Study On Chemical Constituents Of Pulsatilla Cernua(Thunb.)bercht.ex J.presl

As traditional Chinese medicine,Pulsatilla cernua（Thunb.）Bercht.ex J.Presl has the effects of clearing heat,cooling blood,and detoxifying.Modern pharmaceutical research has shown that the main components of P.cernua are lupinane and oleanane triterpene saponins,and have anti-insect,antibacterial and anti-tumor pharmacological activities.With the aim of discovering the diversity of chemical components of P.cernua and enriching the material basis for pharmacological research,this research topic extracts and isolates the compounds from P.cernua from Dandong City,Liaoning Province.P.cernua medicinal material is extracted with 70%Et OH-H₂O solvent and concentrated to obtain a crude extract.First,the crude extract is passed through a D101type macroporous resin column,eluted according to a gradient from large to small polarity,and analyzed by thin-layer chromatography（TLC）and high-performance liquid chromatography（HPLC）to obtain different crude fractions.Separate different fractions according to the characteristics of molecular weight difference,different solubility,similar compatibility,etc.,through HP-20 macroporous adsorption resin column,Sephadex LH-20 gel column,200～300 mesh silica gel column,ODS open chromatographic column and semi-preparative HPLC and other separation means,step by step separation and purification,to obtain the monomer compound.Finally,according to the physical and chemical properties of the monomer compound and ¹H-NMR,¹³C-NMR and MS data,the data attribution and structure identification are carried out.30 compounds were isolated from the 70%Et OH-H₂O extract of P.cernua,and the structures of 22 compounds were determined through structural identification.They were identified as:（+）-pinoresinol（1）、matairesinol（2）、4-ethoxycinnamic acid（3）、p-hydroxy ethyl cinnamate（4）、3-（4’-methoxyphenyl）-2（E）-propenoic acid（5）、methyl 4-hydroxycinnamate（6）、radicol（7）、cryptomeridiol（8）、fraxinellone（9）、diolmycin B2（10）、hederagonic acid（11）、hederagenin（12）、oleanolic acid（13）、3-O-α-L-arabinopyranosyl-oleanolic acid（14）、hederagenin 3-O-α-L-arabinopyranoside（15）、3-O-[α-L-rhamnopyranosyl-（1→2）-α-L-arabinopyranosyl]oleanolic acid（16）、hederasaponin B（17）、kizutasaponin K₁₂（18）、patrinia saponin H3（19）、hederacholichiside F（20）、cernuoside A（21）、cernuoside B（22）.Compounds 1～2 are lignans,3～6 are phenylpropionic acids,7～8 are sesquiterpene,9 is limonoid and 11～22 are triterpene acids and triterpene saponins.Among them,compounds 3～10 were the first compounds isolated from the genus Pulsatilla.On the basis of the chemical composition research,the filter paper method was used to test the antibacterial activity of compounds 11～22 in vitro against Escherichia coli and Staphylococcus aureus.The preliminary screening did not yield positive results under the conditions of 30μL and 50μL at the concentration of 1 mg/m L.