Extraction, Separation, Antitumor Activity, Structure And Effect Of Deguelin From The Roots Of Derris Trifoliate And Its Analysis

Deguelin, as one of natural rotenoids, is consisted in Derris Lour and Tephrosia Pers. In the paper, its extraction, purification, stability, chromatogram, spectrum, anti-tumour activity and structure were studied, respectively. In chapter one, a detailed review is presented in the paper of advances in the research of the structure, extraction, separation, bioactivity, nature resource tissue culture techniques supercritical extraction and molecule simulation. In chapter two, three methods were applied to extract deguelin from Derris trifoliate Lour roots. Different solvents were used to separate rotenoids, such as deguelin, rotenone and tephrosin. The weight of total extract and the relative deguelin extraction efficiency were determined. It is clearly shown that the total amount of the extracted substance assisted by using ultrasound is a little lower than the Sohxlet or vibrating extraction method. However, deguelin extraction rate by using the ultrasound-assisted extraction is the highest. Furthermore, with respect of rotenoids separation, the solvent mixture of petroleum ether:chloroform:acetone at the ratio of 6:1:1 is found to be the most effective. In chapter three, super critical CO₂. extraction was used to extract deguelin from the roots of Derris trifoliata in different sample size, time, modifiers, modifiers quantity, modifiers adding modes, press, temperature and flux of CO₂. In order to extract deguelin from 10g roots of Derris trifoliate, the rational conditions are sample size of 30 mesh, time of extraction before dynamic extracting 30min, adding 9ml methanol as modifier into extraction tube, press 30MPa, temperature 55℃and flux of CO₂ 25kg/h. In the condition, the maximum deguelin is extracted, which is 1.6 sizes higher than that is extracted by chloroform. In chapter four, HPLC, UV, MS, FT-IR and 1H NMR were used to analyze deguelin. It is clearly that the mass of deguelin is 394 and the maximum spectrum value of deguelin is 271.6. Chapter five studies the anti-tumour research of deguelin. And the results show deguelin is a wanderful substance to restrain the propagation of K562 and A549 cells, but take a little effect on BEL-7402 cells. Chapter six shows that the solubility of tephrosin, duguelin and dehydrodeguelin in variety of solvents was compared, and the effect of light, heat, microwave and pH on tephrosin, deguelin and dehydrodeguelin was determined by HPLC. Results show that tephrosin, deguelin and dehydrodeguelin are well dissolved in methanol, ethanol, glycol, acetonitrile, acetone, chloroform, carbon tetrachloride and ethyl acetate. Deguelin is decompounded by light while tephrosin and dehydrodeguelin are relatively steady. Tephrosin, deguelin and dehydrodeguelin are unchanged by heat and microwave. As pH is lower than 7, tephrosin duguelin and dehydrodeguelin are stability. However, the structure of duguelin is changed distinctly when pH is higher than 7. In chapter seven, HyperChem was used to study the energy which is absorbing or releasing in variety of reactions. This chapter may be usefully to improve the synthesis deguelin in the future study. In chapter eight, molecular mechanics geometry optimization was used to optimize the structure of deguelin. And the optimized structure was simulated by Molecular Dynamics and CNDO/2 methods of Semi-empirical. The simulation pays attention to the change of energy in different conditions and the distributing of charge in deguelin. As the results are shown, deguelin can form H-bonds with water in super-low temperature but not in room temperature. The C=O of deguelin is easy to be replaced by nucleophilic reactivity. In chapter nine, Absorption isotherm and absorption dynamic curve of Derris trifoliate Lour roots chloroform extract is studied. The results show that the maximal q ofsilica gel is 310.29, K_d is 44.19 and adsorption velocity value is 1.9972 for concentration1.0mg/mi. As for deguelin, the maximal q of silica gel is 260.29, K_d is 40.67 and adsorption velocity value is 1.6913 for concentration 0.39mg/ml.