荧光偏振免疫分析方法快速检测沙拉沙星残留  

Development of a Fluorescence Polarization Immunoassay for Rapid Determination of Sarafloxacin in Milk and Pig Urine

作　　者：宋佩[1] 孟萌[1] Sergei A Eremin 张太昌[1] 田溪[1] 薛虎寅[1] 张昱[1] 尹永梅[1] 郗日沫[1] 

机构地区：[1]南开大学药学院,南开大学药物化学生物学国家重点实验室,天津市分子药物研究重点实验室,天津300071 [2]Faculty of Chemistry,M.V.Lomonosov Moscow State University,Moscow 119991,Russia

出　　处：《分析化学》2012年第8期1247-1251,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.81173017;31101277);中央高校基本科研业务费项目(No.65011751)资助

摘　　要：以异硫氰酸荧光素(FITC)标记沙拉沙星合成荧光标记物,采用薄层色谱法提纯,优化了反应时间、标记物和抗体的工作浓度,建立了沙拉沙星的快速荧光偏振免疫分析法(FPIA)。本方法测定沙拉沙星在缓冲液中的半数抑制浓度(IC50)为43.2μg/L;检测范围为5.7～327μg/L,可以达到国家规定的动物性食品中兽药最高残留限量(80μg/kg)的要求。本研究考察了FPIA测定沙拉沙星的动力学过程及对其它4种喹诺酮类药物的交叉反应。结果表明,环丙沙星、恩诺沙星、加替沙星及氧氟沙星的交叉反应率分别为3.3%,1.8%,1.7%和0.7%。在牛奶和猪尿中沙拉沙星的回收率分别在71%～94%和74%～102%之间。本方法操作简单快捷,整个检测过程只需5 min、而且灵敏度较高、特异性强,适用于动物性食品中沙拉沙星残留的快速筛选检测。To develop a rapid and sensitive fluorescence polarization immunoassay（FPIA） for the determination of sarafloxacin（SAR）,fluorescein-labelled tracer（SAR-FITC） was synthesized and purified by TLC.The reaction time,tracer and polyclonal antibody concentration were optimized,and the FPIA method showed a dynamic range from 5.7 to 327.6 μg/L with IC50 value of 43.23 μg/L to SAR in buffer.The specificity of the FPIA for SAR was investigated using other 4 quinolones and the cross-reactivity for ciprofloxacin,enrofloxacin,gatifloxacin,ofloxacin were 3.3%,1.8%,1.7%,0.7%,respectively.The recoveries in milk samples ranged from 71%-94%,and those of pig urine samples were in the range of 74%-102%.The FPIA developed in this study is a rapid and convenient method,which is suitable to be used as a screening method to detect residues of sarafloxacin.

关 键 词：荧光偏振免疫分析 沙拉沙星 喹诺酮类药物 荧光标记 

分 类 号：R927.2[医药卫生—药学] O657.3[理学—分析化学]