ABSTRACT
Objective To optimize the extraction method of Coptidis Rhizoma protein; To compare and analyze the differences of protein in different parts of wild and cultivated Coptidis Rhizoma. Methods SDS-PAGE gel electrophoresis and Coomassie brilliant blue method were used to compare the differences of the protein of Coptis chinensis from the 3 extraction methods (water extraction, Tris-HCl and ammonium sulfate precipitation), and the protein in different parts of the wild and cultivated Coptis chinensis, and analyzed and evaluated the differences. Results The optimum extraction method of Coptis chinensis protein is Tris-HCl method, and the protein content of Rhizoma Coptidis is the highest, and the content of protein is the highest in every gram of medicinal material. The differences among different parts of wild and cultivated Coptidis Rhizoma were obvious, and the ranks for protein abundance and protein content per gram were: rhizome of cultivated Coptis chinensis＞ rhizome of wild Coptis chinensis＞ stem and leaf of cultivated Coptis chinensis＞ stem and leaf of wild Rhizoma Coptidis＞ fibrous roots of cultivated Coptidis Rhizoma. Cluster analysis showed that the correlation between protein of wild and cultivated rhizome of Coptidis Rhizoma was obvious. Conclusion The optimum Tris-HCl method can extract the protein from Coptidis Rhizoma, and the protein content of roots of Coptidis Rhizoma is significantly higher than other parts.
ABSTRACT
OBJECTIVE:To analyze the content of water-soluble proteins in Euryale ferox from different regions. METHODS:The water-soluble proteins in E. ferox were extracted by cryogenic grinding. With the reference of bovine albumin,Coomassie bril-liant blue G-250 was used for coloration,visible spectrophotometry was conducted to determine the content of water-soluble pro-teins in E. ferox from different regions and clustering analysis was used to analyze the results. RESULTS:The linear range of bo-vine albumin was 0.005 01-0.150 3 mg/ml(r=0.999 2)with the average recovery of 98.95%(RSD=1.76%,n=6).RSDs of preci-sion,stability and reproducibility tests were no more than 2.04%. The average value of content of water-soluble proteins in samples was 0.457 7 mg/g. Cluster analysis showed that the content of water-soluble proteins in E. ferox from Yangtze River basin was high-er,followed by the southward and the northward was the least. CONCLUSIONS:The method is simple,reliable and reproducible, and suitable for the content determination of water-soluble proteins in E. ferox.
ABSTRACT
Objective: To investigate the feasibility of four methods for macromolecule protein determination in Chinese materia medica (CMM) injection. Methods: The sensitivity, specificity, and protein test results of sulfosalicylic acid, Coomassie brilliant blue, Lowry, and size exclusion chromatograph (SEC) methods on five kinds of CMM injections were compared. Results: Sulfosalicylic acid method had low sensitivity, Coomassie brilliant blue and Lowry methods had poor specificities, and SEC method had high sensitivity and good specificity. No protein was detected in five kinds of CMM injection by SEC. Conclusion: SEC is suitable for the determinatoin of macromolecule protein in CMM injection.