超高效液相色谱-四极杆/静电场轨道阱高分辨质谱对药酒中乌头类生物碱的快速筛查与测定

Rapid screening and determination of toxic alkaloids in medicinal liquor by ultra-high-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry

目的 建立快速筛查与定量测定药酒中乌头类生物碱的方法。方法 样品经水稀释,HSS T3色谱柱(100 mm×2. 1 mm,1. 8μm)分离,以含0. 1%甲酸的甲醇-5 mmol/L乙酸铵为流动相梯度洗脱,全扫描/数据依赖二级质谱扫描(Full MS-dd MS2)模式分析,利用自建生物碱数据库对药酒样品检索匹配定性,并对筛查出的乌头类生物碱以目标离子选择性扫描(Targeted-SIM)模式分析,外标法定量。结果 筛查出的生物碱信息与标准生物碱信息比对一致,并与临床中毒症状吻合,考察的7种乌头类生物碱在5μg/L~100μg/L内线性关系良好,相关系数均> 0. 99,方法检出限(LOD)为0. 5μg/L~1. 5μg/L,定量限(LOQ)为1. 5μg/L~4. 5μg/L,在5μg/L、40μg/L、100μg/L 3个水平下,加标回收率为76. 3%~119. 1%,相对标准偏差为4. 3%~13. 4%。结论 该筛查方法具有快速、准确、分析通量高等优点,结合定量方法能够用于药酒中毒样品中乌头类生物碱的快速筛查与定量测定。

Objective Ultra-high-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC/Q-Exactive)was established for rapidly screening and quantitatively analyzing aconitum alkaloids in medicinal liquor.Methods The sample was diluted with water and then separated by HSS T3 column(100 mm × 2.1 mm,1.8 μm)with gradient elution using Methanol-5 mmol/L ammonium acetate(containing 0.1% formic acid)as mobile phase.After screened by Full MS-ddMS2 mode,the aconitum alkaloids in samples were qualitatively analyzed through searching and matching using the self-built alkaloids database.The qualitative alkaloids were quantitatively analyzed by Targeted-SIM mode using external standard method.Results The alkaloids screened in samples were consistent with the alkaloids standards and the relevant clinical symptoms of the alkaloids poisoning.The seven aconitum alkaloids in this study showed a good linear relationship in the concentration range of 5μg/L-100μg/L,and the correlation coefficients were above 0.99.The method's limits of detection(LODs)were within 0.5μg/L-1.5μg/L,the limits of quantification(LOQs)were within 1.5μg/L-4.5μg/L,the average recovery rates were within 76.3%-119.1%,and the relative standard deviations were within 4.3%-13.4%.Conclusion The screening method has the advantages of fast,accurate,and high analytical throughput.Combined with the quantitative method,the screening method can be used for the rapid screening and the quantitative analysis of aconitum alkaloids in the poisoning samples of medicinal liquor.