PMID: 3903413Nov 11, 1985

Measurement of acetylcholine and choline in brain by HPLC with electrochemical detection

Life Sciences
F P BymasterD T Wong

Abstract

A simple and rapid method for measuring acetylcholine and choline using high performance liquid chromatography (HPLC) with electrochemical detection is presented. Acetylcholine and choline were first separated using reverse-phase chromatography; acetylcholine was then hydrolyzed post-column to choline by acetylcholinesterase. Choline was oxidized enzymatically by choline oxidase to betaine and hydrogen peroxide, and the peroxide was detected electrochemically. Changes in methodology from previous procedures include a different mobile phase, controlled heating of chromatography column and post-column reaction coil, and a different extraction method for quaternary amines. The changes resulted in less inhibition of derivatizing enzymes by mobile phase, narrow and consistent elution of peaks, and a rapid and efficient extraction of quaternary amines. Measurement of acetylcholine and choline in brain tissue was found to be replicable, and the levels agreed with literature values.

Related Concepts

Choline
Amines
Hydrogen Peroxide
Alcohol Oxidoreductases
Enzymes, antithrombotic
August Rats
Chromatography
Lentiform Nucleus Structure
ACHE
Rapid Technique

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