Research On The Correlation Analysis Of The Preparation Process And Quality Characterization Of Lianqiaoyin Drug Based On The Drug System

This paper includes six chapters.The first chapter:studies on the extraction technology of the medicinal-preparation of LianqiaoYin based on the medicinal system;The second chapter:studies on the purification technology of the medicinal-preparation of LianqiaoYin based on the medicinal system;The third chaper:quality representation of the medicinal-preparation of LianqiaoYin based on the total content;The fourth chapter:quality representation of the medicinal-preparation of LianqiaoYin based on the effective component content;The fifth chapter:quality-representation correlation of medicinal-preparation of LianqiaoYin based on medicine-efficacy;The sixth chapter:summary and discussion.The first chapter:studies on the extraction technology of the medicinal-preparation of LianqiaoYin based on the medicinal systemThe big rat were given LianqiaoYin extract orally after being infected by Staphylococcus aureus.To study PK-PD-DI to find and select out antimicrobial medicinal-system of LianqiaoYin,providing basis for preparation.To choose the total Phenol,Terpenoid and representative indexes as indexes to comprehensively guide the preparation technology of LianqiaoYin.Study the extraction technology of the medicinal-preparation of Lianqiao Yin.Using L9(34)orthogonal test,which were valued by extraction quantity of total Terpenoids,total Phenols and the effective component.To analyse all the experiments results synthetically,the best extraction technology of the medicinal-preparation of LianqiaoYin is A2B3C3D2,the A2 alcohol extract B3 times,C3 hour for one time,and the quantity of solvent is D2 times of the medical material.3 batches of samples of extraction technology shows:the average extraction ratio is 31.18%;the average content of total Phenol was 22.55%,and the average extraction ratio was 97.45%;the average content of total Terpenoids was 5.00%,and the average extraction ratio was 91.74%;the average content of Gardenoside(GD)was 3.58%,and the average extraction ratio was 95.17%;the average content of Forythoside A(FTA)was 0.32%,and the average extraction ratio was 82.55%;the average content of Glycyrrhizic acid(GZA)was 1.19%,and the average extraction ratio was74.01%;the average content of Phillyrin(Phil)was 0.11%,and the average extraction ratio was 86.17%;the average content of Cimifugin(CN)was 0.12%,and the average extract ratio was 94.70%.The method were steady and reliable.The second chaper:studies on the purification technology of the medicinal-preparation of LianqiaoYin based on the medicinal systemStudy the purification technology of the the medicinal-preparation of LianqiaoYin.After investigated the absorptive property of 6 kinds of macroporous adsorptive resin,D resin was selected due to its best absorptive and separative property of total Terpenoids,total Phenols.We investigated the loading capacity,the ratio of diameter and height,concentration of solution,absorptive rate,purified solvent,velocity of flow,eluting solvent and et al.The parameter of the purification was considered,and optimized condition of resin application and enriching was as follows:Decompression enrichment the extract prepared in accordance with the optimum technology,then disperse solution by water to make the concentration get N1(content of medicinal herbs in solusion per mL),subject the concentrated solution to D resin column(diameter/height=J2),the ratio of loading capacity(corresponded dried medicinal herb gram)to resin volume was B3,the adsorbing current velocity V2,SB3 times volume of resin of water to wipe off impurity with SV2 velocity of flow,then M times volume of TC2 alcohol to elute with the flow of TV3,reclaim the solvent of TC2 alcohol,vacuum drying,weighting,the total Terpenoids fraction was obtained.Then N times volume of FC4 alcohol to elute with the flow of FV3,reclaim the solvent of FC4alcohol,vacuum drying,weighting,the total Phenols fraction was obtained.3 batches of samples of extraction technology shows:The average extraction ratio of TC2 is 4.12%.The average content of total Terpenoids in the total Terpenoids fraction was 34.22%,and the transferring ratio was 87.95%;The average content of Gardenoside(GD)in the total Terpenoids fraction was 25.55%,and the transferring ratio was 92.18%;The average content of total Terpenoids in the total Phenols fraction was 35.09%.The average extraction ratio of FC4 is 6.29%.The average content of total Phenol in the total Phenols fraction was 64.71%,and the transferring ratio was 77.58%;The average content of