HPLC Determination of Zearalenone with Purification by Immunoafinity Chromatographic Column

Zearalenone (ZEN in short) was extracted from sample (e.g.wines,drinks,cereals etc.) with equal volume of acetonitrile (for liquid sample) or with 100 mL of mixture of acetonitrile and H2O (90+10) (for 20 g of solid sample).The extract obtained was diluted and filtered on glass fibres.The filtrate was purified by passing through the Zearala Test column (an immuno affinity chromatographic column).After rinsing with buffer solutions and water successively,ZEN was eluted from the column with 1 mL of methanol which was used for HPLC determination.The ZORBAX SB-C18 column (150 mm×4.60 mm,5 μm) was used as separation column,and a mixture of acetonitrile,water and ethanol (mixed in the ratio of 46 to 46 to 8 by volume) was used as mobile phase at the flow-rate of 1.0 mL·min-1.Fluorescence detection at wavelengths of (λex) 274 nm and (λem) 440 nm was adopted in the determination.Linear relationship between values of peak area and mass concentration of ZEN was obtained in the range from 10-500 μg·L-1.Detection limit (3S/N) found was 10 μg·kg-1.Tests for recovery and precision of the method were made,giving values of recovery in the range of 95.3%-98.9%,and values of RSD′s (n=6) less than 1.5%.