e c t i o n s of each sample were r un . C o n c e n t r a t i o n s of t r yp tamines in the samples were c a l c u l a t e d based on t h e i r peak he igh t r a t i o s wi th r espec t to the i n t e r n a l s t a n d a r d , us ing the c a l i b r a t i o n curves c o n s t r u c t e d us i ng t ryptamine s t anda rds . Th i s gave the c o n c e n t r a t i o n of t r yp tamines in the samples in terms of mg/ml, which was e q u i v a l e n t to mg/g dry wt, s i n c e the sample c o n s i s t e d of 1.0 g dry wt/ml.. The M y r i s t i c a c e o u s paste and snu f f samples had not been evapora ted to dryness and t h e r e f o r e the a l k a l o i d f r a c t i o n s from known dry weights of p l a n t m a t e r i a l were d i s s o l v e d in approx imate l y known volumes of methano l . These samples were heated b r i e f l y over a steam b a t h , f i l t e r e d through a co t t on-plugged Pas teur p i p e t t e and brought to a known volume wi th a d d i t i o n a l methano l . Thus the c o n c e n t r a t i o n of the sample c o u l d be expressed in terms of g dry wt/ml, e . g . , " 2 . 0 g in 4 m l " . At t h i s p o i n t 1 ul of sample was i n j e c t e d i n t o the GC to es t imate the d i l u t i o n r e q u i r e d to b r i n g the sample w i t h i n the range of the c a l i b r a t i o n c u r v e s . An a l i q u o t r e p r e s e n t i n g an a p p r o p r i a t e p o r t i o n of the t o t a l dry wt of the sample was d i l u t e d wi th an equa l a l i q u o t of me thano l /py r i d i ne 9:1 c o n t a i n i n g gramine (0.5 mg/ml) to g i ve a f i n a l c o n c e n t r a t i o n of gramine of 0.25 mg/ml. Peak he igh t r a t i o s wi th r espec t to the i n t e r n a l s tandard were 1 30 c a l c u l a t e d , and the c o n c e n t r a t i o n s of t r yp tamines in the samples were es t ima ted as d e s c r i b e d above. F i v e r e p l i c a t e i n j e c t i o n s (1 M D were made fo r each sample; f o r the paste samples , d u p l i c a t e samples were prepared as d e s c r i b e d and 5 r e p l i c a t e i n j e c t i o n s of each were made. I I I . R e s u l t s and D i s c u s s i o n The r e s u l t s are p resen ted in Tab l e s IX, X, X I , and F i g s . 8, 9, 10 and 11. A. D e t e c t i o n and I d e n t i f i c a t i o n of A l k a l o i d s in M y r i s t i c a c e o u s Samples 1. Bark and l e a f samples The base compos i t i on of the M y r i s t i c a c e o u s bark and l e a f samples v a r i e s c o n s i d e r a b l y (Table IX ) ; the v a r i a t i o n observed extends both to d i f f e r e n t p a r t s of the same p l a n t , and to d i f f e r e n t c o l l e c t i o n s of the same s p e c i e s . These o b s e r v a t i o n s are c o n s i s t e n t wi th a s i m i l a r recent survey of M y r i s t i c a c e o u s s p e c i e s [6] whose au thors a l s o remarked upon the v a r i a t i o n found in the base compos i t i on of d i f f e r e n t p l a n t s and a l s o in d i f f e r e n t p a r t s of p l a n t s . In the p resen t s tudy , a l k a l o i d s were de t e c t ed in 13 of the 27 c o l l e c t i o n s . Among those which were nega t i v e f o r a l k a l o i d s were s e v e r a l c o l l e c t i o n s of V . p a v o n i s , and 6 I r y an the ra spp . Of the I r y an the ra s p e c i e s examined in the p resen t su rvey , on ly one (I_. u l e i ) was i n c l u d e d i n the survey of Holmstedt et a l . [ 6 ] , These i n v e s t i g a t o r s d e t e c t e d a t r a ce (0.013%) of 5-MeO-DMT in I_. u l e i . The V. pavon i s samples , of 131 which 4 c o l l e c t i o n s were examined, were a l l nega t i ve fo r a l k a l o i d s wi th the excep t i on of DMK-30 which con ta ined a low l e v e l (0.07 mg/g dry wt) of DMT in the l eaves and shoot t i p s . T h i s c o l l e c t i o n has been t e n t a t i v e l y determined (W. Rodr igues 1983) as V. pavon is on the b a s i s of s t e r i l e m a t e r i a l , however the presence of DMT tends to c a s t doubt on t h i s d e t e r m i n a t i o n . An inde te rmina te V i r o l a spec i e s from B r a z i l (Plowman 12,218) the bark of which r e p o r t e d l y i s smoked by w i t ch d o c t o r s as an a d d i t i v e to tobacco (Plowman, T . , p e r s . comm., 1983) was a l s o nega t i ve fo r a l k a l o i d s . S ix c o l l e c t i o n s of V. e longa ta were ana l yzed and a l l were p o s i t i v e fo r a l k a l o i d s . F i v e of these were found to c o n t a i n t ryptamine bases , v i z . , DMT, 5-MeO-DMT, NMT; a l l th ree compounds were de t e c t ed in some c o l l e c t i o n s whi le o the r s con t a i ned on ly one or two out of the t h r e e . A l l samples in which DMT was de tec t ed a l s o con ta ined l e s s e r amounts of N-methy l t r yp t am ine , an o b s e r v a t i o n which p robab l y r e f l e c t s the p o s i t i o n of t h i s compound as the penu l t ima te s t ep in the b i o s y n t h e s i s of DMT. No s i m i l a r a s s o c i a t i o n of e i t h e r DMT or NMT wi th 5-MeO-DMT was found . DMK-35, 36, 37 were a l l c o l l e c t e d at the same s i t e , and a l l had a s i m i l a r base c o m p o s i t i o n ; a l though the voucher m a t e r i a l fo r these c o l l e c t i o n s i s s t e r i l e , they p robab l y r epresen t V. e l o n g a t a . DMK-45, determined as V. e l o n g a t a , possessed a comp le t e l y anomalous a l k a l o i d p r o f i l e ; on ly t r a c e s of E h r l i c h - p o s i t i v e compounds c o u l d be de t e c t ed by TLC . The major bases are appa r en t l y f l u o r e s c e n t compounds s i m i l a r to /3-carbol ines . The f l u o r e s c e n t c o n s t i t u e n t s de t e c t ed d i d not match the Rfs and mass s p e c t r a of the a v a i l a b l e /3-c a r b o l i n e s t anda rds , however. The l i m i t e d amount of p l a n t TABLE IX - DETECTION OF ALKALOIDS IN MYRISTICACEOUS BARK AND LEAF SAMPLES name & Col 1ect ion H a l k a l o i d reagent tests:* D M V TLC : t UVfluor. UVabs. Ehr1 i ch's react ion:* compounds detected^ DMK-30 V i r o l a pavonis leaves (3.0 g d wt.) twigs (3.0 g) ++ + / - ++ + + + ++ _ 4-4-+ (2 ) + (2) DMT.NMT DMT.NMT DMK-32 V. pavonis leaves (3.0 g) _ _ _ - - -twigs (3.0 g) . _ _ . . . DMK-34 V. pavonis leaves (3.0 g) twigs (3.0 g) bark (3.0 g) V - +/- V -+/- +/- v- + 4-+ - -DMK-35 V i r o l a sp. leaves (3.0g) + + - + 4- - 4- + (2) DMT.NMT DMK-36 V i r o l a sp. leaves (3.0 g) - 4- + (2) DMT.NMT DMK-37 V i r o l a sp. leaves (3.0 g) + + - 4-4- - 4- + (2) DMT,NMT DMK-40 V. sebifera leaves (3.0 g) bark (3.0 g) + - + 4-4- 4- + 4-+ 4-+ ( 1 ) + (3) NMT DMT.NMT.5Me0--DMT DMK-41 V. elongata bark (3.0 g) + + + + 4- - 4- + (2) NMT.DMT DMK-44 Iryanthera l o n g i f l o r a bark (3.0 g) - - 4- - -DMK-45 V. elongata bark (3.0 g) * + + - + + + 4- + + /-( 1 ) -DMK-46 V. c a l l o p h y l l a bark (6.8 g) 4- 4 - / - 4. - 4- + (3) NMT,5MeO-DMT, , TA DMK-47 Iryanthera macrophylla bark (3.0 g) - - 4- - -DMK-48 I. u l e i bark (3.0 g) _ - 4- - -Table IX (cont'd) name & Col 1ectIon H D a l k a l o i d reagent tests:* M V TLC : t UVfluor. UVabs. Ehr1 i ch's react ion:1 compounds detected^ DMK-49 I. bark (2.3 crass 1fol1 a 9) - - - - - - -DMK-50 I. juruensls bark (3.0 g) - - - - + - -OMK-51 I. bark (3.0 paraensis g) - - - - - - -OMK-52 V i r o l a multinervia bark (3.0 g) _ _ _ DMK-56 V.callophylla seeds & f r u i t s (3.0 g) + + + + - + + (2) DMT,NMT DMK-59 V. bark (3.0 leaves (3 elongata 9) 0 g) + + + + ++ + + + + (2) + (1) NMT.5-MeO-DMT NMT,DMT DMK-63 V. bark (3.0 pavon i s 9) - - - - + - -DMK-67 V. leaves (3 bark (3.0 elongata 0 g) g) + /-+ + +/-+ + + (1) 5MeO-DMT DMK-68 V. leaves (3 bark (3.0 elongata 0 g) 9) ++ + + + + + + ( 1 ) 5MeO-DMT DMK-69 V. leaves (3 bark (3.0 elongata 0 g) 9) + + + /-+ + + */-+ + + ( D 5-MeO-DMT DMK-75 V. leaves (3. 1oretens i s 0 g) - - - - - - -DMK-78 Osteophloem platyspermum leaves (3.0 g) ++ + + + - + + (1) N-methyltryptophan methyl ester T a b l e IX ( c o n t ' d ) name & C o l l e c t I o n # a 1ka1o i d t e s t s : * D reagent M V TLC : t U V f l u o r . UVabs. Ehr1 i ch's r e a c t i on:$ compounds detected)) DMK-82 V. a l b i d i f l o r a bark (3.0 g) - - - . . _ Plowman 12218 V i r o l a sp. bark (2.0 g) - - - - + - -* r e a c t i o n to a l k a l o i d p r e c i p i t a t i o n reagents. D=Dragendorff's reagent; M=Meyer's r e a g e n t : V=Valser's reagent. C o m p o s i t i o n r e a g e n t s i s a c c o r d i n g to Farnsworth [ 8 ] , t R e s u l t s of TLC of b a s i c f r a c t i o n s : presence/absence of UV-f1uorescent or UV-absorbing s p o t s i s i n d i c a t e d . $ Presence/absence of E h r 1 1 c h - p o s i t 1 v e spots ( d i a g n o s t i c f o r tr y p t a m i n e s ) on TLC p l a t e i s i n d i c a t e d . Numbers in p a r e n t h e s e s i n d i c a t e n of E h r 1 i c h - p o s i t i v e s p o ts d e t e c t e d . b I n d i c a t e s major compounds i d e n t i f i e d i n sample, u s i n g a combination of methods i n c l u d i n g TLC, GC, HPLC, and i n some c a s e s GC/MS. TABLE X - DETECTION OF ALKALOIDS IN MYRISTICACEOUS PASTE AND SNUFF SAMPLES name & amt. a l k a l o i d reagent TLC: t C o l l e c t i o n # e x t r a c t e d t e s t : * of s o u r c e p l a n t (g d wt. ) D M V UV f1uor . UVabs. E h r l i c h ' s r e a c t ion: i compounds detected)) A. O r a l l y A c t i v e Paste Samples:* 1. DMK-40 V. s e b i f e r a 2.0 +++ A l f r e d o Moreno n\ - Puco U r q u i l l o + + + + + - + + (2) NMT.DMT, 5-MeO-DMT 5-MeO-NMT 2. DMK-67 V. e l o n g a t a 2.0 +++ A l f r e d o Moreno *2 - Puco U r q u i l l o + + + - + + ( 1 ) NMT.DMT, MTHpC (GC/MS) 3. DMK-68 V. e l o n g a t a 2.0 +++ A l f r e d o Moreno *3 - Puco U r q u i l l o + + + + + - + + ( 1 ) NMT 4. DMK-69 V. e l o n g a t a 2.0 +++ A l f r e d o Moreno #4 - Puco U r q u i l l o + + 4- + + - + + ( 1 ) NMT 5. DMK-34 V. p a v o n i s 2.0 - - - -J o r g e Churay - Puco U r q u i l l o 6. DMK-59 V. e l o n g a t a 5.4 +++ Marcos F l o r e s - B r i l l o Nuevo + + + + + - + + (2) 5MeO-DMT. 5Me0-NMT 7. no voucher 2.0 + + + "oo'-koey" La C h o r r e r a . Colombia (1971) + + + + + - + + (2) DMT. NMT. MTHpC. DMTHpC (GC/MS) B. Yanomamo s n u f f s (Venezuela. 1972).• 1. no voucher 1.7 ++ "buhenak + mashahara" = J u s t i c i a p e c t o r a l i s + + + unknown spec i es? + + - coumarin. umbel 1 i f e r o n e (TLC.GC/MS) 2. no voucher 3.0 ++ "mashahari" = J u s t i c i a p e c t o r a l i s + + + + + + (3) DMT. 5-MeO-DMT. NMT. umbe11iferone. coumarin (TLC.GC/MS) 3. no voucher 3.0 ++ + + + + + + + (2) DMT,5MeO-DMT "caraknak" = V i r o l a sp. c a r b o n i z e d r e s i n ? 4. no voucher 4.0 +++ + + + + + + "yakuana-sagona" = V i r o l a sp. or complete s n u f f + (2) NMT, 5Me0-DMT * r e a c t i o n to a l k a l o i d p r e c i p i t a t i o n r e a g e n t s . D=Oragendorff's reagent; M=Meyer's reagent; V=Valser's reagent. C o m p o s i t i o n of r e a g e n t s a c c o r d i n g to Farnsworth [ 8 ] . t R e s u l t s of TLC of b a s i c f r a c t i o n s . U V f l u o r . I n d i c a t e s presence/absence of f l u o r e s c e n t s p o t s : UVabs. I n d i c a t e s p r e s e n c e / a b s e n c e of UV-absorbant s p o t s . * Presence/absence of E h r 1 i c h - p o s i t i v e s p o ts ( d i a g n o s t i c f o r t r y p t a m i n e s ) on TLC. Numbers i n p a r e n t h e s e s i n d i c a t e P of Ehr 1 1 c h - p o s i t i v e spots d e t e c t e d b I n d i c a t e s major compounds i d e n t i f i e d i n sample, u s i n g combination of methods i n c l u d i n g TLC, GC. HPLC. & GC/MS. H F i r s t l i n e i n d i c a t e s c o l l e c t i o n n and i d e n t i t y of s o u r c e - p l a n t , where known; second l i n e i n d i c a t e s name of p e r s o n m a n u f a c t u r i n g sample and v i l l a g e of o r i g i n * No vouchers a v a i l a b l e f o r Yanomama snu f f samples. Second l i n e l i s t s Yanomama name of sample and p r o b a b l e b o t a n i c a l i d e n t i t y based on names c i t e d i n S c h u l t e s [ 2 1 . 1 37 m a t e r i a l p r e c l u d e d f u r t h e r i n v e s t i g a t i o n . DMK-45 may represent a gene t i c v a r i a n t of V. e l onga t a in which t r yp tamine b i o s y n t h e s i s has been r e - d i r e c t e d to the s y n t h e s i s of 0 - c a r b o l i n e s , or i t may r ep resen t a m i s i d e n t i f i c a t i o n of the c o l l e c t i o n . Cassady et a l . [7] r epo r t ed the i s o l a t i o n of 6-MeO-tetrahydroharman, 6-MeO-harmalan , and 6-MeO-harman as the major bases of V i r o l a  c u s p i d a t a ; these i n v e s t i g a t o r s d i d not de tec t t r yp tamines in t h i s s p e c i e s . The most recent taxonomic monograph of the genus V i r o l a [10] does not r e cogn i ze V. c u s p i d a t a as a l e g i t i m a t e s p e c i e s , c o n s i d e r i n g i t e q u i v a l e n t to V. e l o n g a t a . T h i s i s not the on ly i n s t ance in which c o n f u s i o n has a r i s e n in the phy tochemica l and e t h n o b o t a n i c a l l i t e r a t u r e as a r e s u l t of the i l l - d e f i n e d s p e c i e s concepts in the genus V i r o l a . Numerous p u b l i c a t i o n s [ 1 , 2 , 3 , 5 , 6 , 1 1 , 1 2 ] r e f e r to the use of V i r o l a t h e i o d o r a in the p r e p a r a t i o n of h a l l u c i n o g e n i c s n u f f s and o r a l l y - i n g e s t e d p a s t e s ; yet t h i s spec i e s i s not r e cogn i zed by Rodr igues [10] and i s t r e a t e d as equ i v a l en t to V. e l onga ta or V. c a l o p h y l l a . For i n s t ance the s p e c i e s l i s t c i t e d in [6] c o n t a i n s 2 e n t r i e s fo r V. c u s p i d a t a , 5 e n t r i e s fo r V. e l o n g a t a , and 2 e n t r i e s f o r V i r o l a t h e i o d o r a ; i f Rod r igues ' r e v i s i o n of the genus i s f o l l o w e d , a l l of these are p r o p e r l y des i gna t ed by the b i nomia l V. e l o n g a t a . Inasmuch as the m a j o r i t y of the M y r i s t i c a c e o u s voucher c o l l e c t i o n s examined in the p resen t study were determined by D r . Rod r i gues , the spec i e s concepts e s t a b l i s h e d in [10] have been f o l l o w e d . Tab le IX shows s e v e r a l o ther d i f f e r e n c e s from the survey r e p o r t e d in [6] which deserve comment. Holmstedt et a l . , examined th ree c o l l e c t i o n s of V i r o l a m u l t i n e r v i a , 2 of which 1 38 con t a i ned DMT in the bark wh i le the t h i r d was a l k a l o i d n e g a t i v e . A s i n g l e specimen of t h i s s p e c i e s was examined in the present survey and was a l k a l o i d n e g a t i v e . In [ 6 ] , no a l k a l o i d s were de t e c t ed in the l eaves of V. seb i f e r a and t h i s was the on ly t i s s u e examined; in our su rvey , t h i s s p e c i e s c o n t a i n e d s u b s t a n t i a l amounts ( c f . Tab le XI) of DMT, 5-MeO-DMT, and NMT in the bark and t r a ce s of NMT in the l e a v e s . Our c o l l e c t i o n of t h i s s p e c i e s (DMK-40) was the sou rce-p lan t fo r one of the o r a l l y -inges ted pas te samples ob ta ined at Puco U r q u i l l o . C o r o t h i e and Nakano [13] r epo r t ed the i s o l a t i o n of DMT as the s o l e base from the bark of V. seb i f e r a . Holmstedt et a l . , [6] examined 3 c o l l e c t i o n s of Osteophloem platyspermum ( i n c o r r e c t l y c i t e d in Tab le I of [6] as Osteophloem p l a t y p h y l l u m ) . DMT, 5-MeO-DMT, and 5-hydroxy-DMT were de tec t ed in one of the samples , wh i le the o the r s were a l k a l o i d - n e g a t i v e . 0. platyspermum was r ep resen ted in the p resen t survey by a s i n g l e sample of l eaves (DMK-78) which con t a i ned a s i n g l e E h r l i c h p o s i t i v e base not c o r r e s p o n d i n g to any of the a v a i l a b l e t ryp tamine s t anda rds . T h i s compound gave Rf=0.6 in the TLC so l v en t used ( c f . M a t e r i a l s and Methods) and a GC r e t e n t i o n time of 10.1 min under the c o n d i t i o n s s t a t e d . The compound was i s o l a t e d by c o l l e c t i n g the peaks from HPLC runs . UV s p e c t r a l a n a l y s i s was t y p i c a l of u n s u b s t i t u t e d i n d o l e s ( F i g . 8) w i th a b s o r p t i o n maxima at 289, 279, and 273 nm and minima at 286. The mass spectrum ( F i g . 9) matched that of a r e f e r e n c e spectrum of N-methyl-L-tryptophan methy l e s t e r ; the base peak was 130 mass u n i t s and the parent ion was m/z 232. The compound N,N-dimethy1-L-tryptophan has an i d e n t i c a l mo l e cu l a r weight and base peak, however N-methyl-L-tryptophan methyl e s t e r can be 100-1 130 DMK 78 BASE M/ 130 CH3 M+232 • f r rrNH COOCH3 H 102 [ *T 1 f ' 1 1' t 1 1 173 U—r—i—| r !—• |-l i i—r—i—i— r ' > ••—r 232 ' l' •—1—i—|—i—l—i—l—'—l—'—i—'—1—•—l—i—l—'—r m/e F i g u r e 9 - Mass Spectrum of N-methyl-tryptophan methyl e s t e r from leaves of Osteophloem platyspermum 1 4 0 d i s t i n g u i s h e d from i t by the presence of a peak at 173 mass u n i t s which r ep resen t s the l o s s of the methyl c a r b o x y l a t e fragment (COOMe) from the parent ion (M-59). T h i s peak i s absent from the mass spectrum of N ,N-d imethy l-L- t r yp tophan . The compound de t e c t ed in 0. platyspermum c l e a r l y shows an M-59 peak at m/z 173, thus p r o v i d i n g s t rong ev idence that i t i s in f a c t N-methy l - l - t r yp tophan methyl e s t e r . T h i s compound has not been r epo r t ed in the M y r i s t i c a c e a e a l t hough i t has been found in s e v e r a l s p e c i e s in the Leguminosae [14,15] and in S ida  c o r d i f o l i a (Malvaceae) [16 ] . The mass s p e c t r a l data fo r our sample i s c o n s i s t e n t in a l l r e s p e c t s wi th that r epo r t ed in [16 ] . I d e n t i f i c a t i o n of an N-methylated t ryptophan in a M y r i s t i c a c e o u s genus i s i n t e r e s t i n g from a b i o s y n t h e t i c s t andpo in t as i t i n d i c a t e s tha t the pathway to t r yp tamines in t h i s f am i l y may i n v o l v e me thy l a t i on of the s i de cha in n i t r o g e n p r i o r to deca rboxy l a t i o n . 2. Snuff and pas te samples The a l k a l o i d c o n s t i t u e n t s of a number of samples of M y r i s t i c a c e o u s drug p r e p a r a t i o n s were a l s o i n v e s t i g a t e d (Table X ) . These samples i n c l u d e d one sample of o r a l l y - i n g e s t e d pas te c o l l e c t e d at La C h o r r e r a , Co lomb ia , in 1971, and 6 pas te samples c o l l e c t e d in 1981 at the Wi to to/Bora v i l l a g e s of Puco U r q u i l l o and B r i l l o Nuevo on the R io Ampiyacu in Pe ru . Four samples of V i r o l a snu f f from Venezeu la were a l s o s c r eened . A l l but one of the M y r i s t i c a c e o u s pas te samples con t a i ned s u b s t a n t i a l c o n c e n t r a t i o n s of a l k a l o i d (Table X and X I ) . In most cases the base compos i t i on of the pas tes r e f l e c t e d the base compos i t i on of the source p l a n t s used to p repare them; the 141 c o n c e n t r a t i o n s of t r yp tamines in the pas tes were u s u a l l y 1 to 2 o rde r s of magnitude g rea t e r than the c o n c e n t r a t i o n in the crude p l a n t m a t e r i a l . )3-carbol ines were found in on l y two of the samples and in both i n s t a n c e s were present in such low c o n c e n t r a t i o n s that mass s p e c t r a l a n a l y s i s was the on ly method capab le of d e t e c t i n g them. The second sample prepared by A l f r e d o Moreno (DMK-67, V. e longata ) c o n t a i n e d low c o n c e n t r a t i o n s of DMT, NMT and a t r a c e c o n s t i t u e n t wi th a mass spectrum c l o s e l y match ing tha t p u b l i s h e d [5] f o r 2-methy l-tetrahydro-j3-carbol i n e . In t h i s sample the /3-carboline component was so c l o s e to the l i m i t of d e t e c t i o n that a poor spectrum was o b t a i n e d . However the sample from La C h o r r e r a , f o r which no voucher i s a v a i l a b l e , c on t a i ned h igh l e v e l s of NMT and DMT, and t r a c e s of two j3-c a r b o l i n e s ; one of these c o u l d be u n e q u i v o c a l l y e s t a b l i s h e d as £-methyl-TH/3C based on i t s i d e n t i t y wi th the p u b l i s h e d mass spectrum [5] (M+=186, m/z 143=100%, a d d i t i o n a l peaks at m/z 130, 115, 89, 77) wh i l e the o t h e r , e l u t i n g immediate ly a f t e r 2-Me-TH/3C, had M+ = 200, m/z 185=100%, and s t rong peaks at m/z 157, 144, 129, 118, 92, 77 ( F i g . 10) . The f ragmenta t ion p a t t e r n i s s i m i l a r to 2-Me-TH0C except tha t a l l major peaks are 14 to 15 mass u n i t s h i g h e r . L i ke 2-Me-THp"C, t h i s compound a l s o shows a prominent peak at M-43, co r r e spond ing to the l o s s of the C H 3 -N=CH2 f ragment , a l though the base peak at m/z 185 (M-15) p robab l y r e p r e s e n t s the f a c i l e l o s s of a methyl from C, and the fo rmat ion of a s t a b i l i z e d i n t e rmed i a t e wi th p o s i t i v e charge on the p i p e r i d i n e N. The prominent peak at m/z 157 i s produced by l o s s of the p i p e r i d i n e N and the ad jacent carbon 3, a t y p i c a l f r agmenta t ion p a t t e r n f o r te t rahydro-/3-carbo l ines [17 ] . A BASE M/Z 143 M*186 3 rlOO H -50 42 I li I S SI SB M i |l> i nl n 89 i l j l j , , lmil|ll h. ,a 2 3 .Mi.. ,|i m M e F i g u r e 10 - Mass Spectrum of MTH/3C from La Chor re ra Oo '-koey  BASE M/Z 185 M+200 -100 F i g u r e 1 1 - Mass Spectrum of DMTH/3C from La Cho r r e r a Oo '-koey 143 reasonab le s t r u c t u r e can be p o s t u l a t e d fo r t h i s compound, v i z . , 1,2-dimethyl t e t rahydro-/3-ca rbo l ine . A s i m i l a r compound, 6-MeO-1 ,2-dimethyl-THj3C, was i s o l a t e d by A u g u r e l l and coworkers [ 1 8 ] from the seeds of Anadenanthera p e r e g r i n a (Leguminoseae) . The f a i l u r e to de tec t /3-carbol ines in a l l of the paste samples and the extremely low l e v e l s found in two samples r a i s e s some i n t e r e s t i n g ques t i ons wi th regard to the p o s t u l a t e d mechanism of o r a l a c t i v i t y of these p a s t e s . The h a l l u c i n o g e n i c a c t i v i t y of the M y r i s t i c a c e o u s pas tes a lmost c e r t a i n l y r e s u l t s from the h igh c o n c e n t r a t i o n s of psychotomimet ic t r yp tamines which they c o n t a i n , yet these compounds are not o r a l l y a c t i v e by themse lves , r e q u i r i n g an MAO i n h i b i t o r to p r o t e c t them from i n t e s t i n a l and hepa t i c d e g r a d a t i o n . The j3-carbol ines are e f f e c t i v e MAO i n h i b i t o r s and t h e o r e t i c a l l y c o u l d o r a l l y p o t e n t i a t e the t ryp tamines in the p a s t e s , p rov i ded they were present in s u f f i c i e n t c o n c e n t r a t i o n to e f f e c t i v e l y i n h i b i t MAO. T h i s mechanism has been p o s t u l a t e d [11 ,12 ,6 ] to account fo r the o r a l a c t i v i t y of the M y r i s t i c a c e o u s pas tes as i t p robab ly a l s o does fo r ayahuasca ( c f . Chapter IV a l s o [ 1 8 , 1 9 , 2 0 ] ) . The aromat ic /3-carbol ines found in h i g h c o n c e n t r a t i o n in ayahuasca ( c f . Chapter IV) are c o n s i d e r a b l y more e f f e c t i v e as MAO i n h i b i t o r s than the te t rahydro-/3-carbo l ines d e t e c t e d in the V i r o l a pas tes ( c f . Chapter V I , a l s o [21 ] ) . S ince not a l l of the pas te samples c o n t a i n /3-carbol ines , and those which do c o n t a i n t r a ce amounts of the l e s s e f f e c t i v e t e t r ahydro-/3-ca rbo l i nes , i t seems ext remely u n l i k e l y that these c o n s t i t u e n t s c o u l d have any pha rmaco log i c a l s i g n i f i c a n c e in terms of o r a l l y p o t e n t i a t i n g the t r yp tamines i n the p a s t e s . In f a c t , the q u e s t i o n of whether the 144 M y r i s t i c a c e o u s pas tes are o r a l l y e f f e c t i v e as h a l l u c i n o g e n s remains open ( c f . -Chap te r I I I ) . In any c a se , i t appears that t h e i r o r a l a c t i v i t y , i f p r e s e n t , must depend on some mechanism other than /3-carboline mediated i n h i b i t i o n of MAO. The low l e v e l s of /3-carbol ines d e t e c t e d in the p a s t e s , and the f a i l u r e to de tec t s i m i l a r compounds in the s o u r c e - p l a n t s used fo r the d r u g s , a l s o l eaves open the p o s s i b i l i t y that they may be formed as a r t i f a c t s e i t h e r du r i ng the cook ing and c o n c e n t r a t i o n of the V i r o l a r e s i n or d u r i n g the work-up of the a l k a l o i d e x t r a c t . T h i s p o s s i b i l i t y has a l s o been r a i s e d by Holmstedt et a l . [ 6 ] . Te t rahydro - /3-carbo l ines of the type found in the pas te samples can be r e a d i l y formed from t ryp tamine d e r i v a t i v e s such as NMT by a ldehyde condensa t ion fo l l owed by c y c l i z a t i o n [22 ] . Four non-Myr i s t i c a ceous c o l l e c t i o n s u t i l i z e d as admixtures to the o r a l l y - i n g e s t e d pas tes were screened f o r a l k a l o i d s us ing p r e c i p i t a t i o n t e s t s . A l l were a l k a l o i d - n e g a t i v e . The use of these s p e c i e s ( v i z . . : R inora racemosa (Mart . & Zucc . )Kuntze, (DMK-38), Anemia s p . (DMK-39), Theobroma subinacum Mar t . (DMK-43) , T\ b i c o l o r H_;_ & (DMK-65), and Ph i lodendron nervosum ( S c h u l t . & S c h u l t . ) K u n t h (DMK-64)), as admixtures to the V i r o l a pas tes i s d i s c u s s e d more f u l l y in Chapter I I I . In a d d i t i o n to the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s samples , four M y r i s t i c a c e o u s snuf f samples c o l l e c t e d among the Yanomama Ind ians of Venezeu la were a n a l y z e d . These samples represen t v a r i o u s components used in the manufacture of the s n u f f s , but are not accompanied by voucher spec imens . They were c o l l e c t e d by D r . E r n e s t o M i g l i a z z a , an e t h n o l i n g u i s t who conducted f i e l d r e s e a r c h on Yanomama language and grammar from 1970-72. They 1 45 were g iven to the author in 1974 by D r . M i g l i a z z a , who at tha t ' t ime was on the f a c u l t y of the Department of Anthropo logy at the U n i v e r s i t y of Mary l and , C o l l e g e Park . Dr . M i g l i a z z a c o l l e c t e d the samples in connec t i on wi th h i s l i g u i s t i c s t u d i e s ; a l t hough no vouchers were c o l l e c t e d , the c o r r e c t na t i v e name of each of the samples was noted by D r . M i g l i a z z a . By comparing these names w i th those r epo r t ed by S c h u l t e s [ 2 ] , I was ab l e to a r r i v e at the p robab le b o t a n i c a l i d e n t i t y of each sample. Subsequent chemica l ana l y se s tended to c o n f i r m these t e n t a t i v e i d e n t i f i c a t i o n s . Two of the samples are a romat ic powdered l eaves of an herbaceous p l a n t which i s a lmost c e r t a i n l y J u s t i c i a p e c t o r a l i s J a c q . Va r . s t e n o p h y l l a Leona rd . One of these samples i s l a b e l l e d "mashaha r i " wh i le the o ther i s l a b e l l e d "buhenak + mashahara " . S c h u l t e s [2] s t a t e s that the K a r a u e t a r i I nd i ans , a Yanomama subgroup on the R io Cauabur i in B r a z i l , app ly the name mashahari to J u s t i c i a p e c t o r a l i s v a r . s t e n o p h y l l a , wh i le Ind ians of the R io T o t o t o b i , B r a z i l , have two names fo r J u s t i c i a p e c t o r a l i s : masha-hara-hanak ( hanak means " l e a f " ) and boo-hanak ; t h i s l a t t e r term i s s u f f i c i e n t l y c l o s e to "buhenak" tha t the terms are p robab l y synonymous. Another p o s s i b i l i t y i s tha t the Yanomama may r e cogn i ze more than one k ind or v a r i e t y of J u s t i c i a  p e c t o r a l i s and app ly d i f f e r e n t terms to each t y p e . Chemica l ana l y se s of the samples l ends suppor t to t h i s l a t t e r s u p p o s i t i o n . The sample l a b e l l e d "mashahara + buhenak" c o n t a i n e d no a l k a l o i d s ( c f . Tab le X) but d i d c o n t a i n the benzopyran d e r i v a t i v e s coumarin and u m b e l l i f e r o n e . These compounds were found in the Pe ruv ian c o l l e c t i o n s of Jus t i c i a p e c t o r a l i s made by the author (DMK-1, [ 23 ] ) ; t h e i r i d e n t i t y has been e s t a b l i s h e d by 146 compar ison of t h e i r mass s p e c t r a and TLC Rf v a l ues w i th a u t h e n t i c s t anda rds . Based on t h i s e v i dence , i t seems reasonab le to specu l a t e tha t the sample l a b e l l e d "buhenak + mashahara" c o n s i s t s of powdered Just i c i a p e c t o r a l i s l eaves wi thout any admix tu res . The s i t u a t i o n i s l e s s c l e a r w i th r espec t to the sample l a b e l l e d " m a s h a h a r i " . A l though the l a b e l i n d i c a t e s that on l y one i n g r e d i e n t ( J u s t i c i a p e c t o r a l i s ) i s p r e s e n t , chemica l a n a l y s i s (Table X & XI ) has de tec t ed DMT, 5-MeO-DMT, and NMT in t h i s sample i n a d d i t i o n to the coumarin d e r i v a t i v e s c h a r a c t e r i s t i c of Just i c i a p e c t o r a l i s . S e ve ra l e x p l a n a t i o n s may account fo r t h i s d i f f e r e n c e but u n f o r t u n a t e l y the i s sue cannot be dec ided in the absence of b o t a n i c a l voucher spec imens. One e x p l a n a t i o n i s that the l a b e l s have been exchanged on the two samples , and the sample l a b e l l e d "mashahar i " i s r e a l l y "buhenak + mashahara" ; i f t h i s i s the c a s e , then the i n g r e d i e n t "buhenak" may not r epresen t Just i c i a p e c t o r a l i s, but i n s t e a d may be a term a p p l i e d to another i n g r e d i e n t , p robab ly a V i r o l a s p e c i e s , which does c o n t a i n t r yp tamines and i s used in the manufacture of Yanomama s n u f f . An a l t e r n a t i v e e x p l a n a t i o n i s tha t t h i s sample does in f a c t r ep resen t J u s t i c i a p e c t o r a l i s or a r e l a t e d s p e c i e s , which s y n t h e s i z e s t r yp tamines as we l l as coumar ins . There e x i s t r e p o r t s [24, P rance , G. T . , p e r s . comm., 1983] that o c c a s i o n a l l y the Yanomama do prepare an i n t o x i c a t i n g snu f f us ing J u s t i c i a  p e c t o r a l i s as the so l e i n g r e d i e n t ; these r e p o r t s need to be c o r r o b o r a t e d by f u r t h e r e t h n o b o t a n i c a l and chemica l i n v e s t i g a t i o n s . A t h i r d sample c o n s i s t s of b l a c k , p a r t i a l l y cha r r ed and c a r b o n i z e d p l an t m a t e r i a l , l a b e l l e d wi th the Yanomama term " c a r aknak " . T h i s term does not co r respond to any 1 4 7 of those r e p o r t e d by S chu l t e s [ 2 ] , but may c o n s i s t of p a r t i a l l y c a r b o n i z e d V i r o l a r e s i n . S c h u l t e s , d e s c r i b i n g the p r e p a r a t i o n of the snuf f on the Rio T o t o t o b i [ 2 ] , mentions tha t a p o r t i o n of the r e s i n becomes c a r b o n i z e d in the p rocess of b o i l i n g the r e s i n ; t h i s c a r b o n i z e d m a t e r i a l i s powdered s e p a r a t e l y from the r e s t of the r e s i n to form a b l a c k i s h brown powder. The remainder of the unca rbon ized r e s i n i s ground i n t o a l i g h t c o f f e e - c o l o r e d powder, which i s then mixed w i th the c a r b o n i z e d m a t e r i a l to produce the f i n a l s n u f f . Based on t h i s d e s c r i p t i o n , i t seems p robab le tha t the p a r t l y c h a r r e d m a t e r i a l l a b e l e d " ca raknak" c o n s i s t s of c a r b o n i z e d V i r o l a r e s i n . The low l e v e l s of DMT and 5-MeO-DMT which t h i s sample con t a i ned ( c f . Tab le X I ) support t h i s i d e n t i f i c a t i o n . The f o u r t h s a m p l e ' i s l a b e l l e d "yakuana-sagona" and c o n s i s t s of dark r e d d i s h brown powdered bark m a t e r i a l . Both i t s appearance and i t s h igh a l k a l o i d content ( c f . Tab l e X I ) make i t v i r t u a l l y c e r t a i n that t h i s sample c o n s i s t s e i t h e r of powdered, concen t r a t ed V i r o l a r e s i n or of the f i n i s h e d snu f f c o n t a i n i n g V i r o l a r e s i n p l u s o ther admix tu res . A c co rd ing to S c h u l t e s [ 2 ] , the term a p p l i e d to the snu f f on the R io T o t o t o b i i s nyakwana, which i s reasonab ly c l o s e to yakuana. De te rmin ing the exact equ i v a l ence of these na t i v e terms i s c o m p l i c a t e d by the f a c t tha t the Yanomama l i n g u i s t i c f am i l y i n c l u d e s at l e a s t four major l anguages , each wi th numerous d i a l e c t s ; many of these d i a l e c t s are mutua l l y u n i n t e l l i g i b l e [ 25 ] . In view of t h i s i t i s remarkable tha t the names recorded by M i g l i a z z a fo r the v a r i o u s snu f f components can be matched so c l o s e l y w i th those recorded by S c h u l t e s . 1 48 B. Q u a n t i t a t i v e Ana l yses of M y r i s t i c a c e o u s Samples The q u a n t i t a t i v e data fo r the t r yp tamines found in the v a r i o u s M y r i s t i c a c e o u s drug samples (Table XI) show that the V i r o l a snu f f ( "yakuana-sagona") and the v a r i o u s pas te samples c o n t a i n s i m i l a r amounts of a l k a l o i d s on a dry wt b a s i s . The snu f f sample con ta ined 19.7 mg/g 5-MeO-DMT, a f i g u r e of the same order of magnitude as that r epo r t ed by Holmstedt et a l . [5] who found 7.15 mg/g in a sample of " epena " snuf f from the R io C a u a b u r i . A sample of nyakwana snu f f from the R io T o t o t o b i was r epo r t ed by Holmstedt et a l . [5] to c o n t a i n more than 110 mg/g DMT and 5-MeO-DMT, or more than 11% a l k a l o i d s . In l i g h t of the o ther q u a n t i t a t i v e work r epo r t ed in t h i s and o ther s t u d i e s [ 5 , 6 ] , t h i s f i g u r e i s open to q u e s t i o n . It i s p o s s i b l e that an e r r o r was made e i t h e r in the measurement of the a l k a l o i d content of the sample, or in the c a l c u l a t i o n of the expe r imen ta l d a t a . The other components of the Yanomama s n u f f s which were found to c o n t a i n t r yp tamines ( "mashahar i " and " ca raknak " ) c on t a i ned q u i t e low l e v e l s compared to the "yakuana" sample. Tryptamine c o n c e n t r a t i o n s found in the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s drugs were g e n e r a l l y of the same order of magnitude as those in the s n u f f . For i n s t a n c e , the f i r s t batch of paste (des igna ted o o ' - koey in Wi to to ) ob ta ined from A l f r e d o Moreno at Puco U r q u i l l o , c on t a i ned 18.8 mg/g a l k a l o i d , 70% of which was 5-MeO-DMT. The second , t h i r d , and f o u r t h batches ob ta ined from A l f r e d o Moreno con t a i ned anomalously low l e v e l s of a l k a l o i d . These batches were d e r i v e d from DMK-67,68, and 69, r e s p e c t i v e l y , a l l of which have been determined as V. e l o n g a t a . Comparison of the voucher spec imens , and the q u a l i t a t i v e and q u a n t i t a t i v e s i m i l a r i t i e s of TABLE XI - QUANTITATIVE ANALYSIS OF TRYPTAMINES IN MYRISTICACEOUS SAMPLES Sample name Spe c i e s & a 1ka1o i ds & o r i g i n Col l e c t i o n .# detec ted mg/g d wt. % A. O r a l l y a c t i v e M y r i s t i c a c e o u s p a s t e s : "oo'-koey" no voucher DMT 1 .58 14 La C h o r r e r a . NMT 9 . 43 86 Colombia T o t a l : 1 1 .01 "oo'-koey" DMK-40 DMT 3 .8 20 Puco Urqu i11o V. s e b i f e r a 5-MeO-DMT 13 . 2 70 A l f r e d o Moreno #1 NMT 1 . 78 10 T o t a l : 18 . 78 "oo'-koey" DMK-67 DMT 0 .065 7 Puco Urqu i11o V. e l o n g a t a NMT 0 . 86 93 A l f r e d o Moreno #2 To t a l : 0 .92 "oo'-koey" DMK-68 DMT 0 . 25 10 Puco Urqu i11o V. e l o n g a t a NMT 2 . 2 90 A l f r e d o Moreno #3 T o t a l : 2 . 45 "oo'-koey" DMK-69 DMT 0 . 164 10 Puco U r q u i 1 l o V. e l o n g a t a NMT 1 .43 90 A l f r e d o Moreno #4 T o t a l : 1 59 "ku' -ru-ku" DMK-59 5-MeO-DMT 15 . 72 100 B r i l l o Nuevo V. e l o n g a t a Marcos F l o r e s #1 B. Yanomama s n u f f s 8> s n u f f admixtures: "mashahar i " = J u s t i c i a p e c t o r a l i s DMT 0. 09 15 Venezuela (no voucher) 5-MeO-DMT 0. 52 85 T o t a l : 0. 61 "caraknak" = V i r o l a sp. DMT 0. 064' 17 Venezuela c a r b o n i z e d r e s i n 5-MeO-DMT 0. 32 83 (no voucher) T o t a l : 0. 384 "yakuana-sagona" = V i r o l a sp. ( s n u f f ) 5-MeO-DMT 19.7 Venezuela (no voucher) 100 T a b l e XI (cont'd) Sample name S or i g i n S p e c i e s & Co 11ect i on # a 1ka1o i ds d e t e c t e d mg/g d wt. % C. V i r o l a bark & l e a f samples: V i r o l a sp. DMK-30 1 eaves twigs DMT NMT DMT NMT 0.08 t r a c e 0.085 t r a c e 100 100 V i r o l a sp. DMK-35 1 eaves DMT NMT 0.097 t r a c e 100 V i r o l a sp. DMK-36 1 eaves DMT , NMT 0. 106 t r a c e 100 V i r o l a sp. DMK-37 1 eaves DMT NMT 0.097 t r a c e 100 V i r o l a s e b i f e r a DMK-40 1 eaves bark NMT DMT 5-MeO-DMT NMT T o t a l : t r a c e 0.078 0.181 t r a c e 0.259 30 70 V i r o l a e l o n g a t a DMK-41 bark DMT NMT To t a l : 0.063 0. 102 0. 165 38 62 V i r o l a c a l l o p h y l l a DMK-46 bark DMT 0. 56 100 V i r o l a c a l l o p h y l l a DMK-56 seeds & f r u i t DMT NMT 0. 185 t r a c e 100 V i r o l a e l o n g a t a DMK-59 bark 1 eaves 5-MeO-DMT NMT DMT NMT 0. 23 t race 0.17 t r a c e 100 100 151 the a l k a l o i d p r o f i l e s of the bark , l e a f and pas te samples ob ta ined from these th ree specimens ( c f . Tab l es IX, X, XI) make i t q u i t e c l e a r that these three pas te samples were a l l d e r i v e d from the same t r e e . The v a r i a t i o n seen in the l e v e l s of a l k a l o i d in the o r a l l y i nges t ed samples (Table XI) ranged from no a l k a l o i d to s u b s t a n t i a l c o n c e n t r a t i o n s of a l k a l o i d ( e . g . A l f r e d o Moreno #1). T h i s q u a n t i t a t i v e v a r i a t i o n i s accompanied by c o n s i d e r a b l e v a r i a t i o n in the k ind and number of t r yp tamine bases p r e s e n t . For example the sample p repared from DMK-40 con t a i ned measurable l e v e l s of DMT, 5-MeO-DMT, and NMT, wh i le that p repared from DMK-59 con t a i ned on l y 5-MeO-DMT in any s i g n i f i c a n t q u a n t i t y . S i m i l a r d i f f e r e n c e s in compos i t i on were found- in the snu f f samples . G e n e r a l l y the base compos i t i on of the pas tes r e f l e c t e d that of the s o u r c e - p l a n t s , at l e a s t in those i n s t a n c e s where the source-p l a n t s were a v a i l a b l e fo r examinat ion (Table IX & X ) . The a l k a l o i d c o n c e n t r a t i o n s in the p repared pas tes were u s u a l l y 60 -200 t imes g r ea t e r than i n the source p l a n t s . T h e r e f o r e i t seems that the techno logy of p r e p a r i n g the d r u g , which b a s i c a l l y i n v o l v e s e x t r a c t i n g , c o o k i n g , and c o n c e n t r a t i n g the r e s i n , does not r e s u l t in a d r a s t i c change in the base compos i t i on compared to the s o u r c e - p l a n t s ; however minor c o n s t i t u e n t s , such as the te t rahydro-/3-carbol i n e s , may be a r t i f a c t s of the cook ing p r o c e s s . The f a c t tha t these compounds have a l s o been i s o l a t e d from "raw" un t r ea ted V i r o l a bark argues a g a i n s t t h i s , however. D i s r e g a r d i n g fo r the moment the q u e s t i o n of whether the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s drugs r e q u i r e a MAOI in o rder to p o t e n t i a t e t h e i r a c t i v i t y , we can use the q u a n t i t a t i v e data from 1 52 Tab le XI to es t imate the amount of drug which would be r e q u i r e d to e l i c i t psychotomimet ic e f f e c t s t y p i c a l of t r y p t a m i n e s . Szara [26] r e p o r t e d in s e l f - e x p e r i m e n t s that the lower t h r e s h o l d dose fo r DMT adm in i s t e r ed i .m. was 0.2mg/kg, i . e . , 15 mg fo r a 75 kg a d u l t , w i th op t ima l e f f e c t s reached at 0.7 - 1.0 mg/kg, i . e , , between 50 and 75 mg. Thus from Tab le XI i t i s c l e a r that for samples wi th the h ighes t a l k a l o i d content a t h r e s h o l d dose would r e q u i r e the i n g e s t i o n of somewhat l e s s than 1 gram of paste or s n u f f , wh i le an " o p t i m a l " dose would r e q u i r e the i n g e s t i o n of 4 - 5 g dry wt of p l an t mater ia l .- However i f 5-MeO-DMT ins t ead of DMT were the major c o n s t i t u e n t then approx imate l y one-tenth as much p l an t m a t e r i a l would be an adequate t h r e s h o l d dose ( c f . Tab le I ) . The i n g e s t i o n of s e v e r a l grams of the o r a l l y - i n g e s t e d pas t e s would not be a problem but the a b s o r p t i o n of 4 grams of V i r o l a snu f f through the mucous l i n i n g of the nose and th roa t might p resen t some mechan ica l d i f f i c u l t i e s . The most a c t i v e pas t e s and s n u f f s may be those that c o n t a i n a r e l a t i v e l y g rea te r p r o p o r t i o n of 5-MeO-DMT; c e r t a i n l y t h i s was a c h a r a c t e r i s t i c of the two pas te samples which showed the g r e a t e s t a c t i v i t y in the b i o a s s a y s ( c f . Chapter III and Tab le X I ) . In f a c t , the a c t u a l p h a r m a c o l o g i c a l a c t i v i t y of these p r e p a r a t i o n s i s p robab ly de termined by more than a s imple dose-response r e l a t i o n s h i p ; the p resence or absence of MAO i n h i b i t o r s , the route of i n g e s t i o n , the d i e t , body we ight , and gene ra l h e a l t h of the person i n g e s t i n g the d r u g , the f requency of use and whether or not t o l e r a n c e i s deve loped , are a l l f a c t o r s which c o u l d a f f e c t the p h y s i o l o g i c a l a c t i o n of these d r u g s . The v a r i a b i l i t y in response which was observed in s e l f - e x p e r i m e n t s wi th some samples 1 53 examined in the p resen t study ( c f . Chapter I I I ) a l s o tends to i n d i c a t e that the a c t i v i t y / i n a c t i v i t y of these M y r i s t i c a c e o u s p r e p a r a t i o n s i s a f u n c t i o n of numerous v a r i a b l e s , not a l l of them b o t a n i c a l or chemica l in n a t u r e . IV. Summary A l k a l o i d c o n s t i t u e n t s in M y r i s t i c a c e o u s bark and l e a f samples and in p u r p o r t e d l y h a l l u c i n o g e n i c p r e p a r a t i o n s d e r i v e d from M y r i s t i c a c e o u s sources were q u a l i t a t i v e l y and q u a n t i t a t i v e l y ana l yzed us ing TLC, GC, a l k a l o i d p r e c i p i t a t i o n t e s t s and GC/MS. S i x t een of the twenty e i gh t bark and l e a f samples ana l yzed c o n t a i n e d d e t e c t a b l e amounts of a l k a l o i d s . The major bases were DMT and/or 5-MeO-DMT; much sma l l e r amounts of t r yp tamine and/or NMT were a l s o u s u a l l y p r e s e n t . D e t e c t a b l e l e v e l s of /3-carbol ines were not found in the bark or l e a f samples . C o n s i d e r a b l e v a r i a t i o n in a l k a l o i d p r o f i l e s was found , ex tend ing to d i f f e r e n t c o l l e c t i o n s of the same s p e c i e s . Four teen of the e igh teen V i r o l a samples con t a i ned a l k a l o i d s ; none of the s i x I r y an the ra s p e c i e s had d e t e c t a b l e a l k a l o i d s . A s i n g l e sample of Osteophloem platyspermum c o n t a i n e d an i n d o l i c base , i d e n t i f i e d as N-methyl-tryptophan methyl e s t e r on the b a s i s of UV and mass s p e c t r a l c h a r a c t e r i s t i c s . Seven samples of an o r a l l y - i n g e s t e d drug made from V i r o l a spp . were a n a l y z e d . A l l except one con t a i ned s u b s t a n t i a l amounts of t r y p t a m i n e s , but the types and p r o p o r t i o n s of t r yp tamines p resen t v a r i e d g r e a t l y between samples . Samples of V i r o l a snu f f i n c l u d i n g v a r i o u s admixtures were ana l yzed and a l l components but one con t a i ned 1 54 d e t e c t a b l e l e v e l s of t r y p t a m i n e s . The drug samples hav ing the h ighes t c o n c e n t r a t i o n s of a l k a l o i d s con t a i ned 15-20 mg/g dry wt whi le the M y r i s t i c a c e o u s bark and l e a f samples had much lower c o n c e n t r a t i o n s rang ing from 0.05 to 0.26 mg/g dry wt. Only two of the M y r i s t i c a c e o u s drug samples con t a i ned d e t e c t a b l e amounts of /3-carbol ines which had mass s p e c t r a co r r e spond ing to MTH/3C and DMTH/3C. The /3-carbol ines were t r a ce c o n s t i t u e n t s , however. These r e s u l t s i n d i c a t e tha t the i n a c t i v a t i o n of p e r i p h e r a l MAO by /3-carbol ines i s u n l i k e l y to account fo r the o r a l a c t i v i t y of the M y r i s t i c a c e o u s p a s t e s , s i n ce the amounts de t e c t ed are p robab ly not p h a r m a c o l o g i c a l l y s i g n i f i c a n t . 155 V. L i t e r a t u r e C i t e d 1. R. E. S c h u l t e s . (1979) E v o l u t i o n of the I d e n t i f i c a t i o n of the M y r i s t i c a c e o u s H a l l u c i n o g e n s of South Amer i ca . J ou rna l of  Ethnopharmacology 1:211-239 2. R. E. S c h u l t e s , £< B. Holmstedt (1968) The V e g e t a l I ng red i en t s of the M y r i s t i c a c e o u s Snu f f s of the Northwest Amazon. Rhodora '70:11 3-160 3. R. E. S chu l t e s & A. Hofmann. (1980) The Botany and Chemist ry  of H a l l u c i n o q e n s (2nd Ed. ) C h a r l e s C . Thomas, P u b l i s h e r s . S p r i n g f i e l d , 111 4. 