Study on the Protein Extraction Method in Angelicae sinensis Seed and Its SDS-PAGE Technical System / 中国药房

ABSTRACT

OBJECTIVE:To establish the technical system that is suitable for protein extracting in Angelicae sinensis seed and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),and provide technical support for detecting the protein quality and variety purity. METHODS:Using protein content and number of electrophoretic bands as indexes,8 methods,includ-ing voncentrated gel method,salt-soluble protein method,electrode buffer method,dimercaptosyl alcohol (DTT) method,urea method,mercaptoethanol method,trimethylolaminomethane (Tris) method,and acetone precipitation method,were conducted to extract the protein in A. sinensis seed and screen the optimal extraction method. Then based on optimal extraction method,effects of different materials-lipid ratios,sample dilution times(sample volume)and separate gel concentration on SDS-PAGE were investi-gated. RESULTS:Mercaptoethanol method extracted the highest protein contents(29.931 mg/g),with many electrophoretic bands and clear background. When mercaptoethanol method was used as optimal extraction method,electrophoresis effects were the best in the conditions of materials-lipid ratio of 110,sample volume of 5 times,separate gel concentration of 15%,which obtained 18 bands totally. CONCLUSIONS:Established protein extraction method and SDS-PAGE technical system are suitable for detecting the purity of A. sinensis seed.