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

Indirect Competitive Chemiluminescence Enzyme Immunoassay for Furaltadone Metabolite in Metapenaeus Ensis

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

Corresponding author: WANG Hong, SUN Yuan-Ming,
Received Date: 16 March 2012
Available Online: 6 June 2012
Fund Project: 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)

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.
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沈阳化工大学材料科学与工程学院沈阳 110142