0. R. G o t t l i e b . (1979) Chemica l S tud i e s on M e d i c i n a l M y r i s t i c a c e a e from Amazonia . J ou rna l of Ethnopharmacology 1:309-323 5. S. A g u r e l l , B. Ho lms ted t , & J . E. L i n d g r e n . (1969) A l k a l o i d s in C e r t a i n Spec ies of V i r o l a and Other South American P l an t s of E thnopharmaco log ic I n t e r e s t . Acta Chemica S cand inav i ca 23:903-916 6. B. Ho lmstedt , J . E. L i n d g r e n , T . Plowman, L. R i v i e r , R. E. S chu l t e s & O. Tova r . (1980) Indo le A l k a l o i d s in Amazonian M y r i s t i c a c e a e : F i e l d and Labora to ry Resea rch . Harvard B o t a n i c a l Museum L e a f l e t s 28:215-234 7. J . M. Cassady , G. E. B l a i r , R. F. Ra f f au f & V . E. T y l e r . (1971) The I s o l a t i o n of 6-methoxyharmalan and 6-methoxyharman from V i r o l a c u s p i d a t a . L l o y d i a 34:161-162 8. R. M a r t e l l o & N. Fa rnswor th . (1962) Obse r va t i ons on the S e n s i t i v i t y of Seve ra l Common A l k a l o i d P r e c i p i t a t i n g Reagents . L l o y d i a 25:176-185 9. D. B. Repke, D. T . L e s l i e & G. Guzman. (1977) Baeocys t i n in P s i l o c y b e , Conocybe, and Panaeo lus . L l o y d i a 40:566-78 10. W. A . Rod r i gues . (1980) Rev isao Taxonomica das E s p e c i e s de V i r o l a Aub le t ( M y r i s t i c a c e a e ) do B r a s i l . Ac ta Amazonica 1:1-123 1 56 11. R. E. S chu l t e s (1969) V i r o l a as an O r a l l y - a d m i n i s t e r e d H a l l u c i n o g e n . Harvard B o t a n i c a l Museum L e a f l e t s 22:229-40. 12. R. E. S chu l t e s & T . Swain. (1976) Fu r the r Notes on V i r o l a as an Ora l H a l l u c i n o g e n . J ou rna l of P s y c h e d e l i c Drugs 8:317-324. 13. E. C o r o t h i e & T . Nakano. (1969) C o n s t i t u e n t s of the Bark of V i r o l a s e b i f e r a . P l an ta Medica 17:184-88 14. S. R. Johns , J . A. Lamberton, & A. A. S i oumis . (1971) The Major A l k a l o i d of Aotus subg lauca (Leguminoseae) , S-(+)-N -methy l t r yp tophan methyl e s t e r . A u s t r a l i a n J o u r n a l of  Chemis t ry 24:439-40 15. N. Mandava, J . D. Anderson & S. R. Dutky. (1974) Indole P lant-growth I n h i b i t o r from Abrus p r e c a t o r i u s Seeds. Phy tochemis t ry 13:2853-2856 16. S. G h o s a l , R. B a l l a v , P. S. Chauhan k R. Mehta. (1975) A l k a l o i d s of S ida c o r d i f o l i a . Phy tochemis t ry 14:830-832 17. R. T . C o u t i s , R. A. Locock & G. W. A. S lywka. (1970) Mass Spec t ra of S e l e c t e d /3-carbol ines [ /3-9H-pyr ido ( 3 , 4-b ) i n d o l e s ] . Organ ic Mass Spect rometry 3:879-889 18. S. A g u r e l l , B. Ho lmstedt , J . E. L i n d g r e n , & R. E. S c h u l t e s . (1968) I d e n t i f i c a t i o n of Two New /3-carboline A l k a l o i d s in South American H a l l u c i n o g e n i c P l a n t s . B i o chemica l  Pharmacology 17:2487-88 19. S c h u l t e s , R. E. (1972) E t h n o t o x i c o l o g i c a l S i g n i f i c a n c e of A d d i t i v e s to New World H a l l u c i n o g e n s . P lant Sc i ence < B u l l e t i n 18: 34-41 20. Der M a r d e r o s i a n , A. H . , H. V. P i n k l e y , & M. F. Dobbins IV. 91968) Na t i ve Use and Occur rence of N ,N-d imethy l t ryptamine in the Leaves of B a n i s t e r i o p s i s rusbyana . American J ou rna l  of Pharmacy 140:137-147 21. N. S. Buckho l tz & W. 0. Boggan. (1976) E f f e c t s of Te t rahydro-/3-carbo l ines on Monoamine Oxidase and Se ro ton in Uptake in Mouse B r a i n . B i o chem i ca l Pharmacology 25:2319-2321 1976 157 22. R. G. Taborsky & W. M. M c l s a a c . (1964) The S yn thes i s and P r e l i m i n a r y Pharmacology of Some 9H-Py r i do [3 ,4-b ] i ndo l e s (/3-carbolines) and Tryptamines Re la ted to S e ro ton in and M e l a t o n i n . J ou rna l of M e d i c i n a l Chemist ry 7:135-141 23. D. MacRae & G. H. N. Towers (1984) J u s t i c i a p e c t o r a l i s , a Study of the Bas i s f o r i t s Use as a V i r o l a Snuf f Admix tu re . J ou rna l of E thnopharmacology. Submi t ted . 24. Chagnon, N . , P. LeQuesne, & J . M. Cook (1971) Yanomamo H a l l u c i n o g e n s : A n t h r o p o l o g i c a l , B o t a n i c a l , and Chemica l F i n d i n g s . Cur rent Anthropo logy 12:72-74 25. E. C. M i g l i a z z a . (1972) Yanomama Grammar and I n t e l l i g i b i l i t y . PhD. T h e s i s , Ind iana U n i v e r s i t y . 26. S. Szara (1957) The Comparison of the P s y c h o t i c E f f e c t of Tryptamine D e r i v a t i v e s wi th the E f f e c t s of Mesca l i ne and LSD-25 in S e l f - e x p e r i m e n t s . pp . 460-467 in S. G a r a t t i n i & V. G h e t t i ( eds . ) P s y c h o t r o p i c Drugs E l s e v i e r , Amsterdam. 1 5 8 CHAPTER V I : ORALLY-ACTIVE MALPIGHIACEOUS AND MYRISTICACEOUS HALLUCINOGENS AND THEIR CONSTITUENTS AS MONOAMINE OXIDASE INHIBITORS I. I n t r o d u c t i o n The m i t o c h o n d r i a l l y - l o c a l i z e d enzyme monoamine ox idase (MAO) p l a y s an important r o l e in mammalian metabo l i sm. MAO c a t a l y z e s the o x i d a t i v e deaminat ion of b i o g e n i c monoamines i n c l u d i n g t y ramine , t r yp t am ine , s e r o t o n i n , n o r e p i n e p h r i n e , dopamine, and other monoamines, a c c o r d i n g to the gene ra l r e a c t i o n equa t ion [ 1 ] : RCH 2 NH 2 + 0 2 + H 2 0 --MAO—> RCHO + NH 3 + H 2 0 2 The enzyme i s w ide ly d i s t r i b u t e d throughout v a r i o u s t i s s u e s in v e r t e b r a t e s and i n v e r t e b r a t e s , i n c l u d i n g the l i v e r , b r a i n , sma l l i n t e s t i n e , b l ood plasma and p l a t e l e t s , h e a r t , and lungs [ 2 ] , The most s i g n i f i c a n t metabo l i c f u n c t i o n of MAO i s r e l a t e d to i t s a b i l i t y to i n a c t i v a t e endogenously produced monoamines such as s e r o t o n i n or dopamine, which f u n c t i o n as CNS n e u r o t r a n s m i t t e r substances [ 3 ] , C o n s i d e r a b l e exper imenta l ev idence has been accumulated [ 2 ] which i n d i c a t e s tha t the v i s c e r a l MAO system f u n c t i o n s as a d e t o x i f i c a t i o n mechanism s e r v i n g to p r o t e c t the nervous and c a r d i o v a s c u l a r systems from t o x i c b i o g e n i c amines i nges t ed in the d i e t and formed as a r e s u l t of a romat i c amino a c i d d e c a r b o x y l a t i o n . Ev idence based on s u b s t r a t e s p e c i f i c i t y and s e l e c t i v e s e n s i t i v i t y to c e r t a i n MAO i n h i b i t o r s i n d i c a t e s that MAO c o n s i s t s of two s p e c i e s , de s i gna t ed MAO-A and MAO-B [ 4 , 5 , 6 , 7 ] . Houslay & T i p t o n [ 7 ] c a r r i e d out a k i n e t i c e v a l u a t i o n of the two s p e c i e s us ing mixed s u b s t r a t e assays and conc luded 159 that the s u b s t r a t e s fo r s p e c i e s A a c t i v i t y were s u b s t i t u t e d 0 -pheny le thy l amines and 0-pheny le thano lamine d e r i v a t i v e s hav ing a f r ee p-hydroxy l g roup ; and 5-hydroxyt ryptamine . Subs t r a t e s fo r s p e c i e s B a c t i v i t y i n c l u d e d s u b s t i t u t e d benzylamine and 0 -pheny le thy l amine d e r i v a t i v e s , but not s u b s t i t u t e d 0 -pheny l e thano l amines . T r yp tamine , 5-methoxytryptamine, dopamine, and the m-O-methyl d e r i v a t i v e s of dopamine are s u b s t r a t e s fo r both MAO-A and MAO-B. Nef f et a l . , [8] p o i n t out that substances which i n t e r a c t wi th MAO-A such as s e r o t o n i n and no rep ineph r i ne have more p o l a r a romat ic r i n g s than substances which i n t e r a c t w i th MAO-B ( e . g . , 0-pheny l e thy l amine , benzy l am ine ) . In gene ra l add ing a p o l a r hyd roxy l group to 0-pheny le thy l amines ( e . g . tyramine) or removing one from s e r o t o n i n to form t ryp tamine r e s u l t s in a common s u b s t r a t e . The s p e c i f i c metabol ism of DMT has not been i n v e s t i g a t e d , but i f i t conforms to the above r u l e i t would be a s u b s t r a t e fo r both MAO-A and MAO-B s i n c e i t l a ck s a romat i c s u b s t i t u e n t s . Fu r the r ev idence f o r the e x i s t e n c e of two s p e c i e s of MAO comes from exper iments us ing s e l e c t i v e MAO i n h i b i t o r s [ 5 ] . Most of the MAO i n h i b i t o r s which have been the focus of b i o chem i ca l and c l i n i c a l i n v e s t i g a t i o n s be long to one of four c l a s s e s , v i z . , hyd raz ine d e r i v a t i v e s , pheny l c y c l op ropy l am ine d e r i v a t i v e s , N-benzyl-N-methyl p ropa rgy l am ine , or 2-methyl-3-p i p e r i d i n o p y r a z i n e [ 9 ] . These are s y n t h e t i c compounds which are not known to have c o r r e s p o n d i n g ana logues in n a t u r e . The major excep t i on i s the 0 - c a r b o l i n e d e r i v a t i v e s , a group of t r i c y c l i c i n d o l e a l k a l o i d s which are widespread in nature [10] ( c f . F i g . 2 ) . These compounds are b i o s y n t h e t i c a l l y d e r i v e d from t ryptamine 1 60 and i t s d e r i v a t i v e s and can be r e a d i l y s y n t h e s i z e d from t ryp tamine d e r i v a t i v e s [11 ] . Uden f r i end et a l . , [12] were the f i r s t to demonstrate tha t harmal ine and r e l a t e d compounds are ext remely potent r e v e r s i b l e i n h i b i t o r s of monoamine o x i d a s e . These i n v e s t i g a t o r s showed that the f u l l y a romat i c 0 - c a r b o l i n e s were the most e f f e c t i v e i n h i b i t o r s , and that a c t i v i t y decreased wi th i n c r e a s i n g s a t u r a t i o n of the p i p e r i d i n e r i n g ; t e t r ahyd ro - / 3 -c a r b o l i n e s s t i l l showed s i g n i f i c a n t a c t i v i t y , however. Subsequent l y , exper iments by F u l l e r et a l . [ 4 ] showed that harmal ine s e l e c t i v e l y i n h i b i t e d o x i d a t i o n of s e r o t o n i n , i n d i c a t i n g that i t was a s p e c i f i c i n h i b i t o r of MAO-A. The c a p a c i t y of 0 - c a r b o l i n e s to i n h i b i t MAO has been suggested to be the mechanism r e s p o n s i b l e fo r the o r a l a c t i v i t y of ayahuasca [13 ,14 ,15 ,16 ] and p o s s i b l y a l s o f o r the a c t i v i t y of o ther o r a l l y i nges ted drugs d e r i v e d from V i r o l a spp . [ 1 7 , 1 8 , 1 9 ] . Both ayahuasca and the o r a l l y - i n g e s t e d h a l l u c i n o g e n i c pas tes d e r i v e d from V i r o l a spp. c o n t a i n psychotomimet ic t r yp tamines such as N ,N-d imethy l t ryptamine (DMT), 5-methoxy-N,N-d ime thy l t r yp tamine (5-MeO-DMT) and r e l a t e d t r yp tamine d e r i v a t i v e s , as we l l as 0 - c a r b o l i n e d e r i v a t i v e s ( c f . Chapters IV & V ) . Ayahuasca c o n t a i n s s u b s t a n t i a l amounts of the 7-s u b s t i t u t e d 0 - c a r b o l i n e s harmine, ha rma l i ne , and te t rahydroharmine as we l l as DMT d e r i v e d from the admixture p l a n t P s y c h o t r i a v i r i d i s , commonly used in the p r e p a r a t i o n of ayahuasca ( c f . Chapter IV ) . The o r a l l y - i n g e s t e d V i r o l a p a s t e s , which are prepared from the cambia l sap of v a r i o u s V i r o l a s p e c i e s , c o n t a i n p s y c h o a c t i v e t r yp tamines as the major a l k a l o i d s ; in a d d i t i o n some samples c o n t a i n t r a c e amounts of 161 t e t r a h y d r o - 0 - c a r b o l i n e s ( c f . Chapter V and [ 16 ,19 ] ) . The t r yp tamines DMT and 5-MeO-DMT are potent h a l l u c i n o g e n s , but are not o r a l l y a c t i v e , presumably due to deaminat ion by p e r i p h e r a l MAO; in the presence of 0 - c a r b o l i n e s , however, the compounds may be p r o t e c t e d from deaminat ion and thus rendered o r a l l y a c t i v e . A l though t h i s mechanism has been proposed to under l y the o r a l a c t i v i t y of ayahuasca and the V i r o l a paste p r e p a r a t i o n s , the i n h i b i t i o n of MAO by these drugs or t h e i r a l k a l o i d f r a c t i o n s has not p r e v i o u s l y been i n v e s t i g a t e d . In the present s tudy , the a c t i v i t y of v a r i o u s 0 - c a r b o l i n e and t ryptamine d e r i v a t i v e s as s p e c i f i c i n h i b i t o r s of MAO-A was i n v e s t i g a t e d . The s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of a s e l e c t e d sample of d e r i v a t i v e s from each c l a s s were de te rm ined . A c t i v i t y measured wi th i n d i v i d u a l compounds was compared wi th the a c t i v i t y of mix tures of compounds which approximated the compos i t i on of the a l k a l o i d f r a c t i o n s of ayahuasca and the o r a l l y - i n g e s t e d V i r o l a p a s t e s . The mix tu res of s tandards were assayed in o rder to determine whether combina t ions of a l k a l o i d s e x h i b i t s y n e r g i s t i c a c t i v i t y as MAOI. The MAOI a c t i v i t y of two ayahuasca samples and the bas i c f r a c t i o n s of th ree V i r o l a pas te samples were i n v e s t i g a t e d . The a c t i v i t y of one V i r o l a pas te sample l a c k i n g a l k a l o i d s and a crude l i g n a n f r a c t i o n i s o l a t e d from V. e l onga t a was a l s o i n v e s t i g a t e d [26 ] . I I . M a t e r i a l s and Methods A. P r e p a r a t i o n of r a t - l i v e r c y t o s o l R a t - l i v e r , ob ta ined from mature female W is ta r r a t s , was 162 used as the source of monoamine ox idase fo r the a s s a y s . The r a t s were s a c r i f i c e d by s tunn ing f o l l owed by c e r v i c a l d i s l o c a t i o n . The abdominal c a v i t y was opened by removal of the b e l l y sk i n and per i toneum and the l i v e r pe r fused wi th c o l d 0.1 M sodium phosphate b u f f e r , pH 7.2 [20] by i n j e c t i o n i n t o the hepa t i c ve in w i th a beve led #22 s t a i n l e s s s t e e l s y r i n g e . The pe r fu sed l i v e r was removed from the abdominal c a v i t y and immediate ly p l a ced in c o l d b u f f e r . P o r t i o n s of l i v e r were t r a n s f e r r e d to a g l a s s homogenizer and homogenized in c o l d b u f f e r to a smooth paste (± 1 g l i v e r / 3 ml b u f f e r ) . The homogenate was c e n t r i f u g e d at 3000 x g fo r 20 min to remove nuc l ea r f ragments . The supernatant was. c o l l e c t e d and d i l u t e d 1:1 wi th b u f f e r . The d i l u t e d c y t o s o l f r a c t i o n was s o n i c a t e d fo r 10 s e c . T h i s d i l u t e d whole c y t o s o l f r a c t i o n was kept on i ce fo r the d u r a t i o n of the exper iment , and was used without f u r t h e r c e n t r i f u g a t i o n . B. P r e p a r a t i o n of r e a c t i o n mixture and a d d i t i o n of l a b e l l e d s u b s t r a t e The assay was conducted at 3 7 . 5 ° C in 13 x 100 mm d i s p o s a b l e t e s t t ubes . Volume of the r e a c t i o n mix ture was 0.5 m l . Compos i t ion of the r e a c t i o n mixture in c o n t r o l s l a c k i n g added i n h i b i t o r c o n s i s t e d of 375 ul b u f f e r , 100 ul whole c y t o s o l , and 25 ul l a b e l l e d s u b s t r a t e . The s u b s t r a t e c o n s i s t e d of 5-hydroxytryptamine ( s i d e - c h a i n - 2 - 1 " O c r e a t i n i n e s u l f a t e (Amersham). The l a b e l l e d s u b s t r a t e was d i l u t e d wi th c o l d c a r r i e r 5HT to a s p e c i f i c a c t i v i t y of 0.835 mCi/mmole and a c o n c e n t r a t i o n of 2 x 10~ 3 M. At t h i s d i l u t i o n , a d d i t i o n of a 25 ul a l i q u o t to the r e a c t i o n mixture r e s u l t s i n a f i n a l volume of 1 63 500 M1 and a 5HT c o n c e n t r a t i o n of 0.1 mM, c o n t a i n i n g a t o t a l of 0.04 vCi of l a b e l l e d 5HT. A l l components of the r e a c t i o n mixture except the l a b e l l e d subs t r a t e were added to the r e a c t i o n tubes which were ma in ta ined at 0 ° C in an i ce ba th . I n a c t i v a t e d c o n t r o l b l a n k s , i d e n t i c a l in compos i t i on to the a c t i v e c o n t r o l s , were p repa red by immersing the tubes in b o i l i n g water fo r 10 min. P r i o r to i n i t i a t i o n of the a s say , the r e a c t i o n tubes were immersed in a 3 7 . 5 ° C water bath and e q u i l i b r a t e d f o r 5 min . The assay was i n i t i a t e d by adding a 25 nl a l i q u o t of the d i l u t e d s u b s t r a t e to the tubes , mix ing fo r 1-2 sec on a vor tex mixer , and r e p l a c i n g in the water b a t h . F o l l o w i n g a d d i t i o n of the s u b s t r a t e the assay tubes were incuba ted fo r 30 min at 3 7 . 5 ° C . A f t e r 30 min the r e a c t i o n was te rmina ted by add ing 1.0 ml of 1.0 M c i t r a t e . T h i s was fo l l owed by the a d d i t i o n of 0.5 ml s a tu r a t ed N a C l , and 2.0 ml e t h y l a ce t a t e to each tube . Tubes were then vo r t exed f o r 5 sec each and c e n t r i f u g e d at 2000 rpm for 10 min to separa te the o rgan i c and aqueous l a y e r s . In t h i s method, [21] the l a b e l l e d r e a c t i o n p roduc ts are s o l u b l e in the o rgan i c (upper) phase whi le the p ro tona ted subs t r a t e remains in the lower aqueous phase . F o l l o w i n g c e n t r i f u g a t i o n , a 1.0 ml a l i q u o t of the upper phase was t r a n s f e r r e d to a s c i n t i l l a t i o n v i a l c o n t a i n i n g 9.0 ml Aquasol-2 (New England N u c l e a r ) . S c i n t i l l a t i o n v i a l s were counted fo r 10 min in a Sea r l e Isocap 300 L i q u i d S c i n t i l l a t i o n Counte r . C. Assays us ing t ryptamine and 0 - c a r b o l i n e s tandards Stock s o l u t i o n s of v a r i o u s 0-ca rbo l i ne and t ryptamine d e r i v a t i v e s , d i s s o l v e d in 0.1 N H C l , were prepared so that 1 64 a d d i t i o n of a 50 a l i q u o t to a r e a c t i o n mixture would r e s u l t in an i n h i b i t o r c o n c e n t r a t i o n of 10~ 3 M and a t o t a l volume of 500 u l . The t ryptamine d e r i v a t i v e s 5-Methoxy-d i i sopropy l t r yp tamine and 3 - [ 2 - ( 2 , 5 - d i m e t h y l ) p y r r o l y l e t h y l ] - i n d o l e were p o o r l y s o l u b l e in 0.1 N HCI so these compounds were d i s s o l v e d in 10% Tween-80. Stock s o l u t i o n s fo r lower c o n c e n t r a t i o n s were prepared by making s e r i a l 1:10 d i l u t i o n s , over a c o n c e n t r a t i o n range from 10~ 3 to 10" 1 0 M. Assay m ix tu res c o n t a i n i n g i n h i b i t o r c o n s i s t e d of 325 ul b u f f e r , 100 ul whole c y t o s o l , 50 ul i n h i b i t o r s o l u t i o n of the a p p r o p r i a t e c o n c e n t r a t i o n , and 25 ul s u b s t r a t e . The i n h i b i t o r s o l u t i o n was added j u s t be fo re the assay tubes were t r a n s f e r r e d to the warm water ba th , so that the i n c u b a t i o n time of the c y t o s o l p l u s i n h i b i t o r at 3 7 . 5 ° C was 5 min . B o i l e d b lanks c o n t a i n i n g denatured enzyme were i n c l u d e d in each r u n . In a d d i t i o n c o n t r o l s c o n t a i n i n g a c t i v e enzyme but no i n h i b i t o r were a l s o assayed s imu l t aneous l y ( i n h i b i t o r was r e p l a c e d e i t h e r with b u f f e r , 10% Tween-80, or w i th 50 ul 0.1 N HCI ) . There was no a p p r e c i a b l e d i f f e r e n c e between c o n t r o l s in which b u f f e r r ep l a ced the i n h i b i t o r and those in which 0.1 N HCI or 10% Tween-80 r ep l a ced the i n h i b i t o r . The c o n t r o l s l a c k i n g i n h i b i t o r r ep resen ted 0% MAO i n h i b i t i o n ; and the % i n h i b i t i o n of tubes c o n t a i n i n g v a r i e d c o n c e n t r a t i o n s of i n h i b i t o r was c a l c u l a t e d r e l a t i v e to these c o n t r o l s , by d i v i d i n g the amount of l a b e l l e d r e a c t i o n p roduc t s r ecove red (measured as cpm) by the amount r ecove red from the c o n t r o l t u b e s . The background count r ep resen ted by the a c t i v i t y de t e c t ed in the b o i l e d b lanks ( u s u a l l y 200-300 cpm) was s u b t r a c t e d from both c o n t r o l counts and i n h i b i t o r c o u n t s . 165 D. P r e p a r a t i o n and q u a n t i t a t i o n of ayahuasca samples fo r MAO assay The ayahuasca samples used in the assay were Don F i d e l #1 and Don Juan #2. F i v e ml a l i q u o t s of each were d i l u t e d to 50 ml w i th c h i l l e d methanol and the white n o n a l k a l o i d a l p r e c i p i t a t e removed by f i l t r a t i o n . The f i l t e r e d s o l u t i o n was then evaporated to d ryness under reduced p ressu re and r e s o l u b i l i z e d in 5 ml 0.01 N HCl to r e c o n s t i t u t e the o r i g i n a l volume. A 1:1 d i l u t i o n of t h i s r e c o n s t i t u t e d s o l u t i o n was used to q u a n t i f y the a l k a l o i d s p resent in the mixture us ing the HPLC method d e s c r i b e d in Chapter IV. T h i s q u a n t i f i c a t i o n showed the r e c o n s t i t u t e d samples to c o n t a i n 3.5 mg/ml and 4.8 mg/ml t o t a l a l k a l o i d , r e s p e c t i v e l y . S e r i a l 1:10 d i l u t i o n s of the r e c o n s t i t u t e d samples , rang ing from 0 to 1 0 ~ 7 , were made us i ng 0.01 N H C l . F i f t y M1 a l i q u o t s of the a p p r o p r i a t e l y d i l u t e d ayahuasca s o l u t i o n s were added to the r e a c t i o n mixture in the same manner as the i n h i b i t o r s , and % i n h i b i t i o n was c a l c u l a t e d r e l a t i v e to the c o n t r o l s . I n h i b i t o r assays were a l s o conducted us i ng mix tu res of s tandards which approximated the compos i t i on of ayahuasca . M ix tu res hav ing the f o l l o w i n g compos i t i ons were a s sa yed : a . Equimolar mix ture of harmine, ha rma l i ne , and t e t r ahyd roha rm ine ; t o t a l a l k a l o i d c o n c e n t r a t i o n of u n d i l u t e d s tock s o l u t i o n : 2.115 mg/ml ( 1 0 " 3 M, based on average MW). b. Equimolar mixture of harmine, ha rma l i ne , t e t r ahyd roha rm ine , and DMT; t o t a l a l k a l o i d c o n c e n t r a t i o n of u n d i l u t e d s o l u t i o n : 2.16 mg/ml (10~ 3 M, based on average MW). c . "ayahuasca analogue #1" - 69% harmine, 4.6% ha rma l i ne , and 26% t e t r ahyd roha rm ine ; o v e r a l l c o n c e n t r a t i o n e q u i v a l e n t to the 1 66 equ imolar m i x t u r e . d . "ayahuasca analogue #2" - 65% harmine, 6% ha rma l i ne , 22% t e t r ahyd roha rm ine , and 7% DMT; o v e r a l l c o n c e n t r a t i o n e q u i v a l e n t to the•equ imolar m i x t u r e . E. P r e p a r a t i o n and q u a n t i t a t i o n of M y r i s t i c a c e o u s paste samples f o r MAO assay The M y r i s t i c a c e o u s paste samples used fo r the assay were A l f r e d o Moreno sample #1, p repared from V i r o l a s e b i f e r a , (DMK-40) , Don Marcos sample #1, (V. e l o n g a t a , DMK-59) and the "oo-'-koey" from La C h o r r e r a , Colombia (no vouche r ) . A l i q u o t s of the methanol e x t r a c t s of each pas te sample equ i v a l en t to 0 .2-0.4 g dry wt of the o r i g i n a l sample were evaporated to dryness and r e s o l u b i l i z e d in 2 ml methano l . One ml was removed, d i l u t e d 1:1 w i th methano l , and f i l t e r e d through a co t ton-p lugged Pasteur p i p e t t e . Twenty ul a l i q u o t s of t h i s d i l u t e d sample were i n j e c t e d onto the HPLC in order to q u a n t i t a t i v e l y determine the a l k a l o i d conten t of the sample. The q u a n t i t a t i v e methods and a n a l y t i c a l c o n d i t i o n s were i d e n t i c a l to those used in the q u a n t i t a t i o n of the ayahuasca samples (cf Chapter IV ) . The molar c o n c e n t r a t i o n of t r yp tamines in each sample was c a l c u l a t e d from the q u a n t i t a t i v e data (Table X I I ) . An "ana logue " of each sample was p repared us i ng t ryptamine s t a n d a r d s . Each paste " ana logue " c o n s i s t e d of a mixture of t r yp tamine s tandards in the same p r o p o r t i o n s and c o n c e n t r a t i o n s as were measured in the pas te samples . Each pas te sample and i t s analogue were s imu l t aneous l y assayed in the monoamine ox idase system. If the MAOI a c t i v i t y of the pas te sample and the analogue were e q u i v a l e n t , t h i s would T a b l e X II: A l k a l o i d C o n c e n t r a t i o n i n M y r i s t i c a c e o u s P a s t e Samples Assayed f o r MAOI A c t i v i t y * Sample Name A l k a l o i d s A l k a l o i d D e t e c t e d C o n c e n t r a t i o n (mg/ml) Mo 1 ar C o n c e n t r a t i o n (M) La C h o r r e r a "oo'-koey" (no voucher) NMT DMT To t a l 1 . 