Studies On The Basic Constituents Of Anti-depression Effects Of Xiao Chai Hu Decoction And The Chemical Constituents Of Pinellia Ternata

Xiao Chai Hu Decoction was first listed in "Treatise on Febrile Diseases" by Zhong-Jing Zhang of the Han Dynasty and it has also been used to treat Shao Yang symptom.Xiao Chai Hu Decoction is a well-known Traditional Chinese Medicine Formula used in Asia and is consists of seven crude drugs:Radix Bupleuri,Panax ginseng,Scutellaria baicalensis,honey fried Glycyrrhizae Radix,Rhizoma Pinelliae preparata,Ziziphus jujuba and Zingiber officinale.In recent years,it has been used to treat patients with chronic liver disease and has also been demonstrated to have anti-depressants effects.In these settings,Xiao Chai Hu Decoction has attracted the attention of practitioners of Western medicine.However,characterization methods used in the past lacked sensitivity and the absence of a reference substance has hindered progress;Thus,the nature of the active constituents of XCHD remains unclear.Bioactivity-guided fractionation and LC-ESI-Q-TOF-MS/MS were applied for separation and identification of the basic biological active components in XCHD.D101 macroporous resin was used to enrich and separate the material base into four fractions,XCHD-1,XCHD-2,XCHD-3,and XCHD-4;Each fraction was then evaluated for their antidepressant effect using depression-related parameters.By using various chromatographic techniques,40 compounds have isolated from XCHD-3.On the basis of analysis of UV,NMR and MS data and measuring of physic-chemical characters,their structures were elucidated as follows:isoliquiritin(1),neoisoliquiritin(2),5,7,2',6'-tetrahydroxy-dihydroflavonol(3),chrysin-8-C-?-D-glucopyranoside(4),liquiritin(5),5,2',6-trihydroxy-7,8-dimethoxyflavone 2'-O-?-D-glucopyranoside(6)?viscidulin ?(7),baicalin(8),luteolin-7-O-gentibioside(9),5,7-dihydroxy-6-methoxyflavone-7-glucuronide methyl ester(10),viscidulin ?(11),3,5,7,2'6'-pentahydroxy flavone(12),wogonoside-7-O-?-D-glucuronide methyl ester(13),wogonoside(14),glucoliquiritin apioside(15),wogonin(16)E 6,7-dimethoxy-4-hydroxy-l-naphthoic acid(17),myricetin(18),quercetin(19),martynoside(20),5,6'-dihydroxy-7,8-dimethoxy-2,-O-?-D-glucopyranosylflavone(21),ononin(22),syringing(23),3-(4-hy droxyphenyl)propionic acid methyl ester(24),apiol(25)chrysin-6-C-glucosyl-8-C-arabonoside(26),chrysin-6-C-arabinosyl-8-C-glucoside(27),liquiritigen-7-O-?-D-glucoside(28),apigenin 7,4'-O,O-diglucoside(29),3-isobutylpyrrolopiperazine-2,5-dione(30),vicenin-2(31),apigenin 6,8-di-C-arabinopyranoside(32),rutin(33),scutellarin(34),schafloside,(35),isoschafloside(36),kaempferol-3-O-?-D glucopyranoside(37),quercetin(38),kaempferol(39),vanillic acid(40).Identification of the constituents was carried out by comparing their HPLC retention times and accurate masses with those of available standards.When no standard was available,we investigated the fragmentation behaviors of reference standards,and then applied rules for the structural elucidation of their derivatives with the same basic skeleton.As a result,79 compounds including polysaccharides(inulin,sucrose,fructose/glucose,stachyose),flavonoids(liquiritigenin-7,4'-diglucoside,liquiritin-7-O-glucoside-4'-O-glucosyl-apioside,vicenin-2,schaftoside,isoviolanthin,chrysin-6-C-glucosyl-8-C-arabonoside,chrysin-6-C-arabinosyl-8-C-glucoside,liquiritin apioside,isoacteoside,liquiritin,scutellarin,acteoside,5,6'-dihydroxy-7,8-dimethoxy-2'-O-?-D-glucopyranosylflavone,chrysin-8-C-?-D-glucopyranoside,5,2',6'-trihydroxy-6,7,8-trimethoxyflavone 2'-O-glucoside,licuraside,cistanoside D,5,8-dihydroxy-6-methoxy-7-O-glu acid,isoliquiritin,cosmosiin,neoisoliquiritin,baicalin,5,6-digydroxy-7-glucuronic acid flavanone,galengin 7-O-glucuronide,5,2'-dihydroxy-6-methoxy-7-O-glu acid,oroxylinA-7-O-?-D-glu acid,chrysin-7-O-?