虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立

引用本文: 杨武英, 董洁娴, 沈玉栋, 杨金易, 王弘, 徐振林, 杨星星, 孙远明. 虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立[J]. 分析化学, 2012, 40(12): 1816-1821. doi: 10.3724/SP.J.1096.2012.20268 shu

Citation: YANG Wu-Ying, DONG Jie-Xian, SHEN Yu-Dong, YANG Jin-Yi, WANG Hong, XU Zhen-Lin, YANG Xing-Xing, SUN Yuan-Ming. Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis[J]. Chinese Journal of Analytical Chemistry, 2012, 40(12): 1816-1821. doi: 10.3724/SP.J.1096.2012.20268 shu

虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立

杨武英 ^1,2, ,
董洁娴 ^1, ,
沈玉栋 ^1, ,
杨金易 ^1, ,
王弘 ^{1,
,} ,
徐振林 ^1, ,
杨星星 ^1, ,
孙远明 ^{1,
,}

通讯作者: 王弘,孙远明; 王弘,孙远明

基金项目:
本文系国家自然科学基金(No.30871755) (No.30871755)

广东省科技计划项目(No.1211150100102) (No.1211150100102)

广东省食品质量安全重点实验室(华南农业大学)开放基金(No.09003566)资助 (华南农业大学)开放基金(No.09003566)

摘要: 采用对碘苯酚增强的HRP-鲁米诺-H₂O₂化学发光体系,建立了虾肉中呋喃它酮代谢物5-吗啉甲基-3-氨基-2-恶唑烷基酮(AMOZ)残留的间接竞争化学发光酶免疫分析 (icCLEIA) 检测方法。检测所用抗体是基因重组的抗AMOZ衍生物单链抗体。优化的icCLEIA最佳工作条件为:AMOZA-OVA包被浓度62.5 μg/L,抗AMOZ衍生物单链抗体最佳稀释度为1:10,竞争免疫反应时间45 min,HRP酶标记羊抗鼠抗体最佳稀释度为1:10000,孵育时间50 min。本方法的IC₅₀为1.38 μg/L;灵敏度为0.09 μg/L;线性范围为0.26-9.08 μg/L (IC₂₀-IC₈₀);批内和批间相对标准偏差均小于15%;抗AMOZ衍生物单链抗体与其它硝基呋喃类抗生素及其代谢物均没有交叉反应,特异性良好。4个不同添加量的AMOZ加标样品的平均回收率分别为72.2%,73.4%,72.6%和78.6%。与HPLC-MS/MS法测定值进行比较发现,两种方法相关性良好(R²=0.9997)。本方法可用于水产品中AMOZ残留的快速检测。

关键词:

English

Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis

1.
¹ Key Laboratory of Food Quality and Safety of Guangdong Province/College of Food Science, South China Agricultural University, Guangzhou 510642, China;

Corresponding author: 王弘,孙远明; 王弘,孙远明

Received Date: 16 March 2012
Fund Project:
本文系国家自然科学基金(No.30871755)

广东省科技计划项目(No.1211150100102)

广东省食品质量安全重点实验室(华南农业大学)开放基金(No.09003566)资助

Abstract: An indirect competitive chemiluminescence enzyme immonoassay (icCLEIA) for the quantitative analysis of furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ) in shrimp sample was established with the highly sensitive luminol-H₂O₂-horseradish peroxidase(HRP)-4-iodophenol chemiluminescence reaction detection system. The icCLEIA method was based on a single-chain variable fragment (scFv) antibody against AMOZ derivative. The optimized assay conditions were 62.5 μg/L of coating antigen (5-morpholinomethyl-3-(glyoxalamino)-2-oxazolidone-ovalbumin, AMOZA-OVA), the dilute time of scFv antibody was 1:10, the immunoreaction time was 45 min, the dilute times and incubation time of HRP-goat anti mouse IgG were 1:10000 and 50 min, respectively. The icCLEIA results showed IC₅₀ value, limit of detection (LOD) and linear range were 1.38 μg/L, 0.09 μg/L and 0.26-9.08 μg/L, respectively. Inter-assay and intra-assay RSD was both below 15%. The scFv antibody showed high specificity. The average recoveries of four spiked level of AMOZ were 72.3%, 73.4%, 72.6% and 78.6%, respectively. Compared with HPLC-MS/MS, there was a good correlation between the two methods (R²=0.9997). Thus, the established icCLEIA method could be further used for detecting AMOZ in aquatic products samples rapidly and efficiently.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立

通讯作者: 王弘,孙远明; 王弘,孙远明

English

Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis

Corresponding author: 王弘,孙远明; 王弘,孙远明

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立

通讯作者: 王弘,孙远明; 王弘,孙远明

English

Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis

Corresponding author: 王弘,孙远明; 王弘,孙远明

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs