Studies On Quality Control Of Gansu Granules

Penthorum chinense Pursh belongs to genus Penthorum, Saxifragaceae family. Thechemical investigation on P. chinense revealed the occurrence of flavonoids andpolyphenols, which were considered as possessing multiple biological activities. Gansu, aChinese prepared medicine, was derived from the extracts of P. chinense and has been usedin clinics as a remedy for hepatoprotection and anti-fibrosis. To explore the therapeuticmaterial basis and quality evaluation of Gansu Granules, systemic researches about GansuGranule have been performed in this current study.1. Study on the chemical composition of Gansu GranulesOur research group previously focused on the isolation of EtOAc and part ofn-butanol fractions of Gansu Granule extract. In current study, the isolation andpurification of the chemical constituents from the petroleum ether fraction and the B-3/B-4part of n-butanol fraction from Gansu Granule extract were carried out silica gel, MCI gel,ODS C18and Sephadex gel column chromatography. Thirty-two compounds were isolatedfrom these two parts and their structures were identified based on physical, chemicalmethods and spectroscopic analysis (UV, IR, MS,1D-NMR and2D-NMR), includingtwelve phenolic acids: PC-05(gallic acid)，PC-06(methyl gallate), PC-07(ellagic acid),PC-08(ethyl brevifolincarboxylate), PC-09(brevifolin-carboxylic acid), PC-10(methylbrevifolincarboxylate), PC-11(9,9’-O-Diferuloyl-(-)secoisolariciresinol), PC-12(2,6-dihydroxyacetophenone-4-O-β-D-glucoside), PC-13(pinocembrin-7-O-[4",6"-(S)-hexahydroxydiphenoyl]-β-D-glucose), PC-14(pinocembrin-7-O-[-3"-O-galloyl-4",6"-(S)-hexahydroxydiphenoyl]-β-D-glucose)，PC-15(thonningianin A)，PC-16(thonningianinB); sixteen flavonoids: PC-17(apigenin), PC-18(kaempferol), PC-19(pinocembrin-7-O-β-D-glucopyranoside)， PC-20(quercitrin), PC-21(quercetin-3-O--L-arabinoside),PC-22(kaempferol-3-O--L-rhamnopyranoside), PC-23(2’,4’,6’-trihydroy-dihydrochalcone-4’-β-D-glucoside)， PC-24(2’,6’-dihydroydihydrochalcone-4’-O-[3"-O-galloyl]-β-D-glucoside)， PC-25(qercetin), PC-26(alpinetin-7-O-β-D-glucopyranoside), PC-27(naringenin), PC-28(chrysin), PC-29(isoqueritrin), PC-30(hispidulin7-(6-E-p-coumaroyl-β-D-glucopyranoside))， PC-31(rutin), PC-32(apigenin-7-O-β-D-apiofuranosyl (1â†'2)-β-D-glucopyranoside); and four other compounds:PC-01(β-sitosterol), PC-02(β-daucosterol), PC-03(glyoeryl nonopalmitate), PC-04(linoleic acid). 2. Anti-liver fibrosis study on Gansu GranulesThe anti-fibrosis activities of Gansu extract, extracted fraction, and isolates weretested on HSC-T6cells induced by TGF-β1. The result showed that the EtOAc andn-BuOH possess a remarkable anti-fibrosis activity, while phenols also have adistinguished anti-fibrosis activity among the isolates.3. Studies on fingerprint and quality evaluation of Gansu GranulesIn our research, the chemical fingerprint and main content evaluation of23sampleswere carried out by a method of high performance liquid chromatography DAD UVdetector (HPLC-DAD). Meanwhile12common peaks have been identified based onLC-MS, including gallic acid (Peak1), brevifolin-carboxylic acid (Peak4),2,6-dihydroxyacetophenone-4-O-β-D-glucoside (Peak5),(-)-epicatechin-3-O-gallate(Peak6), kaempferol-3-O--L-arabinoside (Peak7), quercitrin(Peak8), qercetin(Peak11),pinocembrin-7-O-β-D-glucopyranoside(Peak12), pinocembrin-7-O-[4",6"-(S)-hexa-hydroxdiphenoyl]-β-D-glucose (Peak13), thonningian A (Peak14),thonningian B(Peak15), PGHG(Peak17). Also, nine in the twelve identified commonpeaks have been content evaluated, including1，4，5，8，11，12，13，15，17, whichprovide the optimal harvest of P.chinense.In addition, this subject has defined thirteen characteristic common peaks on accountof the HPLC-DAD fingerprint of GANSU GRANULES. Through HPLC-MS comparisonof above thirty-two constituents with the common peaks, elven common peaks were finallyidentified to be: gallic acid(Peak1); brevifolin-carboxylic acid(Peak2);2,6-dihydroxyacetophenone-4-O-β-D-glucoside(Peak3); quercitrin (Peak5); qercetin(Peak6); pinocembrin-7-O-β-D-glucopyranoside(Peak7);2’,4’,6’-tri-hydroydihydrochalcone-4’-O-β-D-glucoside (Peak8);2’,6’-dihydroydihydro chalcone-4’-O-[3"-O-galloyl]-β-D-glucoside (Peak9); pinocembrin-7-O-[4",6"-(S)-hexa-hydroxydiphenoyl]-β-D-glucose (Peak10);thonningian A (Peak12), PGHG (Peak13). Atthe same time, this reseach has established the methods for the content determination of1，2，3，5，7，8，10，12，13in GANSU GRANULES.