LC-MS法同时测定3种脑立清制剂中6种甾体

Simultaneous determination of six steroids in three Naoliqing preparations by LC-MS

目的建立LC-MS法同时测定脑立清丸、脑立清胶囊、脑立清片(磁石、赭石、牛膝等)中β-蜕皮甾酮、25 R-牛膝甾酮、25 S-牛膝甾酮、牛磺猪去氧胆酸、甘氨猪去氧胆酸、甘氨鹅去氧胆酸的含量。方法3种制剂内标(黄芪甲苷)提取液的分析采用Waters ACQUITY UPLC HSS C_(18)色谱柱(2.1 mm×100 mm,1.8μm);流动相乙腈(含0.1%甲酸)-水(含0.1%甲酸),梯度洗脱;体积流量0.4 mL/min;柱温40℃;电喷雾离子源;正离子检测;多反应监测模式。结果3种制剂中6种甾体在各自范围内线性关系良好(r>0.9990),平均加样回收率分别为99.06%~104.19%、99.08%~103.78%、99.24%~103.38%,RSD分别为1.35%~4.68%、0.94%~4.54%、1.23%~2.91%。16家厂家88批样品中各甾体含量差异明显。结论该方法简便准确,重复性好,专属性强,可用于3种脑立清制剂的质量控制。牛膝、猪胆粉质量应引起关注。

AIM To establish an LC-MS method for the simultaneous content determination ofβ-ecdysterone,25 R-inokosterone,25 S-inokosterone,taurohyodeoxycholic acid,glycohyodeoxycholic acid and glycochenodeoxycholic acid in Naoliqing Pills,Naoliqing Capsules and Naoliqing Tablets(Magnetitum,Haematitum,Achyranthis bidentatae Radix,etc.).METHODS The analysis of internal(astragaloside A)extracts of three preparations was performed on a 40℃thermostatic Waters ACQUITY UPLC HSS C_(18)column(100 mm×3.0 mm,2.7μm),with the mobile phase comprising of acetonitrile(containing 0.1%formic acid)-water(containing 0.1%formic acid)flowing at 0.4 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive ion detection with multiple reaction monitoring mode.RESULTS Six steroids in the three preparations showed good linear relationships within their own ranges(r>0.9990),whose average recoveries were 99.06%-104.19%,99.08%-103.78%and 99.24%-103.38%with the RSDs of 1.35%-4.68%,0.94%-4.54%and 1.23%-2.91%,respectively.The contents of various steroids demonstrated obvious differences in eighty-eight batches of samples from sixteen manufacturers.CONCLUSION This simple,accurate,reproducible and specific method can be used for the quality control of three Naoliqing preparations.The qualities of Achyranthis bidentatae Radix and Suis Fellis Pulvis should be paid attention to.