Study On Separation And Derivation Of Deguelin From The Roots Of Derris

Deguelin was one of the most common rotenoids in plant. experimental studies demonstrated deguelin has a significant inhibition of cancer cell proliferation, improve symptoms and other effects, and minimal side effectshas effect in the treatment of malignant tumors, especially lung cancer.This topic select the Derris Derris root for the study, to explore the optimum extraction and purification of deguelin; improve and optimize the detection means of deguelin; using high speed countercurrent chromatography and other means of new technology to enhance deguelin extraction rate, to provide material for further toxicology studies, and to analysis the stability and restore of deguelin. The main results are as follows:1. Optimization of Derris extraction conditionsUsing the methed of Orthogonal design of extract liquid ratio, ultrasonic processing time, extraction time on extraction of rotenone to find the best extraction process. Ultimately determine the optimal extraction process of total flavonoids were: pharmaceutical raw materials with 30 times the 4:1 mixed solvent of chloroform and methanol as solvent, extraction 3 times by ultrasonic, each time of ultrasonic treatment was 500S.2. Optimization of HPLC analysis of total flavonoids in Derris rootThe extract of Derris root mainly contained rotenone and deguelin, the two were very similar and difficult to separate and testing.This method improved the rotenone and deguelin HPLC Detection. The HPLC figure of crude extract of Derris root.Conditions:column: Waters Symmetry C18 column(250 mm×4.6 mmI.D.5μm);mobile phase: methyl cyanide ~0.1% trifluoroacetic acid(methyl cyanide:0-8min(55%-65%);8-11min(65%-70%);11-15min(70%-90%));flow rate:1mL/min; column temperature:25.0℃,detection wavelength:270.5nm.. This method of detection avoided the tail of chromatographic peaks and the problem of low substance, can be accurately determinated the content and purity of deguelin.3. Optimization of HSCCC separation conditionTwo rotenoids, rotenone and deguelin were successfully isolated and purified from the roots of Derris by High-speed Counter-current Chromatography(HSCCC) with a two-phase solvent system composed of n-hexane-ethylacetate-methanol–water (7:0.25:5:3, v/v) on a preparative scale.The lower phase was used as the mobile phase and the upper phase was used as the stationary phase.Within 3 hours,6.4mg of rotenone and 23.4mg of deguelin with their purities of 96.60% and 97.87% were obtained from 50mg of the crude extract of the roots of Derris in one-step elution under the conditions of a flow rate of 2.0 mL/min,the revolution speed of 850r/min and the detection wavelength of 254nm.This method shorted the separation time,had no loss of sample, solvent recovery was high.4. Determine the stability of deguelinDeguelin is poor in Light resistance, it should be stored. Deguelin has Strong ultrasonic of heat resistance and long time heating and ultrasonic treatment were not changed its structure. Deguelin has good Acid Resistance, but the molecular structure of deguelin rapid change in the alkaline conditions.5. Optimization of reducing conditionDeguelin can be reduced to hydroxyl deguelin by sodium borohydride, increasing temperature can accelerate the reaction process, the reaction time is shortened under the ultrasonic, appropriately increase the ultrasonic power and reduce ultrasonic frequency can complete the reaction in a very short time . the reduction time of deguelin is the shortest in the ultrasonic frequency of 30kHz, and power of 450W.