Evalution On The Quality And Analgesic Efficacy Of Vinegar Corydalis Yanhusuo Under Different Sterilization Methods

ObjectiveCorydalis yanhusuo,also known as "the holy medicine for pain relief,"possesses analgesic activity.Its efficacy is enhanced after vinegar processing,and vinegar-processed products are widely used in clinical practice.The growth and proliferation of microorganisms can affect the taste and quality of traditional Chinese medicinal materials,leading to a decrease in therapeutic efficacy and even posing a threat to the health of users.Sterilization is an important method to control the microorganisms in medicinal materials.It is crucial to evaluate the quality and biological activity of medicinal materials after sterilization.In this study,various methods were employed to sterilize vinegar Corydalis yanhusuo medicinal materials.The quality of the sterilized medicinal materials was evaluated,and the central and peripheral analgesic effects were investigated.The feasibility of using quantitative analysis of multi-components by single marker,fingerprint analysis,and orthogonal design-multivariate linear regression analysis was explored for the quality evaluation and analgesic efficacy of vinegar Corydalis yanhusuo medicinal materials under different sterilization methods.MethodsCorydalis yanhusuo medicinal materials were prepared using a standardized processing method.vinegar Corydalis yanhusuo was subjected to seven sterilization methods:radiation sterilization,ultraviolet sterilization,microwave sterilization,dry heat sterilization,high-pressure sterilization,circulating steam sterilization,and ethanol sterilization.Quantitative analysis of multi-components by single marker method was established to simultaneously determine the content of five alkaloids in vinegar Corydalis yanhusuo:protopine,berberine hydrochloride,palmatine hydrochloride,tetrahydropalmatine and corydaline,.This method was applied to determine the alkaloid content in vinegar Corydalis yanhusuo after sterilization.HPLC fingerprint analysis,FTIR fingerprint analysis,and second derivative spectra analysis were conducted to analyze the impact of different sterilization methods on the quality of vinegar Corydalis yanhusuo medicinal materials.A mouse S180 tumor compression model was constructed,with five alkaloids as influencing factors,orthogonal experimental design,and injection through tail vein.And the use of HPLC-FLD method to measure the amino acid content in mouse brain,the ratio of excitatory amino acids to inhibitory amino acids was used as an index.Orthogonal design-multivariate linear regression analysis was performed to determine the contribution rate of the five alkaloids to analgesia.Based on the prediction results,radiation sterilization,dry heat sterilization,and ethanol sterilization were selected as the sterilization methods to investigate the central analgesic efficacy of vinegar Corydalis yanhusuo under different sterilization methods.A mouse acetic acid-induced writhing model was established,with the inhibition rate of writhing as an index,to investigate the peripheral analgesic efficacy of vinegar Corydalis yanhusuo under different sterilization methods.ResultsThe results of the quantitative analysis of multi-components by single marker method showed a general decrease in the content of the five alkaloids in vinegar Corydalis yanhusuo under different sterilization methods.The method was accurate,feasible,and yielded reliable results for the quantitative determination of alkaloid components in vinegar Corydalis yanhusuo under different sterilization methods.The similarity results of HPLC fingerprint analysis and FTIR fingerprint analysis indicated that the similarity values of unsterilized Corydalis yanhusuo,radiation sterilization,ultraviolet sterilization,and microwave sterilization were above 0.96,indicating good similarity.The similarity values of dry heat sterilization,circulating steam sterilization,and high-pressure sterilization were above 0.90,while ethanol sterilization had a similarity value below 0.90,indicating poor similarity.The results of the second derivative spectra analysis demonstrated that the characteristic regions of unsterilized Corydalis yanhusuo,radiation sterilization,ultraviolet sterilization,and microwave sterilization were similar and could differentiate the differences between vinegar Corydalis yanhusuo medicinal materials under different sterilization methods.The fitted multivariate linear regression equation was Y=1.351-0.045X1-0.035X2-0.009X3-0.112X4-0.221X5.The orthogonal design multivariate linear regression analysis indicated that the contribution rates of the five alkaloids to analgesia were tetrahydropalmatine>corydaline>protopine>berberine hydrochloride>palmatine hydrochloride.The determination results of amino acid content in mouse brain indicated that the central analgesic effects were in the order of radiation sterilization>dry heat sterilization>ethanol sterilization,which was consistent with the prediction results of orthogonal design-multivariate linear regression analysis.The results of the acetic acid-induced writhing test showed that sterilized vinegar Corydalis yanhusuo could reduce the number of writhing responses in mice,indicating certain peripheral analgesic effects.ConclusionsQuantitative analysis of multi-components by single marker method was established,with tetrahydropalmatine as the internal reference,to simultaneously determine the content of protopine,berberine hydrochloride,palmatine hydrochloride and corydaline.By combining HPLC fingerprint analysis for accurate quantification and FTIR fingerprint analysis for precise identification,the quality evaluation method for vinegar Corydalis yanhusuo under different sterilization methods was improved.Orthogonal design multivariate linear regression analysis was used to predict the central analgesic efficacy of vinegar Corydalis yanhusuo under different.The acetic acid-induced writhing test showed that sterilized vinegar Corydalis yanhusuo had certain peripheral analgesic effects.