程序可变波长双标法同时测定参益安神片中5种有效成分的含量

Simultaneous determination of five active ingredients in Shenyi Anshen tablets by program variable wavelength method with dual markers

目的:利用HPLC-DAD程序可变波长功能,建立一种同时测定参益安神片中人参皂苷Rb_1、酸枣仁皂苷A、五味子醇甲、五味子甲素和五味子乙素5种有效成分含量的双标检测法。方法:采用Hypersil ODS2色谱柱(4.6 mm×250 mm,5μm),柱温30℃;乙腈(A)-水(B)为流动相,流速为1 mL·min^(-1),梯度洗脱;检测波长:203 nm(检测人参皂苷Rb_1,酸枣仁皂苷A)、217 nm(检测五味子醇甲、五味子甲素、五味子乙素);建立酸枣仁皂苷A和内参物人参皂苷Rb_1之间的相对校正因子;建立五味子甲素和五味子乙素与内参物五味子醇甲之间的相对校正因子。结果:在90 min内参益安神片中人参皂苷Rb_1、酸枣仁皂苷A、五味子醇甲、五味子甲素和五味子乙素5种有效成分可完全分离;峰面积与其浓度呈良好的线性关系;双标法计算结果与外标法实测值之间没有显著性差异,实验所得相对校正因子可信。结论:利用程序可变波长高效液相色谱法,结合一测多评技术建立了一种可靠的同时测定参益安神片中5种有效成分含量的检测方法。

Objective： To establish a simultaneous determination method of ginsenoside Rbl, Jujuboside A, schisandrin, deoxyschizandrin and γ-schizandrin in Shenyi Anshen tablets, using variable wavelength function of HPLC-DAD and dual markers. Methods： The separation was performed on a Hypersil ODS2 column （ 4.6 mm × 250 mm, 5 μm ） and the column temperature was set at 30 ℃. The mobile phase was composed of acetonitrile （ A ） and water （ B ） with gradient elution at a flaw rate of 1.0 mL· min-1. The detection wavelength was set at 203 nm for ginsenoside Rbl and Jujuboside A, and at 217 nm for schisandrin, deoxyschizandrin and γ-schizandrin. Ginsenoside Rb1 was chosen as the internal standard for Jujubosides A while schisandrin was chosen as the internal standard for deoxyschizandrin and γ -schizandrin and the relative correction factors were determined. Results： The five active ingredients could be separated well in 90 min and the linear relationship between their peak areas and concentration was good. Results determined by dual markers method and external standard method showed no significant differences, which indicated that the relative correction factors were credible. Conclusion： A reliable simultaneous determination method of 5 active ingredients in Shenyi Anshen tablets was established by HPLC using program variable wavelength combined with QAMS （ quantitative analysis of multi-components by single marker ） technology.