Preparation of Anti-Zearalenone IgY and Development of an Indirect Competitive ELISA Method for the Measurement of Zearalenone in Post-Fermented Tea
Author:
Qiu Taotao1, Zhang Huayi1, Lei Hongtao2ORCID, Zhang Lin1, Zhang Yaqiong2, Shen Xing2ORCID, Xu Biyun1, Zhu Jialin1ORCID, Xiao Wentao1, Zheng Jixu1, Chen Jiahong2
Affiliation:
1. College of Physical Education and Health, Guangxi Normal University, Guilin 541004, China 2. Guangdong Provincial Key Laboratory of Food Quality and Safety, National-Local Joint Engineering Research Center for Machining and Safety of Livestock and Poultry Products, College of Food Science, South China Agricultural University, Guangzhou 510642, China
Abstract
Post-fermented tea (PFT) is one of the most commonly consumed beverages worldwide. Rapid microbial growth and significant changes in the microbial composition of PFT during processing and storage pose a potential risk of contamination with mycotoxins such as zearalenone (ZEN). Screening for ZEN contamination in a simple, rapid, and inexpensive manner is required to ensure that PFT is safe for consumption. To monitor ZEN in PFT, ZEN was conjugated with bovine serum albumin to prepare egg yolk immunoglobulins (IgY). A specific indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on IgY was developed and validated. ZEN was extracted with acetonitrile and water (50:50, v/v) containing 5% acetic acid and purified using a mixture of primary and secondary amines and graphitized carbon black to remove matrix interference from the PFT samples. Under optimal conditions, the linear range of this assay was 13.8−508.9 ng mL−1, the limit of detection was 9.3 ng mL−1, and the half-maximal inhibitory concentration was 83.8 ng mL−1. Cross-reactivity was negligible, and the assay was specific for ZEN-related molecules. The recovery rate of ZEN in the control blanks of PFT samples spiked with a defined concentration of ZEN of 89.5% to 98.0%. The recovery and accuracy of the method were qualified for PFT matrices. No significant differences were evident between the results of the actual PFT samples analyzed by high-performance liquid chromatography and ic-ELISA. The collective data indicate that the developed ic-ELISA can be used for the rapid and simple detection of ZEN in PFT products.
Funder
Innovation Project of Guangxi Graduate Education Guilin Science and Technology Development Program National Key Research and Development Program of China Guangxi Natural Science Fundation
Subject
Plant Science,Health Professions (miscellaneous),Health (social science),Microbiology,Food Science
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