目的:建立UPLC-MS/MS法测定大蓟中7个吡咯里西啶生物碱(印美定、石松胺、凌德草碱、N-氧化凌德草碱、N-氧化印美定、N-氧化石松胺和克氏千里光碱)的含量,并根据测定结果进行初步的风险评估。方法:采用ACQUITY UPLC HSS T3色谱柱(100 mm×2.1 mm, 1.8 μm),以含0.05%甲酸和2.5 mmol·L-1甲酸铵的水溶液(A)及含0.05%甲酸和2.5 mmol·L-1甲酸铵的乙腈溶液(B)为流动相,梯度洗脱;电喷雾离子源,正离子多反应模式监测,外标法测定。结果:印美定、石松胺、凌德草碱、N-氧化凌德草碱、N-氧化印美定、N-氧化石松胺和克氏千里光碱质量浓度分别在1.024～102.4、1.032～103.2、1.016～101.6、1.022～102.2、1.056～105.6、0.958～95.8和0.988～98.8 ng·mL-1范围内与峰面积呈现良好的线性关系(r>0.999 0);平均加样回收率分别为89.2%、95.2%、88.6%、92.1%、106.2%、91.5%和93.2%,RSD分别为3.2%、3.5%、3.1%、3.8%、3.2%、3.3%和3.9%。10批大蓟样品中印美定、石松胺、凌德草碱、N-氧化凌德草碱、N-氧化印美定、N-氧化石松胺和克氏千里光碱的含量范围分别为1.36～5.32、1.38～5.25、1.12～5.84、1.69～8.52、3.03～25.25、1.25～5.96和1.29～4.25 μg·kg-1。结论:本研究所建立的方法可同时测定大蓟中7个主要吡咯里西啶生物碱含量。初步风险评估显示,该药材存在一定的用药安全风险。
Objective: To establish an UPLC-MS/MS method for the simultaneous determination of seven main pyrrolizidine alkaloids (imtermedine, lycopsamine, rinderine, rinderine N-oxide, imtermedine N-oxide and lycopsamine N-oxide and senkirkine) in Cirsii Japonici Herba, and to carry out preliminary risk assessment according to the research results. Methods: The assay was performed on an ACQUITY UPLC HSS T3 column (100 mm×2.1mm, 1.8 μm) and the sample was eluted with a gradient mobile phase consisting of 0.05% formic acid and 2.5 mmol·L-1 ammonium formate in water (A)-0.05% formic acid and 2.5 mmol·L-1 ammonium formate in acetonitrile (B). The MS system was operated by using electrospray ionization (ESI) in the positive ion mode, and the scan mode was in multiple reactions monitoring (MRM) mode. External standard method was used in the experiment. Results: The linear ranges of imtermedine, lycopsamine, rinderine, rinderine N-oxide, imtermedine N-oxide and lycopsamine N-oxide and senkirkine were 1.024-102.4, 1.032-103.2, 1.016-101.6, 1.022-102.2, 1.056-105.6, 0.958-95.8 and 0.988-98.8 ng·mL-1, respectively (r>0.999 0). The average recoveries were 89.2%, 95.2%, 88.6%, 92.1%, 106.2%, 91.5% and 93.2% with RSD of 3.2%, 3.5%, 3.1%, 3.8%, 3.2%, 3.3% and 3.9%, respectively. The content ranges of the above-mentioned 7 pyrrolizidine alkaloidsin the 10 batches of Cirsii Japonici Herba were 1.36-5.32,1.38-5.25, 1.12-5.84, 1.69-8.52, 3.03-25.25, 1.25-5.96 and 1.29-4.25 μg·kg-1, respectively. Conclusion: The method can be used for the determination of 7 pyrrolizidine alkaloids in Cirsii Japonici Herba, providing a scientific basis for the quality evaluation and safe medication of this herb.
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