Development Of IAC-HPLC Method To Determine Ctrinin In Red Rice

Citrinin (CIT) is a nephrotoxin which is mainly produced by numerous species of Penicillium Aspergillus and Monascus. Contaminations of citrinin were reported in a number of agricultural commodities, foods, feedstuffs. So far, commonly used methods to analyze citrinin are thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) with UV or fluorescence detection (FLD), and enzyme-linked immunosorbent assay (ELISA).A method (IAC-HPLC) for determination of CIT in red rice was development. 41 samples of red rice were detected by the IAC-HPLC.The main content and results of the study are descripted as follow:1 Preparation of Citrinin immunoaffinity columnThe Citrinin immunoaffinity column (IAC) was prepared by coupling CNBr-activated Sepharose 4 Fast Flow (4FF) with the anti-CIT monoclonal antibody which was purified by the caprylic acid ammonium sulfate method. The coupling reaction was identified by UV-absorbance measurements and the IAC to be prepared was evaluated by indirect-competitive ELISA and HPLC. The column capacity was determined to be 0.35μg when using 0.5 mL CNBr activated Sepharose 4FF and 0.50 mg purified anti-CIT monoclonal antibody.2 Development of IAC-HPLC to quantify citrinin in red riceAverage recoveries of CIT from red rice spiked at levels of 100μg/kg～600μg/kg ranged from 74.2%～87.14%, with relative standard deviations from 2.85 %～13.12%. The detection limit was 100μg/kg based on a signal-to-noise ratio of 3:1. 3. Determination of 41 red rice samples from market and our laboratory by IAC-HPLC and ELISA.41 red rice samples were detected by HPLC. The result of IAC-HPLC: 25 red rice samples were polluted by CIT, the content of CIT was ranged from 100μg/kg～986μg/kg.