The Separation And Purification Of Oxymatrine And Matrine From Crude Extract Of Sophora Flavescens

Alkaloids are the main activity ingredients of natural product and otherpharmaceutical plants. It is important to establish a method of simple and efficient toanalysis and purify the natural product of modern industrial Chinese herb. Oxymatrineand Matrine as the main ingredients of Sophora alkaloids were found not only have aneffect on anti-inflammatory, anti-fibrotic, anti-virus, but also have an extensivepharmacological effect on nervous system and cardiovascular system. In the paper,the separation and purification of oxymatrine and matrine from crude extract ofsophora flavescens were investigated to explore a suitable procedure. The resultsacquired could be used to provide a reliable basis of industrialization.A method was developed to determination oxymatrine and matrine of crudeextract of sophora flavescens by HPLC. AichromBond-AQ C18column （4.6mm×250mm,5μm）was used as the solid phase. Methanol-0.05%triethylamine solution（70:30, v/v）was taken as the mobile phase. The flow rate was1.00mL/min. The UVdetection wavelength was215nm.The calibration curves of matrine and oxymatrinewere linear in the range of0.02~160mg/L and0.04~100mg/L, respectively.The adsorption behavior of oxymatrine and matrine on Purolite Ca ion exchangeresin in system of water, hydrochloric acid, ammonia were determined respectively bystatic method. The processes of separation and purification were based on it.Regarding yield of matrine and oxymatrine, The optimal conditions of separationand purification of oxymatrine and matrine within crude extract of sophora flavescenson Purolite Ca ion exchange resin were obtained by orthogonal design experiment:sample amount-to-solvent ratio was1:16, injection volume was0.5mL, height todiameter ratio was20:1,5%NH3·H2O-40%CH3CH2OH was50:50（V/V）, thecolumn was eluted at the rate of1.5ml/min. The yield of oxymatrine and matrine was49.49%and55.28%, respectively.Based on the data of the orthogonal design experiment, Artificial NeuralNetworks was used to further optimize the process parameters of separation andpurification of oxymatrine and matrine. The optimal conditions of separation andpurification was simulate experiment: sample amount-to-solvent ratio was10:1,injection volume was0.2mL, height to diameter ratio was10:1, the column waseluted at the rate of1.0ml/min to obtain5%NH3·H2O-40%CH3CH2OH was50:50（V/V）, the yield of oxymatrine and matrine was51.50%and59.44%, respectively.The actual yield was52.98%and60.71%, respectively. When sample amount-to-solvent ratio was8:1, injection volume was0.2mL, height to diameterratio was10:1, the column was eluted at the rate of1.0ml/min,5%NH3·H2O-40%CH3CH2OH was50:50（V/V）, the yield of oxymatrine was64.95%, actual yields63.88%. When the sample amount-to-solvent ratio was16:1, injection volume was0.2mL, height to diameter ratio was10:1, the column was eluted at the rate of4.0ml/min,5%NH3·H2O-40%CH3CH2OH was50:50（V/V）, the yield of matrine was64.45%, actual yields was66.27%. By the method of solvent extraction, the purity ofcrystalline product of oxymatrine and matrine were92.36%,95.72%, respectively.