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An investigation into the cascara content of the wood and bark of Rhamnus Purshiana Gillie, Kenneth Beresford 1923

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AJT IHVESTIGATIOI IITO THE CASCARA GOMTEM! OF THE WOOD AED BARK OF RHAMFUS PURSHIAIA by Kenneth Beresford G i l l i e A Thesis submitted f o r the Degree of MASTER OF APPLIED SCE3NCE i n the Department of CHEMISTRY TEE UNIVERSITY OF BRITISH COLUMBIA APRIL, 1923 AN INVESTIGATION INTO THE GASCARA CONTENT OF THE WOOD AND BARK OF RHAMNUS PURSHIANA. CONTENTS;-Introduction. Objects of the Present Work. Procedures Used i n This Investigation* Explanations of Tables* Tabulated P a r t i c u l a r s of Samples including Results of Physiological Tests. Tabulated Results of Determinations of Physical and Chemical Properties. Meteorological Reports. Conclusions. Bibliography. (1) A! HYESTIGATIOH IHIO THE CASCARA. CONTEIT OF THE WOOD AID BARE OF RHA1MUS PURSHIAIA, INTRODUCTION. Gascara Sagrada, the bark of Rhamnus Purshiana, a tree native to the P a c i f i c slope of North America, contains a purgative drug used to a considerable extent i n medicine; e s p e c i a l l y i n th© treatment of chronic constip-ation. I t s action i s superior to that of most purgatives, i n that i t i s mildly laxative and has a tonic e f f e c t on the system. The name i s Spanish and translated means, "Sacred Bark", The aborigines of the region i n which the tree grows used the bark for medicinal purposes for many years; but i t was only i n 1887 that i t attracted any not-ice from the medical profession, on being investigated by l r , J.H. Bundy (Ref, Lab. of the Inland Revenue Dept. B u l l , l o , 586 Page &). Since that time i t s use has become very general, and great quantities of the bark are consumed every year i n the manufacture of various laxative prepar-ations both l i q u i d and s o l i d . The res u l t of t h i s popular-i t y and of the r e s t r i c t e d area i n which the trees are found has caused a depletion of the world's known supply, f h i s has not as yet appreciably affected th© priee paid f o r th© bark; ana the work of indiscriminate e x p l o i t a t i o n goes on unchecked. A few people are beginning to not© the (2) danger of the s i t u a t i o n , and the cry f o r the conservation and repletion of the supply i s growing continually louder. The supply of hark i n the United States i s ra p i d l y being exhausted, and that of B r i t i s h Columbia, the only other one available, i s now being reduced i n the same reckless manner. I t i s estimated that 4,000 tons of dried bark are being shipped annually from B.C. When one consid-ers that the average tree produces only between ten and twenty pounds of bark, i t i s easy to r e a l i z e the r a p i d i t y with which t h i s valuable natural resource i s being consumed. The c o l l e c t i n g of the bark i s carr i e d on between May and August; and consists of f e l l i n g the trees, trimming o f f the branches, and stripping the bark from the trunks and larger limbs. The bark i s then l a i d out to dry s u f f i c i e n t l y to prevent i t s decaying when i t i s sacked for shipment. Mr. Hope, a landowner i n the Eraser Valley, who has been interested i n t h i s industry f or several years, and who was one of the f i r s t to note the approaching short-age, states that the stumps of trees which are s u f f i c i e n t l y exposed to sunlight w i l l send up shoots, which w i l l , i n the course of time, develop, into trees of commercial dimens-ions. In those locations which are not exposed to the direct rays of the sun, as i s generally the case, he claims that the stumps w i l l die. A l l t h i s goes to show that the present methods w i l l r a p i d l y exhaust the supply; but precautions can be taken to avoid or at least delay such a (3) catastrophe. The active p r i n