| ObjectiveCompound compatibility is the main form of traditional Chinese medicine, The compatibility of drugs is the feature and core of compound medicine. Chinese medicine compound compatibility allows the active ingredient in the quality or quantity changed,Produce pharmacodynamics in multi-component interaction. Compound medicine been mostly studied nowadays focuses on the pharmacology and pharmacodynamics, few emphasis is laid on the study of chemical substances, compatibility mechanism and quality evaluation of multi-component in compound traditional Chinese. In particularly,the quality control method which only an individual component is needed to limit its content of compound traditional Chinese medicine is very popular.So it was absolutely insufficient to evaluate the quality and pharmacodynamics thoroughly. In order to investigated the target of the main component of principal drug-epimedium and ministerial drug-achyranthes of gushuling.Study on main component contents Under different compatibility group of principal drug and ministerial drug,in order to Provide scientific basis for the drug’s prescription compatibility and development.MethodThe Simple and rapid High performance liquid chromatography(HPLC) method was established to determine the main component contents of icariin and β-ecdysterone in the epimedium and achyranthes of GSL. And orthogonal design of GSL were demolished party grouping.The content of main component in different groups is measured, analysis the results combined with statistical analysis.HPLC method was established to analyze 10 batches of epimedium herbs, HPLC fingerprint analysis of Epimedium is established, Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(China Pharmacopoeia Committee 2010 Version) was used to conduct similarity evaluation and determine the characteristic peak of epimedium and determine the content of epimedium characteristic peak in the decoction solution of different compatibility. the results were analyzed by orthogonal design and statistical analysis method.ResultsTO investigate the methodological for Determination of icariin in GSL by HPLC,The results showed that the sample concentration of icariin between 10~250μg/m L have good linear relationship with the peak area;the lower limit of quantitation was10.00μg/m L;The range of RSD for inter-day and intra-day was 1.49%~1.70% and1.79%~2.78%;The result of RSD for Repeatability test was 1.90%; The average recovery was 99.41%(n=6), The result of RSD for average recovery was 1.94%. The results showed that the established method is suitable the requirements of the experimental analysis. Determined icariin contents in the formulation of Gushuling grains under different compatibility groups are analyzed by adopting orthogonal design and statistical software,The results of statistics show that: astragalus has statistical significance to total content of epimedium(P <0.05); oyster and achyranthes have no statistical significance to total content of epimedium(P >0.05) and that no matter which one(including astragalus+oyster, astragalus+achyranthes or oyster + achyranthes) is compatible with epimedium herb, there is no statistical significance to total content of epimedium.TO investigate the methodological for Determination of β-hydroxyecdysone in GSL by HPLC, The results showed that the sample concentration of β-hydroxyecdysone between 10~250μg/m L have good linear relationship with the peak area;the lower limit of quantitation was 10.00μg/m L;The range of RSD for inter-day and intra-day was1.49%~1.83% and 2.33%~2.87%c,;The result of RSD for Repeatability test was 1.67%;The average recovery was 98.02%(n=6), The result of RSD for average recovery was2.61%. The results showed that the established method is suitable the requirements of the experimental analysis. Determined β-hydroxyecdysone contents in the formulation of Gushuling grains under different compatibility groups are analyzed by adopting orthogonal design and statistical software, and results show that: epimedium herb,astragalus and oyster have no statistical significance to β-hydroxyecdysone(P >0.05), and interaction between the two also has no statistical significance.Through the research of external characteristic fingerprint under different compatibility groups, results show that: there are significant differences between two P values. P value of influence of astragalus on epimedium(peak 8) is 0.034, while that on Icarisid II(peak 9) is 0.013. P values of influence of astragalus on peak 3 and 5 are separately 0.034 and 0.023, so significant differences exist. Therefore, astragalus may reduce contents of peak 3, 5, 8(epimedium) and peak 9(icarisid II). Difference between astragalus and separated peak 1, 2, 4, 6 and 7 is not large; difference between oyster and separated peak 1~peak 8 is not large(P >0.05); difference between achyranthes and peak 1~ peak 8 is also not large(P>0.05). Influences of interaction among astragalus,oyster and achyranthes on peak 1~ peak 8 are non-significant.ConclusionFor established GSL, determination of main contents of sovereign and ministerial drug optimizes quality evaluation criterion of the drug and provides references for establishing quality standard. The research on main contents and fingerprint of sovereign and ministerial drugs in vitro and in vivo under different compatibility groups of GSL provides reference basis for studying compatibility theory, chemical component and pharmacodynamic material basis. |
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