Abstract
High pressure liquid chromatography has been applied for the quantitative determination of vicine and convicine, two glucosides present in fababeans. The following method has been developed: extraction of vicine and convicine by 0.1N NaOH, precipitation of proteins by adjusting the pH to 4.2 with 1N HC1, adequate dilution of the extract before its injection into a Waters high pressure liquid chromatograph. A LiChrosorb NH2 column and a mixture of H2O/acetonitrile in the ratio of 30/70 (v/v) at a flow rate of 2mL/min were used for the separation. Vicine, convicine and α-galactosides were detected by an ultraviolet detector (λ=273nm) and a differential refractometer, respectively. Identification and quantitative estimation were based on known reference samples. Treatment of fababean extracts with β-glucosidase led to a disappearance of the glucoside peaks that correspond in retention time to those of reference glucosides. The proposed method is reliable, rapid, and may be used as a technique for analysis of glucosides and α-galactosides for plant breeding and processed fababean products.
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