Study On The Chemical Constituents And Biological Activity Of The National Medicine Dijiao

The ethnodrug of Thymus quinquecostatus Celak,which the aerial part used as the medicinal parts,belongs to the genus Thymus of family Lamiaceae.The plants mainly distributed in Ningxia,Inner Mongolia,Shan'xi,Qinghai and Gansu etc.,and are widely used for detoxification and cancer treatment in multiple nationalities of Northwest China.Currently,the research of T.quinquecostatus are focused on its chemical constituents of the volatile oils,but studies on different origin of chemical components of volatile oils and its anticancer pharmacology,as well as the chemical constituents of alcohol extract,quality evaluation and so on are in the blank.In view of the above problems,the theses include the following sections:Part 1 Literature ReviewCheck out 55 references that related the plants of genus Thymus at home and abroad to review the chemical constituents and pharmacological activities,as well as the plant resources and its utilization that have reported for further research.To investigate the chemical constituents,bioactivity and its application in civil and clinical to provide a foundation for further research and development of plant T.quinquecostatus.Part 2 Chemical composition and biological activity studies of essential oils of T.quinquecostatus.The studies of anticancer activity show that the essential oils have significant inhibitory activity on BGC-823 and HepG2 cells.Among of them,the essential oils from qingyang and jingbian district are with higher inhibition rate then other two districts.The DPPH and ABTS free radical scavenging experiments shows that the four samples are with free radical scavenging activities.The DPPH radical scavenging capacity followed by T.quinquecostatus(yulin)>T.quinquecostatus(Jingbian)>T.quinquecostatus(qingyang)>T.quinquecostatus(longde).The ABTS radical scavenging activity from stronger to weaker orders T,quinquecostatus(qingyang)>T.quinquecostatus(yulin)>T.quinquecostatus(jingbian),the essential oils from longde shows no antioxidant activity.Studies on the chemical constituent of essential oils from yulin,jingbian,qinyang and longde district shows that the compounds are mainly aromatic constituents.,but there difference in both kinds and contents,only 1,8-cineole(1.84%-10.6%),linalool(1.22%-12.8%),terpinen-4-ol(1.03%-10.56%)and y-terpinene(0.15%-11.11%)are identified as the common constituent of four district.Result indicates that there are significant differences between the types of chemical constituents and their content.Part 3 Studies on chemical constituents and biological activities as well as multicomponent determination.To study the chemical constituent of T.quinquecostatus,13 compounds are separated and identificated.Seven of them are flavonoids,include 5,6-dihydroxy-4'-methoxy-7-O-n-alkyl flavonoid,scutellarein 7-O-glucuronide(scutellarin),scutellarein,luteolin-7-O-glucoside(luteoloside),luteolin,apigenin,rutin;four phenolic acid derivatives,include chlorogenic acid,vanillic acid,tanshinol and 3,4-dihydroxybenzoic acid;other two compounds,include oleanolic acid and daucosterol.Studies on inhibition of BGC-823 cell proliferation activity of different extract parts showed the ethyl acetate part with significant inhibition activity on human gastric cancer cell.The different parts of DPPH free radical scavenging activity are orders ethyl acetate part>n-butanol part>chloroform part>petroleum ether part>water part.The different parts of ABTS free radical scavenging activity are orders ethyl acetate part>n-butanol part>water part>petroleum ether part>chloroform part.The result means that T.quinquecostatus has significant anticancer activity and free radical scavenging activity.Use the antioxidant effect as indicators,by using the response surface analysis method to optimal alcohol extraction conditions.The curve fitting is DPPH=0.093-0.008X1-0.01X2+0.0007X3-0.003X1X2-0.0003X1X3+0.006X2X3-0.014X12-0.02X22-0.016X32,the fitting is in good condition,can accurately predict experimental results.The correction factor R2=0.8396 can be used to explain 83.96%variability of test data,the noise-signal ratio is 9.289,indicating a sponger signal.It can be concluded from the experiment that when ethanol concentration was around 45%,solid to liquid ratio is 20:1 and extract time is 1.5 h,the extract can possess highest antioxidant capacity.Established the HPLC quantitative analysis methods of 6 compounds,include danshensu,vanillic acid,chlorogenic acid,luteolin glycosides,scutellarin and apigenin.6 components are with good linear relationship in 4.68-150.0?g/mL,0.4-12.9 ?g/mL,6.25-100.0 ?g/mL,1.25-20.0?g/mL,6.25-200.0 ?g/mL,0.625-20.0 ?g/mL,respectively.Recoveries are within reasonable range of 95%-105%,RSD values are smaller than 2%within the reasonable range.Result shows that this method is simple,accurate,stable and viable,as a determination of compounds in T.quinquecostatus,and could as a quality evaluation of the different harvest stage.Part 4 The inhibit cancer cell proliferation synergy research of T.quinquecostatusIn view of the biological activities of essential oils and ethyl acetate part,the study aims to explore the synergy effect of anticancer activity of T.quinquecostatus.Result shows that two mix part can significantly increase the inhibition rate of BGC-823 cells,when ethyl acetate(mg):volatile oil(?L)=20.16:60,the concentration is 100 the inhibition rate can be up to 92.24%,indicate that the T.quinquecostatus works as a result of multipart synergy.Through the study on chemical constituents and biological activity of essential oils and alcohol extract part,to evaluate its bioactivity,study the type of chemical constituents of the volatile oils and the relationship between producing area and content,to clarify the anti-cancer ingredients,establish quality control method and undertake a study on quality of different harvest times.Part 5 Summary and discussionInnovation1 The first isolation and identification of the chemical components of plant T.quinquecostatus,19 compounds was separated.Seven of them are flavonoids,5,6-dihydroxy-4'-methoxy-7-O-n-alkyl flavonoid,scutellarein 7-O-glucuronide(scutellarin),scutellarein,luteolin-7-O-glucoside(luteoloside),luteolin,apigenin,rutinum;three phenolic acid derivatives,include chlorogenic acid,vanillic acid,tanshinol,3,4-dihydroxybenzoic acid;other tow compounds include oleanolic acid and daucosterol.2 First to carry out the comparative analysis of chemical constituents and biological activity of essential oils in different areas.3 First to carry out the study of chemical constituents and biological activity of different extract parts.4 First to establish the method to obtain the best antioxidant extract.5 First to establish a HPLC method for simultaneous determination the content of danshensu,vanillic acid,chlorogenic acid,luteolin glycosides,scutellarin and apigenin,to provide the foundation for a comprehensive and accurate quality evaluation of T.quinquecostatus.6 First to use the scutellarin as the index constituents to evaluate the quality of T.quinquecostatus that obtained from different seasons.7 First to study the synergy anticancer activity of the various active components of T.quinquecostatus.