丁草胺荧光偏振免疫分析——示踪物的合成与鉴定  

Development of Fluorescence Polarization Immunoassay for the Herbicide Butachlor——Synthesis and Identification of Tracers

作　　者：薛钢[1] 雷红涛[1] 沈玉栋[1] 刘细霞[1] 吴青[1] 孙远明[1] 王弘[1] 

机构地区：[1]华南农业大学食品质量安全研究所/广东省食品质量安全重点实验室,广州510642

出　　处：《中国农业科学》2008年第11期3651-3655,共5页Scientia Agricultura Sinica

基　　金：国家自然科学基金(20377016;30700663);广东科技攻关项目(2007A020100006-10);国家"863"计划现代农业技术项目(2006AA10Z447)

摘　　要：【目的】为建立除草剂丁草胺的荧光偏振免疫检测方法,对关键试剂示踪物进行合成与鉴定。【方法】以异硫氰酸荧光素、乙二胺和己二胺为原料,合成了异硫氰酸荧光素乙二胺(EDF)和异硫氰酸荧光素己二胺(HDF)两种中间体,薄层层析法分离出目标产物,电喷雾电离质谱法鉴定;然后将5-氨基荧光素(AF)、EDF、HDF分别与丁草胺半抗原(BMPA)连接,得到3种不同间隔臂的示踪物(BMPA-AF,BMPA-EDF和BMPA-HDF),利用抗血清稀释曲线测定抗血清效价,竞争抑制曲线测定不同示踪物检测灵敏度。【结果】3种示踪物合成成功,抗血清稀释曲线和竞争抑制曲线显示3种示踪物的抗血清效价分别为1/600、1/1500和1/600,半抑制浓度分别为1292、433和298ng·ml-1。【结论】合成出3种不同间隔臂的丁草胺示踪物,对示踪物的性质进行初步研究,为下一步建立丁草胺荧光偏振免疫分析方法奠定了基础。[ Objective ] To establish fluorescence polarization immunoassay method （FPIA） for the herbicide butachlor, the key reagent （tracer） was synthesized and identified. [ Method ] Using fluorescein isothiocyanate （FITC）, ethylenediamine and hexylenediamine as materials, fluoresceinthiocarbamyl ethylenediamine （EDF） and fluoresceinthiocarbamyl hexylenediamine （HDF） were obtained, which were purified by thin layer chromatography and identified by ESI-MS. Various tracers （BMPA-AF, BMPA-EDF and BMPA-HDF） with different lengthes of bridge between the hapten molecule and 5-Aminofluorescein （AF）, EDF, HDF, were synthesized to achieve high-sensitivity FPIA. The titers of antiserum and the sensitivity of various tracers were estimated by dilution curves and calibration curves, respectively. [ Result ] Three tracers were synthesized successfully, the titers of antiserum of three tracers were 1/600, 1/1500 and 1/600, and the ICs0s of butachlor FPIA of three tracers were 1292, 433 and 298 ng-ml^-1. [Conclusion] The arm length of tracers affects effect the characters of FPIA, the tracer with longer arm is more sensitive to butachlor FPIA than one with shorter arm. This research is of benefit to further optimization of FPIA.

关 键 词：丁草胺 荧光偏振免疫分析 示踪物 异硫氰酸荧光素 

分 类 号：S482.4[农业科学—农药学]