HPLC-QAMS同时测定屏风生脉胶囊中9个指标成分含量

Simultaneous Determination of Nine Active Components in Pingfeng Shengmai Capsules by HPLC-QAMS

目的:建立高效液相色谱一测多评法(HPLC-QAMS)同时测定屏风生脉胶囊中毛蕊异黄酮葡萄糖苷、芒柄花苷、9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、芒柄花素、升麻素苷、升麻素、亥茅酚苷、麦冬甲基黄烷酮A及甲基麦冬二氢高异黄酮B含量的方法。方法:采用AgilentZORAXStableBond C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相乙腈-0.1%甲酸溶液,梯度洗脱。多波长切换检测(0~36 min,254 nm,毛蕊异黄酮葡萄糖苷、芒柄花苷、9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、芒柄花素、升麻素苷、升麻素和亥茅酚苷;36~60 min,296 nm,麦冬甲基黄烷酮A和甲基麦冬二氢高异黄酮B),进样量10μL。以升麻素为内参物,建立其他8个指标成分的相对校正因子(RCF),并计算各成分含量。采用外标法(ESM)测定屏风生脉胶囊中9个指标成分含量对QAMS计算结果的准确性进行验证。结果:测定了3个厂家12批屏风生脉胶囊中9个指标成分的含量,9个指标成分分别在2.68~67.00(r=0.9994)、1.76~44.00(r=0.9991)、2.15~53.75(r=0.9992)、4.37~109.25(r=0.9997)、6.58~164.50(r=0.9995)、2.49~62.25(r=0.9992)、1.88~47.00(r=0.9998)、1.36~34.00(r=0.9992)、1.09~27.25μg·mL^(–1)(r=0.9993)有良好的线性关系。平均加样回收率及RSD分别为99.34%(1.40%)、97.25%(1.13%)、98.73%(1.33%)、100.09%(0.68%)、100.04%(0.77%)、98.46%(1.26%)、97.94%(1.06%)、96.89%(1.03%)和98.90%(0.72%)。QAMS与ESM所测结果差异无统计学意义。结论:该方法操作便捷、结果准确,可用于屏风生脉胶囊中9个指标成分含量的同时测定,实现多指标质量控制。

Objective:To establish a high performance liquid chromatography-quantitative analysis of multi-components by single-marker(HPLC-QAMS)method for simultaneous determination of calycosin 7-O-β-D-glucopyranoside,ononin,9,10-dimethoxy-pterocarpane-3-O-β-D-glucoside,formononetin,prim-O-glucosylcimifugin,cimifugin,sec-O-glucosylhamaudol,methylophiopogonanone A,and methylophiopogonanone B in Pingfeng Shengmai Capsules.Methods:HPLC conditions are as follows:Agilent ZORAX StableBond C_(18) column(250 mm×4.6 mm,5μm),gradient elution with the mobile phase of acetonitrile-0.1%formic acid solution,injection volume of 10μL,detection wavelength of 254 nm(0-36 min)for calycosin 7-Oβ-D-glucopyranoside,ononin,9,10-dimethoxy-pterocarpane-3-O-β-D-glucoside,formononetin,prim-O-glucosylcimifugin,cimifugin,and sec-O-glucosylhamaudol,and 296 nm(36-60 min)for methylophiopogonanone A and methylophiopogonanone B.With cimifugin as an internal standard,the relative correction factors(RCFs)of the other 8 components were established and the content of each component was calculated.In order to validate the accuracy of QAMS,we also calculated the content of the 9 components with the external standard method(ESM).Results:The content of 9 effective components in 12 batches of Pingfeng Shengmai Capsules from three manufacturers was determined,and they showed good linear relationships within the ranges of 2.68-67.00(r=0.9994),1.76-44.00(r=0.9991),2.15-53.75(r=0.9992),4.37-109.25(r=0.9997),6.58-164.50(r=0.9995),2.49-62.25(r=0.9992),1.88-47.00(r=0.9998),1.36-34.00(r=0.9992),and 1.09-27.25μg·mL^(-1)(r=0.9993),respectively.Their average recoveries(RSDs)were 99.34%(1.40%),97.25%(1.13%),98.73%(1.33%),100.09%(0.68%),100.04%(0.77%),98.46%(1.26%),97.94%(1.06%),96.89%(1.03%),and 98.90%(0.72%),respectively.There was no significant difference between the results yielded by HPLC-QAMS and ESM.Conclusion:The established HPLC-QAMS method is convenient and accurate,which allows the simultaneous determination of 9 active components in Pingfeng Shengmai Capsules and the multi-index quality control.