38 0. 1 1 . 48 7.9 x 1 0 " 0.53 x 1 0 " 8 . 43 x 1 0 " Don Marcos #1 (DMK-59) 5-MeO-DMT 2 .03 9.3 x 1 0 " A l f r e d o Moreno *'1 5-MeO-DMT 1.19 (DMK-40) DMT 0.3 To t a l 1.49 5 . 48 x 1 0 " 1 .54 x 1 0 " 7.02 x 1 0 " * HPLC q u a n t i t a t i o n c a r r i e d out on samples r e s o l u b i 1 i z e d i n 10% Tween-80. Cf M a t e r i a l s and Methods. 168 i n d i c a t e that enzyme i n h i b i t i o n by the pas tes was a t t r i b u t a b l e s o l e l y to the t r yp t am ines , which are c o m p e t i t i v e s u b s t r a t e s of 5HT fo r MAO-A. If the MAOI a c t i v i t y of the pas te exceeded the a c t i v i t y of the ana logue , however, t h i s would i n d i c a t e tha t some c o n s t i t u e n t s _in a d d i t i o n to the t r yp tamines were c o n t r i b u t i n g to the o v e r - a l l i n h i b i t o r y a c t i v i t y . F o l l o w i n g HPLC q u a n t i t a t i o n , the paste samples were d i l u t e d w i th an e q u i v a l e n t amount of 10% Tween-80 and the methanol was removed under n i t r o g e n . If necessa ry a d d i t i o n a l Tween-80 was added to r e s t o r e the sample to i t s o r i g i n a l volume. The paste " ana l ogues " were p repared from stock s o l u t i o n s of t r yp tamine s tandards d i s s o l v e d in 10% Tween-80. The a l k a l o i d - f r e e paste sample ( de r i v ed from DMK-34, V . pavon is ) and the l i g n a n f r a c t i o n from V. e l onga t a (DMK-59) were a l s o p repared in 10% Tween-80. C o n c e n t r a t i o n s of the u n d i l u t e d s tock s o l u t i o n s of each was 3.0 g dry wt/ml and 10 mg/ml, r e s p e c t i v e l y . I I I . R e s u l t s and D i s c u s s i o n A. 0 - c a r b o l i n e s and Tryptamines as MAOI: S t r u c t u r e / A c t i v i t y R e l a t i o n s h i p s 1. 0 - c a r b o l i n e s In order to a ssess the i n f l u e n c e of s t r u c t u r a l v a r i a t i o n of the /3-carboline ske l e ton ( c f . F i g . 2) on the MAO i n h i b i t o r y a c t i v i t y of these compounds, a s e l e c t e d group of s tandards (Table XI I I ) were assayed i_n v i t r o u s i ng the r a t - l i v e r c y t o s o l f r a c t i o n as the source of enzyme and 1 " C - l a b e l l e d 5-169 hydroxyt ryptamine c r e a t i n i n e s u l f a t e as s u b s t r a t e . Tab le XIII l i s t s 1 5 0 va lues f o r the present experiment and a l s o i n c l u d e s the va lues r epo r t ed by Mclsaac Es tevez [22] and B u c k h o l t z , et a l . , [23] f o r compar i son . The I 5 0 va lue co r responds to the molar c o n c e n t r a t i o n of i n h i b i t o r at which enzyme a c t i v i t y i s 50% i n h i b i t e d wi th respec t to c o n t r o l s l a c k i n g i n h i b i t o r . Mclsaac & Es tevez [22] used a c a l f - l i v e r m i t o c h o n d r i a l f r a c t i o n and 1 a C tyramine as s u b s t r a t e , wh i le Buckho l tz et a l . [23] u t i l i z e d mouse whole b r a i n homogenate and 1 " C - t r y p t a m i n e as s u b s t r a t e . S ince the degree of MAO i n h i b i t i o n measurable _in v i t r o i s p a r t i a l l y determined by the t i s s u e source and p r e p a r a t i o n of the enzyme, and p a r t i a l l y by the p a r t i c u l a r s u b s t r a t e used , the I s o va lues r epo r t ed in Tab le XIII fo r the present study are not d i r e c t l y comparable to those r epo r t ed in [22] and [23 ] . In g e n e r a l , however, the c o n c l u s i o n s suggested by the present study r ega rd ing the s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of /3-carbol ines as monoamine ox idase i n h i b i t o r s are in agreement wi th the p r e v i o u s s t u d i e s . Thus , fo r example, the f u l l y a romat ic and dihydro-/3-carbo l ines are s i g n i f i c a n t l y more potent i n h i b i t o r s than ana logues in which the p i p e r i d i n e r i n g i s f u l l y s a t u r a t e d ( c f . harmine, ha rma l i ne , and t e t r ahyd roha rm ine ) . I n h i b i t o r y potency i s rough ly e q u i v a l e n t between the 7-methoxylated f u l l y a romat i c and d ihyd ro d e r i v a t i v e s harmine and ha rma l i ne , but of the 6-methoxylated ana logues the f u l l y a romat i c compound i s more p o t e n t . The 7-subs t i t u t ed /3-carbol ines were g e n e r a l l y more potent i n h i b i t o r s than the co r r e spond ing 6 - s u b s t i t u t e d analogues in t h i s study and that of Buckho l tz et a l . [23] but Mclsaac & Es tevez [22] found 6-methoxy-harman was equ ipo ten t wi th harmine 170 TABLE XIII STRUCTURE/ACTIVITY RELATIONSHIPS OF /3-CARBOLINES AS MAO INHIBITORS I n h i b i to r I 50* I s o Mclsaac et a l . t I s o Buckhol et a l . * Harmine 1 .26 X 10" 8 1.5 X 1 o - 8 8 . 0 X 1 0 " 8 Harmal ine 1 .58 X 10" 8 1.0 X 1 o - 6 6 . 0 X 1 0 " 8 THH 1 . 77 X 10- 6 not t e s t e d 1 . 4 X 1 0 " 5 6MeO-harmalan 1 . 2 0 X 10" 6 4 .5 X 1 0 - 7 1.8 X 1 0 - 5 6MeO~harman 7 .08 X 10" 7 1 .5 X 1 0 " 8 3.1 X 1 0 " 5 Harmol 5 . 00 X 10- 7 2 . 7 X 1 o - 8 5.8 X 1 0 " 6 Harman 4 . 4 7 X 10" 7 5 . 0 X 1 0 ' 9 3 . 3 X 1 0 - 6 Norharman 3 .55 X 10" 6 7 .5 X 1 0 ' 1 0 2 . 0 X 1 0 " 5 6-MeO-MTH/3C 3 .98 X 10- 7 not t e s t e d not t e s t ed * I 5 0 = molar c o n c e n t r a t i o n of i n h i b i t o r equa l to 50% i n h i b i t i o n of a c t i v i t y w i th r espec t to c o n t r o l s (p resent expe r imen t ) . Va lues g i ven are means of at l e a s t three separa te d e t e r m i n a t i o n s , t R e s u l t s r epo r t ed by Mc lsaac et a l . [22] u s i ng c a l f - l i v e r m i tochond r i a & tyramine as s u b s t r a t e . $ R e s u l t s r epo r t ed by Buckho l t z et a l . [23] us ing mouse whole b r a i n homogenate and t r yp tamine as s u b s t r a t e . 171 TABLE XIV - MAOI ACTIVITY OF AYAHUASCA SAMPLES AND MIXTURES OF /3-CARBOLINES I n h i b i t o r 5 0 % i n h i b i t i o n at 1 0" 3 M Harmine + THH + Harmal ine-equ imolar mix #1 Harmine + THH + Harmal ine -equ imolar mix #2 Harmine + THH + DMT + Harmal ine -equimolar mix #3 ayahuasca " ana logue " #1* ayahuasca " ana logue " #2+ ayahuasca samples :$ Don F i d e l #1 Don Juan #2 3.16 x 10' 2.51 x 10 - 8 7.1 x 10- 8 3.98 x 1 0 ' 7 2.82 x 1 0 " B 1.58 x 10- 5 <1.0 x 1 0 ' 7 94 99.6 99.2 99.3 99.3 * M ix tu re of /3-carbolines approx imat ing p r o p o r t i o n s found in ayahuasca , v i z . , 69% harmine, 26% THH, and 4.6% ha rma l i ne . t M ix tu re of 0-ca rbo l i nes + DMT approx imat ing p r o p o r t i o n s found in ayahuasca , v i z . , 65% harmine, 6% ha rma l i ne , 22% THH, and 7% DMT. $ F i g u r e s g iven fo r ayahuasca r ep resen t d i l u t i o n f a c t o r w i th r e spec t to u n d i l u t e d samples . 1 72 wh i le 6-methoxy-harmalan was a more a c t i v e i n h i b i t o r than h a r m a l i n e . Hydroxy l s u b s t i t u t i o n on the aromat ic r i n g r e s u l t s in a l e s s a c t i v e compound than the co r r e spond ing methoxy-s u b s t i t u t e d compound ( c f . harmine and ha rmo l ) . In the present study harmane, l a c k i n g s u b s t i t u e n t s on the a romat i c r i n g , was l e s s a c t i v e than the 7-methoxylated analogue but approx imate l y equ ipo ten t to the 6-methoxylated f u l l y a romat ic compounds, r e s u l t s which agree wi th those of B u c k h o l t z , et a l . [ 23 ] . Mc lsaac et a l . , [22] r epo r t ed harmane to be somewhat more a c t i v e than e i t h e r harmine or 6-methoxy-harman. Lack of a methyl group at C i (cf harmine and norharmane) r e s u l t e d in reduced a c t i v i t y in these assays and in those of B u c k h o l t z , et a l . [23] but Mc lsaac and Es tevez [22] r epo r t ed somewhat g r e a t e r a c t i v i t y for norharmane than fo r harmane. I found 6-methoxy-2-methyl-tet rahydro-/3-carbol i n e , a compound r epo r t ed from V i r o l a s p p . , [16] to be in the same gene ra l range of potency as harman and ha rmo l . Thus 6-MeO-MTH(3C e x h i b i t e d the g r e a t e s t i n h i b i t o r y a c t i v i t y of any of the 6-methoxylated /3-carbol ines t e s t e d , and was approx imate l y an order of magnitude more a c t i v e than THH. The g r ea t e r a c t i v i t y of t h i s compound wi th r e spec t to THH and the o ther 6-methoxylated ana logues may be due p r i m a r i l y to the m e t h y l a t i o n of the p i p e r i d i n e n i t r o g e n . 6-MeO-MTH/3C was not i n v e s t i g a t e d by e i t h e r Mc lsaac & Es tevez [22] or B u c k h o l t z , et a l . [ 2 3 ] ; the former a u t h o r s , however, r epo r t ed that a c e t y l a t i o n of the p i p e r i d i n e n i t r o g e n r e s u l t e d in a compound l a c k i n g i n h i b i t o r y a c t i v i t y . T h i s s u b s t i t u t i o n would comp le t e l y a b o l i s h the b a s i c i t y of the p i p e r i d i n e n i t r o g e n , which i s p robab l y a s t r u c t u r a l requirement fo r MAOI a c t i v i t y . 173 2. Tryptamines A l though the s t r u c t u r e / a c t i v i t y r e l a t i o n s h i p s of the 0-c a r b o l i n e d e r i v a t i v e s as MAO i n h i b i t o r s has been s y s t e m a t i c a l l y i n v e s t i g a t e d [22,23] s i m i l a r i n v e s t i g a t i o n s of the MAOI a c t i v i t y of t r yp tamine d e r i v a t i v e s have not been conduc ted . Barlow [24] i n v e s t i g a t e d the e f f e c t s of DMT and s e v e r a l r e l a t e d methy la ted t r yp tamines as i n h i b i t o r s of amine ox idase in gu inea-p ig l i v e r homogenates and found a g r ea t e r degree of i n h i b i t i o n when tyramine and 5HT were used as s u b s t r a t e s than when t ryptamine was the s u b s t r a t e . In a more recent study [25] MAO i n h i b i t i o n by 5-subs t i t u t ed d i m e t h y l t r y p t a m i n e s , a-methy l t r yp tamines , and gramines was examined. These i n v e s t i g a t o r s c o u l d f i n d no c o r r e l a t i o n of i n h i b i t o r y a c t i v i t y w i th the s t e r i c and e l e c t r o n i c c h a r a c t e r i s t i c s of the 5-pos i t i on s u b s t i t u e n t s . The DMT d e r i v a t i v e s were more a c t i v e than the gramine d e r i v a t i v e s as MAOI and both were g e n e r a l l y l e s s a c t i v e than the a-methy l t r yp t am ines . Of the 4 DMT d e r i v a t i v e s t e s t e d , g r e a t e s t i n h i b i t o r y a c t i v i t y was shown by DMT, a 1 mM c o n c e n t r a t i o n g i v i n g 50% i n h i b i t i o n ; 5-MeO-DMT was 3.4 t imes l e s s a c t i v e on a molar b a s i s . The present study was undertaken to a s sess the i n f l u e n c e of v a r i o u s i n d o l e r i n g s u b s t i t u e n t s and m o d i f i c a t i o n s of the a l k y l s i d e - c h a i n on the a c t i o n of t r yp tamines as MAOI. Presumably the a c t i v i t y e x h i b i t e d by the v a r i o u s d e r i v a t i v e s i s a l s o a r e f l e c t i o n of t h e i r a c t i v i t y as c o m p e t i t i v e s u b s t r a t e s of MAO-A. Comparison of the I 5 0 v a lues f o r the t ryp tamine d e r i v a t i v e s (Table XV) w i th the I 5 0 v a lues of the g-ca rbo l i ne s (Table XI I I ) i n d i c a t e s that most of the t r yp tamines assayed are one to two TABLE XV - MAOI ACTIVITY OF TRYPTAMINE DERIVATIVES AND MYRISTICACEOUS PASTES Compound or sample 150* % I n h i b i t i o n at Assayed 1 x 10~ ' M Tryptamine d e r i v a t i v e s : 5-hydroxy-tryptophan 6 . 31 x 10" ' 56 Tryptam1ne 2 . 82 x 1 0 - 5 89 5-Me0-1ryptam i ne 3 . 98 x 1 0 - ' 89 5-MeO-N-acety1-tryptamine (meiaton i n) 3 . 98 x 1 0 - • 70 N-methy1 -tryptam i ne 2 . 24 x 10" s 90 N,N-d i methy 1tryptam i ne 1 . 58 x 10" ! 99 5-Me0-N,N-dimethyltryptami ne 4 . 47 x 1 0 - 5 98 4-hydroxy-N.N-dimethy 1 -tryptamine ( p s i l o c i n ) 2 . 82 x 10" ' 96 4-phosphoryl-N,N-dimethyl-t r y p t a m i n e ( p s i l o c y b i n ) > 1 .00 x 10" ' 35 5-Me0-N,N-d i i sopropy1 -tryp t a m i n e >1 .00 x 10" 1 40 3-{2(2.5d1methyl )-p y r r o 1 y 1 - e t h y 1 ] - i ndole) 2 . 51 x 1 0 - 5 91 T a b l e XV (cont'd) Compound or sample I s o * % I n h i b i t i o n at Assayed h i g h e s t c o n c e n t r a t i o n M y r i s t i c a c e o u s p a s t e s and p a s t e analogues: La C h o r r e r a "oo'-koey" 1 .51 X 10' 5 96 (8 . 43 X 10" ' M)t La C h o r r e r a analogue 1 .90 X 10" 5 94 (8. .43 X 10" ' M) A l f r e d o Moreno sample tt\ 8 .91 X 10" * 96 (7 .02 X 10" * M) A. Moreno analogue 6 . 30 X 10" 6 97 (7 .02 X 10" 1 M) Marcos F l o r e s sample #1 2 .95 X 10" 5 92 (9 . 30 X 10" ' M) Marcos F l o r e s analogue 2 . 34 X 10" 5 92 (9 . 30 X 10" * M) N o n - a l k a l o i d a l samples: Jor g e Churay sample #1 (DMK-34. V. P a v o n i s ) 1200 „g d wt/ml* 88 (15 x 10 1 „g d wt/ml) Crude l i g n a n f r a c t i o n (DMK-59. V. E l o n g a t a ) 20 „g d wt/ml 64 (50 „g d wt/ml) * Molar c o n c e n t r a t i o n at 50% i n h i b i t i o n of enzyme a c t i v i t y t F i g u r e s g i v e n are % i n h i b i t i o n at h i g h e s t molar c o n c e n t r a t i o n of t r y p t a m i n e s i n sample or analogues. t F i g u r e s g i v e n are c o n c e n t r a t i o n s e x p r e s s e d as ^g dry weight/ml. 176 o rde r s of magnitude l e s s e f f e c t i v e than the B-carbo l ines as i n h i b i t o r s of MAO. The major excep t i on i s N,N-d i m e t h y l t r y p t a m i n e ; w i th the lowest I 5 0 va lue of a l l t ryptamine d e r i v a t i v e s t e s t e d , t h i s compound has an I 5 0 comparable to t e t rahydroharmine and 6-MeO-harmalan (Table X I I I ) . A l l o ther t r yp tamine d e r i v a t i v e s are one to s e v e r a l o rde r s of magnitude l e s s e f f e c t i v e than DMT as MAOI, and t h i s r e d u c t i o n in a c t i v i t y may be r e l a t e d to the r e l a t i v e s i z e and p o l a r i t y of r i n g and s i d e - c h a i n s u b s t i t u e n t s and a l s o to the b a s i c i t y of the s i de-cha in N. Thus 5-MeO-DMT i s some one and one-ha l f o rde r s of magnitude l e s s a c t i v e than DMT as MAOI, and the co r r e spond ing 5-MeO-d i i sop ropy l analogue i s e s s e n t i a l l y w i thout s i g n i f i c a n t MAOI a c t i v i t y at the h i g h e s t c o n c e n t r a t i o n used . The i n f l u e n c e of bu lky r i n g s u b s t i t u e n t s i s we l l i l l u s t r a t e d by compar ison of p s i l o c i n and p s i l o c y b i n ; the 4-hydroxy s u b s t i t u t e d compound p s i l o c i n shows reduced but s t i l l s i g n i f i c a n t MAOI a c t i v i t y compared to DMT, but s u b s t i t u t i o n w i th the l a r g e phosphory l e s t e r ( p s i l o c y b i n ) e s s e n t i a l l y a b o l i s h e s the a c t i v i t y . The e f f e c t of v a r i o u s a l k y l s i d e - c h a i n s u b s t i t u e n t s appears to be more complex than s imply r e l a t i v e s i z e ; f o r i n s t ance 3-[2-(2,5-d i m e t h y l ) - p y r r o l y l e t h y l ] - i n d o l e e x h i b i t s approx imate l y the same i n h i b i t o r y a c t i v i t y as t r yp t am ine , even though i t has by f a r the l a r g e s t a l k y l - N s u b s t i t u e n t ; in t h i s case the l i p o p h i l i c nature of the N-subs t i tuen t may permit a c o n s i d e r a b l e degree of i n t e r a c t i o n wi th l i p o p h i l i c r e s i d u e s of the enzyme d e s p i t e i t s s i z e . A c e r t a i n degree of b a s i c i t y of the s i d e cha in n i t r o g e n appears to be a requi rement fo r MAOI a c t i v i t y , presumably because the p ro tona ted amine i n t e r a c t s w i th an a n i o n i c moiety at 177 the a c t i v e s i t e of the enzyme. S u b s t i t u t i o n s which a b o l i s h t h i s b a s i c c h a r a c t e r , e . g . the z w i t t e r i o n i c 5-hydroxy-t ryptophan, and the N-ace ty l a ted compound m e l a t o n i n , appear a l s o to a b o l i s h the MAOI a c t i v i t y . Presence of non-polar s u b s t i t u e n t s on the bas i c n i t r o g e n may enhance i n t e r a c t i o n s w i th a secondary l i p o p h i l i c s i t e ; t h i s would e x p l a i n why DMT i s a more a c t i v e MAOI than t r yp tamine or NMT. In the case of the d i i s o p r o p y l and h igher homologs, however, a c t i v i t y may be d i m i n i s h e d due to s t e r i c f a c t o r s even though the s u b s t i t u e n t s are non-po l a r . B. A c t i v i t y of ayahuasca and ayahuasca ana logues as MAOI Once the r e l a t i v e i n h i b i t o r y a c t i v i t y of a r e p r e s e n t a t i v e sample of 0 - c a r b o l i n e s had been eva lua t ed in our assay system, the next s t ep was to eva lua te the a c t i v i t y of a p p r o p r i a t e l y d i l u t e d ayahuasca samples . The a c t i v i t y e x h i b i t e d by the ayahuasca samples was compared to tha t shown by m ix tu res of harmine, ha rma l i ne , and t e t r ahyd roha rm ine , which a re known to be the major /3-carbol ines found in ayahuasca . Four types of s t andard mix tu res were compared. One was an equ imolar mixture of harmine , ha rma l i ne , and te t rahydroharmine wh i le the second c o n t a i n e d the 3 /3-carbol ines + DMT at an equ imolar c o n c e n t r a t i o n ; DMT was i n c l u d e d in the mixture i n o rder to determine whether i t i n t e r f e r e d wi th or enhanced the MAO i n h i b i t i o n by the /3-carbol ines . The t h i r d and f o u r t h mix tures c o n t a i n e d approx imate l y the same molar c o n c e n t r a t i o n of a l k a l o i d s as the equ imolar m i x t u r e s , but the p r o p o r t i o n s r e f l e c t e d the p r o p o r t i o n s found in the ayahuasca brews; DMT was omi t ted from analogue mix ture #3, but was i n c l u d e d in analogue 178 mixture #4 ( c f . M a t e r i a l s and Methods ) . Two of the ayahuasca samples from P u c a l l p a , Don Juan sample #2 and Don F i d e l sample # 1 , were ana l yzed us ing q u a n t i t a t i v e HPLC in o rder to determine the c o n c e n t r a t i o n of a l k a l o i d s in the u n d i l u t e d samples . T o t a l a l k a l o i d c o n c e n t r a t i o n was determined to be 4 . 8 mg/ml and 3 . 5 mg/ml, r e s p e c t i v e l y . The samples were then sub j ec t ed to s e r i a l 1 : 1 0 d i l u t i o n s so that the most d i l u t e s o l u t i o n was 1 x 1 0 " 7 as c o n c e n t r a t e d as the u n d i l u t e d sample. 5 0 M1 a l i q u o t s of the a p p r o p r i a t e d i l u t i o n s from each sample were assayed fo r MAO i n h i b i t o r y a c t i v i t y us ing the j_n v i t r o ra t-l i v e r c y t o s o l system ( F i g . 1 2 , Tab l e X IV ) . F i g . 1 2 shows c l e a r l y that both ayahuasca samples are ex t reme ly e f f e c t i v e MAO i n h i b i t o r s ; Don F i d e l ' s #1 s t i l l showed > 4 0 % i n h i b i t i o n of the enzyme at 1 0 " 5 f u l l s t r e n g t h , wh i l e Don J uan ' s #2 exceeded 5 0 % i n h i b i t i o n even at one t e n - m i l l i o n t h ( 1 0 ~ 7 ) the c o n c e n t r a t i o n of the u n d i l u t e d brew. These in v i t r o r e s u l t s i n d i c a t e that ayahuasca i s a c t i v e as an MAO i n h i b i t o r even when d i l u t e d by many o rde rs of magni tude. These o b s e r v a t i o n s c o n s t i t u t e the f i r s t e m p i r i c a l demonst ra t ion of the e f f e c t of ayahuasca on MAO and p rov i de ev idence fo r the h y p o t h e s i s that the h a l l u c i n o g e n i c p r o p e r t i e s of ayahuasca are due to i t s i n a c t i v a t i o n of v i s c e r a l MAO and consequent o r a l p o t e n t i a t i o n of the DMT in the p r e p a r a t i o n . It shou ld be emphas ized, however, tha t the p resen t in v i t r o s tudy r ep r e sen t s on ly the f i r s t s t ep toward unders tand ing the pharmacology of ayahuasca . Fu r the r i n v e s t i g a t i o n s are r e q u i r e d , p a r t i c u l a r l y i_n v i v o s t u d i e s of the a c t i o n of ayahuasca i n both an ima ls and humans, in order to f u l l y e l u c i d a t e the pharmacology of t h i s Amazonian d r u g . Another ! 1 1 1 1 1 1 1 0- 1< E - 3- 4. 5* £ • 7* NEGATIVE LOG DILUTION FACTOR F i g u r e 12 - MAOI A c t i v i t y of Ayahuasca + = Don Juan Sample #2 x = Don F i d e l Sample #1 180 p o i n t worth ment ion ing i s the f a c t that on occas ion ayahuasca i s p repared from B. caap i a lone wi thout the a d d i t i o n of any admixture p l a n t s ; in these i n s t a n c e s , DMT would be absent from the p r e p a r a t i o n . T h i s would a l t e r i t s pharmacology and presumably the h a l l u c i n o g e n i c e f f e c t s , i f p r e s e n t , would be due to the |3-carbol ines a l o n e . In t h i s case c o n c e n t r a t i o n s of 0-c a r b o l i n e s c o n s i d e r a b l y g r ea t e r than those measured in our samples would be r e q u i r e d i f a n o n s y n e r g i s t i c mechanism i s assumed. It i s i n f o rma t i v e to compare the % MAO i n h i b i t i o n produced by the m ix tu res of 0 - c a r b o l i n e s tandards wi th the i n h i b i t i o n e l i c i t e d by s i n g l e components of the mixture (Table XIII & XIV, F i g s . 