目的 建立动物药土鳖虫中黄曲霉毒素B1、B2、G1、G2、M1、伏马毒素B1、B2、桔青霉毒素、T-2毒素、呕吐毒素、赭曲霉毒素A及玉米赤霉烯酮等12种真菌毒素的高效液相色谱-串联质谱同时测定方法。方法 土鳖虫样品加入80%乙腈溶液匀浆提取，用水稀释上清浓缩液后使用HLB固相萃取柱净化富集；以乙腈-5mM醋酸铵（pH=3.0）为流动相经C18色谱柱梯度洗脱分离，采用电喷雾（ESI）离子源、多反应监测（MRM）及正负离子模式采集数据。结果12种真菌毒素的线性良好，低、中、高3个水平的加标回收率在68%-110%之间，RSDs为1.5%-13%。抽查4批市售土鳖虫样品，结果 有3批土鳖虫样品黄曲霉毒素有检出，并且超出限量标准。而赭曲霉毒素A、呕吐毒素、玉米赤霉烯酮和伏马毒素类均对土鳖虫有不同程度的污染，其中赭曲霉毒素A和玉米赤霉烯酮的污染较为严重，分别超出限量的30%和60%。结论 土鳖虫易受黄曲霉毒素、伏马毒素以及玉米赤霉烯酮等真菌毒素的污染，本研究以土鳖虫为例建立的检测方法，以期为动物类中药的真菌毒素污染的含量测定提供技术支持及理论依据。

Objective To establish a high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method for determination of 12 kinds of mycotoxins (aflatoxinB1, B2, G1, G2, M1, fumonisinsB1, B2, Citrinin, T-2toxin, deoxynivalenol, ochratoxin A, and zearalenone) in animal drug Eupolyphaga Seu Steleophaga. Methods The samples were extracted with 80% acetonitrile, and then the concentrated liquid supernatant was purified by HLB solid phase extraction column. Acetonitrile-5 mM ammonium acetate (pH=3.0) was used as mobile phase gradient elution and the C18 column was used to separate the mycotoxins. Electrospray ionization (ESI) both in positive and negative modes with multiple reaction monitoring mode were employed to detected the signals. Results The linear regression of the 12 mycotoxins was good in their linear ranges respectively. The recovery rate of the 3 levels of low, medium and high ranged from 68% to 110%, and the relative standard deviations (RSDs) were between 1.5% and 13%. Aflatoxins were detected with over the limit in 3 of the 4 batches market sold Eupolyphaga Seu Steleophaga using the established method. Ochratoxin A, vomitoxin, zearalenone and fumonisin were all detected in different degrees in these samples, exceeding 30% and 60% of the limit respectively. Conclusion The Eupolyphaga Seu Steleophaga are easily infected with the toxins, especially aflatoxins, fumonisins, and zearalenone. The method established in this study can provide technical and theoretical for the detection and content determination of mycotoxin contamination of animal medicine by Eupolyphaga Seu Steleophaga.

湖北省食品药品监督管理局科研项目（201801001）：中药材毒源真菌污染快速筛查关键技术研究及方法建立，负责人：汪波。