Study On The Non-alkaloid Components Of Buxus Microphylla Sieb. Et Zucc. Var. Sinica Rehd. Et Wils. Stems And Branches

Buxus microphylla Sieb.et Zucc.var.sinica Rehd.et Wils.,belongs to Buxus genus,Microphylla family.Its dry stems and branches are used as the medicinal parts.The flavor of it is considered to be bitter but non-toxic,and the nature of it is mild.It possesses the functions of dispelling wind,eliminating dampness,regulating qi and relieving pain.It is commonly used in the folk to treat rheumatic toothache,chest and abdominal inflamation and traumatic injury.It can also be used to treat malaria,syphilis,dermatitis and other diseases.It was reported that B.microphylla mainly contained alkaloids,flavonoids,phenolic acids,and other chemical constituents.The research on the medicine mainly focuses on the pharmacological effects of its main alkaloid,cyclovirobuxine D and its derivatives in protecting cardiovascular and cerebrovascular diseases,while the related studies on flavonoids and phenolic acids are rarely.In order to enrich its pharmacodynamic substances,chromatographic technologies including silica gel,ODS,Sephadex LH-20 column chromatorgraphies and preparative HPLC chromatorgraphy(pHPLC),and spectrometric methods such as 1H,13C,2D NMR,MS,UV,IR,[α]D25 and CD,along with chemical reactions were combined to carry out the systematical chemical constituent study of the non alkaloid parts of B.microphylla var.sinica.As results,64 compounds including 28 new compounds,buxusflavanol A(1),buxusaurones A(2),B(3),buxusauronoside C(4),buxusflavanonoside A(5),buxustannins A-G(6-12),H1-H4(13-16),I(17),J(18),K1(19),K2(20),buxusphenoside A(21),buxusphenols B1(22),B2(23),C-G(24-28)and 36 known ones,(±)-liquiritigenin(29),(±)-7,3’,4’-trihydroxyflavanone(30),(2R/2S)-2-[3-(β-D-glucopyranosyloxy)-4-hydroxyphenyl]-2,3-dihydro-7-hydroxy-4H-1-benz opyran-4-one(31),(1)-eriodictyol(32),(2R*,3R*)-garbanzol(33),(2R*,3R*)-3,7,3’,4’-tetrahydroxyflavanone(34),(2R*,3R*)-dihydroquercetin(35),(2R*,3S*)-3,7,3’,4’-tetrahydroxyflavanone(36),(±)-α,3,4,2’,4’-pentahydroxydihydrochalcone(37),butein(38),7,4’-dihydroxyflavonol(39),fisetin(40),2-(3,4-dihydroxyphenyl)-7-(β-D-glucopyranosyloxy)-3-hydroxy-4H-1-benzopyran-4-one(41),(±)-fustin-3-O-gallate(42),(±)-2,6-dihydroxy-2-[(4-hydroxyphenyl)methyl]-3(2H)-benzofuranone(43),(±)2-benzyl-2,6,3’,4’-tetrahydroxycoumaran-3-one(44),Z-sulfuretin(45),Z-sulfurein(46),cotinignan A(47),1,3-benzenediol(48),p-hydroxybenzoic acid(49),salicin(50),protocatechuic aldehyde(51),3,4-dihydroxybenzoic acid(52),protocatechuic acid methyl ester(53),2,4-hydroxysalicylic acid(54),2-(β-D-glucopyranosyloxy)-4-hydroxybenzoic acid(55),gallic acid(56),di-O-methylcrenatin(57),3,4-dihydroxyphenacyl alcohol(58),α,3,4-trihydroxy-benzeneacetic acid methyl ester(59),(±)-methyl 2-methoxy-2-(3,4-dihydroxyphenyl)acetate(60),methyl 3,4-dihydroxyphenylglyoxylate(61),(8S,8’S,7’R)-(-)-lyoniresinol(62),(2R*,3S*)-2,3-trans-6-[(2R*,3R*,4R*)-2,3-cis-3,4-trans-3’,4’,7-trimethoxyjlavan-4-y1]-8-[(2 R*,3R*,4R*)-2,3-cis-3,4-trans-3’,4’,7-trimethoxyjlavan-4-yl]-3’,4’,5,7-tetramethoxyflavan(63),(2S*,3R*)-2,3-trans-6-[(2S*,3S*,4S*)-2,3-cis-3,4-trans-3’,4’,7-trimethoxyjlavan-4-y1]-8-[(2R*,3R*,4R*)-2,3-cis-3,4-trans-3’,4’,7-trimethoxyjlavan-4-yl]-3’,4’,5,7-tetramethoxyflavan(64)were gained.Among the 36 known isolates,31-39,41-47,50-55,and 57-64 were isolated firstly from Buxus genus;48,49 and 56 were isolated for the plant for the first time.Meanwhile,the 1H,13C NMR data of compounds 41,48,54,59,63,and 64 were reported here firstly.It was found that the non-alkaloids of B.microphylla var.sinica were flavonoids,tannins,phenolic acids,and lignans mainly according to the systematic separation and structural identification of non-alkaloids in it.This experiment would enrich the research work on the chemical constituents of B.microphylla var.sinica,and laid a material foundation for its quality control,pharmacological and biological research.The structural characteristics of flavonoids and tannins were also summarized,which would provide a reference for the rapid structural identification of these components.Since the phenolic components were rich in B.microphylla var.sinica,which will play an important role in the overall efficacy of the medicine,and it should be considered in its clinical medication process.