Determination of sterigmatocystin in barley by acetylation and liquid chromatography

The estimation of sterigmatocystin by fluorescence liquid chromatographic analysis of the acetate derivative has eliminated the background interference normally encountered in analysis of underivatized sterigmatocystin in barley. Barley samples are extracted with acetonitrile-water; the extract is then washed with hexane, transferred to chloroform, and eluted from a silica gel column with cyclohexane-ethyl acetate. The extract is heated with pyridine and acetic anhydride for 3 h to give a stable derivative. Reverse-phase liquid chromatography, using a methanol-water mobile phase gradient and fluorescence detection, is the method of determination. Recoveries from barley samples spiked with 20, 110, 190, and 765 micrograms sterigmatocystin/kg were 31, 69, 75, and 96%, with coefficients of variation between 2.8 and 5.4%. Sterigmatocystin is confirmed by comparing retention times in underivatized extracts of samples and standards, using methanol-water (3 + 2) mobile phase and ultraviolet detection.