大鼠血清和尿液中4种鹅膏肽类毒素代谢特征的超高效液相色谱-四极杆飞行时间串联质谱分析  

Analysis of four amatoxins and phallotoxins content in rat serum and urine by ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry

作　　者：柳洁[1] 黄海燕[2] 曾灼祥[1] 陈效 肖嘉慧 王杨燕 LIU Jie;HUANG Hai-yan;ZENG Zhuo-xiang;CHEN Xiao;XIAO Jia-hui;WANG Yang-yan(Nanshan Center for Disease Control and Prevention,Shenzhen,Guangdong 518054,China)

机构地区：[1]深圳市南山区疾病预防控制中心,广东深圳518054 [2]深圳市疾病预防控制中心,广东深圳518055

出　　处：《中国卫生检验杂志》2018年第16期1941-1946,共6页Chinese Journal of Health Laboratory Technology

基　　金：深圳市科技创新委科技研发资金基础研究项目(JCYJ-20140416095154396)

摘　　要：目的建立血清和尿液中4种鹅膏肽类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,考察和分析4种鹅膏肽类毒素在大鼠体内的代谢特征,为中毒样品的采集提供科学依据。方法血清样品用乙腈+0.1%TFA二氯甲烷提取,残渣用流动相溶解;尿液样品则直接过0.22μm微孔滤膜;样品溶液经HSS T3色谱柱分离后用ESI-QTOF MS进行测定;选取SD健康大白鼠分别用4种鹅膏肽类毒素染毒,按需收集不同时间段的尿液和血液进行分析。结果血清和尿液样品的检出限分别为10 ng/ml和5 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%。大鼠染毒尿液中均检出4种毒素,鹅膏毒肽的含量明显比鬼笔毒肽的含量高,大鼠染毒血清中仅14%检测到毒素的存在。结论检测方法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,采集中毒8 h内的尿液对检出鹅膏毒肽的几率最大。Objective To develop a simple and sensitive method for the determination of 4 amatoxins and phallotoxins in serum and urine by ultra performance liquid chromatography（ UPLC） quadrupole time-of-flight（ Q-TOF） mass spectrometry（ MS）,and to analyze toxins content in rat serum and urine. Methods Serum samples were extracted with acetonitrile ＋0. 1%TFA in dichloromethane,and the residue was dissolved in mobile phase; urine samples directly passed through a 0. 22 μm microporous membrane; sample solutions were separated by HSS T3 column and determined by ESI-Q-TOF MS. SD healthy rats were selected to be infected with 4 kinds of amanita peptide toxoids,and urine and blood were collected and analyzed from different periods. Results The LOD for four toxins in serum sample and urine sample were 10 ng/m L and 5 ng/m L,respectively. The recovery rates of the four toxins in serum sample were in the range of 77. 4%-95. 6% and 73. 8%-103. 8%,with RSDs of 3. 0%-18. 0% and 1. 2%-25. 4%. Four toxins were detected in the urine of the rats,and the content of amanita peptide was significantly higher than that of the ghost peptide. Toxins were detected from 14% of the rats. Conclu sion The present method is suitable for the identification of α-amanitin,β-amanitin,phallacidin and phalloidin in serum and urine samples for poison purposes. The collection of urine sample for analysis within 8 h after poisoning was suggested.

关 键 词：α-鹅膏毒肽 β-鹅膏毒肽 二羟鬼笔毒肽 羧基二羟鬼笔毒肽 超高效液相色谱-电喷雾离子化-四级杆飞行时间串联质谱 大鼠血清 大鼠尿液 

分 类 号：O657.63[理学—分析化学]