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  • Title: Sensitive method for HPLC determination of acetylcholine, choline and their analogues using fluorometric detection.
    Author: Rícný J, Tucek S, Vins I.
    Journal: J Neurosci Methods; 1992 Jan; 41(1):11-7. PubMed ID: 1578898.
    Abstract:
    A method is described for reversed-phase HPLC separation of acetylcholine and choline and of their homologues in tissue extracts or perfusion fluids, combined with postcolumn enzymatic derivatization and fluorometric quantification. The separation occurs on a polymeric resin derivatized with hydrophobic moiety and the mobile phase consists of Na2HPO4, 3-(p-hydroxyphenyl)propionic acid and sodium dodecylsulphate; postcolumn enzyme reactor contains immobilized acetylcholinesterase, choline oxidase, and peroxidase. The limits of detection are 1 pmol choline and 3 pmol acetylcholine per sample. The method is free of interferences encountered with electrochemical detection and well suited for non-attended automatic operation.
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