Internal standard method for the measurement of choline and acetylcholine by capillary electrophoresis with electrochemical detection |
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Authors: | Wise Dana D Barkhimer Tatyana V Brault Pierre-Alexandre Kirchhoff Jon R Messer William S Hudson Richard A |
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Affiliation: | Department of Chemistry, College of Arts & Sciences, University of Toledo, Toledo, OH 43606-3390, USA. |
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Abstract: | An internal standard method has been developed for the determination of the neurotransmitter acetylcholine and/or its metabolic precursor choline. This approach couples the high separation efficiency of capillary electrophoresis with the sensitivity and selectivity of electrochemical detection at an enzyme-modified electrode. Indirect electrochemical detection is accomplished at a 25 microm platinum electrode modified by cross-linking the enzymes choline oxidase and acetylcholinesterase with glutaraldehyde. Although in this simple form of electrode fabrication there is a gradual loss of response from the electrochemical detector with time, accurate quantitation is achieved by the addition of butyrylcholine, which is also a substrate for acetylcholinesterase, as an internal standard. A linear response is achieved between 0 and 125 microM with a limit of detection of 2 microM (25 fmol). The utility of this method was demonstrated by monitoring the kinetics of choline uptake in synaptosomal preparations. |
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Keywords: | Choline Acetylcholine |
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