The Study On Green Separation And Purification Technology Of Alkaloids In Sophora Flavescens Ait. And Comparative Explore Of Anti-lung Cancer LETP-α-2 Activity By MAT And OMT

Sophora flavescens Ait is a traditional Chinese medicine herb,its main active ingredient is alkaloid. Among total alkaloid in Sophora flavescens Ait, oxymatrine and matrine whose pharmacological activity is extensive and significant,has been used in the treatment of hepatitis B currently,furthermore,their antitumor activity were significant.However,the current extraction and separation process of matrine has some defects,such as high consumption,serious pollution and low efficiency. Based on the comprehensive analysis of the existing kinds of alkaloid extraction and separation methods, a low consumption,less pollution,high efficient green technology in extraction and separation has been designed.The detection, extraction and purification of total alkaloids, separation and purification of the main alkaloids have been studied in detail.The main research contents are as follow:1.Detection is essential for extraction and separation. Thin Layer Chromatography(TLC) were selected out for detection of alkaloids in Sophora flavescens Ait. Based on the previous result, the condition of TLC were optimized by response surface methodology.The data were treated and analysised,and the regression equation was established.The optimized condition is that EAc:C2H5OH:NH3-H2O (5.2:0.7:0.6).2.In the process of total alkaloids extraction, percolation were selected out for extraction of total alkaloid in Sophora flavescens Ait. After discussion, crush size, leaching solution pH, solid to liquid ratio, percolation rate was considered as the main factor,and the rang of factor which will be studied were limited.The process were optimized by response surface methodology also. The data were treated and analysised,and the regression equation was established. The optimized condition is that pH of percolation solution is 1.53,crush size is 65-80,the flow rate is 3.39mL·min-1,5.86 times the volume of drug solution percolation will be collected.3.In the process of alkaloids purification by macroporous resin The process were optimized by single factor experimental design,and the desorption solution were dealed. The purity of the total alkaloids were improved,and is above 50%.4.in the process of separation of three alkaloid monomers,three alkaloid monomers,whose purity is above 90%, were gained with silica gel column chromatography by TLC condition.Eventually, the effect of Matrine on human lung cancer cell strain LETP-a-2 in vitro has been detected, which meets the demand of clinical medication and could provide some references for MAT and OMT served as a new medicine.In MTTtest, LETP-α-2 were treated with 0.5g·L-1,1.0g·L-1,2.0g·L-1,3.0g·L-1,4.0g·L-1 MAT and OMT, and cultured for 24h,48h,72h and 96h respectively. The result showed a linear relationship of effect-time and effect-concentration. Survival rate gradually decreased with prolonging affeced time and increasing concentrations.matrine inhibited more significant than oxymatrine on the LETP-a-2 cells proliferation...