The identification of acetylcholine and choline in oat seedlings by gas chromatography and nuclear magnetic resonance (NMR)

Abstract

Four methods of isolation and purification of choline esters from green 7-day-old oat ( <em>Avena sativa</em> L. cv. Diadem) seedlings were tested The results showed that the best recovery of acetylcholine and choline from plant tissues was obtained using an extraction solution composed of 15% 1N formic acid and 85%, acetone followed by precipitation of both these substances with ammonium reineckate. The presence of acetylcholine and choline in the plant extracts was confirmed by nuclear magnetic resonance (NMR) and gas chromatography. In the case of gas chromatography, after isolation and purification of the studied compounds from the plant material, estrification of choline followed by N-demethylation of acetylcholine and estrified choline were performed The demethylation reaction was conducted in a reaction mixture of 50 mM sodium thiophenolate and 25 mM thiophenol in anhydrous acetone. After its completion, the mixture was removed with pentanone and the demethylated esters were extracted into chloroform.