6种动物类药材中黄曲霉毒素污染的液质联用检测

Detection of Aflatoxin Contamination in 6 Traditional Chinese Animal Medicines by LC-MS/MS

目的：建立一种快速检测动物类药材污染黄曲霉毒素B1（AFB1）和黄曲霉毒素G1（AFG1）的液质联用方法,并用于水蛭等6种动物类中药材污染情况的分析。方法：药材样品加84%乙腈提取,提取液挥干后用甲醇溶解,采用高效液相串联质谱检测方法,C18色谱柱（2.1 mm×100 mm,3μm）,以甲醇-4.0 mmol·L^-1乙酸铵-0.1%甲酸水溶液为流动相梯度洗脱,质谱使用ESI离子源,正离子扫描模式,选择反应检测（SRM）模式进行定量。结果：6种药材中AFB1,AFG1的基质效应介于82%~119%;以水蛭作为代表性基质,进行加样回收率试验,按低、中、高3个浓度水平添加时,AFB1与AFG1的提取回收率、绝对回收率介于80%~117%。方法学考察精密度RSD 1.8%~7.5%,重复性RSD 5.1%~14%。将该方法应用于55份动物药材样品检测,结果显示包括水蛭和土鳖虫在内的4份样品受到污染,其中土鳖虫污染率达33.3%,污染水平处于1.26~26.3μg·kg^-1。结论：土鳖虫等动物类药材受黄曲霉毒素污染的情况应当引起足够的重视。针对药材生产的各个环节,采取恰当的措施降低黄曲霉毒素污染的风险是十分必要的。

Objective： To establish the liquid chromatography-mass spectrometry（LC-MS） method for rapid determination of aflatoxin B1（AFB1） and aflatoxin G1（AFG1） in traditional Chinese animal medicines（TCAM）,and evaluate the contamination of two aflatoxins in the sample of 6 frequently-used TCAM. Method：The sample was extracted with 84% acetonitrile; then the extract was evaporated and the residue was re-suspended in methanol. The separation was carried out on a high performance liquid chromatograph equipped with a C18 column（2. 1 mm × 100 mm,3 μm）,with methanol-4. 0 mmol·L ammonium acetate-0. 1% formic acid as the mobile phase for gradient elution. The quantification was carried out with a tandem quadrupole mass spectrometer by using selective reaction monitoring（SRM） in positive ion mode. Result： The matrix effect for AFB1 and AFG1 in 6 TCAMs was acceptable,ranging from 82% to 119%. Recovery tests were performed with Hirudo as the representative matrix at three different concentration levels（low,middle,and high）. The absolute recoveries and the extraction recoveries of AFB1 and AFG1 were between 80% and 117%. Precision RSD was between 1. 8%-7. 5% and repeatability RSD was between 5. 1%-14%. The developed method was applied to evaluate the contamination of two aflatoxins in 55 samples of 6 TCAMs,and 4 of these 55 samples were contaminated,of which Eupolyphaga Steleophage samples showed a contamination rate of 33. 3% with a contamination level of 1. 26-26. 3 μg·kg^-1. Conclusion： TCAMs like Eupolyphaga Steleophage are susceptible to aflatoxin contamination and attentions should thus be paid to their potential hazards. It is necessary to take appropriate measures to reduce the risk of aflatoxin contamination in various sections during the production of medicinal materials.