Study On Chemical Constituents And Quality Evaluation Of Flos Plumeria Rubra Cv.acutifolia

Objective:Jidanhua is dry flower of Apocynaceae Plumeria rubra Linn cv.acutifolia,it has the effects of "clearing-heat" and "inducing-diuresis".Volume 1(2004 Edition)of "Guangdong province traditional Chinese medicinal material standard" records quality standards of Jidanhua,it only has character identification and still unable to be evaluated advantages and disadvantages by science,so this paper on base of systematical study of its chemical constituents,makes multi index components quantitative analysis method with HPLC and chromatographic fingerprint identification method,in order to provide scientific reference for establishment quality standard of Jidanhua.Method:1.In this experiment,Plumeria rubra Linn cv.acutifolia methanol extraction was isolated by silica gel,Sephadex LH-20,ODS column chromatography and recrystallization as well as other methods,and we confirmed structure of separated compounds by comprehensive physical properties and spectroscopic techniques(including IR,UV,ESI-MS,1H-NMR,13C-NMR,HMBC and NOESY)and comparing with relevant literature data.2.The paper analyzed fingerprint of Jidanhua,With liquid chromatography(HPLC)and(Poroshell 120,SB-C18,2.7 ?m 4.6×100 mm)×2 of chromatography column,flow phase:acetonitrile(A)-methanol(B)-0.3%KH2-PO4+0.2%H3PO4(C),column temperature:25 ?,DVD wavelength at 254 nm.Did similarity analysis,hierarchical cluster analysis and principal component analysis with "traditional Chinese medicine chromatographic fingerprint similarity evaluation software" and SPSS 19.0 version.3.Establish a HPLC method to determination the contents of 15-demethylplumi eride,coumaric acid-4-O-p-D-glucopyranoside,protocatechuate,plumieride,cerberic a cid B,rutin,7',8'(E)-1?-Protodemethyplumericin A,isoquercetin,kaempferol-3-O-P-D-glucopyranosyl-7-O-a-L-Rham-nopyranoSide,coumaric acid,iuteolin-7-O-?-D-gluco pyranoside,luteolin-3'-O-?-D-glucopyranoside,kaempferol-3-O-?-D-glucopyranoside,15-demethylplumieride P-Z-coumarate,Plumieride P-Z-coumarate,quercetin,?-dihidropl umericinic acid,kaempferol in Jidanhua.The conditions of the quantitative analysis:SB-C18(2.7 ?m,4.6×100 mm)×2 analytical column,flow phase:acetonitrile(A)-methanol(B)-0.3%KH2PO4+0.2%H3PO4(C),flow rate:0.6 ml/min,column temperat ure:25?,DVD wavelength at 230 nm and 360 nm.4.This paper inspected and compared if Jidanhua contains cardiac glycosides component with common cardiac glycoside compounds in Apocynaceae plants as control group by physicochemical identification,thin layer chromatography(TLC)and UHPLC-MS.Result:1.53 compounds were isolated from methanol extraction and 50 of them were identified,including 13 iridoids,10 flavonoids,15 organic acids,4 triterpenes,3 phe nolic glycosides,1 coumarin and 4 other compounds.The results were as follows:7',8'(Z)-la-Protodemethyplumericin A(PR-1),7',8'(E)-1?-Protodemethyplumericin A(P R-2),13-O-caffeoy-15-demethyplumieride(PR-3),Plumieride P-Z-coumarate glucoside(P R-4),Plumieride P-E-coumarate glucoside(PR-5)?7-?-Hydroxy-11-methyl forsythide(P R-6),loganic acid(PR-7),15-demethylplumieride(PR-8),plumieride(PR-9),Plumieride P-Z-coumarate(PR-10),13-O-caffeoylplumierides(PR-11),15-demethylplumieride P-Z-coumarate(PR-12),?-dihidroplumericinic acid(PR-13);kaempferol-3-O-?-D-glucopyrano syl-7-O-?-L-Rham-nopyranoSide(PR-14),rutin(PR-15),extraneous rutin(PR-16),isoquer cetin(PR-17),kaempferol-3-O-?