超高效液相色谱-串联质谱法测定野生蘑菇中的6种鹅膏肽类毒素

Determination of 6 kinds of amatoxins and phallotoxins in wild mushrooms by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  鹅膏毒肽  鬼笔毒肽  超高效液相色谱-质谱法  野生蘑菇

英文关键词:amatoxins  phallotoxins  ultra performance liquid chromatography-tandem mass spectrometry  wild mushrooms

中文摘要:

      目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定野生蘑菇中6种鹅膏肽类毒素的含量。方法 干燥后的样品粉末经水提取后, 提取液转移至固相萃取柱净化, 采用ACQUITY UPLC BEH C18色谱柱以5 mmol/L的甲酸铵水溶液和甲醇为流动相进行梯度洗脱, 流速为0.3 mL/min, 柱温40 ℃, 采用多反应监测模式检测。结果 6种待测物在色谱图上全部做到基线分离, 在20~1000 μg/kg的范围内呈现良好的线性关系, 检出限均达到20 μg/kg, 不同水平下的加标回收实验中各待测物的平均回收率为78.2%~95.7%, 相对标准偏差为1.2%~5.2%。结论 该方法操作简单, 灵敏度高, 重复性好, 适用于野生蘑菇中鹅膏肽类毒素的检测。

英文摘要:

      Objective To establish a method for the determination of 6 kinds of amatoxins and phallotoxins in wild mushrooms by ultra performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). Methods After water extraction, the samples were separated with a ACQUITY UPLC BEH C18 colum at the flow rate of 0.3 mL/min by gradient elution using 5 mmol/L ammonium formate aqueous solution and methanol as mobile phases, the temperature of the colum was maintained at 40 ℃, and multiple reaction monitoring mode was adopted for detection. Results The analytes could be separated with a baseline resolution and showed a good linearity in the range of 20-1000 μg/kg. The limits of detection of 6 kinds of amanitins and phallotoxins were 20 μg/kg. The recoveries of analytes in different levels were 78.2%-95.7%, and the relative standard deviations were 1.2%-5.2%. Conclusion This method is simple, sensitive and reproducible, and it is suitable for the determination of peptidotoxin in wild mushroom.