高分辨质谱-非信息依赖数据采集-后靶向筛查策略在不明原因食物中毒鉴定中的应用  

Identification of Unknown Toxicants in Food Poisoning by High Resolution Mass Spectrometry-Data Independent Acquisition-Post Targeted Screening Strategy

作　　者：张美娟 陈佳[1] 林缨[1] 张雅姣[1] 郭磊[1] 徐斌[1] 谢剑炜[1] 

机构地区：[1]军事医学科学院毒物药物研究所,抗毒药物与毒理学国家重点实验室,北京100850

出　　处：《分析化学》2018年第2期157-164,共8页Chinese Journal of Analytical Chemistry

基　　金：本文系全军医学科研重大项目(No.AWS15J007)资助

摘　　要：基于超高效液相色谱-四极杆-飞行时间串联质谱(UPLC-QTOF MS/MS)技术,建立了高分辨质谱-非信息依赖数据采集-后靶向筛查策略(HRMS-DIA-Post targeted screening strategy),并成功应用于不明原因食物中毒患者呕吐物及粪便样本中的有毒化合物筛查鉴定。样品经溶剂提取与净化后,采用5 mmol/L甲酸铵-水(A)和0.1%甲酸-乙腈(B)作为流动相,在反相色谱柱上进行梯度洗脱。采用MS^E模式对样品进行质谱(ESI+)全信息采集,经UNIFI筛查软件进行谱峰识别、谱库检索等数据分析,初筛出疑似有毒化合物α-茄碱,随后结合其化合物特征及体内代谢行为,采用后靶向筛查策略在两种样品中均鉴定出相关化合物(包括水解产物及代谢产物等),如α-卡茄碱与其水解产物β-卡茄碱、γ-茄碱/卡茄碱以及共同代谢产物茄啶。采用单点校正法测得两种样品中α-茄碱含量均约为0.1 mg/kg,采用峰面积归一化法得到两种样品中龙葵素类似物总量分别约为0.5 mg/kg与0.6 mg/kg。本研究建立的高分辨质谱快速筛查策略可为不明原因食物中毒的未知毒物快速筛查鉴定提供有效的技术手段。Based on ultra-high-performance liquid chromatography-quadrupole/time of flight mass spectrometry( UPLC-Q-TOF MS/MS), a post targeted screening strategy using high resolution mass spectrometry under data independent acquisition was established,and successfully applied to screen and identify unknown toxicants in food poisoning patients' vomit and feces. After solvent extraction,the samples were separated and analyzed on a reversed-phase C_(18) column using a gradient elution program of 5 mmol/L ammonium formate and 0.1% formic acid aqueous solution( A) and 0.1% formic acid in acetonitrile solution( B). All-ions MS/MS information of samples was obtained using Q-TOF MS( ESI+) in MSE mode,followed by peak recognition and library searching using the UNIFI Scientific Information System, as a result,suspected toxic compound α-solanine was primarily detected. Furthermore,the related chemicals including hydrolysates and metabolites were identified in both samples based on post-targeted screening strategy,such asα-chaconine,β-chaconine,γ-solanine/chaconine and solanidine. The concentration of α-solanine in both samples was approximately 0.1 mg/kg by one-point calibration method,the total amount of solanine analogues in two samples was about 0.5 mg/kg and 0.6 mg/kg respectively through normalization of peak area. The high resolution mass spectrometry screening strategy established here could provide efficient screening method for rapid detection and identification of toxicants in unknown food poisoning.

关 键 词：后靶向筛查 未知有毒化合物 高效液相色谱-四极杆-飞行时间串联质谱 生物医学样品 龙葵素 

分 类 号：O657.63[理学—分析化学] R155.3[理学—化学]