安宫牛黄丸(体外培育牛黄)中胆酸与牛磺胆酸的含量测定及比例研究

Study on content and ratio of cholic acid and taurocholate acid in Angong Niuhuang pills(Bovis Calculus Sativus)

目的:对标识为体外培育牛黄的安宫牛黄丸中胆酸和牛磺胆酸的含量及比例进行研究。方法:采用YMC-Pack ODS-A(150 mm×3.0 mm,5μm)色谱柱,以乙腈-10 mmol·L^(-1)醋酸铵的0.2%甲酸溶液(40∶60)为流动相,流速为0.4 mL·min^(-1);采用电喷雾负离子模式(ESI-),进行多反应监测(MRM),胆酸选择m/z 407.0→289.0,牛磺胆酸选择m/z 514.0→79.8进行检测,对收集到的5个企业7批安宫牛黄丸(体外培育牛黄)中胆酸、牛磺胆酸的含量进行测定。结果:胆酸和牛磺胆酸进样量分别在0.91~457.2 ng(r=0.999 5)和0.149~373.4 ng(r=0.999 0)范围内线性关系良好;平均回收率分别在99.4%~107.0%和96.0%~103.0%之间,RSD分别在1.3%~4.5%和3.5%~5.7%之间。7批样品中胆酸和牛磺胆酸的含量范围分别为3.3~8.2 mg·g^(-1)和0.06~1.1 mg·g^(-1),胆酸与牛磺胆酸的含量比例主要分布在6.7~14.7之间。结论:某些样品中胆酸与牛磺胆酸含量比例明显偏高。采用指标成分含量比例的方法,为安宫牛黄丸(体外培育牛黄)的质量评价提供了有益的参考。

Objective：To investigate the content and ratio of cholic acid（CA） and taurocholate acid（TCA） in Angong Niuhuang pills containing Bovis Calculus Sativus. Methods：The samples were separated on a YMC-Pack ODS-A C18 column（150 mm×3.0 mm,5 μm）,and eluted with acetonitrile and water（containing 10 mmol·L-1 of ammonium acetate and 0.2% formic acid） at a flow rate of 0.4 mL·min-1. The LC-MS/MS analysis was carried out using negative ESI mode in multiple reaction monitoring（MRM）. Quantification was performed using the following transitions：m/z 407.0 to m/z 289.0 for CA and m/z 514.0 to m/z 79.8 for TCA. The contents of CA and TCA in 7 commercial batches of Angong Niuhuang pills were determined. Results：The calibration curve of CA and TCA was linear in the concentration range of 0.91-457.2 ng（r=0.999 5）and 0.149-373.4 ng（r=0.999 0）. The average recovery of CA and TCA was respectively in the range of 99.4%-107.0%（RSD 1.3%-4.5%） and 96.0%-103.0%（RSD 3.5%-5.7%）. The contents of CA and TCA of Angong Niuhuang pills were from 3.3 mg·g-1 to 8.2 mg·g-1 and from 0.06 mg·g-1 to 1.1 mg·g-1. The content ratio of CA/TCA ranged from 6.7 to 14.7. Conclusion：The content ratio of CA/TCA was significantly high in some samples. The content ratio of CA and TCA in commercial Angong Niuhuang pills could be served as a scientific and effective way for quality research of commercial Angong Niuhuang pills.