广东药科大学学报2023，Vol.39Issue(6)：37-43,7.DOI:10.16809/j.cnki.2096-3653.2023100901
基于QuEChERS样品前处理的间接竞争酶联免疫分析法快速检测薏苡仁中黄曲霉毒素B1
A rapid detection method for aflatoxin B1 in Coix seed based on indirect competitive enzyme-linked immunosorbent assay with QuEChERS sample pretreatment
摘要
Abstract
Objective To address the issue of matrix interference in the detection of aflatoxin B1(AFB1)in Coix seeds by indirect enzyme-linked immunosorbent assay(ic-ELISA),and to develop a convenient high-throughput assay for the rapid screening of AFB1 in Coix seeds.Methods The sample pretreatment process was optimized by evaluating the colorimetric value and inhibition rate of ic-ELISA.Specifically,the effectiveness of QuEChERS technology,as well as different dilution solvents and dilution methods were focused on eliminating matrix effects.The methodology was investigated,and finally,the method was applied to the real samples.The results were confirmed by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Results An ic-ELISA was established for quantitative analysis by solvent standard curve without any correction.The half inhibitory concentration of the method was 0.074 ng/mL,and the linear range was 1.37-20.0 μg/kg.The recoveries of the spiked samples ranged from 75.12%to 84.46%with RSD of less than 6%.In the 122 batches of Coix seeds and related samples,the incidence rate of AFB1 was 20.5%,and the rate of exceeding the standard in positive samples was 24.0%.The contamination level ranged from 1.76 to 27.56 μg/kg.The correlation coefficient(R)between the ic-ELISA and LC-MS/MS results was 0.98.Conclusion The QuEChERS technology is capable of effectively removing interfering components in Coix seeds,thus possessing significant application value in the establishment of rapid sample pre-treatment methods.In combination with the ic-ELISA developed in this study,it provides a convenient and effective detection method for the screening and monitoring of AFB1 in Coix seeds.关键词
QuEChERS技术/间接竞争酶联免疫分析方法/快速检测/薏苡仁/黄曲霉毒素B1Key words
QuEChERS technique/indirect competitive enzyme immunoassay/rapid detection/Coix seeds/aflatoxin B1分类
医药卫生引用本文复制引用
张磊,关凯仪,黄雨心,王淑美..基于QuEChERS样品前处理的间接竞争酶联免疫分析法快速检测薏苡仁中黄曲霉毒素B1[J].广东药科大学学报,2023,39(6):37-43,7.基金项目
国家自然科学基金项目(82374032、81903798) （82374032、81903798）
广东省基础与应用基础研究基金项目(2022A1515012099) （2022A1515012099）
广东省普通高校特色创新类项目(2023KTSCX059) （2023KTSCX059）
