世界中医药

目的:通过高效液相指纹图的检测方法测试不同批次参苓白术散中人参皂苷的含量。方法:进样量为10 μL,采用250 mm×4.6 mm,5 μm的色谱柱(ThermoODS-HYPERSIL柱),选择流动相的条件是在A流动相乙腈和B流动相水中以1 mL/min的流速开展实验,检测波长为203 nm,柱温为30 ℃,根据高效液相(HPLC)(Agilengt-1100高效液相色谱仪)指纹图谱的梯度洗脱程序进行试验,并在Agilengt-1100色谱工作站上进行指纹图谱分析。分别对HPLC方法学的严谨性进行检测,包括专属性试验、精密度试验、稳定性试验、重复性试验、线性回归试验、加样回收率试验等逐步建立方法的科学性;而后进一步采用HPLC对参苓白术散中人参皂苷的含量进行具体的检测和记录。结果:1)HPLC的专属性试验结果显示人参皂苷Rg1、Re及Rb1均有良好的专属性。2)人参皂苷Rg1的线性回归方程是Y=352.681 19X+6.738 50;人参皂苷Re的线性回归方程是Y=292.610 86X+3.703 23;人参皂苷的线性回归方程是Rb1Y=254.203 21X+4.564 71;其相关系数均>0.999,结果显示人参皂苷线性关系良好。3)精密度试验结果显示人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1峰面积的RSD在0.22%～0.29%;稳定性试验结果显示人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1峰面积的RSD在0.06%～1.83%；重复性试验结果显示人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1总量的含量平均值为9.86 mg,RSD 2.5%表明HPLC的精密度、稳定性和重复性均良好。4)结果显示人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1的加样回收率平均值和RSD分别为95.7%、105.7%、100.4%,3.3%、4.8%、3.1%,表明均HPLC测定参苓白术散加样回收率良好。5)不同批次参苓白术散中人参皂苷总的含量存在较大差别。结论:高效液相指纹图谱检测参苓白术散的操作方法较为简便,其专属性、灵敏度和重复性均较好,而不同批次参苓白术散中人参皂苷总的含量存在较大差别,对临床规范控制参苓白术散的质量有试验基础。

English Summary:

To establish a method for the determination of ginsenosides in Shenling Baizhu powder by high performance liquid chromatography fingerprint，so as to improve the quality control of Shenling Baizhu powder.Methods:Chromatographic conditions：ThermoODS-HYPERSIL column (250 mm×4.6 mm，5 micron)，mobile phase (flow rate：1 mL/min)：acetonitrile：water，injection volume：10 microL，column temperature：30 ℃，detection wavelength：203 nm.The standard curve of ginsenoside reference substance series concentration 、blank reference solution were prepared.Shenling Baizhu powder detection solution were carried out on the tested samples，and the liquid chromatogram was processed and analyzed by Agilengt-1100 chromatographic workstation.specific test，precision test，stability test，repeatability test，linear regression test and sample recovery test were done accorded to the procedures，the chromatographic conditions of the high performance liquid chromatography were validated methodologically.Six batches of Shenling Baizhu Powder were processed and prepared for inspected solution.The content of ginsenosides in the samples were determined by the above-mentioned chromatographic conditions.Results:1) HPLC specific test：ginsenoside Rg1，Re and Rb1 have good specificity.2) Linear regression equation of ginsenoside Rg1 was Y=352.681 19X+6.738 50; ginsenoside Re linear regression equation was Y=292.610 86X+3.703 23; ginsenoside linear regression equation was Rb1Y=254.203 21X+4.564 71; the correlation coefficients all exceeded 0.999，and had good linear relationship.3) Precision test：RSD of peak area of ginsenoside Rg1，ginsenoside Re and ginsenoside Rb1 ranged from 0.22% to 0.29%.Stability test showed that RSD of peak area of ginsenoside Rg1，ginsenoside Re and ginsenoside Rb1 ranged from 0.06% to 1.83%.4) Repeatability test：ginsenosition test：ginsenoside Rg1，ginsenoside Re，ginsenoside Rb1 The average content of the total was 9.86 mg，RSD 2.5%，indicated that the precision，stability and repeatability of the method were good.5) The average recovery and RSD of ginsenoside Rg1，ginsenoside Re and ginsenoside Rb 1 respectively were 95.7%，105.7%，100.4%，3.3%，4.8% and 3.1%. 6) The content of ginsenosides in 6 batches of Shenling Baizhu powder：The total content of ginsenosides in different batches of Shenling Baizhu powder were quite different.Conclusion:The method of high performance liquid chromatography fingerprint detection for Shenling Baizhu powder was simple，specific，sensitive and reproducible.The total content of ginsenosides were great differences among different batches of Shenling Baizhu powder，and provided an experimental basis for clinical standard control of the quality of Shenling Baizhu powder is needed.

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