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| Determination of Anaesthetic Eugenol Residues in Aquatic Products by Chemiluminescene Enzyme Immunoassay |
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| DOI： |
| KeyWord:eugenol anaesthetic chemiluminescene enzyme immunoassay polyclonal antibody aquatic product |
| Author | Institution |
| WANG Qiang，WANG Xu-feng，ZHAO Dong-hao，ZHANG Ying-xia，HUANG Ke |
South China Sea Fisheries Research Institute， Chinese Academy of Fishery Sciences; Key Laboratory of Aquatic Product Processing， Ministry of Agriculture and Rural Affairs， Guangzhou ， China |
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| 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（IC50）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，IC10） 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． |
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