A method for simultaneous determination of multiple mycotoxins in Bombyx Batryticatus and Eupolyphaga Steleophaga was established to provide technical support for monitoring and quality control of such medicinal materials. The mildew-prone Bombyx Batryticatus and Eupolyphaga Steleophaga were detected by UPLC-MS/MS technology, and the chromatographic conditions, mass spectrometry conditions, test sample preparation methods, and QuEChERS purification parameters were optimized. Scanning was conducted under multiple reaction monitoring mode(MRM), and quantitative analysis was performed by matrix standard curve method. The detected 14 mycotoxins had good linear relationships in the range of 0.04-500 ng·mL~(-1), with correlation coefficients(r)≥0.99. The limits of detection(LOD, S/N=3) and limits of quantitation(LOQ, S/N=10) were 0.01-0.648 μg·kg~(-1) and 0.022-1.435 μg·kg~(-1). At the high, medium, and low addition levels, the average recovery of the detected toxins was 70.99%-124.7%, and the relative standard deviation(RSD) was 0.44%-8.5%, all of which were in line with the requirements of the 2020 edition of the Chinese Pharmacopoeia. The method was applied to the detection of mycotoxins in 20 batches of Bombyx Batryticatus and Eupolyphaga Steleophaga, and five kinds of toxins including DON, AFB_1, ST, AFG_1, and OTA were detected. Among them, OTA, AFB_1, and AFG_1 were detected in 10 batches of Bombyx Batryticatus samples, and 10.0% of AFs exceeded the standard. Four toxins of AFB_1, AFG_1, ST, and DON were detected in 10 samples of Eupolyphaga Steleophaga, and 10.0% of AFB_1 and AFs exceeded the standard, indicating certain risks and requiring more attention. In this paper, a detection method for multiple mycotoxins in Bombyx Batryticatus, Eupolyphaga Steleophaga, and other animal Chinese herbal medicines was successfully established. This method had a wide range of applications and expanded the narrow application range of existing mycotoxin detection methods. This research can provide a reference for the development of mycotoxin detection methods for other mildew-prone Chinese herbal medicines.
Keywords: Bombyx Batryticatus; Eupolyphaga Steleophaga; QuEChERS; UPLC-MS/MS; mycotoxins.