Extraction, Separation And Purification Of 5-hydroxytrophan From Griffonia Simplicifolia

5-hydroxytrophan,which has many biological functions,is an important mental drug and can be used as raw materials to make health-protecting medicine.It is widely used in clinical treatment.And there will be a bright market prospect due to its great value.Therefore,it is of great use to strengthen the research in extraction, separation,purification and detection of 5-hydroxytrophan.African grain Griffonia simplicifolia contains 5-hydroxytrophan.So we use Griffonia simplicifolia as raw materials to study 5-hydroxytrophan.The production process of 5-hydroxytrophan from natural plants is optimized.The research in extraction,separation,purification and detection methods is carried in the experiment with the aim of lower cost.The achievements are as follows:1 The orthogonal design of four factors and three levels is used to select the optimal extraction method based on the single factor experiments and research in extraction methods and extraction solvent.The best extraction process:Griffonia simplicifolia degreased powder through 20 mesh sieve;temperature is 80℃;the extraction times are two;the ratio of material to liquid is 1:15;the extraction time is 3h.2 A new purification process of 5-hydroxytrophan has been developed in this work.The main steps are contained in this procedure,i.e.,clarification of water extract of 5-hydroxytrophan,macroporous adsorption resin chromatography,active carbon decolourization,crystallization and recrystallization.The optimal condition is determined.The results show that the yield is 6.65%,extraction yield is 59.38%and the purity of 5-hydroxytrophan is high to 97.30%.3 Thin-layer chromatography(TLC) method is employed to make qualitative and quantitative analysis.The developer is normal butanol,glacial acetic acid and water with the ratio 3:1:1.Visualization reagent is 0.2%ninhydrin ethanol solution.R_f is 0.55.Scanning wavelength used in single wavelength reflection zigzag scanning is 570nm.It has good linear relationship in the range of 0.8～1.6μg every dot(r=0.9999). RSD(Relative Standard Divergence) is 1.69%. 4 Determine 5-hydroxytrophan by RP-HPLC,by selecting the ultra-violet detector,with methanol:water =5:95(V/V) as mobile phase.UV examination wave-length is 276nm.The result has shown that it has good linear relationship in the range of 0.8～4μg(r=0.9999).RSD(Relative Standard Divergence) is 0.71%.