Preparation And Application Of Immunoaffinity Chromatographic Column For Purification Of Diniconazole

Diniconazole is a broad-spectrum triazole fungicide and shows excellent efficacy against a variety of fungal diseases of cereals, fruits, vegetables and other field crops when applied both preventively and curatively. Compared with traditional sample pre-treatment methods, the immunoaffinity chromatographic (IAC) technology, having good application prospect, is characterized by simple operation, high selectivity, excellent purification and enrichment capability. For the first time, the anti-diniconazole antibody was encapsulated in a sol-gel matrix to form a IAC column. The performance of this IAC column was systematically evaluated for sample extraction. The main results were as follows:1. The purified anti-diniconazole polyclonal antibody was polymerised to hydorlytic tetramethoxysilane (TMOS) to synthesize the immunosorbent for the IAC column specific to diniconazole. The conditions of IAC column were optimized as follows:water was used as equilibrium and adsorbent medium,30%and50%methanol-water (v/v) as eluent. The results showed that the dynamic column capacity was up to125.4μg/g bed volume. The spiked river water, soil, rice, flour and fruits were cleaned up and enriched by IAC column, and the diniconazole in elute was determined by high performance liquid chromatography(HPLC). The average recoveries of diniconazole from river water were90.36%-100.14%with relative standard deviation (RSD) of2.03%-6.08%, and the average recoveries of diniconazole from soil were89.85%-97.00%with the RSD of2.06%-4.29%. The average recoveries of diniconazole from rice were89.57%-93.95%with the RSD of4.61%-7.35%, and the average recoveries of diniconazole from flour were89.64%-92.07%with the RSD of1.90%-7.32%.The average recoveries from fruits samples were85.55%to94.02%with RSD of3.38%-6.78%. The IAC clean-up procedure provided a new effective analytical method for the determinaion of diniconazole in sample media such as river water and fruit samples.2. With the prepared monoclonal antibody of anti-diniconazole, the high capacity IAC column was established. The conditions of IAC column were optimized as follows:water was used as equilibrium and adsorbent medium,30%and50%methanol-water (v/v) as eluent. The results showed that the dynamic column capacity was up to179.89μg/g bed volume. The spiked river water, soil, rice,flour and fruits were cleaned up and enriched by IAC column, and the diniconazole in elute was determined by HPLC. At different spiked levels (0.01-5.0mg/kg), the recoveries of diniconazole were from80.15%-98.42%with RSD of0.36%-5.92%. The method was satisfied with the requirements of the pesticide residue analysis, and the present study laid the foundation for the production of the commercial IAC column.