血浆中85种有毒生物碱快速筛查方法的建立及其应用  

Establishment and application of a rapid screening method for 85 toxic alkaloids in plasma samples

作　　者：刘小红[1] 李永刚[1] 孔芳 唐琳[1] 何晓宏 杨荣娇 闻胜[1] 罗苹[1] LIU Xiaohong;LI Yonggang;KONG Fang;TANG Lin;HE Xiaohong;YANG Rongjiao;WEN Sheng;LUO Ping(Hubei Provincial Key Laboratory for Applied Toxicology,Hubei Provincial Centre for Disease Control and Prevention,Hubei Wuhan 430079,China;The Center for Disease Control and Prevention of Enshi Tujia&Miao Autonomous Prefecture,Hubei Enshi 445000,China)

机构地区：[1]应用毒理湖北省重点实验室,湖北省疾病预防控制中心,湖北武汉430079 [2]恩施土家族苗族自治州疾病预防控制中心,湖北恩施445000

出　　处：《中国食品卫生杂志》2022年第6期1202-1211,共10页Chinese Journal of Food Hygiene

基　　金：湖北省卫生健康科研基金资助(WJ2021M209);武汉市卫生健康科研基金资助(WY22A02)。

摘　　要：目的 利用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱建立血浆中85种有毒生物碱快速筛查方法。方法 构建85种生物碱高分辨数据库;血浆经乙腈沉淀蛋白进行预处理,采用Acquity Waters BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以5 mmol/L甲酸铵(含0.1%甲酸)和乙腈为流动相进行梯度洗脱,对比了3种数据采集方式下(Full mass/dd-MS~2、Full mass/AIF、Full mass/DIA)加标血浆中85种有毒生物碱筛查情况。结果 血浆在5、50和250 ng/mL 3个浓度添加水平下,Full mass/DIA模式下的筛选准确率均为最高。利用生物碱数据库,选择Full mass/DIA模式采集样品数据信息,建立了血浆中85种有毒生物碱快速筛查方法。80%以上生物碱在血浆中的检出限低于10 ng/mL。利用该方法成功在滇乌头碱毒物代谢动力学实验获得的3份Wistar大鼠血浆中筛选到了目标物滇乌头碱及其可能的代谢产物脱氧乌头碱及印乌头碱。结论 该方法快速、高效,可以在无标准品情况下实现突发性有毒生物碱中毒血浆的快速筛查。Objective To establish a rapid screening method for 85 toxic alkaloids in plasma samples using ultra-high performance liquid chromatography coupled with quadrupole/exactive high resolution mass spectrometry(UHPLC-Q/Exactive).Methods A high resolution mass spectrometry compound database(CDB) of 85 alkaloids was established by UHPLC-Q/Exactive.Acetonitrile was used for protein precipitation as pretreatment in plasma samples.An Acquity Waters BEH C_(18) column(100 mm×2.1 mm,1.7 μm) was applied for gradient elution with 5 mmol/L ammonium formate(containing 0.1% formic acid) and acetonitrile as mobile phases.The screening accuracy of 85 toxic alkaloids in spiked plasma samples was compared in three data collection modes(Full mass/dd-MS~2,Full mass/AIF and Full mass/DIA).Results The screening accuracy of Full mass/DIA was the highest when plasma samples were added at three concentration levels of 5,50 and 250 ng/mL.Combined with the CDB,a rapid screening method for 85 toxic alkaloids in plasma samples was established under the Full mass/DIA data acquisition mode.The limits of detection of more than 80%of the alkaloids in plasma samples were lower than 10 ng/mL.With this method,the target substance-yunaconitine and its possible metabolites,3-deoxyaconitine and indaconitine were successfully screened in the 3 Wistar rat plasma samples obtained from the yunaconitine toxicokinetics experiment.Conclusion The method is fast,efficient,and suitable for the rapid screening of 85 toxic alkaloids in acute toxic alkaloid poisoned plasma samples with the absence of standard substances.

关 键 词：超高效液相色谱-四级杆/静电场轨道阱高分辨质谱 血浆 有毒生物碱 Full mass/DIA 快速筛查 

分 类 号：R155[医药卫生—营养与食品卫生学]