基于长春西汀酸的荧光偏振免疫方法测定磺胺二甲氧基嘧啶  

A Fluorescence Polarization Immunoassay Method Based on Apovincaminic Acid for Determination of Sulfadimethoxine

作　　者：王小勇[1] 龚福春[1,2] 唐连飞 李治章[1] 郑兴良[2] 湛雪辉[2] 

机构地区：[1]湖南科技学院化学与生物系,永州425100 [2]长沙理工大学化学与生物工程学院,长沙410004 [3]湖南省出入境检验检疫局,长沙410004

出　　处：《分析化学》2010年第10期1457-1461,共5页Chinese Journal of Analytical Chemistry

基　　金：湖南省科技计划(Nos.2009FJ4061,2009CK3069)资助项目

摘　　要：合成了荧光标记物长春西汀酸和示踪物磺胺二甲氧基嘧啶-长春胺酸,并采用薄层色谱法对示踪物进行提纯。以磺胺二甲氧基嘧啶为分析对象,利用示踪物与磺胺二甲氧基嘧啶抗体的特异性反应,采用荧光偏振免疫分析方法(FPIA)研究了荧光标记物的荧光光谱特性。此荧光标记物在357nm的激发光下,于453nm处产生一个荧光发射峰;以荧光素为对照,考察了本方法的灵敏性。结果显示,荧光素的检出限仅为2.7μg/L,而本方法的检测范围为0.5～146μg/L,检出限达到0.5μg/L。本方法的RSD为3.0%,重现性好;其它磺胺类药物对磺胺二甲氧基嘧啶的测定不产生明显干扰。运用本方法检测了饲料中磺胺二甲氧基嘧啶,结果与HPLC法相吻合。The fluorescence labeling apovincaminic acid （AVA） and tracers apovincaminic acid-sulfadime-thoxine were synthesized and used in the fluorescence polarization immunoassay method （FPIA） for the analysis of sulfadimethoxine （SMX） .By using AVA as indicating materials,tracers apovincaminic acid-sulfadime-thoxine （AVA-SMX） were prepared,which were purified by thin layer chromatography and identified by FPIA.The fluorescence properties of AVA and the characteristic performance of tracers immunoreacting with SMX antibodies were investigated by using tracers 5-aminofluorescein-sulfadimethoxine conjugates （AF-SMX） as a reference.The results indicated that the detection limit for AF-SMX was 2.7 μg/L,and the dynamic range for AVA-SMX was 0.5-146 μg/L with a detection limit of 0.5 μg/L.The sensitivity of the novel tracers was also estimated by dilution curves and calibration curves,respectively.The RSD of AVA-based FPIA method for SMX determination is 3.0% ,and a good selectivity can be obtained with the solutions in the presence of the other sulfa drugs.The proposed AVA-based FPIA was used to determine SMX in feed and the result is well agreed with that obtained with HPLC.

关 键 词：荧光偏振免疫分析 荧光标记物 长春西汀酸 磺胺二甲氧基嘧啶 

分 类 号：R927.2[医药卫生—药学]