-D-glucopyranoside(PR-18),luteolin-7-O-?-D-g lucopyranoside(PR-19),luteolin-3'-O-?-D-glucopyranoside(PR-20),quercetin(PR-21),k aempferol(PR-22),quercetin-3,3'-dimethylether(PR-23);3-O-coumaroyluquinic acid(PR-24),(Z)-4-coumaric acid-4-O-?-D-glucopyranoside(PR-25)?(E)-4-coumaricacid-4-O-?-D-glucopyranoside(PR-26),caffeic acid(PR-27),protocatechuate(PR-28),methylcerberic acid(PR-29),cerberic acid(PR-30),phydroxy benzoic acid(PR-31),coumaric acid(PR-32),isophthalic acid(PR-33)?4-hydroxy-3-methoxy benzoic acid(PR-34),benzo ic acid(PR-35),salicylic acid(PR-36),stearic acid(PR-37),Oleic acid(PR-38),alpha-Amyrin(PR-39),betulinic acid(PR-40),?-sitosterol(PR-41),oleanolic acid(PR-42),folia chinenosides A2(PR-43),6-Hydro-foliachin-enoside A2(PR-44),foliachinenosides Al(P R-45),6,7-dihydroxy coumarin(PR-46),glucose(PR-47),gentiobioside(PR-48),proto catechu aldehyde(PR-49),linalool(PR-50).2.The similarity of Jidanhua samples fingerprint established in this paper is above 0.9,similarity of Plumeria rubra Linn,Plumeria rubra var alba Linn and Plumeria rubra Linn cv.acutifolia is below 0.8.This method is accurate and reliable,and it can be used to control the quality of Plumeria rubra Linn cv.acutifolia.3.The results of chemical constituents isolation and principal component analys is show that 15-demethyplumieride,15-demethylplumieride P-Z-coumarate,plumieride P-Z-coumarate,kaempferol-3-O-?-D-glucopyranosyl-7-O-?-L-RhamnopyranoSide,isoquercetin and cerberic acid are ingredients of Plumeria rubra Linn cv.acutifolia,which can be Plumeria rubra Linn cv.acutifolia quality evaluation standards.The average contents of 15-demethy plumieride,15-demethylplumieride P-Z-coumarate,plumierideP-Z-coumarate,kaempferol-3-O-?-D-glucopyranosyl-7-O-?a-L-Rham-nopyra noside,isoquercetin and cerberic acid B in Plumeria rubra Linn cv.acutifolia are:5.38 mg·g-1,2.87 mg·g-1,1.40-mg·g-1,0.42 mg·g-1,1.25 mg·g-1,1.22 mg·g-1,8.52 mg·g-1,Therefore,it is suggested that in Plumeria rubra Linn cv.acutifolia,content of 15-demethylation plumieride,15-demethylplumieride P-Z-coumarate,plumieride P-Z-coum arate,kaempferol-3-O-?-D-glucopyranosyl-7-O-?-L-RhamnopyranoSide,isoquercetin and cerberic acid B,shall not beless than 4.3 mg·g-1,2.3 mg·g-1,1.1 mg·g-1,0.3 mg·g-1,1.0 mg·g-1,0.9 mg·g-1,6.8 mg·g-1.In Plumeria rubra Linn cv.acutifolia,average content of 15-demethylation plumieride,15-demethylplumierideP-Z-coumarate,plumierideP-Z-coumarate,kaempferol-3-O-?-D-glucopyranosyl-7-O-a-L-RhamnopyranoSide,isoquercetin and cerberic acid B are higher than that in Plumeria rubra Linn and Plumeria rubra var alba Linn.4.The result shows that there are no steroidal cardiac glycosides compounds in the flowers and leaves of Jidanhua.Conclusion:1.Systematical study of chemical composition shows that iridoids,triterpenes,flavonoids and phenolic acids are the main chemical components of Jidanhua.5 new compounds and 15 known compounds were isolated from Plumeria rubra Linn cv.acutifolia for the first time,which enriches research of material base of Jidanhua.2.The experiment under the same chromatographic conditions,both established Jidanhua fingerprint and simultaneous determination of the contents of 18 components.This method is accurate and reliable,and it can be used to control thequality of Jidanhua.3.Although Jidanhua belongs to Apocynaceae,it has no steroidal cardiac glycolside components.