对照提取物定位结合一测多评法同时测定重楼中9个皂苷的含量

Simultaneous determination of nine saponins in Paridis Rhizoma by reference extract combined with quantitative analysis of multi-components by single marker

目的:建立一测多评法(QAMS法)同时测定重楼中9个皂苷的含量,为其质量标准的研究提供依据。方法:采用Agilent ZORBAX SB-C_(18)色谱柱(250 mm×4.6 mm,5μm);以乙腈与水为流动相,梯度洗脱,流速为1.0 mL·min^(-1),柱温为20℃,检测波长为203 nm。使用重楼对照提取物定位各色谱峰,以重楼皂苷Ⅶ为内参物,建立重楼中重楼皂苷D、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷Ⅰ和重楼皂苷Ⅴ与内参物的相对校正因子(RCFs),并验证所建立方法的可行性和准确性。结果:9个重楼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;重楼皂苷D、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、纤细薯蓣皂苷、重楼皂苷Ⅰ和重楼皂苷Ⅴ的相对校正因子分别为0.8911、0.8969、0.7346、0.9927、0.8803、0.9016、0.8271、0.7687,在不同实验条件下重现性良好。45批云南重楼中9个皂苷的平均总含量为2.211%,35批七叶一枝花中9个皂苷的平均总含量为2.965%。结论:所建立的一测多评法可用于重楼中9个皂苷成分的含量测定,可为重楼的质量控制提供科学理论依据。

Objective:To establish a quantitative analysis of multi-components by single-marker(QAMS)method for simultaneous determination of nine saponins in Paridis Rhizoma,in order to provide basis for studying its quality standards.Methods:The analysis was performed on Agilent Zorbax SB C;column(4.6 mm×250 mm,5μm),with mobile phase composed of acetonitrile-water at a flow rate of 1.0 mL·min^(-1)in gradient elution mode.The column temperature was maintained at 20℃,and the detection wavelengths were set at 203 nm.Paridis Rhizoma reference extract was used to locate chromatographic peaks,Chonglou saponinⅦwas selected as the internal standard,and the relative correlation factors(RCFs)of Chonglou saponin D,Chonglou saponin H,Chonglou saponinⅥ,Chonglou saponinⅡ,dioscin,gracillin,Chonglou saponinⅠand Chonglou saponinⅤwere determined by HPLC,and the feasibility and accuracy of QAMS method were verified.Results:The standard curves of nine saponins in Paridis Rhizoma had good linear relationship in the ranges of the tested concentrations(R;>0.9999).The precision,stability and repeatability complied with the requirements of methodology.The RCFs of Chonglou saponin D,Chonglou saponin H,Chonglou saponinⅥ,Chonglou saponinⅡ,dioscin,gracillin,Chonglou saponinⅠ,and Chonglou saponinⅤwere 0.9063,0.9293,0.7256,0.9890,0.8768,0.8839,0.8221 and 0.8102,respectively,which had good repeatability in different experimental conditions.The total content of nine saponins in 45 batches of Paris polyphylla Smith var.yunnanensis was 2.211%,and in 35 batches of Paris polyphylla Smith var.chinensis was 2.965%.Conclusion:The QAMS method is feasible and accurate for the determination of the nine saponins and is beneficial to the quality control of in Paridis Rhizoma.