The Reduced Pressure Liquid Chromatography Separation And Purification Technology Of Alkaloids In Sophora Flavescens Ait

Sophora flavescens Ait is one of the most widely used traditional Chinese medicine herbs. Previousphytochemical studies revealed that alkaloids are the main chemical components of this plant. Thepharmacological activity of matrine and oxymatrine is extensive and significant, and they have been used forthe clinical treatment of hepatitis B. The anti-tumor activity of Sophora flavescens Ait. alkaloids are widelyrecognized in recent years for they can improve immunity and minimize the damage to the body. In the case ofabundant raw materials and product in great demand, the supply of the crude drug is increasingly tense, andbottleneck which restricting the production is the lack of efficient large-scale industrial technology. In this case,making a large-scale production of Sophora flavescens Ait is an effective method to ease the market demand.In order to investigate the stability of thin-layer chromatography developer which our research group hasbeen established, we studied from the Rfvalue, accuracy, precision, reproducibility, qualitative and quantitativeand many other aspects. The pretreatment method about Sophora flavescens Ait. alkaloids extract precolumnsample, including deoxidate the oxymatrine and others, alkali sinking, and purifying the extract, and theoptimum scheme were discussed. Using macroporous absorption resin and reduced pressure liquidchromatography （vacuum liquid chromatography） separate single Alkaloids provide the technical assistance inorder to achieve industrialization production. At last, Response Surface Methods was used to explore therequirement about preparation of matrine crystallization. Research contents as follows:1. The target content measurement of its extraction, isolation and purification. This study is based onthin-layer developer established by our research group to analysis the isolation and identification effects onfour kinds of alkaloids in Sophora flavescens Ait. Evaluation and analysis the practical application effect withthis developer on the basis of experimental results. Results show that developer ratio（EAc:C₂H₅OH:NH₃·H₂O=5.2:0.7:0.6） has strong practicability on the isolation and identification of alkaloid ofsophora flavescens ait and show better stability.2. In this study, macroporous adsorption resin was choosen to isolate and purify alkaloid of sophoraflavescens ait, and pretreatment of the extract precolumn sample were given: Determined the parameters aboutsodium sulfite reduction the process of oxidation state of alkaloid of sophora flavescens ait, and comparing theweight of impurity respectively using pumping filtration and centrifugation, then the impurity by TLC analysis,the weightiest impurity to be removed when pH13.3.To seperate matrine in the foundation of macroporous adsorption resin combine with vacuum liquid phase separation, three kinds of monomer alkaloids were finally get （MAT:36.3926mg, SC:0.8952mg, SRI:10.4060mg）.4. Using the response surface methodology to find the purification condition of matrine crystallization,purification conditions of matrine crystallization were optimized using response surface methodology.Box-Behnken experimental established quadratic polynomial mathematical model to optimize the level ofinternal factors, and find the optimum value on main factors affect matrine crystallization. The yield of matrinecrystallization up to97.16（Solid-liquid ratio1:28.78, temperature:59.19℃, Time:30min）. In conclusion, this paper choose the way of macroporous adsorption resin combine with vacuum liquidphase separation. It accomplish the effective separation of Akaloid in Sophora flavescens Ait from othernon-alkaloid materials. And also achieve the primary separation of Main monomer in Akaloid in Sophoraflavescens Ait. Meanwhile, this method has the advantages of low processing cost, high processing efficiency,and environmental protection, and will futther the process of industrialization.