UPLC-MS/MS法测定款冬花炮制前后3个吡咯里西啶生物碱的含量

Determination of three main pyrrolizidine alkaloids in crude and prepared Farfarae Flos by UPLC-MS/MS

目的:通过UPLC-MS/MS法比较款冬花蜜炙前后3个吡咯里西啶生物碱(千里光宁碱、N-氧化千里光宁碱和克氏千里光碱)含量的变化。方法:采用CORTECS UPLC C18色谱柱(100 mm×2.1 mm,1.6μm),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:千里光宁碱、N-氧化千里光宁碱和克氏千里光碱质量浓度在1.025~102.5、0.992~99.2和5.040~504.0 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86.5%、92.1%、94.7%,RSD分别为4.1%、3.5%、2.3%。7批款冬花生品中N-氧化千里光宁碱的含量分别为0.942、0.334、0.353、0.536、0.347、0.678、0.552μg·g^-1;克氏千里光碱含量分别为44.72、38.62、35.93、45.14、29.57、38.67、42.31μg·g^-1;千里光宁碱的含量分别0.464、0.115、0.185、0.125、0.468、0.261、0.122μg·g^-1。对应的蜜炙品中N-氧化千里光宁碱含量分别为0.616、0.087、0.237、0.315、0.224、0.553、0.401μg·g^-1;克氏千里光碱含量分别为34.33、29.57、25.01、32.66、20.34、26.45、31.54μg·g^-1;千里光宁碱的含量分别为0.772、0.357、0.314、0.354、0.574、0.379、0.263μg·g^-1。结论:和生品相比,蜜炙后款冬花中N-氧化千里光宁碱和克氏千里光碱呈下降趋势,但千里光宁碱呈上升趋势,N-氧化千里光宁碱部分转化为千里光宁碱。

Objective:To compare the changes of the contents of three main pyrrolizidine alkaloids(senecionine,senecionine N-oxide and senkirkine)in Farfarae Flos before and after stir-baking with honey by UPLC-MS/MS.Methods:The assay was performed on a CORTECS UPLC C18 column(100 mm×2.1 mm,1.6μm)and the sample was eluted with a gradient mobile phase consisted 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 reaction monitoring(MRM)mode.External standard method was used in the experiment.Results:The linear ranges of senecionine,senecionine N-oxide and senkirkine were 1.025^-102.5 ng·mL^-1,0.992-99.2 ng·mL^-1 and 5.040-504.0 ng·mL^-1,respectively(r>0.9990).The average recoveries were 86.5%,92.1%and 94.7%with RSD of 4.1%,3.5%and 2.3%,respectively.In 7 batches crude Farfarae Flos,the contents of senecionine N-oxide were 0.942,0.334,0.353,0.536,0.347,0.678 and 0.552μg·g^-1;the content of senkirkine were 44.72,38.62,35.93,45.14,29.57,38.67 and 42.31μg·g^-1;the content of senecionine were 0.464,0.115,0.185,0.125,0.468,0.261 and 0.122μg·g^-1,respectively.After stir-baking with honey,the corresponding contents of senecionine N-oxide were 0.616,0.087,0.237,0.315,0.224,0.553 and 0.401μg·g^-1;the content of senkirkine were 34.33,29.57,25.01,32.66,20.34,26.45 and 31.54μg·g^-1;the content of senecionine were 0.772,0.357,0.314,0.354,0.574,0.379 and 0.263μg·g^-1,respectively.Conclusion:Compared with crude Farfarae Flos,the contents of senecionine N-oxide and senkirkine decreased,while the content of senecionine increased after processing.Senecionine N-oxide is partially transformed to senecionine.