稻米中三唑磷残留免疫亲合色谱-高效液相色谱分析

中国农业科学 ›› 2006, Vol. 39 ›› Issue (05): 941-946 .

稻米中三唑磷残留免疫亲合色谱-高效液相色谱分析

韦林洪,王莲,刘曙照

收稿日期:2005-08-20 修回日期:1900-01-01 出版日期:2006-05-10 发布日期:2006-05-10

Analysis of Triazophos Residue in Rice by Immunoaffinity Chromatography-High Performance Liquid Chromatography

Received:2005-08-20 Revised:1900-01-01 Online:2006-05-10 Published:2006-05-10

摘要/Abstract

摘要： 【目的】提高三唑磷残留的监测效率。【方法】采用碳酰二咪唑（CDI）将Sepharose CL-4B活化并与纯化的抗三唑磷多克隆抗体共价偶联，合成免疫亲合色谱（IAC）吸附剂并制备对三唑磷具有特异性亲合力的IAC柱。对IAC条件进行了优化，选择0.02 mol·L-1 pH 7.2磷酸盐缓冲液作吸附与平衡介质，60%（体积分数）甲醇水作洗脱剂。【结果】在试验条件下，IAC柱对三唑磷的动态柱容量达1.91 ?g·ml-1床体积，当标样溶液中三唑磷含量为2 ng·ml-1时，经IAC柱富集的效率近250倍。稻米中添加三唑磷0.1 ?g·g-1，提取液以IAC柱分离富集，洗脱液采用高效液相色谱（HPLC）法测定，5次重复测定的平均回收率为102.5%，相对标准偏差4.44%。【结论】成功建立了三唑磷IAC-HPLC分析技术并用于稻米中三唑磷残留的测定。

关键词: 三唑磷残留, 稻米, 免疫亲和色谱（IAC）, 高效液相色谱（HPLC）

Abstract: 【Objective】The purpose of the study is to enhance the efficiency of triazophos residue monitoring.【Method】The purified anti-triazophos antibody was conjugated to the CDI-activated sepharose CL-4B to synthesize the immunosorbent and prepare the immunoaffinity chromatographic (IAC) column which is specific to triazophos. The conditions of IAC were optimized, pH7.2 phosphate buffer was used as equilibrium and absorbent medium, 60% methanol was used as eluent.【Result】Under the experimental conditions, the dynamic column capacity was up to 1.91 ?g·ml-1 bed volume, the efficiency of enrichment by IAC was nearly 250 times when the initial concentration of triazophos was 2 ng·ml-1 in standard sample solution. The extracts of spiked rice were cleaned up and concentrated by IAC, the triazophos in eluates was determined by high performance liquid chromatography (HPLC), and the average recovery of triazophos was 102.5% with relative standard deviation of 4.44% at the spiked level of 0.1 ?g·g-1. 【Conclusion】The IAC-HPLC technology is developed successfully for the analysis of triazophos residue in rice.

Key words: Triazophos residue, rice, immunoaffinity chromatography(IAC), hign performance liquid chromatography(HPLC)

韦林洪,王莲,刘曙照. 稻米中三唑磷残留免疫亲合色谱-高效液相色谱分析[J]. 中国农业科学, 2006, 39(05): 941-946 .

,,. Analysis of Triazophos Residue in Rice by Immunoaffinity Chromatography-High Performance Liquid Chromatography[J]. Scientia Agricultura Sinica, 2006, 39(05): 941-946 .

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