c i p l e contained i n the bark has not yet been i d e n t i f i e d ; but the following substances have been i s o l a t e d i n extracts of the bark :- Emodin, ce r t a i n resins, tannic acid, o x a l i c and malic acids,- c e r t a i n o i l s and wax. (Hef. laboratories of the Dept. of Inland Revenue B u l l e t i n , Ho. 386 "Gascara Sagrada" page 7 ) . A considerable amount of work has been done on Oascara Sagra-da but the r e s u l t s obtained have not added greatly to the exact knowledge of the subject. The reason for t h i s l i e s i n i t s extreme complexity. The active p r i n c i p l e has never been i s o l a t e d and i t s true character i s not known, although much labor has been expended along t h i s l i n e . Dr. Jowett working i n the laboratories of the Burroughs, Wellcome Go.t c a r r i e d out an extremely searching i n v e s t i g a t i o n i n an endeavor to i s o l a t e the active p r i n c i p l e * He found that the active constituent or constituents are contained i n that portion of the lead subacetate p r e c i p i t a t e extracted by ethyl acetate and which i s soluble i n water and i n alcohol (Hef. Chemical Examination of Gascara Bark by H.A.D. Jowett, D.So., The Wellcome Research laboratories, 6 Zing I t . , Snow H i l l , london, E.G. page 8 ) . The present work was undertaken with the object of discovering some means of increasing the world's supply of the raw material f o r the manufacture of extracts of Gascara #agrada and also of fi n d i n g some means of overcoming (4) the griping action claimed to r e s u l t from the use of extracts manufactured from fresh "bark, I take t h i s opportunity of extending my sincerest thanks to th© "Honorary Advisory Council of Research of Canada" for making possible the carrying out of t h i s i n v e s t i g a t i o n by granting f i n a n c i a l a i d , and to Dr. Hutch-of inson, Professor'Botany, and Mr. John Davidson, Assistant Professor of Botany, for t h e i r assistance i n c o l l e c t i n g and i d e n t i f y i n g the samples used. (5) OBJECTS OF THE PRESEflT WORK. This inv e s t i g a t i o n was undertaken with the following detailed objectives i n view To discover whether or not an extract made from the wood of a tree w i l l have s u f f i c i e n t a c t i v i t y to make i t s use as a source of t h i s l a x a t i v e of commercial importance• To f i n d means of curing the extracts chemically which would obviate the necessity of holding the bark f o r a period varying from one to three years before extracting, as i s done at present, i n order to eliminate the griping action which i s claimed to be c h a r a c t e r i s t i c of extracts from fresh bark. To note differences i n the a c t i v i t y of extracts due to varying ages and habitats of trees from which they were made. To determine whether or nat the active p r i n c i p l e i s a glucoside. To discover some physical or chemical property of extracts which may be used i n judging with some degree of accuracy the comparative concentrations of the active constituent. (6) PROCEDURES USES IN THIS INVESTIGATION The extracts from the bark were made i n s t r i c t accordance with the d e t a i l s for the manufacture of F l u i d Extract of Cascara Sagrada, (Ref. U.S.P.TX page 17?) using Type Process D (Ref. U.S.P.TX page 176) f o r percolation. In the case of extracts from wood the only change consist-ed i n the substitution of pulverized wood for bark; the weights taken remained the same. The wood was reduced to powder foirm by planing across the grain with a power planer and then rubbing the dry shavings between the hands. 