Study On Drug Quality Characterization Of Ligustrum Lucidum Series Preparations

The literature review includes the following four aspects of the Fructus Ligustri Lucidi:the origin identification,characters identification,chemical composition,pharmacological action.And the pathological mechanism and treatment of drug research present situation in nervous system diseases(including migraine,alzheimer's disease,insomnia),Fructus Ligustri Lucidi and its active component based on the pharmacological effects of these three diseases were systematically expounded.In the above-referenced documents,the mechanism of action of nervous system diseases and basic chemical constituents of effective substance of Fructus Ligustri Lucidi related preparations were not reported.Purpose:To get the best preparation in the form of Fructus Ligustri Lucidi drugs at different pathogenesis of different pathological models,determine the effective material basis and mechanism of action.To get the pharmaceutical quality characterization method and the quality control way of Fructus Ligustri Lucidi preparations.Method:This paper conducted the drug innovation research,drug system as the guide.Based on the research of drug system,the preparation technology by previous research group,using different pathological conditions neurological disease as an indicator,Using UV,TLC,HPLC,specific chromatogram and other means was prepared for qualitative and quantitative analysis of Fructus Ligustri Lucidi series preparations,systematic correlation analysis of pharmaceutical preparations and pharmacodynamics of Fructus Ligustri Lucidi preparations.The correlation analysis of content of total ingredients and indicators of pieces of a reference and 14 batches of Fructus Ligustri Lucidi will get,and comprehensive accurate evaluate the excellent and good extent of Pieces quality,guidance on how to screening Fructus Ligustri Lucidi Herbal to get better preparations for best efficacy corresponding pathology of the disease.Result:The first section Characterization of Quality of Pharmaceutical Fructus Ligustri Lucidi series preparationsThe preparation of Fructus Ligustri Lucidi parts reference to Lijuan's dissertation,7kg taking Fructus Ligustri Lucidi coarse powder feeding,through extraction,alkalization,macroporous resin enrichment for four preparing products.N on-transformed extractive(FZT),Non-transformed enrichment(FZF),Alkalization extractive(ZT),Alkalization enrichment(ZF),Pure substance(CH)were producted.Fructus Ligustri Lucidi preparing products pilot test experiment was carried out,on its chemical composition has carried on the preliminary analysis.Vanillin-perchloric acid colorimetric method determinate the total terpene content of Fructus Ligustri Lucidi series preparation in our group Zhou Wenjuan dissertation Chapter 4,Sectionl was Optimized.Using H as chemical reference substances,had a favourable linear relationship in the range of 0.0044?0.0219g/mL,the average recovery was 98.88%,RSD was 2.90%.The results showed the total terpene content of FZT,FZF,ZT,ZF,CH is about Z15%?Z14%,Z10%?Z11%,Z8%?Z7%,Z5%?Z6%,Z1%?Z3%.Ferric chloride-potassium hexacyanoferrate color method determinate the total phenols content in the series of preparations in our group Zhou Wenjuan dissertation Chapter 4,Section3 was Optimized.Using A as chemical reference substance,had a favourable linear relationship in the range of 0.001116?0.00558mg/mL,the average recovery was 101.05%,RSD was 2.65%.The results showed that the content of FZT,FZF,ZT,ZF,CH is about f13%?f14%,f12%?f10%,f7%?f8%,f5%?f6%,f1%?f3%.The HPLC method of seven effective component content indexs in Fructus Ligustri Lucidi preparing product drugs are established.The linear regression equation of B is:Y=883894X+1337(r=0.9999),the linear range was 0.0618?g?1.854?g,the average recovery was 101.15%,RSD was 2.20%;The linearregression equation of A was:Y=285631X-26612(r=0.9999),the linear range was0.505?g?20.2?g,the average recovery was 100.61%,RSD was 1.64%;The linear regression equation of C was:Y=694994X-8050(r=0.9998),the linear range was 0.051 ?g?2.04?g,the average recovery was 102.93%,RSD was 1.91%;The linear regression equation of D was:Y=1385037X+2446(r=0.9999),the linear range was 0.121?g?2.904?g,the average recovery was 99.51%,RSD was 2.02%;The linear regression equation of E was:Y=1545488X-16426(r=0.9999),the linear range was 0.0315?g?5.28?g,the average recovery was 101.38%,RSD was 1.96%;The linear regression equation of F was:Y=1633549X-5736(r=0.9998),the linear range was 0.042?g?0.525?g,the average recovery was 98.55%,RSD was 2.46%;The linear regression equation