Studies On The Constituents And Quality Control Of Flemingia Philippinensis Merr. Et Rolfe

Qianjinba,known as "I-Tiao-Gung",is the dried root of perennial herbage Flemingia philippinensis Merr.et Rolfe which belongs to the genus Flemingia of family Leguminosea.It distributes in the south-west,middle-south and east-south China.It is sweet and astringent taste according to the record of Chinese Materia Medica,and it has the functions of dipelling wind and eliminating dampness and treatment for the gynecological diseases.There are little studies about the chemical constituents and pharmacological action of the crude drug,and no method was established to evaluate the quality at the present time.In order to use and develop this drug further,the chemical constituents and quality control were studied systematically,and the pharmacological activities of the crude extract and partial compounds were screened.In addition,the distribution,living environment,medical efficacy,chemical constituent and pharmacological action about the genus Flemingia plants were summarized.Our research results are as follows.Forty-six compounds were isolated from the 75%ethanol extract of the root of famous-region drug Flemingia philippinensis Merr.et Rolfe by using the modern chromatographic techniques.The structures of forty-three ones(FP-1～FP-43) were identified by the physico-chemical properties and modern spectral techniques,including UV,IR,¹H-NMR,¹³C-NMR,HMQC,HMBC,NOESY,COSY,EI-MS,FAB-MS and HR-EI-MS.They contained twenty-eight flavonoids,four anthraquinones,four organic acids and ester,three sterols,one prenylated ketone,one coumarin,one nucleoside and one diolefine compound.These compounds were identified as follows:chrysophanol (FP-1),physcion(FP-2),eriosematin(FP-3),scoparone(FP-4),lupeol(FP-5),betulinic acid(FP-6),3',4'-Dihydroxy-trans-cinamic acid octacosyl ester(FP-7),β-sitosterol (FP-8),flemiphilippinone A(FP-9),monopalmitin(FP-10),emodin(FP-11),islandicin (FP-12),salicylic acid(FP-13),p-methoxyphenylpropionic acid(FP-14),trideca-1, 3-diene(FP-15),lupinifolin(FP-16),flemichin D(FP-17),flemiphilippinin A(FP-18), auriculasin(FP-19),erythrinin B(FP-20),6-C-prenylluteolin(FP-21), 8-(1,1-dimethylallyl) genistein(FP-22),flemiphilippinin E(FP-23),flemiphilippinin F (FP-24),5,7,3',4'-tetrahydroxy-6,8-diprenylisoflavone(FP-25),flemiphilippinin D (FP-26),dorsmaninsâ… (FP-27),osajin(FP-28),6,8-diprenyleriodictyol(FP-29), lupinalbin A(FP-30),genistein(FP-31),3'-O-methylorobol(FP-32),orobol(FP-33), 5,7,2',3',4'-pentahrdroxyflavone(FP-34),the mixture of biochanin A and prunetin (FP-35 and 36),genistin(FP-37),sophororicoside(FP-38),3'-O-methylorobol-7-glucoside(FP-39),the mixture of sissotrin and prunetin 4'-O-β-D-glucoside(FP-40 and 41),adenosine(FP-42) and luteoloside(FP-43,mixture).Among these compounds,FP-9 was a novel skeleton of prenylated ketone,FP-23 and FP-24 were new compounds, FP-34 was a novel natural product,twenty-one were isolated from the Flemingia genus for the first time,and four were isolated from the F.philippinensis for the first time.In addition,anti-oxidative and antitumor activities of seventeen compounds and four crude extract parts were researched.The results indicated that 3',4'-dihydroxy-trans-cinamic acid octacosyl ester,flemiphilippinin A,5,7,3',4'-tetrahydroxy-6,8-diprenylisoflavone,auriculasin and dorsmaninsâ… showed strong activity in free radical-scavenging activity determined with the scavenging effect on DPPH free radical,while flemiphilippinin A showed strong cytotoxicity activity studied with MTT method.At last,the method to control the quality of F.philippinensis was established for the first time.The studies contained:(1) Acorrding to the chemical character and pharmacodynamic analysis of this drug,the quantitative determination of total flavoniod was studied using UV-VIS;(2) The fingerprint of was established using RP-HPLC-DAD comparing 19 samples from different distributing regions;(3) The quantitative analysis of three active isoflavones,genistin,genistein and 5,7,3',4'-tetrahydroxy-6,8-diprenylisoflavone in F.philippinensis was researched using a HPLC-DAD method. Based on above results,a quality standard of Qianjinba was drafted to provide the scientific method for identification and evaluation of the crude drug. In summary,the chemical contituents,pharmacological activities,fingerprint and quantitative analysis were systematical studied in this thesis.Moreover,the method for quality control of Qianjinba was established.These provided the theoretical basis and scientific reference for further study,development and application of Flemingia philippinensis Merr.et Rolfe.