分散固相萃取-超高效液相色谱-串联质谱法同时测定野生菌中6种鹅膏毒肽和鬼笔毒肽毒素

Simultaneous determination of 6 kinds of amatoxins and phallotoxins in wild mushrooms by dispersive solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  野生菌  鹅膏毒肽  鬼笔毒肽  分散固相萃取法  超高效液相色谱-串联质谱法

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

中文摘要:

      目的 建立分散固相萃取(dispersive solid-phase extraction, dSPE)前处理方法和超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速测定野生菌中6种鹅膏毒肽类和鬼笔毒肽类毒素的含量的检测方法。方法 样品经低温干燥, 用甲酸酸化的甲醇超声萃取, 采用石墨化碳黑(graphitized carbon black, GBC)、C18和N-丙基乙二胺(primary secondary amine, PSA)吸附净化, 采用UPLC-MS/MS进行检测。结果 6种毒肽在质量浓度20~1000 μg/kg范围内具有良好的线性关系, 相关系数大于0.998。方法检出限为20 μg/kg, 定量限为50 μg/kg。在加标水平为50、250、500 μg/kg时, 方法的平均回收率为78.9%~115.5%, 相对标准偏差为1.6%~6.8% (n=3)。结论 本方法准确可靠、可操作性强, 适用于野生菌中6种毒肽的检测。

英文摘要:

      Objective To establish a pretreatment method of dispersed solid-phase extraction (dSPE) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the rapid determination of 6 kinds of amatoxins and phallotoxins in wild mushrooms. Methods Samples were dried at low temperature, extracted with methanol acidified with formic acid by ultrasonic extraction, purified by the mixture of graphitized carbon black (GBC), C18 and primary secondary amine (PSA), and then determined by UPLC-MS/MS. Results Good linearities of 6 kinds of toxic peptides were obtained at the concentration of 20-1000 μg/kg, with the correlative coefficients more than 0.998. The limits of detection were 20 μg/kg and the limits of quantitation were 50 μg/kg. When the spiked concentrations were 50, 250, 500 μg/kg, the average recoveries were 78.9%-115.5%, and the relative standard deviations were 1.6%-6.8% (n=3). Conclusion This method is accurate, reliable and operable, which is suitable for the determination of 6 kinds of toxic peptides in wild mushrooms.