分散固相萃取结合超高效液相色谱-串联质谱法快速测定毒蘑菇中的6种野生蘑菇毒素

Rapid determination of 6 kinds of wild mushroom toxins in poisonous mushrooms by dispersive solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  分散固相萃取  超高效液相色谱-串联质谱法  鹅膏毒肽类蘑菇毒素  鬼笔毒肽类蘑菇毒素

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

中文摘要:

      目的 建立一种应用分散固相萃取结合超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速测定毒蘑菇中α-鹅膏毒肽、?-鹅膏毒肽、γ-鹅膏毒肽、二羟基鬼笔毒肽、羧基二羟基鬼笔毒肽、羧基三羟基鬼笔毒肽的分析方法。方法 蘑菇样品混合C18分散剂经甲醇提取后离心, 取上清液+0.2%甲酸水(2:8, V:V)混合, 通过C18色谱柱(2.1 mm×100 mm, 1.7 μm), 以5 mmol/L甲酸铵水溶液和甲醇为流动相进行梯度洗脱, 用超高效液相色谱-串联质谱进行定性、定量分析。结果 α-鹅膏毒肽、?-鹅膏毒肽、γ-鹅膏毒肽、二羟基鬼笔毒肽、羧基二羟基鬼笔毒肽、羧基三羟基鬼笔毒肽在10~100 μg/L浓度范围内具有良好的线性关系, 相关系数(r)均大于0.999, 检出限为0.10~0.60 mg/kg, 定量限为0.30~2.00 mg/kg, 平均回收率为72.9%~101.6%, 相对标准偏差在2.1%~9.0%之间。结论 该方法快速、准确, 能满足蘑菇中6种毒肽类化合物的快速检测要求。

英文摘要:

      Objective To establish an analytical method for the rapid determination of α-amanitin, ?-amanitin, γ-amanitin, phalloidin, phallacidin, phallisacin in poisonous mushrooms by dispersive solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The mushroom samples were mixed with C18 dispersant, extracted with methanol, and centrifuged. The supernatant was mixed with 0.2% formic acid water (2:8, V:V), and passed through a C18 column (2.1 mm×100 mm, 1.7 μm) with gradient elution using 5 mmol/L ammonium formate aqueous solution and methanol as the mobile phase. Qualitative and quantitative analysis was performed by ultra performance liquid chromatography-tandem mass spectrometry. Results α-amanitin, ?-amanitin, γ-amanitin, phalloidin, phallacidin and phallisacin had good linear relationships within the concentration ranges of 10?100 μg/L with the correlation coefficient (r) greater than 0.999, the limits of detection were 0.10?0.60 mg/kg, the limits of quantitation were 0.30?2.00 mg/kg, the average recoveries were 72.9%?101.6%, and the relative standard deviations were 2.1%?9.0%. Conclusion This method is rapid and accurate, and can meet the requirements of rapid determination of 6 kinds of toxic peptides in mushrooms.