ALKALOID CONTENT OF REED CANARYGRASS (Phalaris arundinaceae L.) AS DETERMINED BY GAS-LIQUID CHROMATOGRAPHY

A method using gas-liquid chromatography was developed to measure three alkaloids—gramine, hordenine, and 5-methoxy-N,N-dimethyl tryptamine (5MeO-DMT)—in reed canarygrass. The extraction process involved methanol with ammonium hydroxide, followed by acid and buffer steps, purification with chloroform, and derivatization before separation on a chromatographic column. Recovery of the alkaloids averaged 90%. When applied to five cultivars from two locations, gramine content was higher in Rival than Venture, with others intermediate; hordenine showed no cultivar differences, and no cultivar had detectable 5MeO-DMT. Regrowth had higher gramine than initial growth, but hordenine did not differ between growth stages.

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Abstract

A technique was developed for the quantification of gramine, hordenine and 5-methoxy-N,N-dimethyl tryptamine (5MeO-DMT) in reed canarygrass (RCG) using gas–liquid chromatography. One gram of freeze-dried, ground RCG sample was extracted with methanol containing 1% ammonium hydroxide. Following solvent evaporation, the residue was extracted with dilute hydrochloric acid (0.01 N). The pH of the extract was then raised and stabilized by adding 0.5 M sodium tetraborate-boric acid buffer pH 9. The internal standard solution (0.25 mg mL −1 ) n-nonadecane was added and the extract was purified by partitioning with chloroform. The chloroform was then evaporated and the alkaloids were derivatized with bis(trimethylsilyl)-trifluoroacetamide and effectively separated on a gas chromatographic column packed with 2% OV-7 on Chromosorb W. Derivatization was effective at 100 °C (20 min). Extraction of the sample with methanol was maximal at 30-minute extraction time and the recovery of all the alkaloids studied was shown to average 90%. Development of this procedure provides a means to measure RCG gramine, hordenine and 5MeO-DMT concentrations in the presence or absence of tryptamines or β-carbolines. Initial growth and regrowth of five cultivars of RCG from two locations were evaluated for gramine, hordenine and 5MeO-DMT content using this technique. No differences were detected in hordenine content. Gramine content was higher (P < 0.05) in Rival than Venture, with Castor, Palaton and Vantage being intermediate. None of the cultivars contained detectable levels of 5MeO-DMT. There were no differences (P > 0.05) in hordenine content between initial growth and regrowth; however, gramine content of regrowth was higher (P < 0.05) than initial growth.Key words: Gramine, hordenine, 5-methoxy-N,N-dimethyl tryptamine, reed canarygrass, Phalaris arundinaceae L.