Forythoside A(FTA)in the total Phenols fraction was 1.55%,and the transferring ratio was 71.84%;The average content of Glycyrrhizic acid(GZA)in the total Phenols fraction was 4.84%,and the transferring ratio was 60.88%;The average content of Phillyrin(Phil)in the total Phenols fraction was 0.54%,and the transferring ratio was 81.89%;The average content of Cimifugin(CN)in the total Phenols fraction was 0.56%,and the transferring ratio was 87.12%.The third chapter:quality representation of the medicinal-preparation of LianqiaoYin based on the total contentBased on the medicinal system,the quantitative method of total Phenols by UV-Vis which was carried out Trichloride-Ferricyanatum Kalium Colorimetric reaction was established.The linear regression was Y=0.1307X+0.2721,r=0.9996(n=9),using forsythoside A as indicative constituent,that the concentration(μg/mL)of forsythoside A and absorptance had a favourable linear relationship in the range of 0.1512-4.6872 μg/mL,the average recovery was 98.37%,RSD=2.58%(n=6).The determination results of the contents showed that the methods were steady and reliable and can be used as the determination methods.Using the established quantitative method make the quality representation of the alcohol extraction,resin purification and herbal pieces of LianqiaoYin based on the total content.The content of total Phenols in TC2 fraction of LianqiaoYin is 34.75%(RSD=2.45%);and the content of total Phenols in FC4 fraction of LianqiaoYin is 64.71%(RSD=1.55%);The content of total Phenols in alcohol extraction of LianqiaoYin is 22.55%(RSD=2.57%);The content of total Phenols in herbal pieces of LianqiaoYin is 7.11%(RSD=0.56%).The fourth chapter:quality representation of the medicinal-preparation of LianqiaoYin based on the effective component contentHPLC-DAD method was used in the experiment to determine the effective components in the medicinal-preparation of LianqiaoYin.The linear regression of Gardenoside(GD)was Y=1251050X-13688.4,r=0.9999(n=6),the favourable linear relationship in the range of 0.680μg～4.080μg;The linear regression of Forythoside A(FTA)was Y=610093X-1458,r=0.9993(n=6),the favourable linear relationship in the range of 0.044μg～0.264μg;The linear regression of Glycyrrhizic acid(GZA)was Y=641997X-29482,r=0.9999(n=6),the favourable linear relationship in the range of 0.260μg～1.560μg;The linear regression of Phillyrin(Phil)was Y=512876X-104.13,r=0.9999(n=7),the favourable linear relationship in the range of 0.023μg～0.186pg;The linear regression of Cimifugin(CN)was Y=2447630X-8967.16,r=0.9999(n=6),the favourable linear relationship in the range of 0.030μg～0.247～g;The linear regyression of Prim-O-glucosylcimifugin(PGCN)was Y=1371000X-11693.43,r=0.9999(n=6),the favourable linear relationship in the range of 0.083μg～0.693μg;The linear regression of 4’-O-D-glucosyl-5-O-methylvisamminol(GMV)was Y=1475150X-14767.7,r=0.9999(n=5),the favourable linear relationship in the range of 0.101μg～0.505μg;The linear regression of Liquiritin(LQR)was Y=1716700X-20126.2,r=0.9998(n=5),the favourable linear relationship in the range of 0.101μg～0.505μg.The method had a good linear relationship and qualified average recovery.Taking gardenoside content as the standard and is converted into a same ultraviolet absorption of the total content of the same material.Make the quality representation of drug-preparation of LianqiaoYin based on the total Terpenoids.The average content of GD in TC2 fraction of LianqiaoYin was 26.2948%,The average content of PGCN in TC2 fraction of LianqiaoYin was 2.8349%;The average content of FTA in FC4 fraction of LianqiaoYin was 1.7458%;The average content of GZA in FC4 fraction of LianqiaoYin was 4.8400%;The average content of CN in FC4 fraction of LianqiaoYin was 0.5322%;The average content of Phil in FC4 fraction of LianqiaoYin was 0.4839%;The average content of LQR in FC4 fraction of LianqiaoYin was 2.3230%;The average content of GMV in FC4 fraction of LianqiaoYin was 1.4364%.The average content of GD in alcohol extract of LianqiaoYin was 3.5561%,The average content of PGCN in alcohol extract of LianqiaoYin was 0.4551%;The average content of FTA in alcohol extract of LianqiaoYin was 0.3545%;The average content of GZA in alcohol extract of LianqiaoYin was 1.1900%;The average content of CN in alcohol extract of LianqiaoYin was 0.1258%;The average content of Phil in alcohol extract of LianqiaoYin was 0.1045%;The average content of LQR in alcohol extract of LianqiaoYin was 0.4430%;The average content of GMV in alcohol extract of LianqiaoYin was 0.2897%.The average content of GD in herbal pieces of LianqiaoYin was 1.1400%%,The average content of PGCN in