液相色谱-串联质谱法快速筛查食物中毒样本中28种生物碱  

Rapid screening of 28 alkaloids in food poisoning samples by liquid chromatography-tandem mass spectrometry

作　　者：赵凌国[1] 罗兰 尹臻一 任燕 雷蕾[1] 马志锋 ZHAO Ling-guo;LUO Lan;YIN Zhen-yi;REN Yan;LEI Lei;MA Zhi-feng(Baoan District Center for Disease Prevention and Control,Shenzhen,Guangdong 518101,China;Longhua District Center for Disease Prevention and Control,Shenzhen,Guangdong 518110,China;School of Public Health,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]深圳市宝安区疾病预防控制中心,广东深圳518101 [2]深圳市龙华区疾病预防控制中心,广东深圳518110 [3]重庆医科大学公共卫生学院,重庆400016

出　　处：《中国热带医学》2023年第3期260-266,共7页China Tropical Medicine

基　　金：广东省医学科学技术研究基金项目(No.A2021416)。

摘　　要：目的查明一起食用八宝粥引起中毒事件的原因,建立28种生物碱的液相色谱-串联质谱筛查方法,为此类中毒事件处置提供借鉴。方法采用液相色谱-串联质谱联用技术筛查患者尿液、八宝粥以及食物原料中28种生物碱,对生物碱进行定量检测。样品用0.4%甲酸水溶液提取,用Acquity UPLC BEH C18色谱柱(1.7μm,100×2.1 mm)分离。采用乙腈-0.2%甲酸水为流动相,梯度洗脱。电离方式为电喷雾正电离模式,检测方式采用多反应监测,外标法定量。结果在0~100 ng/mL浓度范围,28种生物碱线性相关系数r≥0.999。尿样中28种生物碱回收率为63.0%~105.0%,相对标准差(relative standard deviations,RSDs)为5.8%~8.6%;八宝粥中28种生物碱回收率为72.0%~109.0%,RSDs为6.3%~9.7%;黑芝麻中28种生物碱回收率为60.0%~95.0%,RSDs为4.8%~8.2%。患者尿液中检出莨菪碱(2380.0 ng/mL)、东莨菪碱(3.6 ng/mL)和山莨菪碱(4.7 ng/mL)。八宝粥中检出莨菪碱(63.3μg/g)、东莨菪碱(5.7μg/g)和山莨菪碱(2.1μg/g)。在原料曼陀罗籽中检出莨菪碱(901.0μg/g)、东莨菪碱(80.0μg/g)和山莨菪碱(30.1μg/g)。曼陀罗籽中东莨菪碱与莨菪碱含量的比率为1∶11,符合曼陀罗(Datura stramonium L.)特征。尿液中,东莨菪碱和山莨菪碱占比为0.15%和0.20%,莨菪碱占比为99.65%。结论种子形态、3种生物碱的含量范围以及比例,均符合曼陀罗特点,结合患者阿托品中毒临床症状,可以确定本次事件是由误食曼陀罗籽引起的一起家庭食物中毒。本研究可以为生物碱中毒患者的临床诊断和治疗提供技术支撑,为相关中毒事件应急检测和处置提供依据。Objective To investigate a poisoning incident caused by eating eight treasure congee,and establish liquid chromatography(LC)-mass spectrometry(MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents.Methods LC-MS/MS was used for screening 28 alkaloids in the urine,eight treasure congee and food raw material,and the detected alkaloids were quantified.Samples were extracted with 0.4%formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column(1.7μm,100×2.1 mm).Acetonitrile-0.2%formic acid aqueous solution was used as the mobile phase and gradient elution was adopted.The ionization mode was electrospray positive ionization mode,and the detection method was multi-reaction monitoring(MRM).Analytes were quantified with the external standard method.Results In the concentration range of 0-100 ng/mL,the linear correlation coefficient r were greater than 0.999 for 28 alkaloids.The recovery of 28 alkaloids in urine sample ranged from 63.0%to 105.0%,and the relative standard deviations(RSDs)were between 5.8%and 8.6%.The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0%to 109.0%,and the RSDs were between 6.3%and 9.7%.The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0%to 95.0%,and the RSDs were between 4.8%and 8.2%.Hyoscyamine(2380.0 ng/mL),scopliamine(3.6 ng/mL)and racanisodamine(4.7 ng/mL)were detected in the patient's urine.Hyoscyamine(63.3μg/g),scopliamine(5.7μg/g)and racanisodamine(2.1μg/g)were detected in eight treasure congee.Hyoscyamine(901.0μg/g),scopliamine(80.0μg/g)and racanisodamine(30.1μg/g)were detected in the seed of Datura stramonium L.The ratio of scopliamine and hyoscyamine in the seed of D.stramonium was 1∶11,which complies with the characteristics of D.stramonium L.In urine sample,the proportion of scopliamine and rac-anisodamine was 0.15%and 0.20%,and hyoscyamine accounted for 99.65%.Conclusion Seed morphology,the content range and proportion of three alkaloids are all in accord with

关 键 词：生物碱 食物中毒 液相色谱-串联质谱法 曼陀罗 莨菪碱 东莨菪碱 山莨菪碱 快速筛查 

分 类 号：R155.3[医药卫生—营养与食品卫生学]