Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay

WANG Qiang; WANG Xu-feng; ZHAO Dong-hao; ZHANG Ying-xia; HUANG Ke

doi:10.19969/j.fxcsxb.21071202

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Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay

Scientific Papers | 更新时间：2022-07-01

- Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay
- Journal of Instrumental Analysis Vol. 41, Issue 3, Pages: 354-360(2022)
- 作者机构：
  
  中国水产科学研究院南海水产研究所，农业农村部水产品加工重点实验室，广东广州 510300
- 作者简介：
- 基金信息：
- DOI：10.19969/j.fxcsxb.21071202
  CLC：
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王强,王旭峰,赵东豪等.化学发光酶免疫分析法检测水产品中残留的麻醉剂丁香酚[J].分析测试学报,2022,41(03):354-360.

WANG Qiang,WANG Xu-feng,ZHAO Dong-hao,et al.Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay[J].Journal of Instrumental Analysis,2022,41(03):354-360.
王强,王旭峰,赵东豪等.化学发光酶免疫分析法检测水产品中残留的麻醉剂丁香酚[J].分析测试学报,2022,41(03):354-360. DOI： 10.19969/j.fxcsxb.21071202.

WANG Qiang,WANG Xu-feng,ZHAO Dong-hao,et al.Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay[J].Journal of Instrumental Analysis,2022,41(03):354-360. DOI： 10.19969/j.fxcsxb.21071202.

摘要

将丁香酚（Eul）与4-溴丁酸叔丁酯进行衍生化反应，合成了半抗原4-（4-烯丙基-2-甲氧基-苯氧基）-丁酸（Eul－4－Tbl）。通过偶联载体蛋白并免疫动物后获取丁香酚多克隆抗体，建立了丁香酚的间接竞争化学发光酶免疫分析方法（ic－CLEIA）。水产样品经乙腈提取，Oasis PRiME HLB固相萃取小柱净化后用于测定。结果表明，ic－CLEIA的最佳反应条件为：包被原质量浓度为0.125 μg/mL，抗体稀释40 000倍，抗体反应时间为30 min，药物稀释液为PBS。该方法的半抑制浓度（IC,50,）为1.28 μg/L，检测线性范围为0.25 ~ 6.43 μg/L，检出限（LOD，IC,10,）为0.11 μg /L。样品中丁香酚的加标回收率为76.6%～108%，相对标准偏差（RSD）为5.9% ~ 12%，测定结果与液相色谱－串联质谱法具有良好的相关性（,r, = 0.990 4），可用于水产品中麻醉剂丁香酚残留的快速测定。

Abstract

Eugenol was derived with tert-butyl 4-bromobutanoate to obtain a hapten 4-（4-allyl-2-methoxy-phenoxy）-butyric acid（Eul－4－Tbl）．The hapten was coupled to carrier protein as immunogen for animal immunization，and a polyclonal antibody against eugenol was raised．Thus，an indirect competitive chemiluminescent enzyme-linked immunosorbent assay（ic－CLEIA）was developed for the determination of eugenol．The samples were extracted with acetonitrile，and then cleaned up with an Oasis PRiME HLB solid-phase extraction column．Results showed that the optimized assay conditions were as follows：coating antigen concentration：0.125 μg/mL，dilution ratio of antibody：40 000，reaction time of antibody：30 min，dilution solution for analyte：PBS．The half inhibition concentration（IC,50,）of the developed ic－CLEIA was 1.28 μg/L，the liner range was 0.25－6.43 μg/L，and the limit of detection（LOD，IC,10,） was 0.11 μg/L．The spiked recoveries of eugenol in aquatic products were in the range of 76.6%－108%，with relative standard deviations of 5.9%－12%．The results obtained by this method exhibited good correlation to those of HPLC－MS/MS（,r ,= 0.990 4）．The developed method was suitable for rapid determination of anaesthetic eugenol residues in aquatic product．

关键词

丁香酚麻醉剂化学发光酶免疫分析多克隆抗体水产品

Keywords

eugenolanaestheticchemiluminescene enzyme immunoassaypolyclonal antibodyaquatic product

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