| Among all of disease threatened human’s life, malignant tumor is one of chief culprits. Some cancer patients are also threatened by the tumor metastasis after the preliminary treatment. Once tumor metastasis occurs, most of the patients will sink into the dilemma of this incurable disease. In this paper, the coumarin in Murraya exotica (L.) was extracted to prevent tumor metastasis. In addition, so we also conducted in-depth study of chromatographic methods of polyamide-amine in the thesis.In chapter 1, the definition, process, mechanisms and various influencing factors of tumor metastasis were introduced firstly. The research progress of the traditional herb Murraya exotica (L.) and some advantages as a metastasis drug were introduced. At the end, the macromolecular material and its application as drug carrier was summarized. Based on this, we proposed the significance of this topic and the main content of this thesis.In chapter 2, the screening and isolation of the active component in Murraya exotica (L.) and the intervention mechanism of tumor cells in the vascular endothelial adhesion were researched. The active component in Murraya exotica (L.) was extract by using the high temperature reflow and purified by liquid-liquid extraction method. The purified product was tested and analyzed by UV spectrum, fluorescence spectrum and high performance liquid chromatography. The possible structures of coumarins were inferred by mass spectrometry and nuclear magnetic resonance (NMR). On the migration ability of HT29 colon tumor cells, Transwell experimental results showed that when the initial concentration of roots and leaves extractions was both 50 p.g/mL the number of HT29 migrating cells of leaves and roots extract group are only 55.9± 18.4% and 15.1 ± 11.4% of blank control group after 24 h, and the effect of root extract significantly better than that of leaf extract. However, leaf and root extract has no effect on the growth of HT29 basic under the same concentration and action time, and relative survival rate of HT29 was 96.2±10.9% and 88.1±16.0% respectively. The key efficacy components was preliminarily locked by comparing the composition and efficacy of the root and leaf extract, and then the semipreparative chromatography method was used to isolate monomer, and determine its structure by the NMR spectroscopy correspondingly, so it can provide powerful basis for the subsequent new drug development research.In chapter 3, a sensitive HPLC assay for quantitative determination of polyam-idoamine dendrimer which is strong polarity was developed. The fluorescein isothiocyanate (FITC) derivation method can improve its hydrophobicity, and the retention on the reversed-phase column also be enhanced accordingly to achieve good separation effect. Although G3-PAMAM-NH2 can lesd to the fluorescence quenching of the FITC, but it is advantageous to detect the G3-PAMAM-NH2 with 455 nm wavelength by ultraviolet-visible absorption spectra detector. The linear range was 10-80μg/mL, and the calibration curve was obtained:A=131.6C/(μg/mL)-295.6 (R2 =0.9901) with the detection limit as 2.7μg/mL. The derivative products were analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS) to further clarified the combination rule of FITC and G3-PAMAM-NH2, the binding ratio of FITC and G3-PAMAM-NH2 was 8:1. |
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