Research On The Quality Evaluation Of Production Process Of Yinhuang Granules Based On Spectrum-Effect Correlation

Objective:To be more scientific,objective to evaluate and control the quality of Yinhuang granule,and to ensure its clinical efficacy,This study used the directly related model between quantitative fingerprint and pharmacodynamic index,and through gray correlation analysis and support vector machine method to establish A complete set of quality control and evaluation system of the whole industry chain applicable to medicinal materials,extracts and preparations.The system ensures the quality of the product is closely related to its efficacy and improves the objectification level of drug quality evaluation.It is of great significance for realizing the quality contral of druy production process and promoting the modernization and internationalization of Chinese medicine.Methods:In this study,50 batches of Astragalus and Honeysuckle herbs were collected from the whole country,and 50 batches of extracts of Astragalus and Honeysuckle were prepared respectively.At least 50 batches of Yinhuang granule were prepared according to the 2015 Chinese Pharmacopoeia.The method of preparation was optimized through Different solid-liquid ratios,different extraction time,the investigation of hot and cold water;The HPLC conditions were optimized by investigating the flow rate,wavelength,column temperature.The HPLC quantitative fingerprints of 250 batches of samples were established by high performance liquid chromatography and selected the common peaks.The relative peak area values of common peak and internal standard peak were calculated as the mathematical expression of quantitative fingerprint;The LC-MS and reference substance were used to analyze the structure of the common peaks.The minimum inhibitory concentration（MIC）and the optimum number of bacteria were determined by microdilution method,and 250 batches of samples were tested for bacteriostasis in vitro to determine the bacteriostasis rate.Through gray correlation analysis and support vector machine method to establish a complete set of quality control and evaluation system of the whole industry chain applicable to Astragalus,Honeysuckle and their extracts and Yinhuang granules.Results:Established the preparation method and SOP（standard operating procedure）of Yinhuang granules,Astragalus membranaceus,honeysuckle medicinal materials,Astragalus membranaceus extract,honeysuckle extract for test solution,the test solution was stable for 30h,and the repeatability test RSD<3%;HPLC quantitative fingerprinting method and SOP were established,HPLC quantitative fingerprinting method and SOP were established,250 samples were measureed,24 common peaks were selected for Yinhuang granules,19 common peaks were selected for Astragalus membranaceus,and Honeysuckle herbs Twenty-one common peaks were selected,10 common peaks were selected from Astragalus membranaceus extract,18 common peaks were selected from honeysuckle extracts,and the chemical constituents of 14 peaks in Yinhuang granules were determined by the attribution of the common peak.Staphylococcus aureus was selected as the strain for in vitro bacteriostasis test,and the concentration of bacteria was determined to be 1×10 ⁵ cfu·mL^-1,In vitro bacteriostasis experiment SOP were established of Yinhuang granules,astragalus,honeysuckle medicinal materials and their extracts.Using gray correlation analysis to analyze 250 batches of samples,the results showed that the significant components of the common peaks of the Yinhuang granules（γ≥0.9）were all derived from the significant or relatively significant components of Honeysuckle herbs,Honeysuckle extract,Astragalus membranaceus,and Astragalus membranaceus extracts.A mathematical model was established for 250 batches of samples by least squares support vector machine method.The results showed that the relative error between the predicted value and the experimental value is less than 7%,and the average relative error is less than5%,the bacteriostatic rate can be accurately predicted by inputting HPLC fingerprint data.Conclusion:The established HPLC quantitative fingerprints method of Yinhuang Granules,Astragalus membranaceus,Honeysuckle medicinal materials,Astragalus membranaceus extract and Honeysuckle extract are stable and reliable.;The standard fingerprints established had large information and high accuracy,It can provide reference for quality evaluation of Yinhuang granule in the process of production;the established bacteriostatic test is accurate and reliable;the established mathematical model of antibacterial spectrum-effect correlation of Yinhuang granule can be used to determine the antibacterial effect of Yinhuang granule,medicinal materials and intermediates by quantitative fingerprint analysis.So as to evaluate the quality of the drug from the result of efficacy.