1 gm. of wood or bark made 1 ec. of extract containing 25% alcohol, by volume. I t should here be mentioned that the percolation was done i n two ways. In one case the b o i l i n g water was added to the powdered bark and allowed to percolate through i t into a f l a s k which was connected to the percolator by a tube; when s u f f i c i e n t of the l i q u o r had found i t s way into the f l a s k heat was applied to t h i s vessel and the percolate evaporated the steam condensing i n the percolator and when the volume became small i n the f l a s k the flame was removed and the l i q u o r allowed to run back into the f l a s k , addit-i o n a l water was added to the percolator from time to time to keep up the volume. In the other method b o i l i n g water was added to the contents of the percolator and collected i n a large container while fresh water was continually (7) added u n t i l the extraction was complete when the whole volume was evaporated down to the required concentration, the f i r s t method was much faster than the second as the l i v e steam acted as a mechanical s t i r r e r f o r the contents of the percolator and at the same time the temperature was kept at a higher point; i n addition there was l e f t a much smaller volume to be evaporated i n the next step of the manufacture of the extracts. Sine© i t appeared to be common b e l i e f that extracts made from fre s h l y c o l l e c t e d bark gave a vio l e n t g r i p i n g action when administered to patients, (Hef. Chemistry and Analysis of Drugs and Medicines, by Henry 0. F u l l e r , B.S., page 372 and also Parke, Davis & Go. of Detroit, and the H.Z. l u l f o r d Co. of Philadelphia) an attempt was made to chemically cure the extracts i n order to overcome t h i s action and thus avoid the present practice of holding the bark for a period of from one to three years. The d i f f i c u l t y that arose i n t h i s connection was most unexpect-ed. I t lay i n the fact that from a l l the samples examined only one was found to produce the gr i p i n g effect f o r cer t a i n . This l e d to the manufacture of extracts from the bark of trees which were i n a diseased state and growing i n widely varying l o c a l i t i e s . The proposed chemical treatment of the extracts consisted i n adding to them, during the evaporation of the (8) percolate to the required volume, one pound of 3$ hydrogen peroxide and continuing the evaporation i n the usual manner. The reasoning behind t h i s treatment was that since present commercial practice consists i n storing th© bark for a long period, the only possible reaction taking place must have been one of oxidation due to the oxygen of the a i r . j>00 cc. of extraet were made i n each of these cases. I t was thought possible that some of the physical or chemical properties (Ref. Lab. of the Inland Revenue Dept. B u l l e t i n 386 "Gascara Sagrada") might have some direct r e l a t i o n s h i p to the a c t i v i t y of the extracts. I f t h i s proved to be the cawe a more or l e s s easy method of comparative analysis of the extracts would be possible. The following were th© properties with the methods of deter-mination used :-(1) S p e c i f i c Gravity - to be determined by means of a S p e c i f i c Gravity b o t t l e . (2) % Solids - to be determined by the evaporation of 10 cc. of extract at 100° C., to constant weight, a f t e r dessieation over calcium chloride. Th© formula used for th© c a l c u l a t i o n of the percentage s o l i d s was 1 x 100 r % Solids - where "W" i s V x S p e c i f i c Gravity the weight of the residue i n grams and Y i s the volume of the extract taken, i n eubie centimeters. (3) % Ash - to be found by i g n i t i n g and c a l c i n i n g the residue from procedure $2 to constant weight using the (?) following expression f o r the c a l c u l a t i o n of the r e s u l t s : -Wi x 100 - % Ash V x Specific Gravity Where i s the weight of ash i n grams. (4) The Manganese Number - to be derived by determin-ing c o l o r i m e t r i e s l l y the amount of manganese present i n the ash from procedure §5 by leaching i t with 10 cc. of one to one n i t r i c acid and