13 & 14). In the f i r s t a s say , an equimolar mixture of harmine, ha rma l i ne , and te t rahydroharmine was assayed in p a r a l l e l w i th "ayahuasca analogue #1" - hav ing the same o v e r a l l c o n c e n t r a t i o n as the equ imolar mixture but w i th the i n d i v i d u a l components p resent in approx imate l y the p r o p o r t i o n s found in ayahuasca , v i z . : 69% harmine , 4.6% ha rma l i ne , and 26% t e t r ahyd roha rm ine . The equ imolar mixture i n h i b i t e d 50% of the enzyme a c t i v i t y at a c o n c e n t r a t i o n of 3.16 x 10~ 7 M. T h i s va lue i s i n t e rmed i a t e between the I 5 0 va lue of the most a c t i v e c o n s t i t u e n t of the mix ture (ha rma l ine , I 5 0 = 1.58 x 1 0 " 8 M) and the l e a s t a c t i v e ( t e t r ahydroha rmine , I 5 0 = 1.77 x 10~ 6 M) i n d i c a t i n g tha t these compounds act a d d i t i v e l y r a the r than s y n e r g i s t i c a l l y wi th r e spec t to t h e i r i n h i b i t i o n of MAO. A s y n e r g i s t i c i n t e r a c t i o n would r e s u l t in I 5 0 v a lues c o n s i d e r a b l y lower than the I 5 0 va lue of any one c o n s t i t u e n t by i t s e l f ; such a r e s u l t i s not obse r ved , i n d i c a t i n g tha t the i n h i b i t o r y 100. -ID* . _ 5-r E G A T I V E LOG ••rCENTRATIQN CM) F i g u r e 13 - MAOI A c t i v i t y of /3-carboline M ix tu r e s harmine + harma l ine + THH: Equimolar M ix tu re x = "Ayahuasca ana logue #1" - 69% harmine , 4.6% ha rma l i ne , 26% THH 182 a c t i v i t y of the three compounds c o l l e c t i v e l y i s not g r ea t e r than the a c t i v i t y of the most e f f e c t i v e compound of the g roup . The I 5 0 va lue of "analogue #1" was n e a r l y i d e n t i c a l w i th the equ imolar mixture (3.98 x 1 0 " 7 M and 3.16 x 1 0 ' 7 M, r e s p e c t i v e l y ) i n d i c a t i n g that the combina t ion of harmine and te t rahydroharmine a lone can account fo r most of the i n h i b i t o r y a c t i v i t y e x h i b i t e d by ayahuasca . A l though harmal ine i s e q u i v a l e n t to or s l i g h t l y s t ronge r than harmine, i t i s e s s e n t i a l l y a t r a ce component in ayahuasca and p robab ly does not c o n t r i b u t e s i g n i f i c a n t l y to the monoamine ox idase i n h i b i t i o n which t h i s drug e l i c i t s . In the second assay ( F i g . 14) an equimolar mix ture of harmine , ha rma l i ne , and te t rahydroharmine was assayed in p a r a l l e l w i th an equimolar mixture of the th ree /3-carbol ines p l u s DMT. T h i s assay was c a r r i e d out in order to determine whether the presence of s i g n i f i c a n t c o n c e n t r a t i o n s of DMT would a f f e c t the a c t i v i t y of the g - c a r b o l i n e s . The I 5 0 v a lues measured (Table XIV) were f a i r l y c l o s e , however i t appears that in t h i s in v i t r o sys tem, the presence of DMT in the equ imolar mix ture may s l i g h t l y m i t i g a t e the e f f e c t i v e n e s s of the /3-carbol ines as MAOI. It must be remembered, however, that in the equ imolar mix ture c o n t a i n i n g DMT the t o t a l c o n c e n t r a t i o n of /3-carbol ines i s l e s s than in the mix ture l a c k i n g DMT; t h e r e f o r e i t i s r easonab le to expect the mixture wi th DMT to have a s l i g h t l y h ighe r I 5 0 v a l u e . "Analogue #2" - c o n t a i n i n g 65% harmine, 6% h a r m a l i n e , 22% t e t r ahyd roha rm ine , and 7% DMT - was a l s o assayed and found to have an I 5 0 va lue of 2.82 x 1 0 " 8 M, q u i t e c l o s e t o the va lue measured wi th the equ imolar mixture c o n t a i n i n g 3-2. 3. A. S> B* 7- B- 9- IO- 11> NEGATIVE LDD CONCENTRATION (M) F i g u r e 14 - MAOI A c t i v i t y of 0-carbol ine/DMT M ix tu res + = harmine + harmal ine + THH: Equ imolar M ix tu re x = harmine+ harmal ine + THH + DMT: Equ imolar M ix tu re = "Ayahuasca analogue #2" - 65% harmine , 6% ha rma l i ne , 22% THH, 7% DMT 184 c a r b o l i n e s a l o n e . A l l of the I 5 0 v a l ues measured in t h i s second group of assays were somewhat lower than those measured in the f i r s t g roup ; the d i s c r epancy i s p robab l y r e l a t e d to ba tch to ba tch v a r i a b i l i t y in the r a t - l i v e r enzyme p r e p a r a t i o n s . C. A c t i v i t y of M y r i s t i c a c e o u s pas tes and paste " ana logues " as MAOI The th ree M y r i s t i c a c e o u s pas te samples which q u a n t i t a t i v e GC a n a l y s i s had p r e v i o u s l y shown to have the h ighes t c o n c e n t r a t i o n s of t r yp tamines (Table XI) were assayed fo r MAOI a c t i v i t y in the _i_n v i t r o r a t - l i v e r enzyme system. The c o n c e n t r a t i o n s and p r o p o r t i o n s of t r yp tamines in the pas te samples were determined by HPLC a f t e r the samples had been prepared fo r use in the a ssay . Paste " a n a l o g u e s " , c o n s i s t i n g of m ix tu res of t ryptamine s tandards in the same c o n c e n t r a t i o n s and p r o p o r t i o n s as in the pas tes themselves were run s imu l t aneous l y w i th each pas te sample ( c f . M a t e r i a l s & Methods ) . The r e s u l t s of these assays (Table XV, F i g . 15-17,) show that the MAOI a c t i v i t y e x h i b i t e d by the o r a l l y - i n g e s t e d pas tes i s p a r a l l e l e d a lmost e x a c t l y by the a c t i v i t y of the pas te ana logues , c o n t a i n i n g on ly t r y p t a m i n e s . In a l l c a s e s , the I 5 0 of the pas te sample i s q u i t e c l o s e to tha t of the analogue mix ture i n d i c a t i n g tha t the MAOI a c t i v i t y of the pas tes i s due s o l e l y to the presence of the t r y p t a m i n e s ; i t seems u n l i k e l y , t h e r e f o r e , that the presence of t r a c e amounts of /3-carbol ines in a d d i t i o n to the t r yp tamines (as i n , e . g . , the La Chor re ra sample) has any s i g n i f i c a n c e in terms of enhanc ing the MAOI a c t i o n of the pas tes or of o r a l l y p o t e n t i a t i n g the t r yp tamines p r e s e n t . The r e l a t i v e MAOI a c t i v i t y 185 1 0 Q . £ • 3- 4 . 5- E* 7-NEGATIVE LOG CONCENTRATION (M) F i g u r e 15 - MAOI A c t i v i t y of M y r i s t i c a c e o u s P a s t e s : La Cho r r e r a Oo '-koey + = La Cho r r e r a Oo '-koey (no voucher ) x = La Cho r r e r a " ana logue " ( c f . T a b l e s XII & XV) 186 of the pas tes appears to be a f u n c t i o n of the types and amount of t r yp tamines p r e s e n t , e . g . , the La Chor re ra sample had an I 5 0 on l y s l i g h t l y lower than the Don Marcos sample ; the former sample con t a i ned NMT + DMT, both of which are more a c t i v e MAOI than 5-MeO-DMT which i s the s i n g l e major t ryptamine in the Don Marcos sample. A l f r e d o Moreno 's sample #1 had the lowest I 5 0 va lue of the t h r e e , which i s c o n s i s t e n t wi th e x p e c t a t i o n s s ince t h i s sample con t a i ned p r o p o r t i o n a t e l y the h ighes t c o n c e n t r a t i o n of DMT, the most e f f e c t i v e MAOI of a l l the t ryp tamine d e r i v a t i v e s t e s t e d . A l though ijn v i v o exper iments would be r e q u i r e d to c o n f i r m these r e s u l t s , the j_n v i t r o ev idence i n d i c a t e s tha t these pas tes do not e x h i b i t MAOI a c t i v i t y beyond what i s due to the t ryptamine bases a l o n e . The re fo r e i f these pas t e s are o r a l l y e f f e c t i v e as h a l l u c i n o g e n s , i t appears that some mechanism other than MAO i n h i b i t i o n must be sought to e x p l a i n t h e i r o r a l a c t i v i t y . The MAOI a c t i v i t y of the pas te samples may a l s o be p a r t i a l l y due to the presence of non-n i t rogenous MAOI in a d d i t i o n to the t r yp tamines and t r a c e s o f /3-carbol ines . In order to i n v e s t i g a t e t h i s p o s s i b i l i t y , a pas te sample ( de r i v ed from DMK-34, V. pavon is ) which was a l k a l o i d - f r e e ( c f . Tab le X, Chapter VI) was assayed in the system; a crude l i g n a n f r a c t i o n o b t a i n e d from V. e l onga ta (DMK-59) was a l s o a s sayed . Some degree of i n h i b i t i o n was observed wi th both samples ( F i g . 18) but was s i g n i f i c a n t on ly at the h i ghes t c o n c e n t r a t i o n s . For the pas te sample , t h i s was 15000 ^g dry wt/ml of crude e x t r a c t , and fo r the l i g n a n sample, the h i ghes t c o n c e n t r a t i o n was 50 /ig dry wt/ml of the crude l i g n a n f r a c t i o n . By c o n t r a s t , the t r yp tamine-1 8 7 10Q* m D L J CK bJ Q_ 9 Q f BQ-7Q; E Q r 5Q: 4 Q r 3Q; S Q r I Q r NEGATIVE LOG CONCENTRATION (M) F i g u r e 16 - MAOI A c t i v i t y of M y r i s t i c a c e o u s P a s t e s : A l f r e d o Moreno Oo'-koey Sample #1 x = A l f r e d o Moreno Oo '-koey Sample #1 (DMK-40, V. s e b i f e r a ) + = A l f r e d o Moreno Sample #1 " ana logue " ( c f . T ab l e s XII & XV) 188 100_i 5 Q . • 1 1 1 1 1 S' 3- A' 5* E> 7. B-NEGATIVE LOG CONCENTRATION (M) F i g u r e 17 - MAOI A c t i v i t y of M y r i s t i c a c e o u s P a s t e s : Marcos F l o r e s Ku '- ru-ku Sample #1 x = Marcos F l o r e s Ku '- ru-ku Sample#l (DMK-59, V. e l onga ta ) + = Marcos F l o r e s Sample #1 " ana logue " ( c f . T ab l e s XII & XV) F i g u r e 18 - MAOI A c t i v i t y of N o n - a l k a l o i d C o n s t i t u e n t s of M y r i s t i c a c e o u s Pas tes n o n - a l k a l o i d a l pas te sample (DMK-34, V. pavon i s ) x = l i g n a n f r a c t i o n (DMK-59, V. e l onga ta ) 190 c o n t a i n i n g pas te samples showed comparable amounts of i n h i b i t i o n at dry wt c o n c e n t r a t i o n s on the order of 3 to 4 ug dry wt/ml of crude e x t r a c t . The i n h i b i t i o n shown by the a l k a l o i d - f r e e samples t h e r e f o r e appears to be f a i r l y n o n s p e c i f i c and p robab l y i s due to a gene ra l dena tu r i ng of the p r o t e i n s in the enzyme p r e p a r a t i o n due to the h igh p h e n o l i c content of the samples . The p o s s i b i l i t y that the M y r i s t i c a c e o u s pas tes may c o n t a i n some h i g h l y s p e c i f i c non-n i t rogenous MAO i n h i b i t o r seems somewhat remote. IV. Summary The a c t i v i t y of a number of t r yp tamine and j3-ca rbo l i ne d e r i v a t i v e s as MAOI was i n v e s t i g a t e d us ing an iji v i t r o enzyme assay and 5-hyd roxy- 1 "C- t r yp t am ine as s u b s t r a t e . A c t i v i t y was measured us ing s i n g l e compounds and mix tu res of compounds and the r e s u l t s were compared to the a c t i v i t y of samples of ayahuasca and samples of o r a l l y - i n g e s t e d M y r i s t i c a c e o u s p a s t e s . The MAOI a c t i v i t y of 0 - c a r b o l i n e d e r i v a t i v e s was found to be s e v e r a l o rde rs of magnitude g r e a t e r than the a c t i v i t y measured w i th t ryptamine d e r i v a t i v e s . M ix tu r e s of j3-ca rbo l i nes were not s i g n i f i c a n t l y more e f f e c t i v e as MAOI than the s i n g l e most a c t i v e compound in the m i x t u r e , i n d i c a t i n g an a d d i t i v e r a t he r than a s y n e r g i s t i c mechanism of a c t i o n . Some s t r u c t u r a l c o r r e l a t i o n s f o r MAOI a c t i v i t y were found f o r both the t r yp tamines and 0 -c a r b o l i n e s . Samples of ayahuasca were found to be h i g h l y a c t i v e as MAOI even when d i l u t e d by s e v e r a l o rde r s of magni tude, and the a c t i v i t y observed was s i m i l a r to tha t measured fo r comparable c o n c e n t r a t i o n s and p r o p o r t i o n s of 0 - c a r b o l i n e 191 m i x t u r e s . Based on t h i s ev idence i t appears l i k e l y that ayahuasca c o u l d f u n c t i o n e f f e c t i v e l y as MAOI i_n v i v o and thus o r a l l y p o t e n t i a t e the DMT which i s p robab l y r e s p o n s i b l e fo r the h a l l u c i n o g e n i c a c t i o n of the d r u g . A l e s s e r degree of i n h i b i t i o n was measured when samples of o r a l l y - i n g e s t e d M y r i s t i c a c e o u s pas t e s were a s sayed . The i n h i b i t i o n e l i c i t e d by the paste samples were c l o s e l y matched by m ix tu res of t r yp tamine s tandards hav ing comparable p r o p o r t i o n s and c o n c e n t r a t i o n s . These o b s e r v a t i o n s i n d i c a t e tha t the MAOI a c t i v i t y of the pas tes i s due main ly to the h igh c o n c e n t r a t i o n s of t r y p t a m i n e s ; the t r a c e s of /3-carbol ines or non-n i t rogenous i n h i b i t o r s p resen t p robab ly do not c o n t r i b u t e s i g n i f i c a n t l y to the t o t a l i n h i b i t i o n . Thus i t appears u n l i k e l y tha t the o r a l a c t i v i t y of the M y r i s t i c a c e o u s pas t e s i s due to the p o t e n t i a t i o n of the t r yp tamines v i a i n h i b i t i o n of MAO by /3-carbol ines ; some mechanism other than MAO i n h i b i t i o n must be invoked to account fo r the o r a l h a l l u c i n o g e n i c a c t i v i t y of the M y r i s t i c a c e o u s pas tes i f they a r e , in f a c t , o r a l l y a c t i v e . 192 V. L i t e r a t u r e C i t e d 1. Yasunobu, K. T . , H. I s h i z a k i , & N. Minamiura (1976) The M o l e c u l a r , M e c h a n i s t i c and Immunological P r o p e r t i e s of Amine O x i d a s e s . Mo l e cu l a r and C e l l u l a r B i o chemis t r y 13:3-29 2. Ma r l e y , E . , & B. B l a c k w e l l (1970) I n t e r a c t i o n s of Monoamine Oxidase I n h i b i t o r s , Amines, & F o o d s t u f f s . Advances in  Pharmacology & Chemotherapy 8:185-239 3. I v e r s o n , L. L . , (1979) The Chemis t ry of the B r a i n . Sc ient i f i c  amer ican Sept . 1979:134-149 4. F u l l e r , R. W., B. J . Warren, & B. B. Mo l loy (1970) S e l e c t i v e I n h i b i t i o n of Monoamine Oxidase in Rat Bra iM m i t o c h o n d r i a . B i o chemica l Pharmacology 19:2934-2936 5. D o n n e l l y , C. H . , E. R i c h e l s o n & D. L. Murphy (1976) P r o p e r t i e s of Monoamine Oxidase in Mouse Neuroblastoma NIE-115 C e l l s . B i ochemica l Pharmacology 25:1629-1643 6. D o n n e l l y , C. H . , & D. L. Murphy (1977) Subs t r a t e- and I n h i b i t o r - r e l a t e d C h a r a c t e r i s t i c s of Human P l a t e l e t Monoamine Ox idase . B iochemica l Pharmacology 26:853-858 7. Hous l a y , M. D. & K. F. T i p t o n (1974) A K i n e t i c E v a l u a t i o n of Monoamine Oxidase A c t i v i t y in Rat L i v e r M i t o c h o n d r i a l Outer Membranes. B i o chemica l J ou rna l 139:645-652 8. N e f f , N. H. And H.-Y. T . Yang (1974) Another Look at the Monoamine Ox idases and the Monoamine Oxidase I n h i b i t o r Drugs . L i f e S c i ences 14:2061-2074 9. P l e t s c h e r , A. (1966) Monoamine Oxidase I n h i b i t o r s . Pha rmaco log i ca l Reviews 18:121-129 10. A l l e n , J . R. F . , & B. Holmstedt (1980) The S imple /3-c a r b o l i n e A l k a l o i d s . Phy tochemis t ry 19:1573-1582 11. R. G. Taborsky & W. M. M c l s a a c . The S yn thes i s and P r e l i m i n a r y Pharmacology of Some 9H-Py r i do [3 ,4-b ] i ndo l e s (/3-carbolines) Re l a t ed to S e ro ton in and M e l a t o n i n . J ou rna l of M e d i c i n a l Chemis t ry 7:135-141 1964 193 12. U d e n f r i e n d , S. , B. Wi tkop, B. G. R e d f i e l d , & H. Weissbach (1958) S tud i e s With R e v e r s i b l e I n h i b i t o r s of Monoamine O x i d a s e : Harmal ine and Re l a t ed Compounds. B i ochemica l  Pharmacology 1:160-165 13. S c h u l t e s , R. E. (1972) E t h n o t o x i c o l o g i c a l S i g n i f i c a n c e of A d d i t i v e s to New World H a l l u c i n o g e n s . P l an t Sc ience B u l l e t i n 18: 34-41 14. S h u l g i n , A. T . (1976) Psychotomimet ic Agen t s . Ch . 4 in Maxwell Gordon (ed. ) P sychopharmaco log i ca l Agents V o l . IV. Academic P r e s s . 15. Der M a r d e r o s i a n , A. H . , H. V. P i n k l e y , & M. F. Dobbins IV. (1968) Na t i ve Use and Occur rence of N,N-dimethy I t ryptamine in the Leaves of B a n i s t e r i o p s i s rusbyana . American J ou rna l of Pharmacy 140:137-147 16. A g u r e l l , S . , B. Ho lmstedt , J . E. L i n d g r e n , & R. E. S c h u l t e s . (1968) I d e n t i f i c a t i o n of Two New /3-carboline A l k a l o i d s in South American H a l l u c i n o g e n i c P l a n t s . B i o chemica l  Pharmacology 17:2487-88 17. S c h u l t e s , R. E. (1969) V i r o l a as an O r a l l y - a d m i n i s t e r e d H a l l u c i n o g e n . Harvard B o t a n i c a l Museum L e a f l e t s 22:229-40 18. S c h u l t e s , R. E. & T . Swain (1976) Fu r the r Notes on V i r o l a as an O r a l H a l l u c i n o g e n . J ou rna l of P s y c h e d e l i c Drugs 8:317-324 19. S c h u l t e s , R. E . , T . Swain, & T . Plowman (1977) V i r o l a as an O r a l H a l l u c i n o g e n Among the Boras of Pe ru . Harvard B o t a n i c a l Museum L e a f l e t s 25:259-272 20. Dawson, R. M. C , D. C. E l l i o t , W. H. E l l i o t , K. M. Jones (eds . ) (1969) Data fo r B i o chemica l Resea rch , 2nd ed . Oxford U n i v e r s i t y P r e s s , London; p. 484 21. T i p t o n , K. F. & M. B. H. Youdim (1976) Assay of Monoamine O x i d a s e . In Monoamine Oxidase And I t s I n h i b i t i o n . C iba Foundat ion Symposium 39 ( N . S . ) . E l s e v i e r , Amsterdam. 1 94 22. M c l s a a c , W. M. & V. Es tevez (1966) S t r u c t u r e - a c t i o n R e l a t i o n s h i p s of 0 - c a r b o l i n e s as Mono-amine Oxidase I n h i b i t o r s . B i o chemica l Pharmacology 26:1625-27 23. B u c k h o l t z , N. S. & W. 0 . Boggan (1977) Monoamine Oxidase I n h i b i t i o n in B r a in and L i v e r Produced by 0 - c a r b o l i n e s : S t r u c t u r e - a c t i v i t y R e l a t i o n s h i p s and Subs t r a t e S p e c i f i c i t y . B i o chemica l Pharmacology 26:1991-96 24. R. B. Bar low. (1961) E f f e c t s on Amine Oxidase of Substances Which Antagon ize 5-hydroxytryptamine More than Tryptamine on the Rat Fundus S t r i p . B r i t i s h J ou rna l of Pharmacology 16:153-162. 25. B. T . Ho, W. M. M c l s a a c , R. An, R. T . H a r r i s , K. E. Walker , & P. M. K r a l i k (1970) B i o l o g i c a l A c t i v i t i e s of Some 5-S u b s t i t u t e d N ,N-d imethy l t r yp tamines , a- me thy l t r yp t am ines , and Gramines . Psychopharmaco log ia 16:385-394 26. D. MacRae. (1984) E t h n o b i o l o g i c a l and Chemica l I n v e s t i g a t i o n s of S e l e c t e d Amazonian P l a n t s . PhD t h e s i s , U n i v e r s i t y of B r i t i s h Co lumbia , Vancouver , B. C. 195 CHAPTER V I I : COMPARATIVE ETHNOPHARMACOLOGY OF MALPIGHIACEOUS AND MYRISTICACEOUS HALLUCINOGENS: SUMMARY AND CONCLUSION I. I n t r o d u c t i o n T h i s t h e s i s p r e sen t s the r e s u l t s of e t h n o g r a p h i c , e t h n o b o t a n i c a l , p h y t o c h e m i c a l , and pha rmaco log i c a l i n v e s t i g a t i o n s of two Amazonian h a l l u c i n o g e n s . The h a l l u c i n o g e n i c d r i nk ayahuasca i s p repared from the l i a n a B a n i s t e r i o p s i s caap i (Ma lp igh iaceae ) and sundry admixture p l a n t s , no tab l y D i p l o p t e r y s cabrerana and v a r i o u s Psychot r i a spp . The cambia l r e s i n of c e r t a i n members of the genus V i r o l a ( M y r i s t i c a c e a e ) i s the source of h a l l u c i n o g e n i c s n u f f s and o r a l l y - i n g e s t e d h a l l u c i n o g e n i c pas tes used by some t r i b e s . A l though d e r i v e d from e n t i r e l y d i f f e r e n t b o t a n i c a l s o u r c e s , s i m i l a r a l k a l o i d s - - t r y p t a m i n e s and /3-carbo l ines—are the a c t i v e c o n s t i t u e n t s of both p r e p a r a t i o n s . In the case of ayahuasca and the o r a l l y - i n g e s t e d V i r o l a p a s t e s , i t has been suggested that monoamine ox idase i n h i b i t i o n - - d u e to the /3-carbo l ines--protec ts the psychotomimet ic t r yp tamines from o x i d a t i v e deaminat ion by p e r i p h e r a l MAO and thus pe rmi t s t h e i r o r a l a c t i v i t y . Expe r imen ta l i n v e s t i g a t i o n s of t h i s p o s t u l a t e d mechanism of o r a l a c t i v i t y — i n v o l v i n g Jjn v i t r o e v a l u a t i o n s of the drugs and t h e i r c o n s t i t u e n t s as MAOI--were one of the pr imary o b j e c t i v e s of t h i s t h e s i s . Other o b j e c t i v e s were the c o l l e c t i o n of e t h n o b o t a n i c a l and e thnograph i c data on the use of these drugs among Ind ian and mes t i zo p o p u l a t i o n s , the c o l l e c t i o n of voucher specimens and m a t e r i a l f o r phy tochemica l a n a l y s i s , and the a n a l y s i s of a l k a l o i d a l c o n s t i t u e n t s in s o u r c e - p l a n t s , admixture p l a n t s , and drug samples . Most of these o b j e c t i v e s have been met, and i t i s 1 96 now a p p r o p r i a t e to summarize and compare the r e s u l t s ob ta ined fo r ayahuasca wi th those found fo r the M y r i s t i c a c e o u s h a l l u c inogens . 