An enzyme-reactor for electrochemical monitoring of choline and acetylcholine: applications in high-performance liquid chromatography, brain tissue, microdialysis and cerebrospinal fluid.
A sandwich-type enzyme reactor in which the enzymes are physically immobilized in a minimal dead space between two cellulose membranes, resulting in improved sensitivity, was developed for the electro-chemical detection of choline (Ch) and acetylcholine (ACh). The reactor contains the enzymes choline oxidase with or without acetylcholine esterase, for the detection of ACh and Ch, respectively. For the HPLC analysis of Ch and ACh the detection system was coupled post column. Levels of Ch and ACh of rat striatum tissue and human cerebrospinal fluid were found to be similar to those determined with published methods. Because of low back pressure--a further advantage of the reactor--the detection system could also be directly coupled to the outlet of a microdialysis device, allowing the on-line real-time measurement of extracellular brain Ch. The versatility of the enzyme reactor for the monitoring of analytes in HPLC eluates, flow injection analysis, with or without prepurification, is emphasized. The usefulness of the reactor-detector system in biomedical applications is illustrated by the measurement of increases of rat striatal extracellular Ch following cardiac arrest.
Flentge
F
F
Department of Biological Psychiatry, University of Groningen, The Netherlands.
Venema
K
K
Koch
T
T
Korf
J
J
D000109:Acetylcholine / Q000032:analysis*; D000110:Acetylcholinesterase / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000818:Animals; D001923:Brain Chemistry; D002794:Choline / Q000032:analysis* / Q000134:cerebrospinal fluid; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003342:Corpus Striatum / Q000737:chemistry; D003956:Dialysis; D004563:Electrochemistry; D004800:Enzymes, Immobilized; D051381:Rats
D004800:Enzymes, Immobilized; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D000110:Acetylcholinesterase; D002794:Choline; D000109:Acetylcholine