UPLC-MS/MS法测定胜红蓟中6个吡咯里西啶生物碱的含量及风险评估  

Risk assessment and determination of six main pyrrolizidine alkaloids in Agerati Conyzoidis Herba by UPLC-MS/MS

作　　者：昝珂[1] 李耀磊[1] 王莹[1] 王赵[1] 金红宇[1] 刘丽娜[1] 马双成[1] ZAN Ke;LI Yao-lei;WANG Ying;WANG Zhao;JIN Hong-yu;LIU Li-na;MA Shuang-cheng(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2021年第7期1169-1175,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：科技部重大新药创制(项目编号:2018ZX09303-024,2018ZX09735-006)。

摘　　要：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,1.8μm),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>0.9990);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5.25-13.36μg·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,1.8μm)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>0.9990).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 5.25-13.36μg·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.

关 键 词：胜红蓟 肝毒性成分 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 N-氧化石胺松 超高效液相色谱-串联质谱 含量测定 风险评估 

分 类 号：R917[医药卫生—药物分析学]