washing the residue with 10 cc. of d i s t i l l e d water which was then added to the n i t r i c acid so l u t i o n . The c h a r a c t e r i s t i c permanganate color was developed by oxid i z i n g the solution with Ammonium Persul-phate using a s i l v e r n i t r a t e solution as a c a t a l y s t , fhese solutions were then compared c o l o r i m e t r i c a l l y with a standard Potassium permanganate Solution which had been reduced and oxidized again by the same method as the n i t r i c a cid solutions i n order to put them i n the same state of oxidation. Then by using the following expression the Manganese number was determined:-Wg x 100000 r Manganese Number. V x Spe c i f i c Gravity Where Wg i s the weight of the Manganese i n grams. (5) The % Reducing Sugars before Hydrolysis - to be determined as dextrose using F e h l i n g ^ Solution (Ref. A. 0, A. C , Methods of Analysis, 1920 e d i t i o n , Page 17, Section 1$) together with the volumetric permanganate (10) method f o r determining the cuprous oxide (Ref. A. 0. A. 0 . , Jfethoas of Analysis, 1920 E d i t i o n Page 80, Section 29) and also A l l i o n ' s fables f o r the determination of Dextrose (Ref. A. 0. A. C*, Methods of Analysis, 1920 E d i t i o n , Pages 91 - 94)• The permanganate sol u t i o n was standard-ized by means of a known solution of Bureau of Standards dextrose and an accuracy of better than 0*5% obtained on ten determinations made on t h i s s o l u t i o n . (6) % Reducing Sugars a f t e r Hydrolysis - to be deter-mined by hydrolysing 2.3 cc. of extract with one drop of concentrated hydrochloric a c i d , by b o i l i n g i t f o r f i v e minutes and allowing i t to cool, and then determining th© percent of reducing sugars as dextrose as i n procedure l o d e f i n i t e information i s to be had concerning the nature of the active ingredient of extracts of ©aseara Sagrada. In order to prove whether or not the active p r i n c i p l e i s a glucoside as claimed by Dohme and Englehart and denied by Jowett; (Ref. "Chemical Examination of Gascara Bark" by H.A.D. Jowett, D.Sc, Th© Wellcome Chem-i c a l Research Laboratories, 6 Zing St*, Snow H i l l , London, E.G., Pages 4 - 8 ) , i t was thought that i f the active p r i n c i p l e were a glucoside complete hydrolysis should destroy i t s a c t i v i t y . In order to test t h i s idea a sample of bark was extracted and the percolate divided into two equal parts. One h a l f was treated i n the usual manner and (11) made up as an ordinary extract for the purpose of checking the other, which was treated by b o i l i n g with a known quantity of hydrochloric acid, and then n e u t r a l i z i n g t h i s with sodium bicarbonate. This sample was then made up as an ordinary extract as regards volume and alcohol content. Hydrochloric rather than sulphuric a c i d was used as i t was desired that the products of n e u t r a l i z a t i o n should have no la x a t i v e e f f e c t s and t h i s could not be claimed for most sulphates, since they have a lax a t i v e action due to the fact that the w a l l s of the stomach and int e s t i n e s act as a semi-permeable membrane to these s a l t s and causes the body f l u i d s to permeate ^nto the stomach and make the contents thereof more f l u i d and thereby f a c i l i t a t e s the evacuation (Ref. "The P r i n c i p l e of Inorganic Chemistry" by Wilhelm Ostwald, translated by Alexander Findlay, M.A., Ph.D., D.Sc, Fourth E d i t i o n , Page 512.) In order to carry out properly controlled and superintended physiological tests on the extracts i t was necessary to make arrangements with the Vancouver General Hospital for t h e i r cooperation i n the work. This favor was r e a d i l y obtained and I wish to take t h i s opportunity to thank the members of the s t a f f for t h e i r ready and e f f i c i e n t a i d . In t h i s connection I desire especially