酶联免疫法检测动物组织及尿液中氟苯尼考与甲砜霉素的残留  

An Indirect Competitive Enzyme-linked Immunosorbent Assay for Simultaneous Determination of Florfenicol and Thiamphenicol in Animal Meat and Urine

作　　者：李然 林泽佳 杨金易[1] 徐振林[1] 王弘[1] 雷红涛[1] 孙远明[1] 沈玉栋[1] LI Ran;LIN Ze-Jia;YANG Jin-Yi;XU Zhen-Lin;WANG Hong;LEI Hong-Tao;SUN Yuan-Ming;SHEN Yu-Dong(Guangdong Provincial Key Laboratory of Food Quality and Safety,College of Food Science,South China Agricultural University,Guangzhou 510642,China)

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

出　　处：《分析化学》2018年第8期1321-1328,共8页Chinese Journal of Analytical Chemistry

基　　金：广州市科技计划项目(No.201704020082);国家重点研发计划国际合作项目(No.2016YFE0106000);广西省科技计划项目(No.2017AB47020);国家自然科学基金项目(No.31371769);广东省高等学校珠江学者岗位计划项目(2017)资助~~

摘　　要：针对食品中氟苯尼考及甲砜霉素多残留污染问题,基于新设计的半抗原制备了系列人工抗原,通过免疫动物获得了可同时特异性识别氟苯尼考与甲砜霉素的多克隆抗体,并在抗体-包被原组合的筛选及反应条件优化基础上,建立了动物组织及尿液中氟苯尼考和甲砜霉素残留同时检测的酶联免疫分析方法。本方法对于氟苯尼考的半数抑制浓度(IC_(50))为1.32 ng/mL,线性检测范围为0.31~5.61 ng/mL,检出限为0.12 ng/mL;对于甲砜霉素的IC_(50)为2.13 ng/mL,线性检测范围为0.41~11.20 ng/mL,检出限为0.15 ng/mL,与氯霉素等多种类似物均无交叉反应,动物组织及尿样中氟苯尼考和甲砜霉素的添加回收率均在77.2%~116.0%之间,相对标准偏差<15%,适用于实际样品中氟苯尼考和甲砜霉素的快速筛查检测。A high-affinity polyclonal antibody was prepared by immunizing animals with haptens FFD and FFM. Under the optimal combination of coating antigen and antibody,an indirect competitive enzyme-linked immunosorbent assay（ icELISA） for simultaneously detecting florfenicol and thiamphenicol residues in animal meat and urine samples was developed. The icELISA showed an IC50 value of 1.32 ng/mL for florfenicol and 2.13 ng/mL for thiamphenicol,respectively. The linear ranges were from 0. 31 to 5. 61 ng/mL with a limit of detection of 0.12 ng/mL for florfenicol,and 0.41 to 11.2 ng/mL with a limit of detection of 0.15 ng/mL for thiamphenicol,respectively. The average recoveries of florfenicol and thiamphenicol in spiked samples ranged from 77.2% to 116.0% with a relative standard deviation of less than 15%. Therefore,this proposed icELISA provided a valid detection for florfenicol and thiamphenicol residues in animal meat product and urine samples.

关 键 词：氟苯尼考 甲砜霉素 抗体 间接竞争酶联免疫吸附分析 

分 类 号：TS207.5[轻工技术与工程—食品科学]