液质联用同时测定喘可治注射液中4种黄酮苷含量

Simultaneous Quantification of Four Flavonoid Glycosides in Chuankezhi Injection Using LC-MS/MS

目的建立同时测定喘可治注射液中朝藿定A,B,C和淫羊藿苷的高效液相色谱-串联质谱（HPLC-MS/MS）方法。方法采用Agilent Eclipse XDB-C18柱（150 mm×2.1 mm,5.0μm）,以乙腈-0.1%甲酸水溶液（35∶65）为流动相进行色谱分离,流速0.22 m L·min^-1,柱温40℃;在负离子检测模式下采用多反应监控（MRM）,以柚皮苷为内标测定朝藿定A,B,C和淫羊藿苷。结果朝藿定A,B,C和淫羊藿苷4种成分在API 3000液质联用系统仪上的定量限均为0.04 ng·m L^-1、检测限均为0.05 ng·m L^-1。4种黄酮成分在1.0-1000.0 ng·m L^-1呈现良好的线性关系（r〉0.9950）,加样回收率在90.11%-100.3%之间,RSD〈2.89%（n=6）,方法重复性及精密度良好;4种黄酮苷在15批喘可治注射液中的含量比较稳定,且符合国家药品标准要求。结论所建立的液质联用定量方法灵敏、准确、稳定,适用于喘可治注射液4种主要黄酮苷的快速测定,可用于喘可治注射液的质量控制。

Objective To develop a quantitative method for simultaneous determination of 4 flavonoid glycosides in Chuankezhi Injection using liquid chromatography-tandem mass spectrometry（LC-MS/MS）.Methods Chromatographic separation was performed on an Agilent Eclipse XDB-C18column（150 mm×2.1 mm,5.0 μm）at40 ℃.Mobile phase consisted of acetonitrile-0.1 % formic acid in water（35∶65）,and the flow rate was 0.22 m L·min^-1.With naringin as the internal standard,epimedins A,B,C and icariin in Chuankezhi Injection were quantified using LC-MS/MS in the multiple reaction monitoring（MRM） system under ESI negative mode.Results Good linearity was obtained for quantitation of 4 flavonoid glycosides in the range of 1.0-1000.0 μg·L^-1（r 0.995）,and recoveries were between 90.11 % and 100.3 % with RSD no more than 2.89 %.The limit of detection of the 4flavonoid glycosides was 0.04 ng·m L^-1and the limit of quantitation was 0.045 ng·m L^-1.The established method showed good reproducibility and precision.Quantitative analysis showed that 15 batches of Chuankezhi Injection contained the four main flavonoid glycosides at stable level and met the requirement of the national drug standards.Conclusion The new developed method is rapid,sensitive,accurate and reliable enough for the quantification of epimedins A,B,C and icariin,and thus can be used to evaluate the quality of Chuankezhi Injection.