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| Establishment of Time-resolved Fluorescence Immunochromatography of Eugenol Based on Eu3+ Fluorescent Microspheres |
| SONG Yan-Ling1, ZHAI Lu1, LI Yin-Lai2, CHEN Qing-Zhou3, LIAO Jie4, JIN Ren-Yao1,* |
1 Institute of Sea Food, Zhejiang Gongshang University, Hangzhou 310012, China;
2 Hangzhou Bioeast Biotech Limited Company, Hangzhou 310018, China;
3 Hangzhou Nankai Biotechnology Limited Company, Hangzhou 310000, China;
4 Zhejiang Huacai Testing Technology Limited Company, Shaoxing 311800, China |
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Abstract Eugenol has been widely used in fishery anesthesia because of its wide source, low cost, good effect and short residual period, but its safety is still controversial over the world, and it is considered that eugenol compounds are carcinogens or potential carcinogens for rodents. In order to provide an efficient and simple method for the on-site rapid detection of eugenol residues in aquatic products, in this study, a time-resolved fluorescence immunochromatographic technique for eugenol and prepared immunochromatographic test strips were established. The fluorescent microspheres and anti-eugenol monoclonal antibody were labeled with Eu3+, and parameters such as microsphere activation time, fluorescent labeling buffer pH, amount of labeled antibody, amount of fluorescent microspheres and T- and C-line encapsulation concentration were optimized, and the immunochromatographic test strips were prepared and evaluated under the optimal conditions. The results showed that the test strips made under the optimal conditions had a good linear relationship between 0.05~0.40 μg/mL with the standard curve equation: y=-2.415lnx+2.589 4, R2=0.994, and the instrumental limit of detection (LOD) of 0.1 μg/mL, which indicated that the strips had a good sensitivity. The recoveries of the spiked samples were betweeen 94.16% and 109.27%, and the coefficients of variation (CV) of intra-batch and inter-batch were from 2.00% to 7.33% and from 2.73% to 6.97%, respectively. The results of the eugenol specimen detection by the newly developed test strips were consistent with those of the traditional high-performance liquid chromatography (HPLC) method. The stability test proved that the test strips prepared in this study had an effective shelf life of at least 1 year. This research is suitable for the rapid detection of eugenol residues in aquatic products and provides a reference for the development of eugenol rapid detection technology.
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Received: 25 September 2023
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Corresponding Authors:
*nodjin@163.com
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