复方丹参片中三七皂苷成分组成的SPE-HPLC-MS法测定

Determination of Saponins in Formulated Danshen Tablets by SPE-HPLC-ESI-MSn Technique

目的研究复方丹参片中的三七皂苷类成分组成,并建立主要成分的含量测定方法。方法采用HPLC-MS分析技术,通过对CID质谱图的解析以及与对照品比较,确定主要的三七皂苷类成分。采用C18SPE预处理方法,分别以水和甲醇为淋洗剂和洗脱剂,甲醇洗脱物用于HPLC含量测定。采用Agela-Venusil C18色谱柱(4.6 mm×250 mm5,μm),以乙腈-水-乙酸为流动相进行梯度洗脱,流速为1.0 ml.min-1,检测波长为202 nm。结果鉴定了7种皂苷成分,其中3种成分首次在复方丹参片中报道。建立了4种主要成分(三七皂苷R1、人参皂苷Rg1、Rb1、Rd)的SPE-HPLC含量测定方法。方法的平均回收率为95.8%～101.3%,RSD小于3.0%。结论该方法操作简便快速,环境良好,结果准确可靠,可为提高复方丹参片质量控制标准的提供有效依据。

Objective To establish a rapid and reliable SPE-HPLC method for simultaneous determination of main saponins in formulated Danshen tablets. Methods The main saponins were identified by the analysis of MSn spectra and compared with reference compounds.On this basis,the saponin extract was pretreated with RP-C18 SPE column,water and methanol were used as the elution solvents.The methanol elute was employed for HPLC assay on an Agela-Venusil C18 column（4.6 mm × 250 mm,5 μm）.A gredient elution was performed with acetonitrile-water-acetate acid as the mobile phase.The flow rate was 1.0 ml·min-1 and the detection wavelength was at 202 nm. Results Seven ginsenosides were detected and identified by comparing MSn spectra with the reference compounds and the literature data.An SPE-HPLC method was established for the assay of four main compounds.The R.S.D.for these saponins was less than 3.0%.The recovery rates of all the saponins varied in the range of 95.8%-101.3%,respectively. Conclusion The method is rapid,easily operated,reliable and can be used for the improvement of quality control for formulated Danshen tablets.