Studies On The Chemical Constituents And Content Determination Of Pyrrosia Petiolosa(Christ) Ching

This paper is divided into four parts to summarize the research work on Pyrrosia petiolosa（Christ）Ching.Firstly,Chapter 1 provides a detailed review of the plant;Chapter 2 investigates the chemical composition;Chapter 3 prese nts the results of content determination of Pyrrosia petiolosa（Christ）Ching;C hapter 4 summarizes the experiments.Pyrrosia lingua mainly includes Pyrrosia lingua（Thunb.）Farwell,Pyrrosi a sheareri（Bak.）Ching and Pyrrosia petiolosa（Christ）Ching,and the whole plant is commonly used by the people as medicine.In folklore,it is mainly us ed to treat major gonorrhea and is a traditional Chinese medicinal plant.It is mainly used in folk medicine for the treatment of ailments,coughs with lung f ever,hot gonorrhea,stone gonorrhea,blood gonorrhea,and lacrimation and astr ingency.We took 200 kg of the herbs of Pyrrosia crushed by reflux extraction wit h 70%ethanol,and the ethyl acetate part of the extract was separated and pur ified by reversed-phase chromatography column(C₁₈),thin-layer chromatography,normal-phase column chromatography silica gel（G100～200 mesh）,dextran gel Sephadex LH-20,and HPLC.At the same time,using spectroscopic and spect ral analysis methods,20 compounds were isolated and identified,In order,Met hyl p-hydroxyphenylpropionate（1）,6-O-[（9Z,12Z,15Z）-octadeca-9,12,15-trienoyl]-β-D-fructofuranosyl-α-D-glucopyranoside（2）,6′-O-linoleylsucrose（3）,（2S）-2,3-O-dioc tadeca-9Z,12Z,15Z-trienoylglyceryl-6’-O-（α-D-galactopyranosyl）-β-D-galactopyranos ide（4）,kaempferol-7-O-β-D-apioside（5）,secoisolariciresinol（6）,（+）-isolariciresinol（7）,Naringenin 7-O-β-D-glucuronide（8）,Eriodictyol-7-O-β-D-glucuronide（9）,4,5-dicaffeoylquinic acid methyl ester（10）,3,4-di-O-caffeoylquinic acid（11）,1,2-O-dicaffeoyl-cychopenta-3-ol（12）,gallic acid（13）,vanillin acid-4-o-β-D-glucoside（14）,neochlorogenic acid methyl ester（15）,chlorogenic acid（16）,kaempferol（17）,kaempferol-3-O-α-L-rhamnopyranoside（18）,kaempferol-7-O-α-L-arabinofuranosi de（19）,and dehydroconiferyl alcohol（20）.Among them,compounds 1-8,10,12-15 were isolated for the first time from Pyrrosia petiolosa（Christ）Ching.To establish an HPLC method for the simultaneous determination of five chemical components from Pyrrosia petiolosa（Christ）Ching,including Chloroge nic acid,Eriodictyol-7-O-β-D-glucuronide,4,5-dicaffeoylquinic acid methyl est er,3,4-di-O-caffeoylquinic acid,and 1,2-O-dicaffeoyl-cychopenta-3-ol,in Pyrro sia petiolosa（Christ）Ching plants.Methods:THE SHIMADZU C₁₈column（250×4.6mm,5μm）was used;acetonitrile（B）-0.2%phosphoric acid aqueous s olution（C）was used as the mobile phase and gradient sequential elution;gradi ent elution at a flow rate of 1.0 m L.min^-1.328 nm is used as the detection w avelength;and the column temperature was at 30℃.Results:The injection vol umes of Chlorogenic acid,Eriodictyol-7-O-β-D-glucuronide,4,5-dicaffeoylquinic acid methyl ester,3,4-di-O-caffeoylquinic acid,and 1,2-O-dicaffeoyl-cychopent a-3-ol were in the ranges of 0.1066～2.6655μg（R²=1.0000）,0.1602～4.0060μg（R²=1.0000）,0.0065～0.1625μg（R²=1.0000）,0.0069～0.1715μg（R²=1.0000）and0.0096～0.2390μg（R²=1.0000）showed good linearity with peak area;good preci sion（RSD<2%）,stability（RSD<2%）and reproducibility（RSD<2%）;The averag e recoveries（n=3）were 101.52%（RSD=0.21%）,100.41%（RSD=0.51%）,99.73%（RSD=0.57%）,97.30%（RSD=0.89%）,101.17%（RSD=1.05%）,respectively;the ranges of five chemical compositions in the four batches were 2.1873～7.4048,7.6936～10.7895,0.3553～0.5113,0.3532～0.5121 and 0.4368～0.7627 mg.g-1,respectively.