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Capillary electrophoresis-mass spectrometry for the analysis of neurochemicals Kowalczyk, Adam
Abstract
In order to quantify low concentrations of dopamine in complex biological matrices, a capillary electrophoresis-mass spectrometry pH stacking method was developed and optimized. The pH stacking method operates by concentrating the analyte on-capillary through manipulation of electrophoretic mobility in regions of different pH. This method allowed for a detection limit of 0.1 ng/mL, approximately 2 orders of magnitude lower than using a more traditional capillary zone electrophoresis (non-stacking) method. The stacking method was determined to be linear from 0.5-1000 ng/mL dopamine with accuracy values within 10% and precision values within 5% across the range. A CE compatible method for the extraction of dopamine from intracellular and extracellular cell culture components was also developed. This method was then applied to the analysis of dopamine in STHdh cells, a model for Huntington’s disease, showing a lower concentration of dopamine in disease cells compared to normal phenotype, consistent with current literature data on Huntington’s disease. In addition, steps were taken toward the development of a CE-MS method for the measurement of uridine diphosphate sugars in cerebrospinal fluid with advantages including short analysis time (<20 mins) and low sample consumption.
Item Metadata
Title |
Capillary electrophoresis-mass spectrometry for the analysis of neurochemicals
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2020
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Description |
In order to quantify low concentrations of dopamine in complex biological matrices, a capillary electrophoresis-mass spectrometry pH stacking method was developed and optimized. The pH stacking method operates by concentrating the analyte on-capillary through manipulation of electrophoretic mobility in regions of different pH. This method allowed for a detection limit of 0.1 ng/mL, approximately 2 orders of magnitude lower than using a more traditional capillary zone electrophoresis (non-stacking) method. The stacking method was determined to be linear from 0.5-1000 ng/mL dopamine with accuracy values within 10% and precision values within 5% across the range. A CE compatible method for the extraction of dopamine from intracellular and extracellular cell culture components was also developed. This method was then applied to the analysis of dopamine in STHdh cells, a model for Huntington’s disease, showing a lower concentration of dopamine in disease cells compared to normal phenotype, consistent with current literature data on Huntington’s disease. In addition, steps were taken toward the development of a CE-MS method for the measurement of uridine diphosphate sugars in cerebrospinal fluid with advantages including short analysis time (<20 mins) and low sample consumption.
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Genre | |
Type | |
Language |
eng
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Date Available |
2022-04-30
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Provider |
Vancouver : University of British Columbia Library
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Rights |
Attribution-NonCommercial-NoDerivatives 4.0 International
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DOI |
10.14288/1.0389803
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
2020-05
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Campus | |
Scholarly Level |
Graduate
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Rights URI | |
Aggregated Source Repository |
DSpace
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Rights
Attribution-NonCommercial-NoDerivatives 4.0 International