| Flavonoids are the kind important natural products. In this study, we first established a reliable method to search a suitable two-phase solvent system. An efficient new model for extraction by ultrasonic-assisted, purification HSCCC, and analysis and identification by HPLC-DAD and HPLC-ESI-MSn were constructed. This model could be used for preparation of flavonoids, construction of chromatography fingerprint and quality control of Chinese herbal medicine. The effects of flavonoids on human hepatocarcinoma cell were also investigated. Our results will accelerate the exploration and application of flavonoids for cancer prevention and anticancer medication.The main research results are as follows,Firstly, optimization of extraction of ultrasonic-assisted by RSM of anthocyanin from Lonicera caerulea L.An efficient extraction was developed through response surface methodology to maximization of the yield of anthocyanin from the berry of Lonicera caerulea L.(L. caerulea). Based on single factor experiments, the effects on extraction conditions of extraction concentration, extraction time and solvent to raw material ratio were determined and optimized by RSM. The results showed that the optimum extraction conditions were as follows:100%methanol concentration,60min extraction time and10:1(v/w) solvent to raw material ratio. Under such conditions, the anthocyanin yield was2.714mg/g.Secondly, isolation and purification of anthocyanins from L. caerulea by HSCCC. HSCCC was successfully used for the isolation and purification of anthocyanins from L. caerulea with a biphasic solvent system composed methyl tert-buty1ether-n-butanol-acetonitrile-water-trifluoroacetic acid (2:2:1:5:0.01, v/v).2.14mg of Cyanidin3,5-O-diglucoside,3.38mg of cyanidin3-O-rutinoside and12.8mg of cyanidin3-O-glucoside were obtained from40mg clean-up extraction by CCC. And the purity of these compounds was92.4%,95.8%and97.6%, respectively, as identified by high-performance liquid chromatography coupled with photodiode array detection (HPLC-DAD) and HPLC electrospray ionization mass spectrometry (HPLC-ESI-MSn). By using this metholds, three bioactive anthocyanins were isolated and identified from material.Thirdly, studies on the chromatography fingerprint of Gentianopsis barbata Ma by HPLC-ESI/MSn Xanthones of Gentianopsis barbata Ma were determined using high-performance liquid chromatography coupled with photodiode array detection (HPLC-DAD) and HPLC electrospray ionization mass spectrometry (HPLC-ESI/MSn). Qualitative and quantitative analysis method for the xanthones composition in different parts of G. barbata from Daxinanling area has been built up for the first time. Totally11kinds of xanthones were detected, of which9kinds were identified in G. barbata for the first time. The contents of main compounds of different parts were evaluated:petal> leaf> lateral branch> stem> root, and the petal contained significantly more xanthones. Based on xanthones constitutions, the control basis of the qualities of medicinal materials were obtained.Finally, anticancer activity of cyanidin-3-O-glucosideThe effects of anthocyanin on human hepatocarcinoma cell using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as well as flow cytometric analysis were also observed. Our study show that, the dominant anthocyanin in L. caerulea, cyanidin-3-O-glucoside was demonstrated strong cytotoxic response of human hepatocarcinoma SMMC-7721cells, and inducing cell apoptotic by flow cytometric analysis。... |
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