目的: 建立UPLC-MS/MS 法测定胜红蓟中6 个吡咯里西啶生物碱（石胺松、凌德草碱、刺凌德草碱、N-氧化凌德草碱、N-氧化刺凌德草碱和N-氧化石胺松）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,流速为0.3 mL·min-1,柱温为40 ℃ ;电喷雾离子源,正离子多反应模式监测,外标法测定。结果: 石胺松、凌德草碱、刺凌德草碱、N-氧化凌德草碱、N-氧化刺凌德草碱和N-氧化石胺松质量浓度分别在0.972~97.2、0.964~96.4、1.016~101.6、1.056~105.6、1.032~103.2 和0.958~95.8 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88.5%、90.2%、92.6%、94.3%、91.8% 和95.7%,RSD 分别为3.1%、3.5%、2.4%、2.1%、2.5% 和3.2%。6 批胜红蓟样品中石胺松、凌德草碱、刺凌德草碱、N-氧化凌德草碱、N-氧化刺凌德草碱和N-氧化石胺松的含量范围分别为1.22~2.87、0.36~0.75、4.23~10.65、1.08~3.65、10.31~24.26 和5.25~13.36 μg·g-1。结论: 本研究所建立的方法可同时测定胜红蓟中6 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 a method for the simultaneous determination of six main pyrrolizidine alkaloids （lycopsamine,rinderine,echinatine,rinderine N-oxide,echinatine N -oxide,and lycopsamine N -oxide） in Agerati Conyzoidis Herba by UPLC-MS/MS. And to carry out preliminary risk assessment according to the research results. Methods： The assay was performed on a ACQUITY UPLC HSS T3 column（ 2.1 mm×100 mm,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 flow rate was 0.3 mL·min-1,the column temperature was 40 ℃. 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 lycopsamine,rinderine,echinatine,rinderine N-oxide,echinatine N -oxide,and lycopsamine N -oxide were 0.972-97.2,0.964-96.4,1.016-101.6,1.056-105.6,1.032-103.2 and 0.958-95.8 ng·mL-1,respectively（r> 0.999 0）. The average recoveries were 88.5%,90.2%,92.6%,94.3%,91.8% and 95.7% with RSD of 3.1%,3.5%, 2.4%,2.1%,2.5% and 3.2%,respectively. The content ranges of lycopsamine,rinderine,echinatine,rinderine N-oxide,echinatine N-oxide,and lycopsamine N-oxide in the 6 batches of Agerati Conyzoidis Herba were 1.22- 2.87,0.36-0.75,4.23-10.65,1.08-3.65,10.31-24.26 and 5.25-13.36 μg·g-1,respectively. Conclusion： The established method can simultaneously determine six pyrrolizidine alkaloids in Agerati Conyzoidis Herba. The preliminary risk assessment showes that there is a certain risk of drug safety.
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