基于特征图谱及一测多评法同时测定重楼药材中9种甾体皂苷的方法研究

Study on the simultaneous determination of the nine steroidal saponins in Paridis Rhizoma based on the characteristic chromatogram and quantitative analysis of multi-components by a single marker(QAMS)

目的建立重楼多种成分同时测定的特征图谱和含量测定方法,对重楼药材和饮片的质量进行有效质量控制。方法采用Agilent SB-C_(18)色谱柱(250 mm×4.6 mm,5μm);以乙腈与0.1%磷酸水溶液为流动相,梯度洗脱,流速为1.1 mL/min,检测波长为203 nm。对野外采集确定基原的17批正品重楼药材中重楼皂苷I、重楼皂苷II、重楼皂苷Ⅲ、重楼皂苷Ⅳ、重楼皂苷V、重楼皂苷VI、重楼皂苷VII、重楼皂苷H、重楼皂苷D等9种甾体皂苷成分,建立特征图谱和含量测定方法,以期对其质量进行评价。结果在特征图谱建立过程中,以相关对照品的出峰时间为标准,9种甾体皂苷分离度均>1.5,其对称因子在0.95~1.05之间,满足定性定量要求。在含量测定方法学建立过程中,9种甾体皂苷在一定的浓度范围内呈良好的线性关系(R^(2)>0.9999);精密度、稳定性、重现性等均符合方法学要求。结论本文建立的重楼特征图谱和9种指标性成分同时进行含量测定的方法,所建立的方法操作简便、结果准确稳定,可用于重楼药材及饮片内在指标性成分含量的检测。

Objective To establish the characteristic chromatogram and content determination method for the simultaneous determination of multiple components of Paridis Rhizoma and to effectively control the quality of the medicinal materials and its decoction pieces.Methods The separation was performed on an Agilent SB-C_(18) column(250 mm×4.6 mm,5μm).Acetonitrile and 0.1%phosphoric acid aqueous solution were used as mobile phases in gradient elution at a flow rate of 1.1 mL/min and with the detection wavelength of 203 nm.In order to evaluate the quality of the 17 batches of Paridis Rhizoma collected in the field,a method for the characteristic chromatogram and content determination was established to analyze the saponin I,saponin II,saponin III,saponin IV,saponin V,saponin VI,saponin VII,styroside H and styroside D in the plant.Results In the process of establishing the characteristic chromatogram,taking the peak time as a standard,the separation degrees of the nine steroid saponins were all more than 1.5,and their symmetry factors ranged from 0.95 to 1.05,which met the qualitative and quantitative requirements.In the process of establishing the content determination method,the nine steroid saponins showed a good linear relationship within a certain concentration range(R^(2)>0.9999),with the precision,stability and reproducibility all meeting the methodological requirements.Conclusion The method established in this study is simple,accurate and stable,and can be used for the content determination of the index components in the medicinal material and decoction pieces of Paridis Rhizoma.