响应面试验优化动物源性食品中呋喃西林代谢物ELISA检测的衍生条件  

Optimization of Derivatization Conditions by Response Surface Methodology for the Determination of Semicarbazide in Animal-Derived Foods by Enzyme-Linked Immunosorbent Assay

作　　者：彭宏威 白瑞樱[2] 陈殿[1] 孙远明[1] 徐振林[1] 曾道平 李段 伍志权 杨金易[1] PENG Hongwei;BAI Ruiying;CHEN Dian;SUN Yuanming;XU Zhenlin;ZENG Daoping;LI Duan;WU Zhiquan;YANG Jinyi(Laboratory of Quality and Safety Risk Assessment in Agricultural Products Preservation, Ministry of Agriculture,Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University,Guangzhou 510642, China;Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang 453003, China;Guangzhou Wanlian Biological Technology Co. Ltd., Guangzhou 510642, China;Guangdong Huixin Agricultural Product Inspection Co. Ltd., Foshan 528200, China)

机构地区：[1]华南农业大学食品学院,农业部农产品贮藏保鲜质量安全风险评估实验室,广东省食品质量安全重点实验室,广东广州510642 [2]新乡医学院生理学与神经生物学教研室,河南新乡453003 [3]广州万联生物科技有限公司,广东广州510642 [4]广东汇信农产品检验有限公司,广东佛山528200

出　　处：《食品科学》2017年第6期287-294,共8页Food Science

基　　金：国家星火计划项目(2012GA780001;2013GA780035);“十二五”国家科技支撑计划项目(2012BAD31B0302);广东省自然科学基金项目(S2013030013338);NSFC-广东联合基金项目(U1301214);广东省科技计划项目(2012A020100002);广州市珠江科技新星专项(2013J2200080);广州市科技计划项目(2014J4200015)

摘　　要：利用响应面法优化酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测呋喃西林代谢物前处理的衍生条件。根据Box-Behnken试验设计原理,在单因素试验基础上,以呋喃西林代谢物回收率为响应值,应用响应面分析法研究衍生剂体积、衍生剂浓度、衍生温度和衍生时间各因素间的交互作用及对呋喃西林代谢物回收率的影响。最终确定ELISA方法线性检测范围为0.13~10.51 ng/m L,IC50为1.17 ng/m L;在选取样品质量为2 g时,最佳衍生条件为衍生剂体积415μL、衍生剂浓度15 mmol/L、衍生温度57℃、衍生时间50 min,实际回收率为98.57%~103.76%;衍生产物与呋喃西林原药交叉反应率为7.63%,与其他结构类似物、功能类似物及衍生剂交叉反应率均小于0.17%;加标回收率为93.45%~103.21%,批内变异系数为1.86%~6.15%,批间变异系数为3.88%~8.84%。该衍生条件高效,检测方法准确可靠,适用于动物源性食品中呋喃西林代谢物的快速筛查。The derivatization conditions for the determination of semicarbazide by enzyme-linked immunosorbent assay(ELISA)were optimized using combination of one-factor-at-a-time method and response surface methodology with Box-Behnken design.The independent variables were volume and concentration of derivatization agent,derivatization temperatureand time and the recovery of semicarbazide was used as response variable.The linear range of ELISA was0.13-10.51ng/mLand the50%inhibitory concentration(IC50)value was1.17ng/mL.The optimal derivatization conditions were determinedas follows:using415μL of15mmol/L derivatization agent and derivative at57℃for50min.Under these conditions,the recovery of semicarbazide was in the range of98.57%-103.76%.The cross-reactivity value of the derivatizationproduct was7.63%with furacilin and lower than0.17%with its structural and functional analogues.The recoveries inspiked negative samples ranged from93.45%to103.21%.The intra-assay and inter-assay coefficients of variation wer e1.86%-6.15%and3.88%-8.84%,respectively.The derivatization conditions were highly efficient and the ELISA methodwas sensitive,accurate,and suitable for the fast screening of semicarbazidee in animal-derived foods.

关 键 词：呋喃西林代谢物 衍生条件 响应面法 动物源性食品 酶联免疫吸附法 

分 类 号：TS207.3[轻工技术与工程—食品科学]