to mention Drs. Strong, Rady and Leeson, under whose immediate charge the work was placed at d i f f e r e n t times. (12) The method employed "by the h o s p i t a l i n te s t i n g th© extracts consisted i n administering doses of one cubic eentimeter to th© patients, without d i s t i n c t i o n as to s©.xt only when a laxative was required by them. Account was kept of the doses given and also of the results obtained, but no notice was taken of the number of motions of the bowels caused by a single administration of the extract. The e f f i c i e n c y was calculated by di v i d i n g the number of eff e c t u a l doses into the t o t a l number of doses given for each extract. (13) EXFLAWATIOUS OJ TABLES. Table X sets f o r t h the time of year at which, and l o c a l i t y from which the samples were coll e c t e d combined with p a r t i c u l a r s of the extracts from the stand point of the materials from which they were made and the re s u l t s of the physiological tests. Table XX gives the re s u l t s of the determinations of physical and chemical properties of the extracts and also the physiological t e s t s . Table 111 gives the average p r e c i p i t a t i o n and temperatures by the month during the three years 1920 -21 and 22. This data was obtained at the request of the Department of Botany In order that they might make botanical deductions from the information obtained i n t h i s i n v e s t i g a t i o n . TABLE J PARTI 0H1ARS OF MATERIALS 3TS0M WHIQH EXTRACTS WERE MADE COMBINED WITH RKSP1TS OJ EMSIOLOSIOAl TESTS. Q a ra Date Oolleoted Looation Q> 60 -4 Diameter Material used for extracts Particulars Times idmlnisterei H 0 a si a o H <D EH <H m O a J 1 Botes on Action Remarks. A October 1981 Eraser River n'r Tano'r. 6-10 Unknown Bark Chemically Treated to prevent Griping. (1) 100 72 72 Ho Sriping. B n n n IT n n 6-10 IT Wood Outer layer of Wood from same tree as nA n 29 18 68 n n 0 " n IT IT IT IT 6-10 Tl Middle layer do. 86 19 73 » IT D n IT n n IT n 6-10 n " Inner layer do. 46 36 78 n il 1 nn IT IT IT IT IT 6 i7 Tl Wood of Whole Trunk 89 48 64 TT IT 2 September ™ Port Haney 25 it IT : " IT n 65 33 60 Tl IT 3 n IT IT IT 65 n n ! n n n 16 4 25 Tl Tl 4 n IT IT n 65 Baric Stripped off a large IJranoh. 20 12 60 1 n 5 Hovember " Tacant lot In Tano'r. 15 Rapid Growth IT Stripped off Trunk. 74 33 44 Muoh " Bo bark l e f t to make ohemioally 6 n n IT IT IT || 14 Unknown Tl Chemically untreated. This tree growing next tn *efi » 7 n n IT n IT IT 14 n IT " treated to frevent griping. (1) 43 30 70 Mo " e Mareh 1922 IT II IT Tl 15 TI Wood ood from same tree (a #5. 29 6 SI Slight " 9 i IT n IT IT IT IT 14 IT lame as #6 -ooarsely Iround. 9 5 56 Sausea & Sriping. Sriping probably psyohologioal sinoa #10 gives none. (1) Treated with Hydrogen peroxide daring evaporatin of the peroolate. TABLE x. - 2 -• o S 3 » r-l 1 CO Sate Colleoted Looatlon <o <l u ID * » to A HhDl 0) Of> •H<H O h OJ OlJ M Particulars <a U ta «D as 1 H a d ;gp S t "« M t» o a ~ 1° -S.fi o T* <H •H notes on Action Remakka. 10 Karon 1922 Vaoant Lot in Vano'r. 14 Unknown Wood Same as #6 - finely ground 28 •81 76 Mo Griping 11 April " Fraser River n'r Vano'r. a l l ages n Bark Uormal Extract. 12 7 58 it n 12 n n n n n it n it n Completely Hydrolysed (2) 12 10 83 it it 13 ft n fl it if n n it n Extracted with Absolute Aloohol. 26 17 65 Ilausea & Griping 14 Sept. 1921 Port Haney 65 n if Trunk of tree -chemic-ally treated to prevent griping. (l) 45 36 80 Slight Griping Griping prdfcbly psyohologioal Since #15 fflvss nnna-16 n it if n 65 it n Trunk of tree same as #14* Ohem. untreated. 21 11 52 No Griping 16 May 1922 Port Haney 22 Hapid Wood Mormal Extract 17 it n ii it 24 Unknown ft Sap not running in this tree. 13 if n n n 22 IT Bark Same tree as # 16. 19 n rr n n 24 n n ft " # 17. SO n IT n n Unknown n if Tree had been ringed i 21 August 1922 Yennadon P.O. 10-12 i i n - 2 " Wood Normal Extract 22 n n it n 10-12 i i " - 2 " Bark n it 23 t( n it n 11-14 2i"-3" Wood — j .