of G was:Y=1134978X+408486,(r=1.0000),the linear range was 0.612?g?51.0?g,the average recovery was 97.53%,RSD was 1.33%.According to the method,the quality effective component content indexs of Fructus Ligustri Lucidi preparing products was measured.An average B content of FZT,FZF,ZT,ZF,CH is b13%?b15%,b10%?b12%,b 7%?b9%,b4%?b6%,b1%?b3%;A is a13%?a15%,al0%?a12%,a7%?a9%,a4%?a6%,a1%?a3%;C is c13%?c15%,c10%?c12%,c7%?c9%,c4%?c6%,c1%?c3%.The quality of D of FZT,FZF is d4%?d6%,dl%?d3%.The quality of E is e4%?e6%,e1%?e3%,The quality of G is g4%?g6%,g1%?g3%;The quality of F in FZT is h1%?h3%.The effective index components D,E,F,Gwas not detected in ZT,ZF,CH,the effective index component F was not detected in FZF.With TLC controlling the Fructus Ligustri Lucidi preparations qualitative quality,divided into four conditions:A,B and C were qualitative analysis with chloroform-Methanol-formic acid(5:1:0.2);D,E and G were qualitative analysis with Ethyl acetate-water-acetic acidformic acid(3:0.15:2:0.2);After pretreatment of spot with 1%iodine-methylene chloride solution,H and M were qualitative analysis with Cyclohexane-methylene chlorideethyl acetateformic acid(20:5:8:0.2);F were qualitative analysis with Methylene chloridemethanol-water-formic acid(15:5:0.3:0.2).High performance liquid chromatography analysis,Affirm characteristic peak by comparing peak area,tR and characteristic UV absorption spectrum,with adding the indicative constituents.6peaks,14peaks,14peaks,36peaks,28peaks are identifiedin CH,ZF,ZT,FZF,FZT.respectively.Select A as a reference peak,the retention time,peak area is set to 1,to calculate the relative retention time and peak area relative to other characteristic constituents,characteristic the features of the quality of each preparation.The second section Characterization study of pharmacodynamic quality of Fructus Ligustri Lucidi preparationsThe series of Ligustrum lucidum Ait preparing products is divided into high,medium and low dose group,accompanied by blank,model,positive group were compared.Purpose of this chapter is to explore the best pharmaceutical drug composition and form the basis of active substances of Ligustrum lucidum Ait under different pathological models.(1)Anti-migraine efficacy experiment is the use of glyceryl nitrate copy hypodermic rat model of migraine,serum 5-HT,CGRP,ET-1 content by biochemical assays,to evaluate the efficacy results of anti-migraine drugs of Ligustrum lucidum Ait.After modeling head redness,nitroglycerin-induced migraine rat serum 5-HT,CGRP,ET-1 content abnormally elevated;after drug intervention,effectively reduce migraine 5-HT content in rat plasma is Purification of high,Alkalization enrichment of high,Non-transformed extractive of high;lowering plasma levels of CGRP in migraine rat is Non-transformed Concentration of high and middle,Alkalization enrichment of high Non-transformed extractive of high;and each high,medium dose group could reduce big murine plasma ET-1 levels.Comprehensive analysis of migraine rat plasma 5-HT,CGRP,ET-1 content of the test results,Alkalization enrichment was better than other preparation administration groups in anti-migraine effect,at 120 mg/kg-240 mg/kg dose,it can effectively regulate the expression levels of related to the neurotransmitter 5-HT,CGRP,ET-1 significantly.(2)Acts of anti-AD efficacy experiments is use intraperitoneal injection of scopolamine hydrobromide copy mouse AD model mice,according to the step-down test results and behavioral AchE activity data for evaluation of pharmacodynamic effect.The results show that the best efficacy of treatment group is purification,its AD model mice brain tissue AchE inhibitory activity on the reduction of AD model mice memory access and memory consolidation phase error times superior to each other than those in administration role group.(3)The efficacy of anti-insomnia insomnia experimental mouse model is replicated by intraperitoneal injection of PCPA,the efficacy results is:the purification>alkalization enrichmention>non-transformed extracts>conversion extracts>non-transformed enrichment.Purification in the range of 120?240 mg/kg has the strongest efficacy in improved 5-HT content and prolonged sleep duration.Alkalization enrichmention at a dose of 90?360 mg/kg,has the strongest efficacy in shorten the time of pentobarbital sodium collaborative sleep.The third section Correlation Analysis the pharmacy-pharmacodynamic of Fructus Ligustri Lucidi series preparationFructus Ligustri Lucidi system drugs has established by the correlation analysis of pharmacy and Pharmacological function.To determine the efficacy of the active