科研之友
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摘要
A sensitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of new herbicide H-9201, and their suitability for the determination of this analyte in environmental water was studied. Starting with phosphorus thiochloride and methanol, an analogs with similar structure of hapten was synthesized, and carboxyl group was conducted by succinic anhydride, then the derivative was conjugated with bovine serum albumin or ovalbumin by means of active ester as immunoagent and coating agent, respectively; IC-ELISA method was developed based on the polyclonal antibody generated in rabbits by immunizing with synthesized hapten. For standard, 50% reduction of the maximum ELISA signal (IC50) was 1. 331 mg/L, whereas the detection limit was 0. 017 mg/L in the competitive standard curve ranged from 0. 05 to 20. 0 mg/L and the combination of the concentrations of coating solution and primary antibody was optimized to 1/512 and 1/18000, respectively. No significant cross-reactivity for related compounds was observed, such as 2,4-dimethyl-6-nitrophenol, p-nitrophenol, Methamidophos, phorate, chlorpyrifos, parathion, the antibody showed negligible cross-reactivity with S-carboxylethyl-O, O-Dimethyl-phosphorodithioate, dimethoate, malathion, but high cross-reactivity with analogs of hapten. Recovery of H-9201 in environmental water was (80.5 +/- 4. 5)%-(95. 2 +/- 3. 2)% based on the developed IC-ELISA method.
- 出版日期2011-2
- 单位南京农业大学