herbal pieces of LianqiaoYin was 0.1547%;The average content of FTA in herbal pieces of LianqiaoYin was 0.1360%;The average content of GZA in herbal pieces of LianqiaoYin was 0.5014%%;The average content of CN in herbal pieces of LianqiaoYin was 0.0404%;The average content of Phil in herbal pieces of LianqiaoYin was 0.0381%;The average content of LQR in herbal pieces of LianqiaoYin was 0.1453%;The average content of GMV in herbal pieces of LianqiaoYin was0.0996%.The average content of Terpenoids in the herbal pieces,alcohol extract and TC2 fraction is 1.60%、4.92%、33.87%.The fifth chapter:quality representation correlation of medicinal-preparation of LianqiaoYin based on the medicine-efficacyThis topic by disc diffusion method,compare different form medicinal-preparation of LianqiaoYin under the same doses,used alone and in combination with different doses of antibiotics for Staphylococcus aureus,the circle of inhibiting bacterium,aims to demonstrate the effectiveness of the medicinal-preparation of LianqiaoYin in terms of efficacy,prove the feasibility of technology.The circle of inhibiting bacterium of water extract was 8.33mm,the alcohol extract was 9.67mm and enrichment was 8.67mm.The three forms of medicinal-preparations all showed antibacterial effect in vitro.In combination with different doses of antibiotics,the circle of inhibiting bacterium of the medicinal-preparation of LianqiaoYin was increased,and the size of antibacterial efficacy function:enrichenment ≥alcohol extract>water extract.This subject adopts the Staphylococcus aureus abdominal cavity infection in mice,observe the survival time of mice and mortality to make a comparison of antibacterial effect among medicinal-preparations in vivo.The mean survival time of water extract was 3.86 days,the mortality was 64.29%;The mean survival time of alcohol extract was 4.64 days,the mortality was 50.00%;The mean survival time of enrichment was 5.14 days,the mortality was 42.86%.Compared with model control group,the mean survival time of medicinal-preparation of LianqiaoYin was lengthened,and the mortality was reduced.The result showed the medicinal-preparation of LianqiaoYin has antibacterial effect in vivo.Make quality-representation of medicinal-preparation based on the content,the relative proportion and the quantity of effective components and based on the extraction rate of medicinal-preparation and to study the quality representation correlation of LianqiaoYin based on the medicinal system to determine the key factors affecting its quality.Based on the content of the effective index components,the relative proportion,the amount of pharmaceutical preparation of a medicinal-preparation of LianQiaoYin,analysis of pharmacological properties,i.e.,the drug content of the drug system,the relative proportion of the amount of the preparation,the preparation rate changes in the magnitude of the drug preparation room,combined vivo antibacterial effect size,carried Pharmacy-quality Characterization of pharmacodynamic correlation analysis.Found relatively decocting,enrichment of LQR,GD,GMV,CN,PGCN,FTA,Phil,GZA content was increased to 9.65 times,9.13 times,8.71 times.7.96 times,7.89 times,7.75 times,7.67 times and 5.83 times.Medicinal preparations between the relative proportions of the different forms of the more obvious changes in the magnitude of the effective index components by size as follows:Phillyrin>forsythiaside A>liquiritin>Geniposide and growth in the enrichment of the biggest changes.According to the medicine-efficacy of correlation analysis,based on the basic composition and extraction rate of medicinal system,increasing the content of the key components and optimizing the proportion is the key to improve efficacy.In the three preparations,especially in enrichnment,its effective components is higher,proportion is unique,extraction rate is lower,dosage is smaller and antibacterial effect is stronger.The enrichnment is the best medicinal-preparation application form of LianqiaoYin.The sixth chapter:summary and discussion1 Firstly established the extraction technology and purification technology of the medicinal-preparation of LianqiaoYin based on the medicinal system.2 Based on the medicinal system,firstly established quantitative determination of medicinal-preparation of LianqiaoYin based on the total and effective component content to make quality-representation of the medicinal-preparation of LianqiaoYin.3 Study the medicine-efficacy of correlation analysis of the medicinal-preparation of LianqiaoYin to find the best form of drug,to provide new model of preparation and quality evaluation for traditional Chinese medicine.