丹磺酰氯衍生-液相色谱-串联质谱法检测野生菌鹅膏蕈氨酸和毒蝇母

Determination of ibotenic acid and muscimol in wild mushroom by dansyl chloride derivatization-liquid chromatography-tandem mass spectrometry

目的建立丹磺酰氯(DNSCl)衍生-液相色谱-串联质谱(LC-MS/MS)检测野生菌中鹅膏蕈氨酸(IBA)和毒蝇母(MUS)法,为毒蘑菇中毒事件的病因鉴定提供技术支持。方法样品用盐酸溶液超声提取10 min,离心后加0.38 mL水、0.5 mL 5%NaHCO3溶液和0.6 mL 30 mmol/L DNSCl丙酮溶液进行双分子DNSCl衍生化,于60℃反应30 min,经乙腈稀释和沉淀无机盐后,采用Waters XBridge^(TM) BEH C18色谱柱分离,LC-MS/MS测定IBA和MUS(多反应监测模式)。结果野生菌中IBA和MUS在0.5~250 mg/kg范围内线性关系良好,相关系数分别为0.997和0.999;检出限分别为0.15 mg/kg和0.1 mg/kg。在0.5、10、250 mg/kg水平下IBA回收率为84.5%~102.0%,相对标准偏差(RSD)为4.7%~8.6%;MUS回收率为88.6%~95.4%,RSD为4.9%~7.5%。结论通过优化提取和双分子DNSCl衍生化反应等前处理条件,采用LC-MS/MS法可以快速准确测定野生菌中的IBA和MUS。

Objective To develop an analytical method of ibotenic acid(IBA)and muscimol(MUS)in wild mushroom by dansyl chloride(DNSCl)derivatization-liquid chromatography-tandem mass spectrometry(LC-MS/MS),and to provide technical support for etiological identification of mushroom poisoning events.Methods The sample was extracted with hydrochloric acid solution,derived by bimolecular DNSCl,diluted and inorganic salts precipitated with acetonitrile.The extract was separated by a waters XBridge^(TM) BEH C18 column and measured by LC-MS/MS.Results The limits of detection for IBA and MUS in wild mushroom were 0.15 mg/kg and 0.1 mg/kg,respectively.Good linear relationship was obtained for IBA and MUS at the range of 0.5-250 mg/kg with the correlation coefficient of 0.997 and 0.999,respectively.The average recoveries at three spiking levels were 84.5%-102.0%with relative standard deviations(RSDs,n=6)of 4.7%-8.6%for IBA.The average recoveries were 88.6%-95.4%with RSDs(n=6)of 4.9%-7.5%for MUS.Conclusion The optimized sample extraction and bimolecular DNSCl derivatization conditions can achieve rapid and accurate analysis of IBA and MUS in wild mushroom poisoning sample.