— n n i 24 if n ft it 11-14 2i"-3" Bark i 25 ff n it n 16 6&" Wood j ff fl fl) Treated with Hydrogen peroxide during evaporation of the percolate. <S) Hydrolysed by boiling with hrimihi this neutralized with sodium bicarbonate. - jarooni orio aold and TABLE r. - 3 -o a CD H J m Date Oolleoted Looatlon a> 60 <•! Diameter Material used for extraots Particulars —IB-ID (H to 10 ID 43 i s iH 0 •rl 1 tOfi gp a-P •rl O <H Pi o a ID _ T I * o • H <H •H Botes on Action Hemaks 26 August 1922 Yennadon, P.O. 16 6 i " Bark Normal Extraot 27 TT n it it 21 8 " Wood T* It 28 II II n it 21 8 " Bark n n 29 n n n n 24 4" Wood n it 30 n IT n n 24 4" Bark W tl 31 n n n it 25 9" Wood TT It 32 fi n it ft 25 9" Bark ff n 33 n n it n 25 10f" Wood tl IT 34 n ft ti n 25 10f" Bark IT TT 35 rt ft it it 36 16" Wood It TT 36 n n it it 36 16" Bark IT IT 37 nil n IT It 55 ei" Wood IT TT 38 it it It IT 55 8 * " Bark fl fl 39 it n IT II 15 5" Wood ff ff 40 n it ft it 15 5" Bark ff ft 41 n n n it 36 16" Bark Same tree as #35 & 36 -oompletely Hydrolysed. TABLE T l . .RESULTS OF DETERMINATIONS OF PHYSICAL AM CHEMICAL PROPERTIES AMD PHYSIOLOGICAL TESTS. Sample Ho. o •*» •H -rt O ^ ft g] P i ft ra e> i Solids •4 >*. % Reducing Sugars "before Hydrolysis % Reducing Sugars after Hydrolysis Manganese Humber % Mangan-ese in Wood or Bark f„ Ash in Wood or Bark to u CD ro-p CD m -rl0 Time Effectual 0 El CD •H •H >H m Remarks A 1.069 13.16 1.50 4.66 5.19 285 .00961 6.394 loo 72 72 Ho Sriping B 0.9825 5.15 O.456 0.507 1.50 374 29 18 62 n 11 C 1.002 3.28 0.285 0.524 1.35 239 26 19 73 li n D O.9865 4.22 0.267 0.675 1.15 201 46 36 78 11 11 1 0.9900 5.63 0.300 1.04 2.22 136 .00616 O.636I 89 48 54 II 11 2 0.9935 6.66 0.254 1.12 1.44 118 .OO638 0.5738 55 33 60 11 n 3 0.9840 4.09 0.2J4 0.668 0.920 253 .00683 0.4295 16 4 25 n tl 4 1.077 26.50 1.00 4.20 5.73 255 .00983 7.558 20 12 60 II ii 5 1.043 18.50 O.958 2.24 5.25 578 .01073 3.235 74 33 44 Mich Sriping 6 I . O 8 5 30.30 1.48 5.82 6.63 663 7 1.084 33.10 1.58 5.74 6.65 666 43 30 70 Ho Sriping 8 0.9913 4.78 0.261 0.861 1.11 368 29 6 21 Slight Sriping ? 0.9927 5.71 0.376 1.27 I .85 539 9 5 56 Ho Sriping 10 O.9898 5.01 0.259 1.145 1.75 406 28 21 75 11 n 11 1.036 18.60 1.68 3.13 3.71 294 12 7 58 n 11 12 1.045 17.80 1.76 5.73 5.77 286 12 10 83 Hausea & Sriping. T A M E T T . - 2. Sample So. Speoifio Gravity % Solids % Seducing Sugars before Hydrolysis f. Reducing Sugars after Hydrolysis CD to <D U "A <D OS rQ bQ & g* f. Mangan-ese in Wood or Bark •H O M to 0 a) «l 0 pq Times Administere1 1 8 3 - H 0 H <H w >. a 0 •H •H •H <H 1 Remarks 13 0.9470 11.90 0.123 2.28 3.56 000 26 17 65 Slight Griping 14 1.033 18.40 0.490 3.73 - 288 .00949 6.081 45 36 80 Ho Griping 15 1.084 22.60 0.736 5.43 - 247 .00949 6.081 21 11 52 16 O.9885 6.72 0.305 I .65 2.05 125 17 0.9845 6.10 0.268 1.31 1.63 280 18 1.077 28.40 1.29 4.74 5.73 677 19 1.086 36.40 0.887 4.78 7.11 317 20 1.080 36.20 0.936 5.35 7.28 720 21 O.9888 4.91 0.297 0.643 1.28 97 22 1.049 22.30 0.861 4.63 5.05 163 23 O.9885 4.51 0.276 0.954 1.50 147 24 I.058 24.5 1.07 5.44 7.08 334 25 0.9872 4.65 0.266 1.23 1.49 124 26 1.075 31.0 0.952 5.15 6.01 262 27 0.9866 3.90 0.251 O.863 1.49 131 28 1.078 29.7 1.02 5.77 8.10 285 29 0.9788 3.09 0.145 0.936 1.59 110 TABLE IT. - 3 -Sample ITo. Specific Gravity % Solids •d ta < > Heduoing Sugars before Hydrolysis % Heduoing Sugars after Hydrolysis 0 m © u a © if 1 a man °£ d © .rl a! a to 0 pq CD O a M •H O M to 0 03 -Horn 1 H <D ta -p © tO irj S ' r l © •H a Time Effectual Efficiency 1 Remarks 3 0 1 . 0 8 5 2 9 . 1 0 . 9 7 5 5 . 5 6 7 . 8 5 265 n 0.?8l2 1 .60 0 . 170 0.73? 1 .26 93 3 2 1.076 3 2 . 9 0 .846 5 . 5 5 8 . 5 2 206 33 O .9909 6.11 0.342 1 . 3 3 l.?3 105 34 1 . 1 0 5 3 6 . 7 0 . 8 4 5 7.41 11.24 2?3 3 5 0 . 9 8 4 8 3 . 2 0 0.227 O.855 1 .36 96 36 1.064 32.6 0 .920 6.18 7 .88 285 37 38 1 .067 30.2 1 . 