【摘要】 目的建立血清和尿液中α-鹅膏毒肽、β-鹅膏毒肽、二羟鬼笔毒肽、羧基二羟鬼笔毒肽毒素的超高效液相色谱-四级杆飞行时间串联质谱测定方法。方法血清样品先用乙腈沉淀蛋白,再用含0.1%TFA的二氯甲烷提取,残渣用含2mmol/L乙酸铵的甲醇-水(30∶70)溶解;尿液样品直接过0.22μm微孔滤膜;样品溶液经HSS T3色谱柱分离后用Q-TOF MS进行测定。结果毒素准分子离子准确质量数及其同位素特征、色谱峰保留时间可作为毒素定性鉴定的依据,血清和尿液样品的检出限分别为10 ng/ml和5 ng/ml,定量限为20 ng/ml和10 ng/ml,加标回收率分别为77.4%~95.6%和73.8%~103.8%,相对标准偏差在3.0%~18.0%和1.2%~25.4%之间。应用该方法测定了大鼠染毒尿液中鹅膏肽类毒素。结论本方法简便、准确、可靠,适用于中毒事件生物样品中4种鹅膏肽类毒素的检测和确证分析。更多还原

【Abstract】 Objective To develop a method for the determination of α-amanitin,β-amanitin,phalloidin and phallacidin in serum and urine by ultra performance liquid chromatography(UPLC) quadrupole time-of-flight(Q-TOF) mass spectrometry(MS).Method For serum sample,the four toxins were extracted with 0.1% TFA in dichloromethane after precipitation of proteins with acetonitrile.The extracted solution was evaporated to dryness and the residue was redissolved in 2 mmol/L ammonium acetate in methanol-water(30∶ 70,v/v).The urine sample was directly filtered through a 0.22μm HPLC syringe filter.The toxins were analyzed on an UPLC-ESI-Q-TOF MS system with HSS T3 column.Result The accurate mass number of the protonated molecular peak together with its isotope peak profiles and its retention time could be used for identification of each toxin by TOF MS.The LOD and LOQ in serum sample were 10 ng/m L and 20 ng/m L,5 ng/m L and 10 ng/m L in a urine sample.The recovery rates of the four toxins in serum sample were in the range of 77.4%-95.6% with RSDs of 3.0%-18.0%.The recovery rates of the four toxins in urine sample were in the range of 73.8%-103.8% with RSDs of 1.2%-25.4%.The rat urine samples after single intraperitoneal administration were successfully analyzed by the proposed method.Conclusion The method is simple,accurate and is suitable for the identification of α-amanitin,β-amanitin,phallacidin and phalloidin in serum and urine samples for poison purposes.更多还原