11• Ayahuasca The contemporary use of ayahuasca in South America occurs p r i m a r i l y w i t h i n the con tex t of mes t i zo f o l k m e d i c i n e , which i s compr ised of an amalgam of many t r i b a l t r a d i t i o n s . A l though most of these t r i b e s have long s i n ce fragmented or d i s a p p e a r e d , much of t h e i r e thnomedica l l o r e has s u r v i v e d and been adopted among m e s t i z o s . Ayahuasca i s among these , and i t o c cup i e s a c e n t r a l and important p o s i t i o n in mes t i zo f o l k med i c i ne . Not on l y i s i t employed as a genera l c u r e - a l l f o r the t reatment of many d i s o r d e r s r ang ing from mental i l l n e s s to p a r a s i t e s , i t i s a l s o the ayahuasquero ' s or h e a l e r ' s passpor t to s u p e r n a t u r a l d imens ions where the s k i l l s i n t r i n s i c to h i s p r o f e s s i o n can be a c q u i r e d . I t enab les the hea l e r to l e a r n the m e d i c i n a l uses of p l a n t s and the songs and chants used in the h e a l i n g ce remon ies ; w i th i t he i s ab l e to d iagnose d i s e a s e s , d i v i n e the s u p e r n a t u r a l causes of i l l n e s s , p r e d i c t the f u t u r e and see and communicate a c r o s s d i s t a n c e s . Whether or not the re i s a r a t i o n a l b a s i s f o r any of these p r a c t i c e s , an impress i ve pharmacopoeia of p l a n t s -many of which do con t a i n h i g h l y biodynamic c o n s t i t u e n t s — a r e (or can be) used in c o n j u n c t i o n w i th ayahuasca . The phy tochemica l i n v e s t i g a t i o n s r epor ted here have found that most mes t i zo ayahuasca brews c o n t a i n s u b s t a n t i a l amounts of /3-carbol ines and N ,N-d imethy l t r yp tamine . The major /3-carbol ines are harmine and t e t r ahyd roha rm ine , and l e s s e r amounts of ha rma l i ne ; on ly t r a c e s 197 of o ther /3-carbolines were d e t e c t e d . The amounts of /3-carbolines in these samples were s e v e r a l o rde r s of magnitude g rea t e r than those r epo r t ed in an e a r l i e r study of ayahuasca prepared by t r i b e s i n h a b i t i n g the upper R io Purfis in southwestern Peru [1 ] . A " t y p i c a l " 100 ml dose of mes t i zo ayahuasca c o n t a i n s between 500 and 800 mg /3-carbolines and 40-80 mg DMT; t h i s i s we l l above the t h r e s h o l d dose fo r DMT and w i t h i n the range at which the /3-c a r b o l i n e s are e f f e c t i v e MAOI; i t i s s t i l l we l l below the t h r e s h o l d l e v e l fo r h a l l u c i n o g e n i c a c t i v i t y of the /3-carbol ines , however. The r e fo r e these data i n d i c a t e that the h a l l u c i n o g e n i c a c t i v i t y of ayahuasca i s p robab l y due to DMT, which i s o r a l l y a c t i v a t e d by some mechanism, presumably the MAO i n h i b i t i o n induced by the h igh c o n c e n t r a t i o n of /3-carbol ines . T h i s h ypo thes i s i s supported by the f i n d i n g that ayahuasca i s a very e f f e c t i v e i n h i b i t o r of MAO jjn v i t r o even when d i l u t e d by many o rde r s of magnitude. A number of ayahuasca samples , brewed by d i f f e r e n t p r a c t i t i o n e r s in d i f f e r e n t p a r t s of Pe ru , were a n a l y z e d . The same a l k a l o i d s were c o n s i s t e n t l y found ; the samples d i f f e r e d main ly in the c o n c e n t r a t i o n and p r o p o r t i o n s of v a r i o u s components. C o n c e n t r a t i o n d i f f e r e n c e s are expected s i n c e t h i s i s dependent on the amount of p l a n t m a t e r i a l e x t r a c t e d and o ther v a r i a b l e s in the p r e p a r a t i o n p rocedu re . P r o p o r t i o n a l d i f f e r e n c e s may be a t t r i b u t a b l e to the use of d i f f e r e n t B. caap i c u l t i v a r s . It i s remarkable t h a t , f aced wi th so many v a r i a b l e s , ayahuasqueros a l l a c ross Peru manage to manufacture a drug hav ing a h igh degree of pha rmaco log i c a l c o n s i s t e n c y from batch to b a t c h ; except fo r c o n c e n t r a t i o n and p r o p o r t i o n a l d i f f e r e n c e s a l l of the ayahuasca samples ana l yzed in t h i s study c o u l d have 198 been taken from the same p o t . A l l of the DMT-conta in ing admixture p l a n t s which were ana l yzed were s i m i l a r ; DMT was the s i n g l e major base in a l l of them; on ly t r a c e s of o ther a l k a l o i d s were d e t e c t e d . The c o n c e n t r a t i o n of DMT was between 1-2 mg/g dry wt. in a l l samples . S i m i l a r c o n s i s t e n c y was not found in the s e v e r a l B. caap i c u l t i v a r s which were a n a l y z e d . More or l e s s the same c o n s t i t u e n t s were present but c o n c e n t r a t i o n s ranged from 1.7 mg/g d wt ( t o t a l a l k a l o i d s ) to 13.6 mg/g dry wt. These d i f f e r e n c e s probab ly are due to env i ronmenta l f a c t o r s ra the r than to g e n e t i c a l l y based b i o c h e m i c a l d i f f e r e n c e s between c u l t i v a r s . I I I . M y r i s t i c a c e o u s H a l l u c i n o g e n s U n l i k e ayahuasca , the use of h a l l u c i n o g e n i c p r e p a r a t i o n s d e r i v e d from V i r o l a spp . or o ther M y r i s t i c a c e o u s genera i s a p r a c t i c e c o n f i n e d to a few ind igenous Amazonian t r i b e s and has never become i n t e g r a t e d i n t o mes t i zo f o l k med i c i ne . As a r e s u l t the use of these M y r i s t i c a c e o u s drugs has d i m i n i s h e d or in some cases d i s appea red as the t r i b a l s o c i e t i e s have fragmented due to o u t s i d e i n f l u e n c e s . T h i s i s p a r t i c u l a r l y t rue of the o r a l l y -i n g e s t e d M y r i s t i c a c e o u s p a s t e s . Use of V i r o l a spp . as an o r a l h a l l u c i n o g e n i s more e t h n o l o g i c a l l y r e s t r i c t e d than i t s use as a s n u f f ; i t has been r e p o r t e d among the Bora , W i t o t o , Muinane, and p o s s i b l y the Maku and a p p a r e n t l y does not occur o u t s i d e these g roups . Even w i th in these t r i b e s , the s o u r c e - p l a n t s , methods of p r e p a r a t i o n , and modes of usage of the o r a l V i r o l a drug are the s p e c i a l i z e d knowledge of the medic ine men and are not known to most members of the t r i b e . Another c o m p l i c a t i n g f a c t o r i s that 199 the Bora and Wi toto p o p u l a t i o n s i n h a b i t i n g the R io Ampiyacu r e g i o n , where the f i e l dwo rk fo r t h i s study was c a r r i e d o u t , are not i nd igenous to that a rea but migra ted the re from no r th of the R io Putumayo in the e a r l y decades of t h i s c e n t u r y , as a r e s u l t of the d i s l o c a t i o n s produced by the rubber i n d u s t r y . T h i s has not c o n t r i b u t e d to the p r e s e r v a t i o n of t h e i r e thnomedica l t r a d i t i o n s nor any other t r i b a l i n s t i t u t i o n s ; the r e s u l t i s that l o c a l knowledge of the o r a l M y r i s t i c a c e o u s drugs has become, in some c a s e s , r a the r i n e x a c t . The work r epo r t ed here has shown tha t there i s a h igh degree of v a r i a b i l i t y in the type and amount of a l k a l o i d c o n s t i t u e n t s , not on ly among d i f f e r e n t V i r o l a s p e c i e s , but even among d i f f e r e n t c o l l e c t i o n s of the same s p e c i e s . The compos i t i on of the o r a l l y - i n g e s t e d pas te samples which were ana l yzed in t h i s study i s s i m i l a r l y v a r i a b l e , both q u a l i t a t i v e l y and q u a n t i t a t i v e l y , and presumably t h i s i s a r e f l e c t i o n of the chemica l v a r i a b i l i t y of the s o u r c e - p l a n t s . Thus , wh i le a l l of the ayahuasca samples had e s s e n t i a l l y the same c o n s t i t u e n t s (with DMT be ing present or absent depending on which P s y c h o t r i a sp . was used as adm ix tu r e ) , not one of the o r a l V i r o l a pas t e s had the same base compos i t i on as any o ther one. Large d i f f e r e n c e s in the c o n c e n t r a t i o n of a l k a l o i d s in v a r i o u s samples were a l s o found . E i t h e r t h i s h igh degree of chemica l v a r i a b i l i t y has always been a f e a tu r e of these o r a l l y - i n g e s t e d p a s t e s , or the c r i t e r i a which were fo rmer l y used to s e l e c t the " b e s t " ( i . e . , s t ronges t ) V i r o l a s p e c i e s to p repare the pas te ( e . g . , t a s t e , s m e l l , and/or v i s u a l appearance of the r e s i n ) have been l a r g e l y f o rgo t t en and the s e l e c t i o n has now become a f a i r l y haphazard p r o c e s s . If the former p o s s i b i l i t y i s t r u e , t h i s may 200 e x p l a i n why the paste was r e s t r i c t e d to the medic ine men and why they u s u a l l y consumed i t a l o n e : i f on l y one out of every three or four samples a c t u a l l y i_s o r a l l y a c t i v e , t h i s i n f o rma t i on was p robab l y kept from the people at l a r g e , in order to p rese r ve t h e i r f a i t h in the e f f i c a c y of the medic ine man and h i s medic i n e s . Another o b j e c t i v e of these i n v e s t i g a t i o n s was to examine the mechanism fo r the (presumed) o r a l a c t i v i t y of these M y r i s t i c a c e o u s p a s t e s ; i s i t due, l i k e ayahuasca , to the o r a l a c t i v a t i o n of the t ryp tamine c o n s t i t u e n t s through the i n h i b i t i o n of v i s c e r a l MAO by 0 - c a r b o l i n e s ? In the p rocess of answer ing some of these q u e s t i o n s t h i s i n v e s t i g a t i o n has c r e a t e d new and even more p u z z l i n g ones . In the f i r s t i n s t ance 0 - c a r b o l i n e s were not c o n s i s t e n t l y found as c o n s t i t u e n t s of a l l of the pas te samples ; even in the two samples c o n t a i n i n g /3-carbo l ines , on ly t r a c e s were d e t e c t e d . Those /3-carbol ines which were found belong to the te t rahydro -/3-carbo l ine t ype , which are poor MAO i n h i b i t o r s compared to the d ihyd ro- and f u l l y a romat ic 0 -c a r b o l i n e s . A f u r t h e r p o i n t i s that presence or absence of 0 -c a r b o l i n e s in the pas te samples appa r en t l y had l i t t l e or no th ing to do wi th t h e i r o r a l a c t i v i t y or i n a c t i v i t y as determined in s e l f - e x p e r i m e n t s ; the La Chor re ra oo ' -koey , which con t a i ned the h i ghes t l e v e l s of 0 - c a r b o l i n e s , was comp le te l y i n a c t i v e in repeated s e l f - e x p e r i m e n t s whi le Marcos F l o r e s ' k u ' - r u - k u , which c o n t a i n e d on ly h igh c o n c e n t r a t i o n s of 5-MeO-DMT in the base f r a c t i o n , was h i g h l y o r a l l y a c t i v e a l though the a c t i v i t y was not t y p i c a l of h a l l u c i n o g e n s . A l though the M y r i s t i c a c e o u s pas te samples d i d e x h i b i t some degree of MAO i n h i b i t i o n , t h i s was 201 shown to be p r i m a r i l y i f not e n t i r e l y due to the t ryptamine c o n s t i t u e n t s ; n o n - a l k a l o i d a l c o n s t i t u e n t s from the samples showed on l y a s l i g h t degree of n o n - s p e c i f i c i n h i b i t i o n . Most of the s y n t h e t i c t ryptamine d e r i v a t i v e s which were assayed as MAOI d i d e x h i b i t some a c t i v i t y but the I 5 0 va lues were u s u a l l y s e v e r a l o rde r s of magnitude lower than those f o r the 0 - c a r b o l i n e d e r i v a t i v e s . I n t e r e s t i n g l y , DMT showed the most MAOI a c t i v i t y of a l l the t r yp tamine d e r i v a t i v e s t e s t e d ; i t s I 5 0 was comparable to THH,' one of the l e a s t a c t i v e of the 0 - c a r b o l i n e s . A l l of t h i s ev idence c o n s i d e r e d toge the r suggests that the M y r i s t i c a c e o u s p a s t e s , when they are o r a l l y a c t i v e , must owe t h e i r a c t i v i t y to some mechanism other than MAO i n h i b i t i o n , whether due to |3-c a r b o l i n e s or other c o n s t i t u e n t s . What a l t e r n a t i v e mechanism might account f o r the o r a l a c t i v i t y of some samples? There are b a s i c a l l y two p o s s i b i l i t i e s worth c o n s i d e r a t i o n ; both would r e q u i r e f u r t h e r expe r imenta l i n v e s t i g a t i o n s to c o n f i r m . The f i r s t i s that the o r a l a c t i v i t y of the M y r i s t i c a c e o u s pas tes i s not due to the t r yp tamines at a l l , but r a the r to some other b i o l o g i c a l l y a c t i v e c o n s t i t u e n t s , such as l i g n a n s . C e r t a i n l y t h i s would he lp to e x p l a i n why the o r a l a c t i v i t y observed in s e l f - e x p e r i m e n t s was a t y p i c a l fo r h a l l u c i n o g e n s . It i s e q u a l l y p o s s i b l e , however, tha t t r yp tamines taken o r a l l y d i f f e r s i g n i f i c a n t l y in t h e i r e f f e c t s from t r yp tamines adm in i s t e r ed p a r e n t e r a l l y . That V i r o l a r e s i n ( i n c l u d i n g the sample assayed) does c o n t a i n b i o l o g i c a l l y a c t i v e l i g n a n s has been e s t a b l i s h e d ; but whether these l i g n a n s are capab le of e l i c i t i n g the spectrum of b i o l o g i c a l responses observed in the b i o a s s a y , i s not known. The second p o s s i b i l i t y , 202 which a l s o c a l l s fo r much f u r t h e r exper imenta l i n v e s t i g a t i o n , i s tha t the o r a l i n a c t i v a t i o n of DMT, 5-MeO-DMT, and r e l a t e d d e r i v a t i v e s i s not due to o x i d a t i v e deaminat ion by p e r i p h e r a l MAO; a l t e r n a t i v e metabo l i c pathways may be r e l a t i v e l y more s i g n i f i c a n t in the deg rada t i on of these compounds in the p e r i p h e r y . Both j_n v i t r o and j_n v i v o s t u d i e s of DMT metabol ism [2 ,3 ] ( c f . Chapter II) suggest that 6-hyd roxy l a t i on and/or N-o x i d a t i o n of DMT occurs more r e a d i l y in p e r i p h e r a l t i s s u e s than deaminat ion by MAO. 6-hyd roxy l a t i on has been shown to occur in p e r i p h e r a l t i s s u e s but appa r en t l y not in b r a i n [ 2 , 3 ] ; s i g n i f i c a n t l y , 6-hydroxy d e r i v a t i v e s of DMT and r e l a t e d compounds are i n a c t i v e as h a l l u c i n o g e n s [ 4 ] . L i t t l e i s known of the h a l l u c i n o g e n i c a c t i o n of DMT-NO, but i f i t f o l l o w s the gene ra l p a t t e r n fo r t e r t i a r y amine N-ox ides , [5] i t would e i t h e r be comp le te l y i n a c t i v e , or ten to one hundred t imes l e s s a c t i v e than DMT. Other s t u d i e s of i_n v i v o DMT metabol ism in the presence of MAO i n h i b i t o r s and microsomal mixed f u n c t i o n ox idase (MFO) i n h i b i t o r s [6] have found tha t wh i le the MAOI i p r o n i a z i d p ro longs plasma and t i s s u e h a l f - l i f e of DMT, the MFO i n h i b i t o r SKF-525A does n o t ; the au thors i n t e r p r e t these r e s u l t s as support f o r the hypo thes i s that DMT i s me tabo l i z ed main ly by MAO in v i v o . A problem wi th most i_n v i v o s t u d i e s of the type r e p o r t e d in [ 6 ] , i s that the DMT i s a d m i n i s t e r e d to the an imal i . p . r a the r than o r a l l y ; thus the compound reaches the c i r c u l a t i o n d i r e c t l y and avo ids " i n t e s t i n a l / h e p a t i c - p o r t a l shunt " metabo l i sm. It i s p o s s i b l e that the metabol i sm of DMT v i a the i n t e s t i n a l / h e p a t i c - p o r t a l shunt may d i f f e r i n important r e s p e c t s from i t s metabol ism when i n t r oduced d i r e c t l y i n t o the 203 b loods t ream or body c a v i t y . In the hepa t i c shunt 6-h y d r o x y l a t i o n , and/or N - o x i d a t i o n , may be r e l a t i v e l y more important than MAO as a c a t a b o l i c route fo r the compound. In any e ven t , _in v i v o me tabo l i c s t u d i e s i n v o l v i n g i . p . or o ther p a r e n t e r a l rou tes of a d m i n i s t r a t i o n a c t u a l l y shed l i t t l e l i g h t on DMT metabol ism f o l l o w i n g o r a l a d m i n i s t r a t i o n . The f a c t i s tha t our unde rs tand ing of the p e r i p h e r a l metabol ism of DMT and r e l a t e d compounds i s f a r from comple te ; a l l that i s known fo r c e r t a i n i s tha t more than type of o x i d a t i v e r e a c t i o n i s i n v o l v e d . MAO may be p a r t i a l l y r e s p o n s i b l e fo r the deg rada t i on of t r yp tamines in the p e r i p h e r y , but microsomal MFOs, which are i n v o l v e d in both the 6-hydroxy l and N-oxide pathways, are p o s s i b l y even more impor tan t . I n t e r e s t i n g l y , i f o r a l l y -a d m i n i s t e r e d DMT i_s a s u b s t r a t e fo r microsomal o x i d a s e s , then a mechanism can be proposed to e x p l a i n the o r a l a c t i v i t y of some V i r o l a p a s t e s , even though they l ack /3-carbol ines and are not s i g n i f i c a n t l y e f f e c t i v e as MAOI. T h i s a l t e r n a t i v e mechanism p o s t u l a t e s that some of the n o n - a l k a l o i d c o n s t i t u e n t s in the pas t e s may possess a n t i - o x i d a n t a c t i v i t y and/or e x h i b i t a c t i v i t y as s p e c i f i c i n h i b i t o r s of microsomal MFOs. In the former c a s e , p resence of h igh c o n c e n t r a t i o n s of n o n - s p e c i f i c a n t i - o x i d a n t s c o u l d scavange a h igh p r o p o r t i o n of the mo lecu la r oxygen in the v i c i n i t y , thus making l e s s of i t a v a i l a b l e as c o - s u b s t r a t e fo r the microsomal enzymes c a t a l y z i n g DMT N-ox ida t ion and 6-h y d r o x y l a t i o n . In the l a t t e r c a s e , c o n s t i t u e n t s in the V i r o l a r e s i n may s p e c i f i c a l l y i n h i b i t h e p a t i c microsomal MFO and thus b lock the o x i d a t i o n ( s ) of DMT caused by these enzymes. In e i t h e r case the compound c o u l d be p r o t e c t e d from o x i d a t i v e 2 0 4 t r a n s f o r m a t i o n in the i n t e s t i n a l / h e p a t i c shunt and thus be taken up i n t o the CNS in the form of the unchanged t e r t i a r y amine. A number of l i g n a n s have been c h a r a c t e r i z e d which e x h i b i t p r o t e c t i v e a c t i v i t y a g a i n s t hepa to tox in s [ 7 ] ; i n h i b i t i o n of hepa t i c MFO has been proposed as the p robab le mechanism. A l l of the a c t i v e compounds possessed a methy lened ioxypheny l mo ie ty , but ana logs l a c k i n g t h i s c o n f i g u r a t i o n d i d not have h e p a t o p r o t e c t i v e p r o p e r t i e s . The methy lened ioxypheny l group has been i m p l i c a t e d in o ther s t u d i e s as the main pharmacophore r e s p o n s i b l e fo r mixed f u n c t i o n ox idase i n h i b i t i o n [ 8 ] , The M y r i s t i c a c e o u s genera V i r o l a and I r yan the ra are both r i c h in c o n s t i t u e n t s i n c o r p o r a t i n g the methy lened ioxypheny l g roup , i n c l u d i n g f a t t y a c i d d e r i v a t i v e s , n e o l i g n a n s , f l a v a n s , and d i a r y l p r o p a n o i d s [ 9 ] . S e ve ra l nove l l i g n a n s hav ing t h i s s u b s t i t u t i o n were c h a r a c t e r i z e d in the bark of DMK-59 (V. e longata ) which was the sou r ce-p l an t fo r Marcos F l o r e s ' pas te sample ; t h i s sample showed the g r e a t e s t degree of o r a l a c t i v i t y in s e l f - e x p e r i m e n t s . A number of other p h e n o l i c compounds, i n c l u d i n g f l a v a n s , f l a v a n o i d s , i s o f l a v o n o i d s , d i a r y l p r o p a n o i d s , and n e o l i g n a n s , have been i s o l a t e d from a number of V i r o l a and I r yan the ra s p p . ; some of these compounds c o u l d ac t as a n t i o x i d a n t s and c o u l d c o n t r i b u t e to the i n a c t i v a t i o n of MFOs v i a t h i s n o n s p e c i f i c mechanism. In any case the p e r i p h e r a l metabol ism of DMT and r e l a t e d compounds, f o l l o w i n g o r a l a d m i n i s t r a t i o n , may be s i g n i f i c a n t l y a l t e r e d in the presence of a n t i o x i d a n t s and/or s p e c i f i c MFO i n h i b i t o r s ; under these c o n d i t i o n s the compounds might we l l reach the CNS in the form of the unchanged t e r t i a r y amine. Fu r the r jjn v i v o and _in 205 v i t r o exper iments would be r e q u i r e d to con f i rm or d i s c o n f i r m t h i s a l t e r n a t i v e mechanism of o r a l a c t i v i t y . In view of the phy tochemica l and pha rmaco log i ca l data accumulated in the present s tudy , i t appears that t h i s a l t e r n a t i v e h ypo thes i s i s at l e a s t as p r o b a b l e , i f not more p r o b a b l e , than MAO i n h i b i t i o n as the mechanism r e s p o n s i b l e fo r the o r a l a c t i v i t y of the M y r i s t i c a c e o u s p a s t e s . IV. C o n c l u s i o n Phytochemica l and pha rmaco log i c a l i n f o rma t i on c o l l e c t e d in the course of t h i s study has been i n s u f f i c i e n t to d e f i n i t e l y e s t a b l i s h the mechanism of o r a l a c t i v i t y in these two Amazonian h a l l u c i n o g e n s ; however i t has p r o v i d e d phy tochemica l data and in  v i t r o pha rmaco log i c a l ev idence which i n d i c a t e s that in the case of ayahuasca the o r i g i n a l h ypo thes i s proposed to e x p l a i n the o r a l a c t i v i t y cannot be d i s p r o v e d . C e r t a i n l y ayahuasca c o n t a i n s h igh enough c o n c e n t r a t i o n s of /3-carbol ines to e f f e c t i v e l y i n h i b i t MAO and by t h i s mechanism the a c t i v e h a l l u c i n o g e n i c c o n s t i t u e n t , DMT, may be p r o t e c t e d from p e r i p h e r a l d e g r a d a t i v e metabo l i sm. The a l t e r n a t i v e mechanism proposed in the above d i s c u s s i o n may a l s o be i m p l i c a t e d in the pharmacology of ayahuasca , but at l e a s t i t i s not necessa ry to invoke t h i s mechanism fo r ayahuasca . In the case of the o r a l l y - i n g e s t e d M y r i s t i c a c e o u s p r e p a r a t i o n s , however, the data r e p o r t e d here i n d i c a t e tha t MAO i n h i b i t i o n - - w h e t h e r due to /3-carbol ines or some o ther c o n s t i t u e n t s — i s a lmost c e r t a i n l y not the mechanism r e s p o n s i b l e f o r t h e i r o r a l a c t i v i t y . The pas tes do not c o n t a i n more than t r a c e s of /3-carbol ines , they show poor a c t i v i t y as 206 MAOI, and t h e i r o r a l a c t i v i t y , when p r e s e n t , i s not c o r r e l a t e d w i th the presence of 0 - c a r b o l i n e s . Some a l t e r n a t i v e mechanism must t h e r e f o r e be invoked to e x p l a i n the o r a l a c t i v i t y of these M y r i s t i c a c e o u s p a s t e s . Two such a l t e r n a t i v e s have been d i s c u s s e d above ; one i s that the o r a l a c t i v i t y i s due to b i o l o g i c a l l y a c t i v e c o n s t i t u e n t s o ther than t r y p t a m i n e s . The other and perhaps more a t t r a c t i v e p o s s i b i l i t y i s that the a c t i v e t r yp tamines are p r o t e c t e d from p e r i p h e r a l deg rada t i on by c o n s t i t u e n t s which i n h i b i t hepa t i c m ixed- func t i on o x i d a s e s , the enzymes r e s p o n s i b l e fo r 6-hyd roxy l a t i on and N-ox ida t ion of t r yp tamine d e r i v a t i v e s . MFO i n h i b i t o r s r e q u i r e the presence of a methy lened ioxypheny l c o n f i g u r a t i o n as the a c t i v e pharmacophore, and V i r o l a spp. are e x c e l l e n t sources of compounds p o s s e s s i n g t h i s mo ie ty . U n f o r t u n a t e l y n e i t h e r a l t e r n a t i v e mechanism can be proven or d i sp roven u n t i l more has been l ea rned ' about the in  v i v o metabol ism of DMT and r e l a t e d compounds. An obv ious p l a ce to s t a r t would be to study the metabol ism of o r a l l y - a d m i n i s t e r e d DMT in the presence of known MFO i n h i b i t o r s . 207 V. L i t e r a t u r e C i t e d 1. R i v i e r , L. & J . l i n d g r e n (1972) Ayahuasca, the South American H a l l u c i n o g e n i c D r i n k : E t h n o b o t a n i c a l and Chemica l I n v e s t i g a t i o n s . Economic Botany 29:101-129 2. B a rke r , S. A . , J . A. M o n t i , and S. T . C h r i s t i a n (1980) Metabo l i sm of the H a l l u c i n o g e n N ,N-d imethy l t ryptamine in Rat B ra in Homogenates. B i ochemica l Pharmacology 29:1049-57 3. S z a r a , S. & J . A x e l r o d . (1959) H y d r o x y l a t i o n and N-demethy l a t ion of N ,N-d imethy l t r yp tamine . E x p e r i e n t i a 15:216-17 4. S h u l g i n , A. T . (1976) Psychotomimet ic Agen t s , chapte r 4 in Maxwell Gordon (ed. ) P sychopharmaco log i ca l Agents V. IV. Academic Press 5. B i c k e l , M. H. (1969) The Pharmacology and B i o chemis t r y of N-o x i d e s . Pha rmaco log i ca l Reviews 21:325-358 6. Shah, N. S. , & M. P. Hedden (1977) Behav iou ra l E f f e c t s and M e t a b o l i c Fate of N ,N-d imethy l t ryptamine in Mice P r e t r e a t e d w i th /3-diethy laminoethy l- D i p h e n y l p r o p y l a c e t a t e (SKF-525-A ) , I p r o n i a z i d , and Ch lo rp romaz ine . Pharmacology,  B i ochemis t r y and Behav iour 8:351-56 7. MacRae, W. D . , & G. H. N. Towers (1984) B i o l o g i c a l A c t i v i t i e s of L i g n a n s . Phy tochemis t r y . In p r e s s . 8. B r a t t s t e n , L. B. (1977) B i o chemica l Defense Mechanisms in He rb i vo r e s Aga ins t P l an t A l l e l o c h e m i c a l s . Chapter 5 in G. A. R o s e n t h a l , and D. H. Janzen (eds . ) H e r b i v o r e s : T h e i r  I n t e r a c t i o n s wi th Secondary M e t a b o l i t e s . Academic P r e s s . 