ABSTRACT
OBJECTIVE To optimize the preparation technology of ethanol extracts from Centipeda minima， and investigate the anti-inflammatory activities of different extraction sites. METHODS Single factor test and response surface methodology were adopted to investigate the effects of ethanol volume fraction， extraction time， solid-liquid ratio and extraction times on the heating reflux extraction technology of total triterpenoids ethanol extract using the extraction rate of total triterpenoids of C. minima as indexes， optimize the extraction technology and carry out validation. Using dexamethasone as positive control drug， the effects of different extraction sites of C. minima （petroleum ether part， ethyl acetate part， n-butanol part， water part） on nitric oxide （NO） production in mononuclear macrophage RAW 264.7 cells of mice induced by lipopolysaccharide （LPS） were compared； the half inhibitory concentration （IC50） was calculated. RESULTS The optimal extraction technology of total triterpenoids ethanol extracts of C. minima was as follows： ethanol volume fraction of 70%， solid-liquid ratio of 1∶40 （g/mL）， extraction time of 2.0 h and extraction times of 3 times. The 3 times of validation tests showed that average extraction rate of total triterpenoids of C. minima was 1.134%， relative error of which with the predicted value was 0.02%. The petroleum ether part and ethyl acetate part of C. minima could inhibit the generation of NO in RAW 264.7 cells induced by lipopolysaccharide to different degrees. IC50 values of NO production were 2.44 μg/mL and 2.22 μg/mL， respectively， and both of them were lower than those of positive control drug dexamethasone （7.65 μg/mL）. CONCLUSIONS The optimized preparation process of ethanol extracts from C. minima is stability and feasibility. The petroleum ether part and ethyl acetate part have obvious anti-inflammatory effects.
ABSTRACT
Objective: To study triterpenes and their anti-inflammatory activity from the ethanol extract of Centipeda minima. Methods: The compounds were isolated and purified by silica gel, Sephadex LH-20, MCI, ODS and RP-HPLC gel column chromatography, and their structures were elucidated by NMR and MS spectroscopic techniques. The inhibitory activity of the compound on the release of inflammatory mediators NO from mouse macrophages (RAW264.7) induced by lipopolysaccharide (LPS) was determined by Griess method, and then the anti-inflammatory activity of the compounds was evaluated Results: A total of 17 compounds were isolated and identified as: 20-oxo-30-nortaraxastan-3β-yl acetate (1), 3β-acetoxytaraxaster-20-en-30-al (2), 3β-hydroxytaraxaster-20-en-30-al (3), taraxasterol (4), arnidiol (5), 3β,21β-dihydroxy-20(30)-en-taraxastane (6), faradiol (7), pseudotaraxasteryl acetate (8), taraxast-20-ene-3β,30-diol (9), 18α-olean-12-ene-3,11-dione (10), maniladiol (11), 3β- hydroxyolean-12-en-11-one (12), coflodiol (13), lupeol (14), 3β,16β-dihydroxylup-20(29)-ene (15), 16β-hydroxylupa-20(29)- en-3-one (16) and garcinielliptone Q (17). Compounds 2, 5-6, 8-10, 12-13, 15-17 displayed moderate inhibitory activity on theoverproduction of NO in LPS-activated RAW 264.7 mouse macrophage cell lines, IC50 values ranging from 11.9 to 27.1 μmol/L. Conclusion: Compound 1 is a new natural product, and its 1H-NMR and 13C-NMR data was first completely assigned on the basis of 1D and 2D NMR spectroscopic evidence. Compounds 2, 3, 6, 8-10, 12, 13, 15 and 16 are isolated from C. minima for the first time. This work provided theoretical basis for clinical application of C. minima
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Objective: To develop an HPLC-DAD method for the simultaneous determination of five active flavonoids (quercetin, kaempferol, apigenin, 3-methoxyl-quercetin, nobiletin) in Centipeda minima (L.) A.Br.et Aschers.Methods: The chromatographic separation was performed on a Diamonsil C18 column (200 mm×4.6 mm,5 μm) with the mobile phase of 0.1% phosphoric acid-acetonitrile with gradient elution at the flow rate of 0.8 ml·min-1.The detection wavelength was set at 360nm,and the column temperature was maintained at 30 ℃.Results: Quercetin, kaempferol, 3-methoxyl-quercetin, apigenin and nobiletin was linear within the range of 0.002 3-0.093 0 μg·μl-1(r=0.999 5) , 0.002 2-0.087 0 μg·μl-1(r=0.999 6),0.002 0-0.079 0 μg·μl-1(r=0.999 8), 0.000 9-0.037 0 μg·μl-1(r=0.999 8) and 0.000 8-0.031 0 μg·μl-1 (r=0.999 9), respectively.The average recovery was 97.66%(RSD=1.17%), 98.33%(RSD=1.16%), 98.63%(RSD=1.10%), 98.40%(RSD=1.52%) and 98.10%(RSD=1.36%)(n=6) , respectively.Conclusion: The method is convenient, accurate and reproducible, which can be used for the quality control of Centipeda minima (L.) A.Br.et Aschers.
ABSTRACT
The active components from Centipeda minima were extracted by water or ethanol, and identified by FTIR spectroscopy and UV-visible spectrophotometer. The molluscicidal effect of aqueous extract and ethanol extract from Centipeda minima against Oncomelania hupensis was determined as referring to the WHO guidelines for laboratory molluscicidal test. Treated with over 2.0 g/L aqueous extract and ethanol extract for five days, the mortality of O. hupensis was up to 100%, and their LC50 for snails was 0.50 g/L and 0.62 g/L, respectively. The molluscicidal activity of aqueous extract was higher than that of ethanol extract. The main components of aqueous extract and ethanol extract were sesquiterpens lactones and sterols.