0 2 5 . 3 2 6 . 5 7 280 3? O .9882 4.14 0 .150 0 . 8 2 5 I . 0 5 88 40 1 .067 2 8 . 7 0 .651 4 .25 7 . 6 6 213 41 " 1 . 106 33.3 1 .36 7 .65 7 . 6 5 273 TABLE TUT METEOROLOGICAL REPORTS. , Av f (Total g. Raindl Preoipit f a l l ;ation) Av'g. , Temperal 1 bures Month 1920 1921 1922 1920 1921 1922 January 8.92 9.39 3.13 36.1 37.5 33.9 February 1.21 6.66 4.75 38.6 40.2 34.6 March 5.20 2.33 3.44 42.9 42.1 40.7 A p r i l 2.51 3.62 2.63 45.6 46.4 46.0 Bay 1.94 2.52 2.46 51.4 53.7 53.8 June 3.06 3.64 .17 57.9 59.5 61.8 July .67 .32 .02 64.9 61.6 64.0 August 2.91 2.84 2.01 64. 61.7 62.3 September 10.37 5.03 5.76 55.6 55- 56.7 October 8.34 10.08 3.26 47.5 50.2 51.0 November 7.14 8.99 2.63 44.3 42.1 41.0 December 11.01 3.36 10.35 41. 34.7 33.8 TOTALS 63.28 61.18 40.63 49.1 48.7 48.3 ( 14) QQHOIiUSIOgS. fable T shows that the wood contains s u f f i c i e n t of the active constituent to make i t a source of commer-c i a l extracts. The average e f f i c i e n c y of extracts from the wood i s J?6f. while that of extracts from the bark i s This shows that a means of increasing the world's supply of material from which to manufacture extracts of Cascara has been obtained* From the experiments c a r r i e d out f o r the purpose of overcoming the g r i p i n g a c t i o n supposedly e x i s t i n g i n extracts made from fresh bark; the f i r s t deduction must be, that t h i s e f f e c t i s f a r from being common. Although reported to be present i n several cases as shown i n Tables T and XT only i n one of these was the evidence considered s u f f i c i e n t to warrant any degree of certainty. In the case of extract #6 a cure was obtained by treating the percolate with hydrogen peroxide. The work done i n t h i s investigation has c l e a r l y shown that the active constituent i s not a glueoside. The a c t i v i t y however may be due to the products of hydrol-y s i s of such a compound since from the increase i n sugar content on hydrolysis, i n every case, i t would appear that glueosides are present i n the extracts. In the oases of hydrolysed samples an increase i n a c t i v i t y was (15) found i n every ease but t h i s may have been due to the v a r i a t i o n bound to arise from physiological methods of t e s t i n g . In the case of an extract made with dry alcohol as a menstruum the a c t i v i t y was greater than that of the check sample but less than that of the completely hydrol-ysed extract. (Ref. fable I Samples No. 11, 12 & 13)* fhe a c t i v i t y of extracts made from samples of bark from trees of various ages shows that age i s to some extent a factor i n the regulation of t h i s property. As the age increases the a c t i v i t y f a l l s o f f somewhat, fhe habitat however has a much more pronounced ef f e c t as may be seen i n the ease of extracts made from trees growing on a vacant l o t i n the c i t y of Vancouver (Ref. fable T Samples No? 5 & % )• None of the physical or chemical properties determined show any direct relationship to the a c t i v i t y of the extracts. BIBLIOGRAPHY Laboratory of the Department of Inland Revenue, B u l l e t i n No, 386, "Cascara Sagrada". Published by the Dominion Government, Chemical Examination of Cascara Bark, by H.A.D. Jowett, D.Sc, The Wellcome Research Laboratories, 6 Zing St., Snow H i l l , London, E. U.S. Pharmacopoeia, T Z . 1916 e d i t i o n . Agents: P. Blakiston* s Son & Company, Philadelphia. Chemistry and Analysis of Drugs and Medicines by Henry C. F u l l e r , B.S., published by John Wiley & Sons, Inc., New York. O f f i c i a l and Tentative Methods of Analysis of the Association of O f f i c i a l A g r i c u l t u r a l Chemists, as compiled by the Committee on Revisions of Methods, 1920 E d i t i o n . Published by the Association of O f f i c i a l A g r i c u l t u r a l Chemists at Washington, D.C» 

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