Bovine Fetuin, And Heparin Sodium Quality

This thesis is mainly discussing on the quality analysis method of bovine fetuin and heparin sodium.One aim of bovine fetuin quality analysis is to set up general methods of sugar chains which will help control the quality of glycoprotein drugs. Another aim is to find out the difference between self-made fetuin and commercial fetuin. Quality analysis of fetuin maily contains determination of purity, molecular weight, isoelectric point, peptide map, ultraviolet absorption spectrum, amino acid composition, monosaccharide composition, oligosaccharide profile and so on, of which the last two projects are more important and difficult. Two methods have been used to determine the purity of three kinds of different fetuin. Fetuin hydrolysis condition was studied and the determination method of monosaccharides composition by HPAEC-PAD was set up. Preparation and profile determination methods of N-glycans and O-glycans were established. The result is that self-made fetuin is much purer than two kinds of commercial fetuin, and the former can be used as a standard glycoprotein. The method of sugar chains analysis can be used for other glycoprotein durgs.Heparin sodium is included in Chinese Pharmacopeia. Its standard is limited in impurity detection. So it is necessary to develop new methods to improve the standard. Multiangle Laser Light Scatter instrument was used to determine the absolute molecular weight and molecular weight distribution of heparin sodium and its analogs for the first time. HPAEC-PAD was used to determine sugar composition. Cellulose acetate electrophoresis was used to separate heparin sodium and its analogs. The result of absolute molecular weight reflects the difference of problem samples and normal samples. According to sugar compositions whether sample contains impurity can be judged. As impurities, Chondroitin sulfate and dermatan sulfate are separated from heparin sodium by Cellulose acetate electrophoresis, whose detection limitations are below 2%. All the data of these three methods evidence the results of NMR and CE.