Studies On The Chemical Constituents And Quality Control Standard Of Rubus Chingii Hu

Rubus chingii Hu,belonging to the family Rosaceae,is distributed widely in Jiangxi,Zhejiang,Fujian and Guangxi Provinces.Fruits are used for treating enuresis with renal asthenia,frequent urination,premature eiaculation,impotence.Rubus chingii Hu is one of the main ingredients of Shenbao mixture,and as the most expensive cost single Chinese herb.Primary standards of Rubus chingii Hu are contained in the �Chinese Pharmacopoeia� 2010 edition of 359 pages.From chloroform part,ethyl acetate part and n-butyl alcohol part of the 70% ethanol extracts of Rubus chingii Hu,18 chemical compounds were isolated with modern chromatography methods,through the various spectroscopic methods identified their structure,they are: 15,18-di-O-?-D-glucopyranosyl-13(E)-ent-labda-7(8),13(14)-diene-3?,15,18�triol(1),15,18-di-O-?-D-glucopyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3?,15,18-triol(2),15-O-?-D-apiofuranosyl(1?2)?-Dglucopyranosyl-18-O-?-D-gluco-pyranosyl-13(E)-ent-labda-8(9),13(14)-diene-3?,15,18-triol(3),Astragalin(4),3-Methoxy-4-hydroxy-benzoic acid(5),4-Hydroxybenzoic acid(6),Gallic acid(7)3,4,5-Trihydroxybenzoic acid ethyl ester(8),(7S,8R)-Dihydrodehydro-diconiferyl alcohol(9),5-[3-hydroxymethyl-5-(3-hydro xypropyl)-7-Methoxyl-2,3-dihydro-benzofuran-2-yl]-2-methoxy-phenol(10),4-[3-hydroxymethyl-5-(3-hydroxy propyl)-2,3-dihydrobenzofuran-2-yl]-2-methoxyphenol(11),1-Oxo-1,2-di-hydro-isoquinoline-4-carboxylic acid(12),ellagic acid(13),Tiliroside(14),kaempferol-3-O-rutinoside(15),13(E)-ent-Labda-8(17),13-diene-3?,15,18-triol(16),13(E)-ent-Labda-8(17),13-diene-3?,15,18-triol(17),Raspberry glucoside F5(18),Compounds 1-3 were new compounds,9-12 were first isolated from this plant.In the 2010 edition of �Chinese Pharmacopoeia� one,the standard of Rubus chingii Hu only includes properties and powder identification.TLC identification of ellagic acid and tiliroside are established by using thin layer chromatography respectively;HPLC was used to determine the contents of ellagic acid and kaempferol-3-O-rutinoside in Rubus chingii Hu.The method for determination of ellagic acid is that the separation was performed in a ODS column(Diamonsil C18 column,250mm�4.6mm,5?m)with a mobile phase of acetonitrile�0.2%phosphoric acid(15:85).The flow rate was 1.0ml/min,and the detecting wavelength was at 254 nm.Results: The linear range of ellagic acid 0.01593~0.09558?g,the peak area and injection volume was a good linear relationship,the regression equation was Y = 9.0�106X-3514.2,with:R2 = 0.9993(n = 6).The average recovery of ellagic acid was 103.1%,with RSD=1.4%;The method for determination of kaempferol-3-O-rutinoside is that the separation was performed in a ODS column(Welch C18 column,250mm�4.6mm,5?m)with a mobile phase of acetonitrile�0.2%phosphoric acid(15:85).The flow rate was 1.0 ml/min,and the detecting wavelength was at 344 nm.Results: The linear range of kaempferol-3-O-rutinoside 0.0789~1.1835?g,the peak area and injection volume was a good linear relationship,the regression equation was Y = 1.0�106X-9996.0,with: R2 = 0.9999(n = 6).The average recovery of kaempferol-3-O-rutinoside was 101.9%,with RSD=0.7%.Using the established HPLC method to determine the contents of ellagic acid and kaempferol-3-O-rutinoside of 10 batches of Rubus chingii Hu,the results show that the method is simple and accurate with good reproducibility can be used as the quantitative analysis method of ellagic acid and kaempferol-3-O-rutinoside of Rubus chingii Hu;We studied on the moisture,total ash,acid insoluble ash,extract and pesticide residues of 10 batches of Rubus Chingii Hu.