QuEChERS-超高效液相色谱-串联质谱法测定野生菌中的15种蘑菇毒素

Determination of 15 kinds of mushroom toxins in wild mushrooms based on QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  野生菌  蘑菇毒素  液相色谱线性离子阱质谱  QuEChERS  MRM-IDA-EPI

英文关键词:wild fungus  mushroom toxin  liquid chromatography-linear ion trap mass spectrometry  QuEChERS  MRM-IDA-EPI

中文摘要:

      【目的】建立野生菌中15种蘑菇毒素的超高效液相色谱串联质谱(UPLC-MS/MS)定量分析方法和基于线性离子阱质谱的信息依赖采集增强子离子扫描(MRM-IDA-EPI)二级谱图筛查方法,为有毒野生菌食源性中毒定量检测和定性筛查提供参考依据。【方法】野生菌样本通过甲酸-甲醇-水-乙腈(1:40:40:19,v/v)混合溶液超声提取,QuEChERS试剂提取净化,采用UPLC-MS/MS和MRM-IDA-EPI方法对15种蘑菇毒素进行定量分析和定性筛查。【结果】通过野生菌样本前处理方法和色谱条件优化,对15种蘑菇毒素进行0.04 mg/kg、0.1 mg/kg和0.4 mg/kg三水平加标回收试验,方法准确度为76.6%～109.2%、精密度(RSD)为0.3%～7.6%；15种蘑菇毒素的线性范围为0.02～1.0 mg/L,线性相关系数(r)在0.998～0.9994之间；其中α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、光盖伞素和鹅膏蕈氨酸的方法检出限(LOD,S/N=3)和定量限(LOQ)分别为10 μg/kg和30 μg/kg,二羟鬼笔毒肽、羧基二羟鬼笔毒肽、羧基三羟鬼笔毒肽、毒蝇碱、蝇蕈醇、甲基裸盖菇素、鹿花菌素、脱磷酸裸盖菇素、奥来毒素和鬼伞菌素的LOD和LOQ分别为20 μg/kg和60 μg/kg,采用QuEChERS前处理方法对野生菌样本进行处理,样本基质效应系数K值在0.91～1.08之间。【结论】所建立的野生菌样本前处理方法对15种蘑菇毒素的定量测定无基质干扰,15种蘑菇毒素的UPLC-MS/MS和MRM-IDA-EPI分析方法结果准确、重现性好、灵敏度高,该方法适用于有毒野生菌引起的食源性中毒定量分析和定性筛查。

英文摘要:

      : Objective:Establish a quantitative analysis method for fifteen mushroom toxins in wild mushrooms using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and a secondary spectrum library screening method based on information-dependent acquisition enhancement ion scanning (MRM-IDA-EPI) using linear ion trap mass spectrometry, providing a reference for quantitative detection and qualitative screening of foodborne poisoning caused by toxic wild mushrooms.Methods:Wild mushroom samples were extracted by ultrasonic extraction using a formic acid-methanol-water-acetonitrile (1:40:40:19, v/v) mixed solution, and purified using QuEChERS reagents. UPLC-MS/MS and MRM-IDA-EPI methods were used for quantitative analysis and qualitative screening of 15 mushroom toxins.Results:Through the optimization of the pre-treatment method and chromatographic conditions for wild mushroom samples, a three-level spiked recovery test was conducted for fifteen mushroom toxins at 0.04 mg/kg, 0.1 mg/kg, and 0.4 mg/kg. The method accuracy was 76.6% to 109.2%, and the precision (RSD) was 0.3%-7.5%. The linear range of the fifteen mushroom toxins was 0.02-1.0 mg/L, and the linear correlation coefficient (r) was between 0.998-0.9994;The detection limit (LOD, S/N=3) and quantification limit (LOQ) of α-amanitin, β-amanitin, γ-amanitin, psilocybine, and ibotenic acid were 10 μg/kg and 30 μg/kg, respectively. The LOD and LOQ of phalloidin, Phallacidin, phallisacin, muscarine, musimol, baeocystin, gyromitra toxins, psilocine, orellanine, and coprine were 20 μg/kg and 60 μg/kg, the sample matrix effect coefficient K value of the wild mushroom samples processed using the QuEChERS pre-treatment method was between 0.91 and 1.08.Conclusion:The established pre-treatment method for wild mushroom samples has no matrix interference for the quantitative determination of fifteen mushroom toxins. The UPLC-MS/MS and MRM-IDA-EPI analysis methods for fifteen mushroom toxins have accurate results, good reproducibility, and high sensitivity. This method is suitable for quantitative analysis and rapid screening of foodborne poisoning caused by toxic wild mushrooms.