食品科学 ›› 2021, Vol. 42 ›› Issue (4): 305-312.doi: 10.7506/spkx1002-6630-20191210-110

• 安全检测 • 上一篇    下一篇

间接竞争化学发光酶联免疫分析方法检测禽肉中金刚烷胺和氯霉素残留

许小炫,苏晓娜,谭庶,钟翠丽,曾道平,张燕,徐振林,王序,杨金易   

  1. (1.华南农业大学食品学院,广东?广州 510642;2.温氏食品集团股份有限公司,广东?云浮 527400;3.广东产品质量监督检验研究院,广东?佛山 528300)
  • 出版日期:2021-02-25 发布日期:2021-02-25
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2016YFE0106000); 广东省自然科学基金重大基础研究培育项目(2018B030314005);广东省重点领域研发计划项目(2019B020211001); 2019年省级乡村振兴战略专项资金第二批(省级项目)入库项目(2019KJ130)

Determination of Amantadine and Chloramphenicol Residues in Poultry Meat by Indirect Competitive Chemiluminescence Enzyme-Linked Immunosorbent Assay

XU Xiaoxuan, SU Xiaona, TAN Shu, ZHONG Cuili, ZENG Daoping, ZHANG Yan, XU Zhenlin, WANG Xu, YANG Jinyi   

  1. (1. College of Food Science, South China Agricultural University, Guangzhou 510642, China; 2. Wens Food Group Co. Ltd., Yunfu 527400, China; 3. Guangdong Product Quality Supervision and Inspection Institute, Foshan 528300, China)
  • Online:2021-02-25 Published:2021-02-25

摘要: 为快速检测禽肉中残留的金刚烷胺和氯霉素,结合高效的前处理方法,建立检测金刚烷胺和氯霉素的间接竞争化学发光酶联免疫分析方法(indirect competitive chemiluminescence enzyme-linked immunosorbent assay,ic-CLEIA)。对包被原与抗体的最佳稀释倍数、竞争时间、酶标二抗稀释倍数进行优化。结果表明:金刚烷胺的IC50为0.33 μg/L,线性范围为0.06~1.77 μg/L;氯霉素的IC50为0.039 μg/L,线性范围为0.010~0.179 μg/L。样品经乙酸乙酯和碳酸钾溶液提取后,取上清液用氮气吹干,净化后进行检测。金刚烷胺和氯霉素的加标回收率分别为92.55%~105.14%和92.00%~112.50%,变异系数均低于10.48%,且实际样品检测结果与仪器方法相关性良好(R2>0.98),表明建立的ic-CLEIA方法具有良好的准确度和可靠性。该方法将为涉及多种药物残留的免疫快速检测方法及相应的前处理技术提供技术依据。

关键词: 禽肉;金刚烷胺;氯霉素;化学发光酶联免疫;前处理

Abstract: An indirect competitive chemiluminescent enzyme-linked immunosorbent assay (ic-CLEIA) was developed for the rapid detection of amantadine and chloramphenicol residues in poultry meat based on efficient sample pretreatment. The dilution factors of coating antigen, antibody and horseradish peroxidase (HRP)-labeled secondary antibody, and competition time were optimized. The results showed that the half-maximum inhibition concentration (IC50) and the linear range were 0.33 μg/L and 0.06–1.77 μg/L for amantadine, and 0.039 μg/L and 0.010–0.179 μg/L for CAP, respectively. Samples were sequentially extracted with ethyl acetate and potassium carbonate solution, and then the extract was blown to dryness with nitrogen gas and purified by n-hexane extraction before being analyzed. The recoveries of amantadine and chloramphenicol from spiked samples were in the ranges of 92.55%–105.14% and 92.00%–112.50%, respectively, and the coefficients of variation were both less than 10.48%. Furthermore, the results determined by ic-CLEIA were highly correlated with those of instrumental analysis method (R2 > 0.98), which demonstrates that the established ic-CLEIA method has high accuracy and good reliability. This method represents a rapid immunoassay for the detection of veterinary drug residues in poultry meat.

Key words: poultry meat; amantadine; chloramphenicol; indirect competitive chemiluminescence enzyme-linked immunosorbent assay; pretreatment

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