9. G o t t l i e b , O. R. (1979) Chemica l S tud i e s on M e d i c i n a l M y r i s t i c a c e a e from Amazonia . J ou rna l of Ethnopharmacology 1:309-323 10. D. MacRae. (1984) E t h n o b i o l o g i c a l and Chemica l I n v e s t i g a t i o n s of S e l e c t e d Amazonian P l a n t s . PhD t h e s i s , U n i v e r s i t y of B r i t i s h Co lumb ia , Vancouver , B. C. 208 APPENDIX I TABLE XVI - BIOLOGICALLY ACTIVE CONSTITUENTS IN AYAHUASCA ADMIXTURES The f o l l o w i n g l i s t of p l a n t s p e c i e s has been compiled from v a r i o u s s o u r c e s and i s i n t e n d e d as a summary of the c u r r e n t s t a t e of p h y t o c h e m i c a l knowledge of those genera and s p e c i e s which are known to be u t i l i z e d as admixtures to ayahuasca. The i n f o r m a t i o n on the use of these s p e c i e s as ayahuasca admixtures i s compiled p r i m a r i l y from r e f e r e n c e s 1,2,3,4,5, and 86; i n f o r m a t i o n on the v e r n a c u l a r names of the p l a n t s used i s compiled from the r e f e r e n c e s c i t e d above and a l s o from 85. The phytochemical i n f o r m a t i o n i s d e r i v e d p r i m a r i l y from a computer s e a r c h of the B i o l o g i c a l A b s t r a c t s Data Base and the American Chemical S o c i e t y Data Base, c o v e r i n g the y e a r s 1970-present. The r e f e r e n c e s c i t e d i n t h i s appendix are not intended to be e x h a u s t i v e but r a t h e r a r e i n t e n d e d as I n d i c a t o r s of the e x i s t e n c e or n o n e x i s t e n c e of i n f o r m a t i o n r e g a r d i n g biodynamic c o n s t i t u e n t s i n the s p e c i e s l i s t e d . In the case of c e r t a i n genera, e s p e c i a l l y Tabernaemontana, Tabebuia. Maytenus. Alc h o r n e a , Ocimum, E r y t h r i na. F i cus. and Uncar i a. the number of a v a i l a b l e r e f e r e n c e s runs well i n t o the thousands; i n these i n s t a n c e s o n l y a few key r e f e r e n c e s have been c i t e d . In most i n s t a n c e s the r e f e r e n c e s c i t e d do not r e f e r s p e c i f i c a l l y to the s p e c i e s used as an ayahuasca admixture, but to some c l o s e l y r e l a t e d s p e c i e s i n the same genus. O f t e n i n f o r m a t i o n i s not a v a i l a b l e on the c o n s t i t u e n t s of the p a r t i c u l a r s p e c i e s used i n c o n j u n c t i o n w i t h ayahuasca, but 1s a v a i l a b l e f o r o t h e r members of the genus. Fam i 1 y : Genus & S p e c i e s V e r n a c u l a r Name Biodynamic C o n s t i t u e n t s R e f e r e n c e s Acanthaceae: T e l i o s t a c h y a l a n c e o l a t a Nees v a r . C r i s p a Nees 1n Mart. "toe negro" none r e p o r t e d 1 . 3 Amaranthaceae: I r e s ine sp. P. Br. - hydroxy-cinnamic a c i d amides 1 , 44 A 1 t e r n a n t h e r a l e h m a n l i I H e r o n " p i c u r u l l a n a - q u i n a " none r e p o r t e d 2 .4.85 Apocynaceae: M a l o u e t l a tamaquarina (Aubl.)A. DC. " c u c h u r a - c a s p l " s t e r o i d a l k a l o i d s , conopharyng1ne 2 ,26,62 Tabernaemontana sp. L. "uchu-sanango" b i s i n d o l e a l k a l o i d s , t e r p e n o i d s , c o r n a r i d i ne 1 . ,2,8.9,31.76,84 Hlmatanthus succuba (Spruce)Woods " b e l l a c o - c a s p i " f l a v o n o i d s , fu1voplurnieron 1 , 32,43 Araceae: M o n t r i c h a r d i a a r b o r e s c e n s S c h o t t . "raya b a l s a " none r e p o r t e d 1 B i g n o n l a c e a e : Mansoa a l l i a c e a e (Lem.)A. Gentry "ajo sacha" none r e p o r t e d 1 Tabebuia heteropoda (DC)Sandw1th. "tahuar1" dibenzoxanthenes. n a p t h o q u i n o n e s . l a p a c h o l 1 , 22,53,73 Tynnanthus p a n u r e n s l s "clavohuasca" none r e p o r t e d 1 (Bur.)Sandwi t h . T a b l e X V I ( c o n t ' d ) F a m 1 1 y : G e n u s & S p e c i e s V e r n a c u l a r Name • B i o d y n a m i c C o n s t i t u e n t s R e f e r e n c e s B o m b a c a c e a e : C e l b a p e n t a n d r a L. " 1 u p u n a " n one r e p o r t e d 1 C a v a m l l e s l a h y l o g e l t o n U l b . " p u c a 1 u p u n a " ? 1 , 37 C a c t a c e a e : O p u n t 1 a s p . M i l l " t c h a i " N -me t h y1 - t y r a m i n e , m e s c a l i n e 5 , 4 0 . 4 2 E p l p h y l l u m s p . Haw. " p o k e r e " n o n e r e p o r t e d 5 C a r y o c a r a c e a e : A n t h o d i s c u s p i l o s u s D u c k e " t a h u a r i " n o n e r e p o r t e d 1 C e 1 a s t r a c e a e : M a y t e n u s e b e n i f o l i a R e i s s " c h u c h u h u a s i " s e s q u i t e r p e n e a l k a l o i d s , n i c o t i n o y l a l k a l o i d s , t n t e r p e n e s , may t e n s i n e . t i n g o n a n e , a n s a m a c r o l i d e s . e t c . 1 , 2 0 , 4 1 , 4 9 5 2 . 7 8 , C y c 1 a n t h a c e a e : C a r l u d o v l c a d l v e r g e n s D r u d e " t a m s h i " n one r e p o r t e d 1 C y p e r a c e a e : C y p e r u s s p . L . " p i r i - p i r i " q u i n o n e s , e s s e n t i a l o i l s s a p o n i n s , s e s q u i t e r p e n e s 1 , 7 , 5 7 G4 C y p e r u s d l g i t a t u s R o x b . " c h i c o r r o " n o n e r e p o r t e d 86 E u p h o r b i a c e a e : A l c h o r n e a c a s t e n i f o l i a (W i 1 I d . ) J u s s . " h i p o r u r u " a l c h o r n i n e , i m a d a z o l e a l k a l o i d s , c o r y n a n t h e - t y p e a l k a l o i d s , a n t i f e e d a n t s , e t c . 1, 6 3 . 70 7 9 , 8 4 , H u r a c r e p i t a n s L. " c a t a h u a " t i g l i o n e d i t e r p e n e s . p i s c i c i d a l c o m p o u n d s , 1 e c t i n s 1 , 4 7 . 5 4 . 6 5 G u t t 1 f e r a e : C l u s i a I n s i g n i s M a r t . " r e n a c o " c l u s i a n o n e , x a n t h o c h y m o l , t r i t e r p e n o i d s 1 . 2 5 . 3 6 . 6 6 L a b l a t e a e : O c i m u m m i c r a n t h u m W i l l d . " p i c h a n a , a b a c a " n e o l i g n a n s , s e s q u i t e r p e n e s , a n t i he 1 mer i t i c s 2 . 1 9 . 3 0 . 5 0 L e c y t h i d a c e a e : C o u r o u p i t a g u i a n e n s i s A u b l . " a y a h u m a " i n d o 1 e a 1 k a 1 o i d s 1 . 1 2 , 6 0 T a b l e X V I ( c o n t ' d ) F a m l l y : G e n u s & S p e c i e s V e r n a c u l a r Name B i o d y n a m i c C o n s t i t u e n t s R e f e r e n c e s L e g u m l n o s a e : C a l l l a n d r a a n g u s t l f o l i a S p r u c e " b o b i n s a n a " i m l n o a c i d s 1 , 3 3 , 3 8 C e d r l l i n g a c a t e n a e f o r m i s D u c k e " h u a i r a c a s p i " n o n e r e p o r t e d 1 P 1 t h e c e l 1 o b i urn l a e t u m ( P o e p p . & E n d l . ) B e n t h . " r e m o c a s p i " p h y t o m i t o g e n s , l u p e o l , s p i n a s t e r o l 1 , 1 3 , 6 9 S c l e r o b l u m s e t i f e r u m D u c k e " p a 1 i s a n g r e " = " p a 1 i s a n t o " ? n one r e p o r t e d 1 , 8 5 E r y t h r i n a p o e p p l g i a n a ( W a l p . ) 0 . F . C o o k " a m a c i z a " = " a m a s i s a " ? E r y t h r i n a a l k a l o i d s , e t c . 1 . 8 5 . 3 4 . 77 V o u c a p o u a a m e r i c a n a A u b l . " h u a c a p u " n o n e r e p o r t e d 1 L o m a r 1 o p s 1 d a c e a e : L o m a r l o p s i s j a p u r e n s i s ( M a r t . ) d . Sm. " s h o k a " n o n e r e p o r t e d 5 L o r a n t h a c e a e : P h r y g 1 1 a n t h u s e u g e n o i d e s v a r . r o b u s t u s G l a z . " m i y a " n one r e p o r t e d 5 P h t l r u s a p y r i f o l i a H . B . K. E l c h l e r " s u e I d a c o n s u e I d a " n one r e p o r t e d 1 M a r a n t a c e a e : C a l a t h e a v e i t c h i a n a V e i t c h . E x H o o k e r "pu1 ma" t r y p t o p h a n 2 , 75 Men i s p e r m a c e a e : A b u t a g r a n d l f o l l a ( M a r t . ) S a n d w i t h . " a b u t a " , " c a i m i t 1 1 l o " , " s a n a n g o " t r o p o l o n e i s o q u i n o l i n e s , o x o - a p o r p h 1 n e s , p a 1 mat i ne 1 . 8 6 . 1 5 . 5 8 , 5 9 M o r a c e a e : C o u s s a p o a t e s s m a n n l i M i l d b r . " r e n a c o " n o n e r e p o r t e d 1 F i c u s r u i z i a n a S t a n d i . F i c u s i n s i p i d a W i l l d . " r e n a c o " " o j e " f u r o c o u m a r i n s . t r i t e r p e n e s , b i p h e n y 1 h e x a -h y d r o i n d o l i z i n e s , p h e n a n t h r o x i n d o l i z i n e s 1 , 2 8 . 2 9 , 71 P o u r o u m a A u b l . " c h u 1 1 a c h a q u i c a s p i " n o n e r e p o r t e d 1 s p . a f f . f o l e a t a M c B r . T a b l e XVI ( c o n t ' d ) F a m l 1 y : G e n u s & S p e c i e s V e r n a c u l a r Name B i o d y n a m i c C o n s t i t u e n t s R e f e r e n c e s M y r i s t i c a c e a e : V i r o l a s p . A u b l . " c u m a l a " d i a r y l p r o p a n o i d s , 2 - m e t h y l k e t o n e s t r y p t a m i n e s . p - c a r b o l i n e s , n e o l i g n a n s 1 . 6 . 2 4 . 6 1 74 V i r o l a s u r i n a m e n s i s W a r b . " c a u p u r i " n e o 1 i g n a n s 1 . 6 Nymph i a c e a e : C a b o m b a a q u a t i c a A u b l . " m u r e r e " = " m u r e r u " ? n o n e r e p o r t e d 1 . 8 5 P o l y g o n a c e a e : T r i p l a r i s s u r i n a m e n s i s M a r t , v a r . c h a m i s s o a n a M e i s n . " t a n g a r a n a " n o n e r e p o r t e d 1 P o n t e d a r i a c e a e : P o n t e d a r i a c o r d a t a L. " a m a r o n b o r r a c h e r o " n o n e r e p o r t e d 2 . 3 P h y t o l a c c a c e a e : P e t i v e r i a a l l I a c e a e L. " m u c u r a " o l i g o s u l f i d e s , t r i t e r p e n e s , t r i t h i o l a n e s 1 , 1 6 . 1 8 . 3 9 R u b i a c e a e : C a 1 y c o p h y 1 1 urn s p r u c e a n u m ( B e n t h . ) H o o k e r " c a p i r o n a n e g r o " n o n e r e p o r t e d 1 G u e t t a r d a f e r o x S t a n d i . " g a r a b a t a " c a n t h e m i n e . h e t e r o - y o h i m b i n e a l k a l o i d s 1 . 1 0 , 4 8 l i n e a r l a g u i a n e n s i s ( A u b l . ) G m e l " g a r a b a t a " s p i r o - o x i n d o l e s . b i s - i n d o l e s . h e t e r o -y o h i mb i n e s 1 . 2 7 , 6 7 . 7 6 , 8 2 , 8 4 S c h i z a e c e a e : L y g o d i u m v e n u s t u m Sw. " t a c h a i d e l m o n t e " a n t f f e r t i 1 i t y a g e n t s 5 . 5 5 S c r o p h u l a r i a c e a e : S c o p a r i a d u l c i s L . " n u c n u p i c h a n a " t r i t e r p e n e s , S - M e O - b e n z o x a z o l i 1 i n o n e 1 , 2 1 . 5 6 . 7 2 T a b l e X V I ( c o n t ' d ) Fam1 1 y : G e n u s & S p e c i e s V e r n a c u l a r Name B i o d y n a m i c C o n s t i t u e n t s R e f e r e n c e s S o l a n a c e a e : D a t u r a s u a v e o l e n s ( W i l l d . ) B r e c h t o l d & P r e s l " t o e " t r o p a n e a l k a l o i d s 1 , 2 . 3 , 4 B r u n f e l s i a c h i r i c s a n a n g o P1owman " c h i r i c s a n a n g o " s c o p a l e t i n , CNS d e p r e s s a n t s , a n t i i n f l a m m a t o r y c o m p o u n d s 1 . 2 , 1 1 , 8 7 . 8 8 I o c h r o m a f u c h s o i d e s M e e r s i n H o o k e r " b o r r a c h e r o " n o n e r e p o r t e d 2 , 3 , 4 J u a n u l l o a o c h r a c e a C u a t r . " a y a h u a s c a " n o n e r e p o r t e d V e r b e n a c e a e : C o r n u t i a o d o r a t a ( P . & E . ) P e o p p . " s h i n g u a r a n a " n o n e r e p o r t e d 8 5 , 8 6 V i t e x t r i f l o r a V a h l . " t a h u a r i " d i t e r p e n e s l a c t o n e s , i r i d o i d g l y c o s i d e s , f l a v o n o i d g l y c o s i d e s 1 , 1 7 . 2 3 . 5 1 . 6 8 214 Re fe rences 1. L u i s E. Luna . The Concept of P l an t s as Teachers Among Four Mes t i zo Shamans of I q u i t o s , Nor theas t Pe ru . Paper p resen ted at the Symposium on Shamanism, X l t h I n t e r n a t i o n a l Congress of A n t h r o p o l o g i c a l and E t h n o l o g i c a l S c i e n c e s , Phase 2. Vancouver , B. C , August 20-23, 1983. 2. R. E. S c h u l t e s . E t h n o t o x i c o l o g i c a l S i g n i f i c a n c e of A d d i t i v e s to New World H a l l u c i n o g e n s . P l an t Sc i ence B u l l e t i n 18: 34-41. 1972. 3. R. E. S c h u l t e s . New Data on the Ma lp igh i a ceous N a r c o t i c s of South America Harvard B o t a n i c a l Museum L e a f l e t s , 2 3 : I 3 7 f f . 1979. 4. H. V. P i n k l e y . P l an t Admixtures to Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k . L l o y d i a 32 :305 . 1969. 5. L. R i v i e r & J . L i n d g r e n . Ayahuasca , the South American H a l l u c i n o g e n i c D r i n k : E t h n o b o t a n i c a l and Chemica l I n v e s t i g a t i o n s . Economic Botany 29:101-129. 1972. 6. L. E. S. B a r a t a , P. M. Baker , 0. R. G o t t l i e b , & E. A. Ruveda. Neo l ignans of V i r o l a Su r i namens i s . Phy tochemis t ry 17:783-86. 1978. 7. R. D. A l l a n , R. J . W e l l s , R. L. C o r r e l l , & J . K, MacLeod. The Presence of Quinones in the Genus Cyperus as an A i d to C l a s s i f i c a t i o n . Phy tochemis t ry 17:263-266. 1978. 8. D. G. K i n g s t o n . P l an t An t i - cance r Agents Par t 6: I s o l a t i o n of Voacang ine , Voacamine and Ep i - voaco r i ne from Tabernaemontana a rborea Sap. J o u r n a l of Pha rmaceu t i ca l S c i ences 67:271-72. 1978. 9. D. G. K i n g s t o n , B. B. G e r h a r t , F. I onescu , M. M. Mangino, & S. M. Sami. P l an t An t i - cance r Agents Par t 5: New b i s- I ndo l e A l k a l o i d s from Tabernaemontana j o h n s t o n i i Stem Bark. J ou rna l of Pha rmaceu t i ca l S c i ences 67 :249-51. 1978. 10. H. P. Husson, C. Kan-Fan, T . Sevenet , & J . P. V i d a l . S t r u c t u r e of Cathenamine, Key In te rmed ia te in the B i o s y n t h e s i s of Indole A l k a l o i d s . Te t rahedron L e t t e r s 22:1889-92. 1977. 215 11. M. Chauba l , & R. P. I y e r . Carbon-13 NMR Spec t r a of S c o p o l e t i n . L l o y d i a 40 :618 . 1977. 12. J . Bergman, B. Eges t ad , & J -O . L i n d s t r o m . The S t r u c t u r e of Some I n d o l i c C o n s t i t u e n t s in Couroup i t a g u a i n e n s i s . Te t rahedron L e t t e r s 30:2625-26. 1977. 13. M. Yadav, V. K. C. Ganaswaran. Phyto-Mitogens of T r o p i c a l Legumes Par t I: I s o l a t i o n from Pa rk i a s p e c i o s a and P i t h e c e l l o b i u m j i r i n g a . Ma lays i an J ou rna l of Sc ience 4:25-26. 1976. 14. C . K. Kokate & K. C. Varma. Pha rmaco log i ca l Study on E s s e n t i a l O i l of Cyperus s c a r i o s u s Part I: E f f e c t on C e n t r a l Nervous System pp. 189-193 in L. D. Kapoor & R. K r i shnan (eds . ) Advances in E s s e n t i a l O i l I ndus t r y . Today & Tomorrow's P r i n t e r s & P u b l i s h e r s , New D e l h i . 1977. 15. J . V. S i l v e r t o n , C. Kabuto, K. T . Buck, M. P. Cava . S t r u c t u r e of Imerubr ine , a Novel Condensed Tropo lone I s o q u i n o l i n e A l k a l o i d . J ou rna l of the American Chemica l  So c i e t y 99:6708-12. 1977. 16. C . Von S z c z e p a n s k i , J . H e i n d l , G-A. Hoyer , & E. S ch roeder . B i o l o g i c a l l y A c t i v e Compounds from P l an t s Part 2: S yn thes i s and A n t i - m i c r o b i a l A c t i v i t y of Some Dissymmetr i c O l i g o S u l f i d e s . European J ou rna l of M e d i c i n a l Chemis t ry 12:279-84. 1977. 17. R. U. Kodanda, R. E. Venka ta , R. D. Venka ta . Pheno l i c C o n s t i t u e n t s of the Bark of V i t e x negundo. Ind ian J o u r n a l  of Pharmacy 39 :41 . 1977. 18. F. P. Segelman, & A. B. Segelman. C o n s t i t u e n t s of P e t i v e r i a a l l i a c e a e (Phy to laccaceae ) Par t 1: I s o l a t i o n of I s o a r b o r i n o l , I s o a r b o r i n o l Ace t a t e and I s o a r b o r i n o l Cinnamate from the Leaves . L l o y d i a 38 :537 . 1975. 19. R. G. Roy, N. M. Madesayaa, R. B. Ghosh, D. V. G o p a l k r i s h n a n , N. N. Murthy , T . J . D o r a i r a , & N. L. S i t a raman . Study on I n h a l a t i o n Therapy by an Indigenous Compound on Plasmodium v i vax and Plasmodium f a l c i p a r u m I n f e c t i o n s : A P r e l i m i n a r y Communicat ion. Ind ian J o u r n a l of M e d i c i n a l Research 10:1451-55. 1976. 216 20. S. M. Kupchan, & R. M. Smi th . M a y t o l i n e , May t ine , and M a t o l i d i n e , Novel N i c o t i n o y l Sesqu i te rpene A l k a l o i d s from Maytenus s e r r a t a . J ou rna l of Organ i c Chemi s t r y 42:115-118. 1977. 21. C-M. Chen & C-T. Chen. 6-Methoxy Benoxazo l inone and T r i t e r p e n o i d s from Roots of S copa r i a d u l c i s . Phytochemist ry 15:1997-99. 1976. 22. R. Y. Wong, K. J . Pa lmer , G. D. Manners, & L. J u r d . The S t r u c t u r e of Guayacanin wi th Acetone of C r y s t a l l i z a t i o n , A N a t u r a l l y O c c u r r i n g Dibenzoxanthene from Tabebuia guayacan. Ac ta C r y s t o l l o g r a p h i c a Sec . B 8:2396-2400. 1976. 23. H. T a g u c h i . S tud i e s on the C o n s t i t u e n t s of V i t e x c a n n i b o l i a . Chemica l & Pharmaceut i ca l B u l l e t i n 7:1668-70. 1976. 24. R. B a r u f f a l d i , E. F e d e l i , & N. C o r t e s i . Study of the Fat of V i r o l a su r inamens i s Part 1: A c i d i c and G l y c e r i d e Compos i t ion and Chemica l Nature of Some of the U n s a p o n i f i a b l e Components. R e v i s t a de Farmacia e B ioqu im ica da Un i v e r s i dade de Sao Paulo 13:91-102. 1975. 25. L. E. M c C a n d l i s h , J . C. Hanson, & G. H. S t o u t . The S t r u c t u r e s of 2 D e r i v a t i v e s of B i - c y c l o - 3 , 3 , 1 - n o n a n e - 2 , 4 , 9 -t i r o n e , a Na tu r a l Product C lus i anone and T r i m e t h y l a t e d C a t e c h i n i c A c i d . Ac ta C r y s t a l l o g r a p h i c a , Sec . B: S t r u c t u r a l  C r y s t a l l o g r a p h y and C r y s t a l Chemist ry 32:1793-1801. 1976. 26. J . D. Medina & R. Bracho. C o n s t i t u e n t s of the Bark of M a l o u e t i a g l a n d u l i f e r a . P l an ta Medica 29:367-69. 1976. 27. Y. Ban, M. Se to , & T . O i s h i . The S yn thes i s of 3 S p i r o Ox indo l e D e r i v a t i v e s Part 7: T o t a l S yn thes i s of A l k a l o i d s Racemic Rhynchophy l l i ne and Racemic I s o r h y n c h o p h y l l i n e . Chemica l & Pha rmaceu t i ca l B u l l e t i n 11:2605-13 1975. 28. Y. I. E i d l e r , G. L. Genk ina , T . T . Shak i r o v . Q u a n t i t a t i v e De te rm ina t i on of Furocoumar ins in F i c u s c a r i c a Leaves . Khimiya P r i r odnykh S o e d i n e n i i 3:349-51. 1975. 29. M. H. A . E l gama l , B. A. H. E l - t a w i l , & M. B. E. F ayez . The T r i t e r p e n o i d C o n s t i t u e n t s of the Leaves of F i c u s n i t i d a . Na tu rw i s senscha f t en 62 :486. 1975. 217 30. S. J . Terhune, J . W. H o f f , & R. W. Lawrence. B i c y c l o s e s q u i p h e l l a n d r e n e and 1-epi B i c y c l o s e s q u i p h e l l a n d r e n e 2 New Dienes Based on the Cadalene S k e l e t o n . Phy tochemis t ry 13:1183-85. 1974. 31. B. T a l a p a t r a , A. P a t r a , & S. K. T a l a p a t r a . Te rpeno ids and A l k a l o i d s of the Leaves of Tabernaemontana c o r o n a r i a . Phy tochemis t ry 14: 1652-53. 1975. 32. R. R. P a r i s , & S. Du re t . The F l a v o n o i d s of V a r i o u s Apocynaceae. P l an tes M e d i c i n a l e s et Phy to the rap i e 8:318-25. 1974. 33. G. A. Dardenne. New Free Amino A c i d s from Leguminosae. Phy tochemis t ry 14:860. 1975. 34. K. I t o , M. Haruna, & H. Fu ruka . S tud i e s on the E r y t h r i n a A l k a l o i d s Part 10: A l k a l o i d s of Seve ra l E r y t h r i n a sp . P l a n t s from S ingapore M a l a y s i a . Yakugaku Zassh i 3:358-62. 1975. 35. S. R. Hemingway, & J . D. P h i l l i p s o n . A l k a l o i d s from South American Spec ies of Unca r i a (Rub iaceae ) . J ou rna l of  Pharmacy and Pharmacology 2 6 ( s u p p l . ) : 1 1 3 p . 1974. 36. D. L. D reye r . Xanthochymol from C l u s i a rosea ( G u t t i f e r a e ) . Phy tochemis t ry 13:2883-84. 1974. 37. J . E. Lopez G u i l l e n & I. K. De C o r n e l i o . M e d i c i n a l P l an t s of Peru pa r t 5. B i o t a 10:76-104. 1974. 38. J . E. Lopez G u i l l e n & I. K. De C o r n e l i o . M e d i c i n a l P l an t s of Peru pa r t 4. B i o t a 10:28-56. 1974. 39. E. K. Adesogan. T r i t h i o l a n i a c i n , a Novel T r i t h i o l a n e from P e t i v e r i a a l l i a c e a . J ou rna l of the Chemica l Soc i e t y ,  Chemica l Communications 21 :906-7. 1974. 40. R. L. Vander Veen & L. G. West. N-methyl Tyramine from Opunt i a c l a v a t a . Phy tochemis t ry 13:866-67. 1974. 41. M. Leboeuf . Maytans ine and M a y t a n s i n o i d s . P l an t e s  M e d i c i n a l e s et Phy to the rap i e 12:53-70. 1978. 218 42. J . P a rdanan i , B. N. Meyer, & J . L. McLaugh l i n . Mesca l i ne and Re l a t ed Compounds from Opunt ia s p i n o s i o r . L l o y d i a 41:286-88. 1978. 43. G. P. Perdue & R. N. B l oms te r . South American P l a n t s Part 3: I s o l a t i o n of F u l v o p l u m i e r i n from Himatanthus sucuuba (Apocynaceae) . J ou rna l of Pha rmaceu t i ca l S c i ences 67:1322-23. 1978. 44. J . Mar t in-Tanguy , F. Cabanne, E. P e r d r i z e i , & C. M a r t i n . The D i s t r i b u t i o n of Hydroxy-c innamic A c i d Amides in F lower ing P l a n t s . Phy tochemis t ry 11:1927-8. 1978. 45. P. K. Mehrot ra & V. P. Kamboj. Hormonal P r o f i l e of C o r o n a r i d i n e H y d r o c h l o r i d e , an A n t i f e r t i l i t y Agent of P lant O r i g i n . P l an ta Medica 33:345-49. 1978. 46. H. Wagner & J . Bu rgha r t . Spermidine A l k a l o i d s and T r i t e r p e n e s from Maytenus h e t e r o p h y l l a s s p . h e t e r o p h y l l a and P l e u r o s t y l a a f r i c a n a : Chemica l C o n s t i t u e n t s of the C e l a s t r a c e a e Part 4. P l an t a Medica 32A:9-14. 1977. 47. F. J . Evans & C. J . Soper . The T i g l i a n e Daphnane and Ingenane D i t e r p e n e s : T h e i r Chemis t r y , D i s t r i b u t i o n , and B i o l o g i c a l A c t i v i t i e s ; A Review. L l o y d i a 4 :193-233. 1978. 48. C. Kan-Fan & H. P. Husson. S t e reochemica l C o n t r o l in the B iomimet ic Convers ion of Heteroyohimbine A l k a l o i d P r e c u r s o r s : I s o l a t i o n of a Novel Key I n t e rmed i a t e . J o u r n a l  of the Chemica l S o c i e t y , Chemica l Communicat ions 14:618-19. 1 978. 49. H. N o z a k i , H. S u z u k i , K-H. Lee , & A. T . M c p h a i l . S t r u c t u r e and S t e reochemis t r y of M a y t e n f o l i c A c i d and M a y t e n f o l i o l , 2 New A n t i - Leukemic T r i t e r p e n e s from Maytenus D i v e r s i f o l i a : X-Ray C r y s t a l S t r u c t u r e s . J ou rna l of the Chemica l Soc i e t y , Chemica l Communications 0:1048-51. 1982. 50. D. G. D e s a i , N. S. Ambade, & R. R. Mane. S y n t h e s i s of Ocimum a New Neo l ignan from Ocimum americanum. Ind ian J o u r n a l of  Chemis t ry S e c t i on B f Organ ic Chemist ry 5:491-92. 1982. 51. C . K. S e h g a l , S. C . T a n e j a , K. L. Dhar , & C. K. A t a l . 2' p Hydroxybenzoy l Musaenos id i c A c i d , a New I r i d o i d G l y c o s i d e from V i t e x negundo . ' Phy tochemis t r y 2:363-66. 1982. 219 52. J . G. Gonza l e s , G. D e l l e Monache, F. D e l l e Manache, G. B. M a r i n i - B e t t o l o . Chuchuhuasha, a Drug Used i n Fo lk Med ic ine in the Amazonian and Andean Areas of South Amer i c a : A Chemica l Study of Maytenus L a e v i s . J ou rna l of  Ethnopharmacology 5:73-78. 1982. 53. D. G. K ings ton & M. M. Rao. I s o l a t i o n , S t r u c t u r e E l u c i d a t i o n and Syn thes i s of 2 New C y t o t o x i c Napthoquinones from Tabebuia c a s s i n o i d e s . P l an ta Medica 3:230-31. 1980. 54. M. Pe re , D. Pe re , & P. Rouge. I s o l a t i o n and S tud i e s of the Phys i cochemica l and B i o l o g i c a l P r o p e r t i e s of L e c t i n s from Hura c r e p i t a n s . P l an t a Medica 41:344-50. 1981. 55. B. B. Ga i tonde & R. T . Mahajan. A n t i f e r t i l i t y A c t i v i t y of Lygodium f l exuosum. Ind ian J ou rna l of Med i ca l Research 72:597-604. 1980. 56. S. B. Mahato, M. C. Das, & N. P. Sahu. The Te rpeno ids of S copa r i a d u l c i s . Phytochemi s t r y 20:171-73. 1981. 57. P. N. S ingh & S. B. S i ngh . A New Saponin from Mature Tubers of Cyperus ro tundus . Phy tochemis t ry 19:2056. 1980. 58. J . W. S k i l e s , J . M. Saa, & M. P. Cava . S p l e n d i d i n e , a New Oxo Aporphine A l k a l o i d from Abuta r u f e s c e n s . Canadian  J ou rna l of Chemis t ry 57:1642-46. 1979. 59. Se tor de F i t o q u i m i c a , INPA, Manaus,Amazonas, B r a z i l . Chemica l Compos i t ion of Amazonian P l a n t s . Ac ta Amazonica 1 :83-86. 1971 . 60. A. K. Sen 6 S. B. Mahato. C o u r o u p i t i n e a New A l k a l o i d from Cou roup i t a g u i a n e n s i s . Te t rahedron L e t t e r s 7:609-10. 1974. 61 . O. R. G o t t l i e b , & A. A . L o u r e i r o . D i s t r i b u t i o n of D i a r y l P ropano ids in Amazonian V i r o l a S p e c i e s . Phy tochemis t ry 12:1830. 1973. 62. F. Khuong-Huu, & M-J. Magde la ine . S t e r i o d A l k a l o i d s from Ma loue t i a b rachy loba and from Ma loue t i a h e u d e l o t i i . Phy tochemis t ry 7:1813-16. 1973. 220 63. F. Khuong-Huu & J . P. Le F o r e s t i e r . A l c h o r n e i n e , I s o a l c h o r n e i n e and A l c h o r n e i n o n e , P roducts I s o l a t e d from A l chornea f l o r i b u n d a . Te t rahedron 28:5207-20. 1972. 64. H. H i k i n o , & K. A o t a . S t r u c t u r e and Abso lu t e C o n f i g u r a t i o n of A lpha Rotunol and Beta R o t u n o l , S e s q u i t e r p e n o i d s of Cyperus rotundus M. Te t rahedron 27:4831-36. 1971. 65. K. Saka ta , & K. Kawazu. S tud i e s on a P i s c i c i d a l C o n s t i t u e n t of Hura c r e p i t a n s D. Part 1: I s o l a t i o n and C h a r a c t e r i z a t i o n of Hura Tox in and i t s P i s c i c i d a l A c t i v i t y . A g r i c u l t u r a l and  B i o l o g i c a l Chemistry 35:1084-91. 1971. 66. S. B. Mathur T r i t e r p e n o i d C o n s t i t u e n t s of C l u s i a rosea D. Phy tochemis t ry 11:1513-14. 1972. 67. M. L a v a u l t , C. M o r e t t i , & J . B runeton . A l k a l o i d s of Unca r i a g u i a n e n s i s . P l an ta Medica 47 :244-5. 1983. 68. A. H. J a ckson , & A. S. Chawla. S tud i e s of E r y t h r i n a A l k a l o i d s . Part IV. GC/MS I n v e s t i g a t i o n s of A l k a l o i d s in the Leaves of E. p o e p p i g i a n a , E. m a c r o p h y l l a , E. b e r t e r o a n a , and E. s a l v i f l o r a . A l l e r t o n i a 3:39-45. 1982. 69. S. P. Gunasekera , G. A. C o r d e l l , & N. R. F a rnswor th . C o n s t i t u e n t s of P i t h e c e l l o b i u m m u l t i f l o r u m . J o u r n a l of N a t u r a l P roducts 45 :651 . 1982. 70. B. L. H. Hank inson . I n v e s t i g a t i o n of C o n s t i t u e n t s and A n t i f e e d a n t A c t i v i t y of A l cho rnea t r i p l i n e r v i a . D i s s e r t a t i o n A b s t r a c t s In te rna t i o n a l Sec . B # DA8217228. 1 982 71. S. R. Venkatacha lam, & N. B. Mu l chandan i . I s o l a t i o n of P h e n a n t h r o i n d o l i z i d i n e A l k a l o i d s and a Novel B i p h e n y l h e x a h y d r o i n d o l i z i n e A l k a l o i d from F i c u s h i s p i d a . Na tu rw i ssenscha f t en 69 :287-8. 1982. 72. J . L i , Y. L i , R. N i e , & J . Zhou. C o i x o l and B e t u l i n i c A c i d of S copa r i a d u l c i s L. Yun-nan Ch ih Wu Yen Ch iu 3:475-7. 1981. 73. L. Prakash & R. S i n g h . Chemica l Examinat ion of the Leaves and Stem heartwood of Tabebuia p e n t a p h y l l a (L inn) Hemsl . ( B i gnon i a ceae ) . J ou rna l of the Ind ian Chemica l S o c i e t y 58:1122-23. 1981. 221 74. O. R. G o t t l i e b . Chemica l S tud i e s on M e d i c i n a l M y r i s t i c a c e a e from Amazonia. J ou rna l of Ethnopharmacoloqy 1:309-323. 1979. 75. W. E. S p l i t t s t o e s s e r & F. W. M a r t i n . The Tryptophan Content of T r o p i c a l Roots and T u b e r s . H o r t s c i e n c e 10:23-24. 1975. 76. G. A. C o r d e l l & J . E. Sax ton . B i s i n d o l e A l k a l o i d s . Chapter 1 in R. H. F. Manske & R. G. A. Rod r i go , ( eds . ) The A l k a l o i d s V o l XX.Academic P r e s s . 1981. 77. S. F. Dyke & S. N. Quessy . E r y t h r i n a and Re l a t ed A l k a l o i d s , Chapter 1 in Manske & R o d r i g o , op_. c i t . , V o l XV I I I . 1981. 78. D. M. Smi th . The C e l a s t r a c e a e A l k a l o i d s . Chapter 4 in R. H, F. Manske (ed. ) The A l k a l o i d s V o l . XVI . 1977. 79. D. S. S e i g l e r . P lant Sys temat i c s and A l k a l o i d s . Chapter 1 in Manske, op_. c i t . , Vo l XVI . 1977. 80. R. L. C l a r k e . The Tropane A l k a l o i d s . Chapter 2 in Manske, op . c i t . , Vo l XVI . 1977. 81 . J . E. Sax ton . A l k a l o i d s of M i t r agyna and Re l a t ed S p e c i e s . Chapter 3 in Manske, pjo. c i t . , V o l . XIV. 1973. 82. J . S. B i n d r a . Ox indo le A l k a l o i d s . Chapter 2 in Manske, op. c i t . , V o l XIV. 1973. 83 . M. Shamma & R. L. Cas tenson . The Oxoaporphine A l k a l o i d s . Chapter 6 in Manske, op_. c i t . , V o l . XIV. 1 973. 84. V . S n i e c k u s . D i s t r i b u t i o n of Indo le A l k a l o i d s in P l a n t s . Chapter 1 in Manske, op . c i t . , V o l X I . 1968. 85. J . Soukup. V o c a b u l a r i o de l o s Nombres Vu lga res de l a F l o r a Peruana C o l e g i o S a l e s i a n a , L ima . 1970. 86. D. McKenna, G. H. N. Towers , & F. S. A b b o t t . Monoamine Oxidase I n h i b i t o r s in South American H a l l u c i n o g e n i c P l a n t s : Tryptamine and 0 - c a r b o l i n e C o n s t i t u e n t s of Ayahuasca .  J ou rna l of E thnopharmacology. 10:195-223. 1984 222 87. Plowman, T . B r u n f e l s i a i n E thnomed ic ine . Harvard B o t a n i c a l Museum L e a f l e t s 25:289-320. 1977. 88. I y e r , R. P., J . K. Brown, M. G. Chaubal & M. H. Malone. B r u n f e l s i a hopeana I. H i p p o c r a t i c Sc reen ing and A n t i -in f lammatory E v a l u a t i o n . J o u r n a l of Na tu r a l P roduc ts 40:356-60. 1c77. 223 APPENDIX II The f o l l o w i n g l i s t of herbar ium voucher specimens i n c l u d e s the c o l l e c t i o n data on a l l of the specimens d i s c u s s e d in the text f o r which herbar ium vouchers are a v a i l a b l e . In a d d i t i o n Appendix II i n c l u d e s any o ther p l a n t c o l l e c t i o n which was s t a t ed by in formants to have a m e d i c i n a l or e t h n o b o t a n i c a l l y s i g i f i c a n t use . Appendix II a l s o i n c l u d e s a l l c o l l e c t i o n s from the M y r i s t i c a c e a e and M a l p i g h i a c e a e , even though these may not have been u t i l i z e d in the p r e p a r a t i o n of drug samples ana l yzed in t h i s work or may not be used in t h i s manner. P l an t c o l l e c t i o n s which were not known to have e t h n o b o t a n i c a l or m e d i c i n a l s i g n i f i c a n c e have been omi t ted from t h i s c o l l e c t i o n l i s t . In i n s t a n c e s where m u l t i p l e c o l l e c t i o n s were made of the same s p e c i e s , the taxonomic a u t h o r i t y i s g iven in the f i r s t c i t a t i o n ; the a u t h o r i t y i s omi t ted i n subsequent c i t a t i o n s . TABLE XVII - LIST OF HERBARIUM VOUCHER COLLECTIONS C o l l e c t i o n # (DMK-) B1nom i a 1 & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access * V172-D u p l i c a t e Vouchers: Remarks: DMK-1 * t J u s t i c i a p e c t o r a l i s J a c q . masha-h1r i T . P1owman(F ) 175 F.USM,MO* a d d i t i v e to V i r o l a s n u f f used to t r e a t c o l d s o r e s DMK-3 B a n i s t e r i o p s i s c a a p i (Spr.ex G r i s e b ) Morton ayahuasca T . P1owman(F) 177 F.USM so u r c e of ayahuasca DMK-16* S a n c h e z i a t i g r i n a Leonard - P . Mateka i t i s ( F ) 189 F.USM,ECON UNAP leav e s smoked or brewed as ha 11uc i nogen DMK-20* P s y c h o t r i a v i r i d i s R. & P. chacruna T . P1owmanf F) 193 UBC o n l y admixture to ayahuasca o r i g . s t o c k f o r DMK-21 OMK-21* P s y c h o t r i a v i r i d i s chacruna T . P1owmanf F) 194 F.USM,ECON. see above MICH.RVH,UNAP DMK-22* T e l i o s t a c h y a l a n c e o l a t a Nees toe negro T . P1owmanf F) 195 F.USM.ECON. UNAP,MO.RVH ayahuasca admixture DMK-23* V i r o l a p a v o n i s (A. DC. )A. C. Smi th cumala W. Rodr iguesfINPA ) 196 F,USM.ECON. UNAP,INPA.RVH DMK-30* V i r o l a p a v o n i s cuma1 a W . Rodr igues(INPA ) 201 F . ECON. INPA UNAP.RVH a l k a l o i d + DMK-3 1 * P a l i c o u r e a sp. - • J . Ruiz(UNAP) 203 F,USM,MO, UNAP,RVH a l k a l o i d ++ DMK-32* V i r o l a p a v o n i s cuma1 a W . Rodrigues(INPA) 204 F.USM,ECON, a l k a l o i d -INPA.UNAP,RVH DMK-34* V i r o l a p a v o n i s cuma1 a: ku'-ru-ku W . Rodr iguesfINPA ) 205 F,USM.ECON, UNAP,MICH. INPA,MO.RVH a l k a l o i d s o u r c e of Bora p a s t e at P U . DMK-35* V i r o l a sp. cuma1 a P . M a t a k a i t i s ( F ) 206 UBC o n l y a l k a l o i d + DMK-36* V i r o l a sp. cumala P . M a t a k a i t i s ( F ) 207 UBC o n l y a l k a l o i d + DMK-37* V i r o l a sp. cuma1 a P . M a t a k a i t i s ( F ) 208 UBC o n l y a l k a l o i d + DMK-38* R l n o r a racemosa (Mart.&Zucc.IKuntze - A . Gentry(MO) 209 F , USM.ECON. UNAP.MO.RVH a l k . -; 1ichen on bark added to V i r o l a p a s t e DMK-39* Anemia sp. - R . G. S t o l z e ( F ) 210 F,USM.ECON, UNAP,RVH a l k . - ; t e a f r . l e a v e s used to cook V i r o l a p a s t e DMK-40* V i r o l a s e b l f e r a Aubl . cumala r o j a ; oo'-koey; p i r i ; W. RodriguesfINPA) 211 F.USM.MICH. INPA,MO.RVH ECON.UNAP a 1k.++:source of D.A. Moreno sample #1 T a b l e XVII (Cont'd) Col l e c t i o n # (DMK-) B1nom i a 1 & a u t h o r i t y V e r n a c u l a r Name Determ1ned by:* UBC Herbarium Access H V172-D u p l i c a t e Vouchers: Remarks: DMK-41* V i r o l a e l o n g a t a (Benth.)Warb. cuma1 a:oo'-koo-na; W. Rodr1gues(INPA) 212 F,USM,ECON, UNAP, INPA, RVH a l k a l o i d s u i t a b l e + + f o r p a s t e DMK-42 P1tyrogramma ca1ome1os(L.)L i nk - R. G. S t o l z e ( F ) 213 F.USM,UNAP RVH j u i c e of used f o r c r o z i e r s c a t a r a c t s DMK-43* Theobroma sub 1nacum Mart. macambo del monte: Bora:a'-he P. M a t a k a l t i s ( F ) 214 F.USM.ECON, UNAP,RVH ashes of added to f r u i t s V i r o l a p a s t e DMK-44* I r y a n t h e r a l o n g l f l o r a Ducke Bora : ehu-ghwa-o-oo-e W. Rodrigues(INPA) 2 15 F.USM a l k a l o i d ECON,UNAP INPA -DMK-45* V i r o l a e l o n g a t a Bora:ugr-pah-1 aye W. Rodrigues(INPA) 216 INPA . a l k a l o i d + DMK-46* V i r o l a c a l o p h y l l a Warb. Bora:kuhr'-re-ko W. Rodr i gues(INPA) 217 F.USM.ECON UNAP a l k a l o i d + ; s u i t a b l e f o r making p a s t e DMK-47* I r y a n t h e r a m a c r o p h y l l a (Bth.)Warb. Bora:hach i u r n e - e W. Rodr i gues(INPA) 219 F .USM.ECON. UNAP. INPA, a l k a l o i d RVH -DMK-48* I r y a n t h e r a u l e l Warb. Bo r a : h a c h i - e W. Rodr igues(INPA) 218 USM,INPA, RVH a l k a l o i d -DMK-49* I r y a n t h e r a c r a s s i f o l l a A. C. Smith Bora:ch i - c h i - c h W. Rodr i gues(INPA ) 220 F,USM.ECON INPA, UNAP, a 1ka1o i d RVH -DMK-50* I r y a n t h e r a j u r u e n s i s Warb. Bo r a : p i j 1 - h a h ' - e h W. Rodrigues(INPA) 221 F.USM.ECON, a l k a l o i d INPA,UNAP.RVH -DMK-51* I r y a n t h e r a p a r a e n s l s Warb. Bora:t i - t i-mueh W. Rodr igues(INPA ) 222 F.USM.ECON, a l k a l o i d INPA.UNAP.RVH -DMK-52* V i r o l a m u l t i n e r v i a Ducke Bora:kat'-so-eh W. Rodr igues(INPA) 223 F.USM.ECON, a l k a l o i d INPA.UNAP RVH -DMK-53 V i r o l a c a l o p h y l l a W1 toto:oo-koo'-na W. Rodr1gues(INPA) 224 F,USM,ECON. UNAP.INPA su i tab 1e f o r p a s t e DMK-54 V i r o l a sp. cuma1 a P. M a t a k a l t i s ( F ) 225 USM.ECON.UNAP,RVH DMK-56* V i r o l a c a l o p h y l l a W 1 toto:ti-ti-mueh W. Rodrigues(INPA) 226 F.USM.ECON INPA.UNAP. RVH a l k a l o i d (seeds & + f r u i t s ) DMK-57 Ban i s t e r i ops i s mur1cata(Cav.)Cuatr. sacha-ayahuasca B . Gates (MICH)- 227 F,USM.ECON, UNAP,MICH. RVH used to make ayahuasca ro T a b l e XVII (Cont'd) Col l e c t i o n H (DMK-) B1nomla1 & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access * V172-Dup1i ca te Vouchers: Remarks: DMK -59* V i r o l a e l o n g a t a cumala w . Rodr1gues(INPA) 229 F.USM,ECON. UNAP,MICH. INPA.MO,RVH s o u r c e of Don Marcos' ku'-ru-ku: a l k a l o i d ++ DMK -63* V i r o l a p a v o n i s cumala b l a n c a ; Bora:ku'-ru-ku w . Rodrigues(INPA) 233 F.USM.ECON. UNAP,MICH a l k . - ; s a i d s u i t a b l e f o r p a s t e DMK -64* P h i l o d e n d r o n nervosum (Schult.cV Schul t . )Kunth - T . Plowman!F) 234 F.USM.ECON, UNAP.MICH, MO,RVH admixture to V i r o l a p a s t e ; a l k a l o i d -DMK -65 Theobroma b i c o l o r H.&B. macambo; Bora:a'-hep T PIowmanfF) 235 F.USM.ECON. UNAP,M0,RVH admixture to V i r o l a p a s t e ; a l k a l o i d -DMK -66* C a l l i a n d r a a n g u s t i f o l i a Spruce - C . Niezgoda(F) 236 F.USM.UNAP, RVH tea used as s e d a t i v e ; admix, to ayahuasca DMK -67* V i r o l a e l o n g a t a cuma1 a; W1 toto:oo'-koo-na w . RodriguesfINPA) 237 F.USM,UNAP, INPA.RVH alk.+;Source of D.A. Moreno sample H2 DMK -68* V i r o l a e l o n g a t a cuma1 a; W i toto:oo'-koo-na w . Rodr iguesfINPA) 238 F.USM.UNAP. INPA,RVH. ECON a l k . + ; s o u r c e of D.A. Moreno sample #3 DMK -69* V i r o l a e l o n g a t a cuma1 a; Wi toto:oo'-koo'-na w. RodriguesfINPA) 239 F.USM.UNAP, INPA,ECON. RVH a1k.+;source of D.A. Moreno sample #4 DMK -70 V i r o l a e l o n g a t a cumala w. RodriguesfINPA) 240 F.USM.ECON. INPA.RVH so u r c e of seeds DMK -71 V i r o l a e l o n g a t a cumala w. Rodr igues(INPA) 241 F.USM.ECON. MO.INPA,UNAP so u r c e of seeds , RVH DMK -73 B a n i s t e r i o p s i s mart i n i a n a ( A d r . J u s s . )Cuatr. v a r . s u b e n e r v i a C u a t r . B. Gates(MICH) 243 F.USM,MO. ECON.MICH, UNAP,RVH DMK -74* Abuta g r a n d i f o l i a ( Mart.)Sandwith abuta,ca1m i 1111o, sanango T . P1owmanfF) 244 F.USM.ECON, UNAP,RVH admixture to ayahuasca; a l k a l o i d ++ DMK -75* V i r o l a l o r e t e n s i s A.C. Smi th cuma1 a W . Rodrigues(INPA) 245 F,USM.ECON. UNAP,MICH. INPA,MO.RVH a l k a l o i d -DMK--78* Osteoph1oem platyspermum (A.DC)Warb. - w. Rodr iguesfINPA) 248 F.USM.ECON, INPA,UNAP, RVH a l k . +; N-Me-tryptophan-m e t h y l - e s t e r i n Ivs. DMK -82* V i r o l a a l b i d i f l o r a Ducke cumala de 1os b r u j o s w. Rodr iguesfINPA) 251 F.USM.UNAP. INPA,RVH a 1ka1o i d -ho T a b l e XVII (Cont'd) Col l e c t i o n # (DMK-) B i nom i a 1 & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access V172-D u p l i c a t e Vouchers: Remarks: DMK-83* Mascagnia s1nemariens 1s ( A u b l . ) G r i s e b . - W . Anderson(MICH) 252 F,USM,UNAP, MICH,MO,RVH DMK-84* Byrsonima p o e p p i g i a n a Adr. J u s s . - W . Anderson(MICH) 253 F,USM.UNAP, MICH,MO,RVH DMK-86 B u r d a c h i a ' pr1smatocarpa Adr. J u s s . v a r . l o r e t o e n s i s Anderson W . Anderson(MICH) 255 F,USM.UNAP, MICH,MO,RVH DMK-87 Mascagnia benthamlana ( G r i s e b . ) A n d e r s o n - W . Anderson(MICH) 25G F,USM,UNAP, MICH.MO,RVH DMK-88 Mascagnia benthamiana - W . Anderson(MICH) 257 F,USM.UNAP. MICH,MO.RVH DMK-89 Mascagnia benthamiana - W . Anderson(MICH) 258 F,USM.UNAP. MICH.MO,RVH DMK-90 H e t e r o p t e r y s o r i n o c e n s i s (H.B.K.)Adr. Juss . w . Anderson(MICH) 259 F.UNAP.MICH, RVH DMK-91 St igmaphy1 Ion hypoleucum Miq - w. Anderson(MICH) 260 F,USM,UNAP. MICH.MO,RVH DMK-92 Byrsonima a r t h r o p o d a Adr. Juss . - w. Anderson(MICH) 261 F,USM.UNAP. MICH.RVH DMK-93 H e t e r o p t e r y s or i nocens1s - w. Anderson(MlCH) 262 F.UNAP.MICH. USM,MO.RVH DMK-94 H e t e r o p t e r y s or 1nocens1s - w. Anderson(MICH) 263 F.UNAP.MICH, RVH DMK-95 H e t e r o p t e r y s o r i n o c e n s i s - w. Anderson(MICH) 264 UNAP,MICH.RVH DMK-96 H e t e r o p t e r y s or 1nocens1s - w. Anderson(MICH) 265 UNAP,MICH.RVH, F,USM,MO DMK-97 B r u n f e l s i a g r a n d i f l o r a D. Don ss p . s c h u l t e s i i P1owman ch1r i csanango T . P1owman(F) 266 F,UNAP,RVH f e b r i f u g e ; a d m i x . to ayahuasca DMK-103 Byrsonima c h r y s o p h y l l a H.B.K. - W. Anderson(MICH) 272 F,USM,UNAP, MICH,RVH DMK-104 H y p t i s suaveolens (L. )Poi t. - C . Ni ezgoda(F) 273 F,USM,MO, used f o r rheumatism UNAP.RVH T a b l e XVII (Cont'd) C o l l e c t I o n n (DMK- ) B i n o m i a l & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access # V172-Dup l i c a t e Vouchers: Remarks: DMK- 107* Davi11a n i tIda ( V a h l . ) K u b . pucahuasca P . Mataka i t i s ( F ) 276 UBC o n l y DMK- 108* P s y c h o t r i a v i r i d i s yage-sem i r uca; c h a c r u n a : s u i j a T . P1owman(F) 277 F.USM,ECON, UNAP,MICH, M0-. RVH a 1k. + ;adm i x . To ayahuasca DMK- 109* P s y c h o t r i a carthagenens1s Jacq. yage;yage-chacruna P . Mataka i t i s(F ) 278 F.UNAP a 1k. -; adm i x. to ayahuasca DMK- 1 10* B a n i s t e r i o p s i s c a a p i c i e l o ayahuasca T . P1owman(F ) 279 F.UNAP ayahausca c u i t i v a r DMK- 1 1 1 B a n i s t e r i o p s i s c a a p i l u c e r o ayahuasca T . P1owman(F) 280 F.UNAP ayahuasca c u l t i v a r DMK- 1 12 B a n i s t e r i o p s i s c a a p i l u c e r o ayahuasca T . Plowman(F) 281 F,UNAP ayahuasca c u l t i v a r DMK- 1 13 Senna o c c i d e n t a l i s ( L. )L i nk retama R. Barnaby(NY) 282 UBC o n l y m e d i c i n a l : use u n c e r t a i n DMK- 1 15 P e t i v e r i a a l l i a c e a e L. mukura P . Matakai t i s ( F ) 284 F,USM.UNAP. MO,RVH used i n the bath; admix, to ayahuasca DMK- 1 16 P i p e r aduncum L. cordonc i11o W . Burger(F) 285 F,UNAP,RVH tea of l e a v e s used to t r e a t asthma S heal bones DMK- 1 17 Ocimum micranthum Wi1 Id. a l b a c a T . P1owman(F) 286 UNAP f o r the stomach; ayahuasca admixture DMK- 1 18 Hippeastrum puniceum (Lam.)Kuntze samangi T . Plowman(F) 287 UNAP f o r dropsy, & sore a r c h e s DMK- 1 19 C o r n u t i a o dorata (P.& E.)Poepp. s h i nguarana T . P1owman(F) 288 UBC o n l y admixture to ayahuasca a l k a l o i d -DMK- 120 Sambucus mexicana s a l c o P r e s l . ss p . b i p i n n a t a ( S . S C . ) S c h w e r . T . P1owman(F) 289 UNAP,RVH treatment of e p i l e p s y , cough, b r o n c h i t u s DMK- 12 1 Chenopod1um a m b r o s i o i d e s L. buseta T . P1owman(F) 290 F.USM.MO. RVH used i n b a t h DMK- 122 Maranta arundinaceae L. sh i mpanpana T . P1owman(F) 291 UNAP.RVH used i n b a t h ro ro T a b l e XVII (Cont'd) Col 1 e c t I o n f (DMK-) B i n o m i a l & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access ff V172-D u p l i c a t e Vouchers: Remarks: DMK- 123 A l p i n i a s p e c i o s a Schum m i sk i-panga P . J . M. Maas(F) 292 F,USM,ECON. MO,UNAP,RVH use u n c e r t a i n DMK- 124* B a n i s t e r i o p s i s c a a p i pucahuasca B . Gates (MICH) 293 UNAP.MICH, RVH ayahuasca c u i t i v a r DMK- 125* B a n i s t e r i o p s i s c a a p i ayahuasca B . Gates (MICH) 294 UNAP.MICH, RVH ayahuasca c u 1 1 i var DMK- 126* B a n i s t e r i o p s i s c a a p i c i e l o ayahuasca B . Gates (MICH) 295 UNAP.MICH, RVH ayahuasca c u i t i v a r DMK- 127* B a n i s t e r i o p s i s c a a p i d e l o ayahuasca B . Gates (MICH) 296 UNAP,MICH, RVH ayahuasca c u 1 1 i var DMK- 128* B a n i s t e r i o p s i s c a a p i rumi ayahuasca T . P 1 owman (F) 297 UBC o n l y ayahuasca c u 1 1 i var DMK- 129 P s y c h o t r i a v i r i d i s chacruna - ' l a hembra' form P . Matakai t i s ( F ) 298 UBC o n l y admix, to c u 1 1 i v a r ayahuasca; DMK- 130 P s y c h o t r i a v i r i d i s chacruna - ' l a hembra' form P . Mataka i t i s ( F ) 299 UBC o n l y adm i x. to c u 1 t 1 v a r ayahuasca; DMK- 131 P s y c h o t r i a v i r i d i s chacruna -- ' e l macho' form P . M a t a k a i t i s ( F ) 300 UBC o n l y admix, to c u 1 1 i v a r ayahuasca; DMK- 132 P s y c h o t r i a v i r i d i s chacruna P . M a t a k a i t i s ( F ) 301 UBC o n l y admix, to c u i t i v a r ayahuasca; DMK- 134 B r u n f e l s i a g r a n d i f l o r a D. Don ch1r 1csanango - 'grande' form T . Plowman(F ) 302 UBC o n l y adm i x. to c u 1 t i v a r ayahuasca; DMK- 135 B r u n f e l s i a g r a n d i f l o r a ssp. s c h u l t e s 1 i P1owman c h i r1csanango -'pequeno' form T . P1owmanfF) 303 UBC o n l y admix, to c u i t 1 v a r ayahuasca: DMK- 136 I r y a n t h e r a l a n c i f o l i a Ducke cuma1 a W . Rodr igues(INPA) 304 INPA s o u r c e of l a r g e seeds DMK- 137 I r y a n t h e r a j u r u e n s i s cuma1 a W. RodriguesfINPA) 305 INPA sou r c e of smal 1 seeds T a b l e XVII (Cont'd) Col l e c t i o n * (DMK-) B i nom1a 1 & a u t h o r i t y V e r n a c u l a r Name Determined by:* UBC Herbarium Access It V172-D u p l i c a t e Vouchers: Remarks: DMK-138* S p l l a n t h u s a l b a L'Her - P . Matakai t i s ( F ) 306 F,USM.MO l e a v e s chewed f o r t o o t h a c h e DMK-139* P s y c h o t r i a v i r i d i s chacruna P . Matakai t i s(F ) 307 UBC o n l y admix, to ayahuasca DMK-142 T r l p o g a n d r a g l a n d u l o s a (Seub.)Rohw. v a r . ( v e l . sp. a f f . ) - B . Faden(US) 308 F.USM,MO use u n c e r t a i n ; s o l d i n market p l a c e s i n Peru DMK-143* Cyperus d i g i t a t u s Roxb. ch1 c o r r o ; c h l c o r r o - p 1 r 1 - p i r i D. Adams(F) 309 USM t u b e r s used as ayahuasca admix.; or smoked as h a l l u c i n o g e n DMK- 144 E l e u t h e r l n e bulbosa (Ml 11)Urb. - T . P1owman(F) 310 UBC o n l y b u l b made i n t o tea to a i d i n c h i l d b i r t h * = Name of taxonomist making d e t e r m i n a t i o n . Herbarium codes a re shown i n p a r e n t h e s e s t - * i n d i c a t e s a d d i t i o n a l m a t e r i a l was c o l l e c t e d f o r phytochemical a n a l y s i s \ - H e r b a r i a where d u p l i c a t e vouchers a r e on d e p o s i t . Herbarium codes a re a c c o r d i n g to Index Herbariorum I (1974): F = F i e l d Museum of N a t u r a l H i s t o r y , Chicago, 111. UBC = Herbarium of U n i v e r s i t y of B r i t i s h Columbia ECON = Oakes Ames Economic Herbarium, Harvard B o t a n i c a l Museum, Harvard U n i v e r s i t y USM = H e r b a r l o San Marcos, Museo de H i s t o r i a N a t u r a l , Lima, Peru MICH = Herbarium of the U n i v e r s i t y of Michigan. Ann Arbor. Mich. INPA = I n s t i t u t o N a c i o n a l de P e s q u i s a s do Amazonia, Manaus, B r a z i l MO = M i s s o u r i B o t a n i c a l Garden, S t . L o u i s , Mo. US = U.S. N a t i o n a l Herbarium. Smithsonian I n s t i t u t i o n . Washington. D. C. UNAP = Herbarium of the U n i v e r s i d a d N a c i o n a l de Amazonense Peruana, I q u i t o s , Peru (not l i s t e d i n Index Herbariorum) RVH = Herbarium of R.V. H e r a c l i t u s , I n s t i t u t e of E c o t e c h i c s , A i x - e n - P r o v e n c e . France (not l i s t e d i n Index Herbariorum) 

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