The Quality Of Preparation And Inula Inula Chemical Markers Studies

Objective: To control the quality of Flos Inulae and its preparations, the chemical constituents were extracted from Flos Inulae, identified and assayed with analytical means. On the basis of the data, they can be used as Flos Inulae reference standards.Method:1 Extraction of chemical compoundsThe Flos Inulae was refluxed for three times, 1 hour per time. The ethanol extract was suspended in water and partitioned with petroleum ether, ethyl acetate, chloroform and n-butanol respectively. 2 Isolation of chemical compoundsThe chloroform extract was subjected to column chromatography on silica gel G using an n-hexane/acetone gradient system(1∶0→0∶1) followed by ethanol to give 5 fractions. Fraction 2 was submitted to repeat column chromatography over silica gel G using petroleum ether: ethyl acetate: ethanol (95%) to yield crude crystals of Ⅰ, which were recrystallizated from ethanol to give the colourless needle crystals; Compound Ⅱ was obtained from fraction 3.3 Determination of the purities of the compoundsⅠ andⅡThe melting point and UV maximum absorption wavelength were detected, the purities of the compounds were determinated by RP-HPLC.4 Identification of the chemical structures of compound Ⅰ andⅡThe chemical structures of the two compounds were identified by UV,IR,1HNMR,13CNMR, 1H-1H COSY and MS . Result:1 Extraction of chemical compoundsThe Flos Inulae was extracted with ethanol by refluence . After filtration, the extract was concentrated and partitioned with different polar solvents , which were removed under pressure to yield the dark-green dopes.2 Isolation of chemical compoundsColourless needle crystals(Ⅰ)0.8g and colourless power (Ⅱ)96mg were obtained from chloroform extract by repeating chromatography on the silica gel G column.3 Determination of the compoundsⅠ andⅡpuritiesThe melting point of compound Ⅰwas 123.9~125.2℃, compound Ⅱ169.0~171.8℃. UV Maximum absorption wavelength was detected as 210nm. Purities of compound Ⅰand Ⅱwere respectively 99.82% and 98.35%.4 Identification of the chemical structures of compound Ⅰ andⅡBy comparing the UV,IR,1HNMR,13CNMR, 1H-1HCOSY and MS spectra data with those of reference materials, the result showed compound Ⅰas 1-O-Acetylbritannilactone and compound Ⅱas Britannilactone which are the known compounds.Conclusions:The two compounds were identified as 1-O-Acetylbritannilactone and Britannilactone. They are seperated from Flos Inulae the first time in China. Their purities conformed to the demand of , the purity of Chinese traditional medicine reference substance is above 98%. Therfore, the two compounds can serve as the index constituents in the quality control of Flos Inulae.Key words: Flos Inulae,column chromatography,purity,1-O-Acetylbritannilactone,Britannilactone...