超高效液相色谱串联质谱法同时测定当归补血汤中6种成分含量

Simultaneous Determination of Six Components in Danggui Buxue Decoction by UPLC-MS/MS

目的建立同时测定当归补血汤中6种成分含量的超高效液相色谱串联质谱(UPLC-MS/MS)法。方法色谱条件,色谱柱为Waters BEH C18柱(100 mm×2.1 mm,1.7μm),流动相为乙腈-0.1%甲酸水溶液(梯度洗脱),流速为0.3 m L/min,柱温为40℃,进样量为2μL;质谱条件,电喷雾离子源(ESI),正负离子模式同时采集,多反应检测模式(MRM),碰撞气为氩气,离子源温度为150℃,毛细管电压为3.0 k V,去溶剂气流量为700 L/h,去溶剂气温度为350℃。结果阿魏酸、藁本内酯、芒柄花素、毛蕊异黄酮葡萄糖苷、毛蕊异黄酮和黄芪甲苷质量浓度分别在0.52~50.62μg/m L(R^(2)=0.9993)、0.82~80.34μg/m L(R^(2)=0.9992)、0.31~30.06μg/m L(R^(2)=0.9996)、0.62~60.23μg/m L(R^(2)=0.9998)、0.34~33.65μg/m L(R^(2)=0.9995)和1.05~102.38μg/m L(R^(2)=0.9996)范围内与峰面积线性关系良好;检测限分别为0.155,0.247,0.092,0.185,0.103,0.315μg/kg,定量限分别为0.518,0.823,0.308,0.617,0.344,1.050μg/kg;精密度、稳定性、重复性试验结果的RSD均小于5.0%;平均加样回收率分别为96.85%,100.08%,97.69%,95.84%,98.83%,101.68%,RSD分别为2.20%,0.74%,0.75%,0.79%,1.50%,0.95%(n=6);样品含量均在均值的±30%之间。结论该方法操作简单、灵敏度高、专属性强,可为当归补血汤的质量标准提升提供参考。

Objective To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of six components in Danggui Buxue Decoction.Methods The chromatographic conditions were as follows:the chromatographic column was Waters BEH C18 column(100 mm×2.1 mm,1.7μm),the mobile phase was acetonitrile-0.1%formic acid aqueous solution(gradient elution),the flow rate was 0.3 mL/min,the column temperature was 40℃,and the injection volume was 2μL.The mass spectrometry conditions were as follows:the multiple-reaction monitoring(MRM)mode was adopted with electrospray ionization(ESI,positive and negative ions polarity model),the collision gas was argon,the ion source temperature was 150℃,the capillary voltage was 3.0 kV,the flow rate of dissolvent gas was 700 L/h,and the dissolvent gas temperature was 350℃.Results The linear ranges of ferulic acid,ligustilide,formononetin,calycosin 7-O-β-D-glucopyranoside,calycosin and astragalosideⅣwere 0.52-50.62μg/mL(R^(2)=0.9993),0.82-80.34μg/mL(R^(2)=0.9992),0.31-30.06μg/mL(R^(2)=0.9996),0.62-60.23μg/mL(R^(2)=0.9998),0.34-33.65μg/mL(R^(2)=0.9995)and 1.05-102.38μg/mL(R^(2)=0.9996),respectively.The limits of detection(LOD)of the six components were 0.155,0.247,0.092,0.185,0.103,0.315μg/kg,respectively.The limits of quantitation(LOQ)of the six components were 0.518,0.823,0.308,0.617,0.344,1.050μg/kg,respectively.The RSDs of precision,stability and repeatability tests were all lower than 5.0%.The average recoveries of six components were 96.85%,100.08%,97.69%,95.84%,98.83%and 101.68%with RSDs of 2.20%,0.74%,0.75%,0.79%,1.50%and 0.95%(n=6),respectively.The content of six components in the samples was within±30%of the mean value.Conclusion This method is simple,sensitive and specific,which can provide a reference for improving the quality standard of Danggui Buxue Decoction.