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Electrophoretically mediated microanalysis for the evaluation of interspecies variation in cholinesterase metabolism
Author(s) -
Moura Joana,
Simplício Ana Luísa
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900769
Subject(s) - acetylthiocholine , microanalysis , hydrolysis , chemistry , chromatography , substrate (aquarium) , enzyme , cholinesterase , horse , metabolism , biochemistry , acetylcholinesterase , aché , biology , endocrinology , organic chemistry , paleontology , ecology
This study describes an electrophoretically mediated microanalysis method, suitable for the preclinical evaluation of the hydrolysis of ester drugs by the serum of different animals and for further characterization of human–animal correlation. Dog, cat, cow, horse, sheep, rat and human serum were diluted (25%) in the appropriate buffer and replaced the enzyme solution usually used in electrophoretically mediated microanalysis methods for the study of enzyme kinetics. They were then compared in terms of the ability to hydrolyze acetylthiocholine and butyrylthiocholine (0.25 mM) by in‐capillary reaction. Human serum afforded the highest conversion rates (52% butyryltiocholine and 34% acetylthiocholine) followed by horse (31 and 35%), dog (26 and 24%), cat (22 and 14%), rat (11 and 15%) and sheep (8 and 8%). Hydrolysis by bovine serum was negligible. The method is fast (under 8 min including rinsing steps), sensitive (under 25 μM substrate could be quantified) and repeatable (RSD≈2%), only requiring minute amounts of sample.