法医学杂志

法医学杂志 ›› 2021, Vol. 37 ›› Issue (5): 646-652.DOI: 10.12116/j.issn.1004-5619.2020.301001

• 论著 • 上一篇    下一篇

血液中5种蘑菇毒素的UPLC-HRMS分析

刘文乔1,2(), 施妍3, 向平3, 于峰1, 谢冰1, 董玫1, 哈婧2, 马春玲1(), 文迪1()   

  1. 1.河北医科大学法医学院 河北省法医学重点实验室 河北省法医分子鉴定协同创新中心 河北医科大学法医鉴定中心，河北 石家庄 050017
    2.河北科技大学化学与制药工程学院，河北 石家庄 050018
    3.司法鉴定科学研究院 上海市法医学重点实验室 司法部司法鉴定重点实验室 上海市司法鉴定专业技术服务平台，上海 200063
  • 收稿日期:2020-10-06 发布日期:2021-10-25 出版日期:2021-10-28
  • 通讯作者: 马春玲,文迪
  • 作者简介:马春玲，女，教授，主任法医师，主要从事法医毒物学研究；E-mail：chunlingma@126.com
    文迪，男，副教授，副主任法医师，主要从事法医毒物学研究；E-mail：wendi01125@126.com
    刘文乔（1990—），女，硕士研究生，主要从事药物分析研究；E-mail：862307735@qq.com
  • 基金资助:
    国家重点研发计划资助项目(2016YFC0800704);司法部司法鉴定重点实验室资助项目;上海市法医学重点实验室资助项目(17DZ2273200);上海市司法鉴定专业技术服务平台资助项目(19DZ2292700)

Analysis of Five Mushroom Toxins in Blood by UPLC-HRMS

Wen-qiao LIU1,2(), Yan SHI3, Ping XIANG3, Feng YU1, Bing XIE1, Mei DONG1, Jing HA2, Chun-ling MA1(), Di WEN1()   

  1. 1.Hebei Key Laboratory of Forensic Medicine, Collaborative Innovation Center of Forensic Medical Molecular Identification, Forensic Identification Center of Hebei Medical University, College of Forensic Medicine, Hebei Medical University, Shijiazhuang 050017, China
    2.College of Chemical and Pharmaceutical Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China
    3.Shanghai Key Laboratory of Forensic Medicine, Key Laboratory of Forensic Science, Ministry of Justice, Shanghai Forensic Service Platform, Academy of Forensic Science, Shanghai 200063, China
  • Received:2020-10-06 Online:2021-10-25 Published:2021-10-28
  • Contact: Chun-ling MA,Di WEN

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摘要： 目的

建立同时快速检测血液中5种蘑菇毒素（α-鹅膏毒肽、羧基二羟鬼笔毒肽、蝇蕈醇、毒蝇碱和赛洛新）的超高压液相色谱-高分辨质谱联用（ultra-high performance liquid chromatography-high resolution mass spectrometry，UPLC-HRMS）分析方法。

方法

血液样品经乙腈-水溶液（V乙腈∶V水=3∶1）提取、PAX粉末净化后，在ACQUITY Premier C18色谱柱上分离，梯度洗脱。采用FullMS-ddMS2/正离子扫描模式对5种蘑菇毒素进行监测，根据一级母离子和二级碎片离子的精确质量数进行定性定量分析。

结果

5种蘑菇毒素在各自线性范围内线性关系良好，决定系数（R2）≥0.99；检出限0.2~20 ng/mL；定量限0.5~50 ng/mL；低、中、高3个添加水平的回收率89.6%~101.4%，相对标准偏差1.7%~6.7%，准确度90.4%~101.3%，日内精密度0.6%~9.0%，日间精密度1.7%~6.3%，基质效应42.2%~129.8%。

结论

本研究建立的方法操作简便快捷、回收率高，可同时对生物检材中多种蘑菇毒素进行快速准确的定性筛选和定量分析，为蘑菇中毒案事件的鉴定提供依据。

关键词: 法医学, 毒物化学, 蘑菇毒素, 超高压液相色谱-高分辨质谱, 血液, 小鼠

Abstract: Objective

To develop a method for the simultaneous and rapid detection of five mushroom toxins （α-amanitin， phallacidin， muscimol， muscarine and psilocin） in blood by ultra-high performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）.

Methods

The blood samples were precipitated with acetonitrile-water solution（Vacetonitril∶Vwater=3∶1） and PAX powder， then separated on ACQUITY Premier C18 column， eluted gradient. Five kinds of mushroom toxins were monitored by FullMS-ddMS2/positive ion scanning mode， and qualitative and quantitative analysis was conducted according to the accurate mass numbers of primary and secondary fragment ions.

Results

All the five mushroom toxins had good linearity in their linear range， with a determination coefficient （R2）≥0.99. The detection limit was 0.2-20 ng/mL. The ration limit was 0.5-50 ng/mL. The recoveries of low， medium and high additive levels were 89.6%-101.4%， the relative standard deviation was 1.7%-6.7%， the accuracy was 90.4%-101.3%， the intra-day precision was 0.6%-9.0%， the daytime precision was 1.7%-6.3%， and the matrix effect was 42.2%-129.8%.

Conclusion

The method is simple， rapid， high recovery rate， and could be used for rapid and accurate qualitative screening and quantitative analysis of various mushroom toxins in biological samples at the same time， so as to provide basis for the identification of mushroom poisoning events.

Key words: forensic medicine, toxicological chemistry, mushroom toxins, ultra-high performance liquid chromatography-high resolution mass spectrometry, blood, mice

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