超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法 检测呕吐物中生物碱

Detection of alkaloids in vomit by ultra-high performance liquid chromatography-quadrupole/electrostatic field orbital trap high resolution mass spectrometry

关键词: 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法  生物碱  呕吐物

Keywords: ultra-high performance liquid chromatography-quadrupole/electrostatic field orbital trap high resolution mass spectrometry  alkaloids  vomit

摘 要: 目的 采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱仪,建立呕吐物中生物碱的检测方法。方法 样品经乙腈提取后,以甲醇-乙酸铵(5 mmol/L)水溶液(含0.1%甲酸)为流动相进行梯度洗脱,采用 HSS T3 色谱柱(100 mm×2.1 mm,1.8 μm)对待测组分进行色谱分离;采用可加热电喷雾离子源(HESI),全扫描/数据依赖二级质谱扫描(Full MS/ddMS²)模式对待测物进行定性分析;在全扫描模式下以外标法对待测物进行定量检测。结果 在5.0~200.0 μg/L 范围内模拟呕吐物中 38 种生物碱均呈现良好的线性关系,相关系数均>0.99,检出限 0.1~0.5 mg/kg,加标回收率 72.9%~111.0%,相对标准偏差 5.6%~16.3%。结论 本方法提供了丰富的化合物特征信息,根据一级质谱和二级质谱碎片离子的精确质荷比,在无相关标准品的情况下可对未知化合物进行结构推断和确证;此方法样品前处理简单,定性分析特异性强,定量测定灵敏度高,可满足突发公共卫生事件快速定性、定量检测要求。

Abstract: Objective Using ultra-high performance liquid chromatography-quadrupole/electrostatic field orbital trap high resolution mass spectrometer,to establish a detection method for alkaloids in vomit.Methods After extraction with acetonitrile,the samples were subjected to gradient elution using a methanol ammonium acetate (5 mmol/L) aqueous solution(containing 0.1% formic acid)as mobile phase, and the components were separated by chromatography using an HSS T3 column(100 mm×2.1 mm,1.8 μm);the heated electric spray ionization source (HESI) was used for qualitative analysis of the analytes in the full scanning/data-dependent secondary mass spectrometry (Full MS/ddMS²) mode;then quantify the test substances using the external standard method in full mass mode (full mass).Results The results showed that all the 38 alkaloids in simulated vomit showed a good linear relationship within the range of 5.0~200.0 μg/L,with correlation coefficients greater than 0.99,the detection limits were 0.1~0.5 mg/kg,the spiked recovery rates were 72.9%~111.0%,and the relative standard deviations were 5.6%~16.3%.Conclusion The results suggested that this method provides a wealth of compound characteristic information,based on the precise mass-charge ratio of fragment ions in primary and secondary mass spectrometry,it can infer and confirm the structure of unknown compounds without relevant standards;inaddition,this method has simple sample preparation,strong specificity in qualitative analysis, and high sensitivety in quantitative determination,which could meet the requirements of rapid qualitative and quantitative detection of public health emergencies.