基于液相色谱-串联质谱技术快速检测6种蘑菇毒素及临床应用探讨  

Rapid detection of six mushroom toxins in human body fluids by LC-MS/MS and its application in clinical poisoning cases

作　　者：阳硕 普山柏 郑粉双 文迪[4] 施妍 Yang Shuo;Pu Shanbai;Zheng Fenshuang;Wen Di;Shi Yan(Academy of Forensic Science,Shanghai 200063,China)

机构地区：[1]司法鉴定科学研究院,上海200063 [2]中国药科大学,江苏南京210009 [3]云南大学附属医院,云南昆明650021 [4]河北医科大学,河北石家庄050017

出　　处：《中国急救医学》2023年第9期673-678,共6页Chinese Journal of Critical Care Medicine

基　　金：“十四五”国家重点研发计划资助项目(2022YFC3302003);云南省科技厅科技计划项目(202301AS070037);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项目(21DZ2270800);上海市司法鉴定专业技术服务平台资助项目。

摘　　要：目的基于液相色谱-串联质谱(LC-MS/MS)技术建立快速检测血液、尿液中6种蘑菇毒素的分析方法,并将其应用于临床实际中毒案例。方法将0.1 mL血液、尿液经蛋白沉淀、氮吹复溶、超声过滤后用ACQUITY UPLC HSS T3液相色谱柱分离,质谱检测采用多反应监测模式(MRM)。结果该分析方法经过验证,符合方法学验证要求,其定量结果准确可靠,并于9例患者血液和(或)尿液样品中检出目标毒素,血液样品中浓度为1.12~5.63 ng/mL,尿液样品中浓度为1.01~9.27 ng/mL,检出物多为γ-鹅膏毒肽(γ-AMA)。新鲜蘑菇样品中检出目标物浓度为1.48~41.54 ng/mg,两份新苦粉孢牛肝菌样品中均检出羧基二羟基鬼笔毒肽(PCD)16.51~41.54 ng/mg。结论本研究中建立的LC-MS/MS方法前处理过程简单快速,具有较高的灵敏度,能同时检测多种蘑菇毒素类化合物,可为临床毒蕈中毒快速诊断提供策略和参考。Objective An analytical method based on liquid chromatography-tandem mass spectrometry(LC-MS/MS)was developed for the rapid detection of six mushroom toxins in blood and urine and applied to clinical poisoning cases.Methods 0.1 mL of blood or urine was subjected to protein precipitation,nitrogen blowing and redissolution,ultrasound filtration and then was separated by a ACQUITY UPLC HSS T3 liquid chromatography column.Multiple reaction monitoring mode was used for mass spectrometry detection.Results The analytical method had been validated and the quantitative results were accurate and reliable.Target toxins had been detected in blood and(or)urine samples of 9 patients.Amatoxins and(or)phallotoxins were identified in each whole blood sample at a range of 1.12-5.63 ng/mL and in urine samples with 1.01-9.27 ng/mL,and most of the detection results wereγ-amanitin.The concentration range of the detected target in fresh mushroom samples was 1.48-41.54 ng/mg.Phallacidin(PCD)was detected in both samples of Tylopilus neofelleus,and the concentration was 16.51-41.54 ng/mg.Conclusions The LC-MS/MS method established in this study has a simple and rapid pre-treatment process,with high sensitivity,and is capable of detecting a variety of mushroom toxin analogues simultaneously,which can be used as a strategy for the rapid diagnosis of mushroom poisoning in clinical emergencies.

关 键 词：鹅膏毒肽 鬼笔毒肽 蘑菇毒素 毒物分析 中毒案例 

分 类 号：R595.6[医药卫生—内科学]