Development of a sensitive indirect competitive enzyme-linked immunosorbent assay for high-throughput detection and risk assessment of aflatoxin B1 in animal-derived medicines.

Animal-derived medicine is an important part of traditional Chinese medicine (TCM). Studies have shown that many animal-derived medicinal products are susceptible to contaminate of aflatoxins, nevertheless, the rapid detection for animal-derived medicine is prone to be ignored. Here we developed a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) for rapid screening of aflatoxin B 1 (AFB 1) in ground beetle , cockroach, silkworm and earthworm. The sensitivity of the icELISA method was significantly enhanced. The IC 50 for the four animal-derived medicinal samples ranged from 0.092 to 0.135 ng mL−1; the limit of detection (LOD) was 0.008–0.020 ng mL−1. To obtain high accuracy, the extraction solution and time were evaluated. By using this method, a total of 138 samples were investigated, and the detection rates of AFB 1 in ground beetle and earthworm samples were 26.6% and 16.7%, respectively. The result was validated by liquid chromatography combined with tandem mass spectrometry, and an excellent correlation was observed between the two datasets, with a R 2 value of 0.999. Our results indicate that the proposed method can be used for the rapid detection of AFB 1 in animal-derived medicine. Furthermore, the quantitative risk assessment was conducted for ground beetle and earthworm based on the results, demonstrating that the intake of AFB 1 in ground beetle had a slight threat to the risk of cancer. [Display omitted] • A sensitive enzyme-linked immunosorbent assay screening of the aflatoxin B 1 in animal-derived medicines was performed. • Compared with previous reports, the limits of detection of aflatoxin B 1 increased by 4–12 times. • The quantitative risk assessment of aflatoxin B 1 was conducted for ground beetle and earthworm. [ABSTRACT FROM AUTHOR]