-D-glucuronopyranoside,5,6-dihydroxy-6-methoxy-7-O-glu acid,wogonoside,5,7-dihydroxy-6-methoxyflavone-7-glucurronide methylester),saikosaponins(acetyl-saikosaponin A or D,SSA,saikosaponin SSb1 or SSb2,acetyl-saikosaponin A or D,diacetyl-Saikosaponin A or D,acetyl-Saikosaponin A or D,SSD),ginsenosides(ginsenoside Rgl,ginsenoside Rf,isomer or ginsenoside Rg2,isomer or ginsenoside Rg2,ginsenoside Rbl,isomer or ginsenoside Rg6 or F4,isomer or ginsenoside Rg6 or F4,isomer or ginsenoside Rg6 or F4,ginsenoside Re),licoricesaponins(licoricesaponin E2,22?-acetoxylglycyrrhizic acid,yungaonoside K2,licoricesaponin G2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,licoricesaponin-J2,licoricesaponin B2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,glycyrrhizin acid or licoricesaponin-H2 or licoricesaponin-K2,licoricesaponin A3)and gingerols(6-gingerol,methy[8]-gingerol,[6]-shogaol,isomer or[6]-paradol)were detected,69 of them were identified or tentatively characterized.Based on our preliminary characterization investigations,polysaccharides,gingerols,and flavonoids in XCHD may contribute to the antidepressant effect of XCHD.In conclusion,the hyphenated method of bioactivity-guided fractionation and LC-ESI-QTOFMS/MS was meaningful for the isolation and preliminary identification of the biological active components in complex matrices of TCM.The chemical constituents of Pinellia ternata is a part of the chemical constituents basic study for Xiao Chai Hu Decoction Pinellia ternata(Thunb.)Breit,a member of the family Araceae,is widely distributed in China.The air-dried and powdered tubers of P.ternata(15 kg)were extracted with 75%EtOH.This study of P.ternata tubers has resulted in the characterization of 40 compounds.On the basis of analysis of 1D,2D NMR analyses,HR-ESI-MS and CD data and measuring of physic-chemical characters,their structures were elucidated as follows:cyclolignanyingoside A(41),isolariciresinol 9-O-?-D-glucopyranoside(42),isolariciresinol(43),verticillatoside A(44),pinoresinol-4'-O-?-D-glucoside(45),medioresinol(46),Pinoresinol(47),burselignan(48),dehydrodiconiferyl alcohol-9-O-?-D-glucopyranoside(49),dehydrodiconiferylalcohol-9'-O-?-D-glucopyranoside(50),dehydrodiconiferylalcohol-4,9-di-O-?-D-glucoside(51),lariciresinol 4-O-?-D-glucopyranoside(52),(7R,8R)-1-(4-hydroxy-3-methoxyphenyl)-2-{4-[(E)-3-hydroxyl-1-propenyl]-2-methoxyphe-noxy}-1,3-propanediol(53),cinnacassoside A(54),neo-olivil(55),americanol A(56),isoamericanol A(57),forsythiayanoside B(58),matairesinoside(59),syringaresinol-4-O-?-D-glucopyrranoside(60),cyclo-Prolyl-Lecine(61),cyclo-Pro-Ile(62),cyclo-Pro-Val(63),uridine(64),adenosine threo-2S-3-O-methyl-guaiacylglycerol(65),erythro-2-R-3-O-methyl-guaiacylglycerol(66),3,4-dihydroxybenzoic acid(67),2,4-dimethoxyphenyl-1-O-?-D-glucopyranoside(68),coniferaldehyde(69),vanillic(70),4-O-?-D-glucopyranosylvanilylalcohol(71),3,4-dimethoxybenzyl-?-D-glucopyranoside(72),3,4-dimethoxybenzyl-?-D-glucopyranoside(73),p-hydroxybenzoic acid(74),4-(2-hydroxyethoxy)-3-methoxy-benzoic acid(75),sachalide 1(76),nikoenoside(77),3-methoxy-4-methylbenzoic acid(78),p-methoxy phenylpropyl aldehyde(79),trans-p-hydroxy cinamic acid(80).The cytotoxic and antioxidant of a part of these isolates were determined.However,all of the selected compounds exhibited no activity on the selected cell lines(Hep-3 B,Bcap-37 and MCF-7).In addition,the selected compound exhibited different scavenging free radical scavenging activity.In addition,the biosynthetic pathways of a part of the isolated lignanoids were summarized.Determination of ten bioactive lignanoids in Pinellia ternate and its adulterants using conversion multiple reaction monitor technology(MRM)were established.5,6 and 10 might be the potent compounds to distinguish P.ternata from adulterants(rhizoma typhonii flagelliformis and rhizoma arisaematis).