substance base and characterize the quality of the system,through analysis of extraction amount,the active ingredient content of the index,based on salidroside content relative ratio,the effective index of the relative amounts of ingredients,based on the relative ratio of the active ingredient content Pieces indicative,change width,preparation ingredient composition of association of series of preparation.A is one of the active ingredients to reduce 5-HT content of migraine in the serum,A,B,C and C analogues are effective components to reduce migraine rats serum content of CGRP and ET-1;B,C and C analogues and the synergies effect of A to reduce CGRP and ET-1 content is one of the mechanism of action.Echinacoside and verbascoside can be reduce CGRP content in serum.The content of A in alkalization enrichment shall not be less than a6%,the B shall not be less than b5%,the C shall not be less than c4%;The range of B with A content ratio is:y1/100.0?y2/100.0,the effective range of C with A content ratio is y3/100.0?y4/100.0.A is the main effective components to reduce AchE activity,adjust the memory disorder mice short-term memory,the cooperate A with B,C and C analogues can adjust the long-term memory of memory impairment in mice.The content of A in Pure substanc shall not be less than al%,the B and C shall not be less than x2%.A is one of the active ingredients to reduce the content 5-HT in hippocampus of insomnia model mice,extend sleep time,the synergy of A with B,C and C analogues has a synergistic effect,which can effectively shorten the sleep time of insomnia mice.The content of A in series preparations shall not be lower than m2%;The range of B with A content ratio in the range of y5/100.0?y6/100.0,the range of C with salidroside content scope is y7/100.0?y8/100.0,in a certain range,the bigger the ratio,the better synergistic effect.The total content of component types and indicators of content 15 batches of Fructus Ligustri Lucidi Pieces was determinatied,based on the reference material(reference Pieces)characterize the quality system as a reference scale test to evaluate the non-relevancy degree and relevancy degree of Fructus Ligustri Lucidi Pieces quality,correlation analysis the content of characteristic index component A,G(can be converted to A),B,C,the relevancy degree of 15 Pieces to screened Fructus Ligustri Lucidi Pieces based on efficacy of Antimigraine,AD,insomnia.Optimum quality in selected Pieces in 15 batches is sample2,3,5,12,the order of priorities is 3>2>12>5.Quality representation and correlation analysis of Ligustrum lucidum Ait is providing the foundation of raw material quality control and application screening.In the preparation of pharmaceutical Fructus Ligustri Lucidi process,based on the optimum drug system,targeted to improve the content of the active ingredient index,optimize its composition ratio is key to enhancing the quality of the drug.The method of correlation analysis on basic drug system characteristic was used for guiding drug preparation and quality evaluation.Based on the complete composition of the drug system,increases the purity of the effective index components,so that the relative ratio of the various types of components more reasonable,to ensure complete quality,quantity properly,in order to obtain ideal Ligustrum drugs.The fourth section Summary and Discussion1.The method of measure content of total composition and ingredients of indicative Fructus Ligustri Lucidi are to be established.The determination method of total terpene and total phenolic content are established.Experiment can be used for determination the content of A,B,C,D,E,F,Grespectively.It aims to quantitative characterization of Fructus Ligustri Lucidi series preparation using the characteristics which are screened by drug system as standard to determine the contents of preparation with UV and HPLC,respectively,established feature maps and thin layer chromatography to achieve the purpose of comprehensive the quality of preparation effectively.2.Correlation Analysis the pharmacy-pharmacodynamic of Fructus Ligustri Lucidi series preparation under different pathological models,Cleared the effective material basis of different pharmacy-efficacy system.3.The newly established quality evaluation model is applied to evaluation the quality of 15 batches of Fructus Ligustri Lucidi.Provide a basis for screening raw material,quality control and application.Establish a system of Fructus Ligustri Lucidi quality control system based on Chinese Pharmacopeia(Edition2015),establish the method for determine the content of total phenols,total terpenoids,index components.