薛康 胡江涛 俞凌云 刘菲 马丽 龚婷婷 华燚 陈佳玥

摘要 [目的]建立一种基于超高效液相色谱-串联质谱法（UPLC-MS/MS）快速测定野生毒蕈中6种鹅膏毒肽和鬼笔毒肽类毒素的分析方法。[方法]将采集的四川野生毒蕈子实体低温烘干，用甲醇超声提取，40 ℃旋转蒸发近干，加水复溶，Oasis HLB固相萃取小柱净化，经Waters HSS T3色谱柱分离，ESI正离子模式下多反应监测（MRM）方式分析。[结果]6种毒肽在50～1 000 μg/kg具有良好的线性关系，相关系数均大于0.99，方法检出限为30 μg/kg。α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、二羟鬼笔毒肽、羧基二羟鬼笔毒肽、羧基三羟鬼笔毒肽在低、中、高3个浓度的平均回收率在93.1%～117.5%，变异系数（CV）在1.49%～7.77%。[结论]该方法能准确、灵敏地测定野生毒蕈中6种毒肽，适用于野生菌中毒肽毒素的测定。

关键词 野生毒蕈;鹅膏毒肽;鬼笔毒肽;超高效液相色谱-串联质谱法

中图分类号 R155.3 文献标识码 A

文章编号 0517-6611（2021）17-0182-04

doi：10.3969/j.issn.0517-6611.2021.17.048

Abstract [Objective]To establish a method for rapid determination of 6 kinds of amanitins and phallotoxins in wild poisonous mushrooms by ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）.[Method]The wild mushrooms samples collected in Sichuan were dried at low temperature， the amanitins and phallotoxins in samples were extracted with methanol by ultrasonic extraction， then 40 ℃ rotary evaporation drying ，water redissolution， purified by HLB solid phase extraction column and separated on Waters HSS T3 column，lastly detected by electrospray（ESI+） in MRM mode. [Result] The 6 kinds of amanitins and phallotoxins had good linear relationships in the range of concentration 50-1000 μg/kg with the correlation coefficient value greater than 0.99.The detection limits of 6 kinds of amanitins and phallotoxins were 30 μg/kg.The average recovery rates of α-amanitin， β-amanitin，γ-amanitin， phalloidin， phallacidin and phallisacin at 3 spiked levels were 93.1%-117.5%. The coefficients of variation were 1.49%-7.77%. [Conclusion]The method can accurately and sensitively determine 6 kinds of amanitins and phallotoxins in poisonous mushrooms， which is suitable for the detection of amanitins and phallotoxins in wild mushrooms.

Key words Wild poisonous mushrooms;Amanitins;Phallotoxins;UPLC-MS/MS

毒蕈即日常所说的毒蘑菇，是指不同类型的含有毒素的大型野生真菌，由于有些毒蕈在外观上与食用菌没有明显区别，导致误食毒蕈而引發中毒的事件时有发生，且在群体性食物中毒中，毒蕈中毒的致死率极高，其中肝损伤型毒蕈中毒致死率高达90%～100%[1]。据统计，我国毒蕈中毒事件中，95%是由鹅膏菌属引起的[2]。因为鹅膏菌中含有化学性质稳定且毒性极强的鹅膏多肽类毒素，该类毒肽耐高温、干燥和酸碱环境，一般的烹调加工不会破坏其毒性，不幸误食后，毒肽就会通过消化系统进入肝脏，对肝脏造成严重的损伤，最终引发多器官功能衰竭而死亡[3-5]。……