Optimization Of The Conditions Of Extracting And Degrading Isatis Polysaccharide And Determination Of Its Monosaccharide Composition By Cze

Isatis has antibacterial and antiviral in vitro antibiotic and antivirus effect, detoxification, improve the immume system, antitumor, treatment of leukemia, and other effects. Isatis polysaccharide is one kind of various chemical constituents in Isatis. Isatis polysaccharide has good biological activity, which is closely related with the improving the immume system. At present, determination of polysaccharide composition of Isatis by capillary zone electrophoresis(CZE) has no report. In this thesis, CZE was used to analyze and determine the monosaccharide composition of Isatis polysaccharide, which is provide a scientific basic for the quality control of traditional Chinese medicine Isatis. There are six chapters in this thesis:Chapter one: A review mainly concerned with the efficacy of Isatis, research situation, polysaccharide extraction and degradation method, and expounded the purpose and methods of this thesis.Chapter two: Using the monosaccharide reference substances as the test samples. CZE is used for the analysis method,α-naphthylamine and 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatives methods were investigated. By comparing the chromatograms, the derivative steps and the electrophoresis conditions of the two derivative methods, we finally selected PMP derivative method in our article.Chapter three: CZE is used for the analysis method to investigate the different extraction methods of Isatis polysaccharide. The result showed that Isatis polysaccharide content was 16% when extracted by ultrasonic extraction, and the content was 78% when extracted by water extracting-alcohol precipitating. Finally, we selected water extracting- alcohol precipitating methed. The best extracting condition of Isatis polysaccharide was 200 mL, 100 mL and 100 mL water added for 120 min, 60 min and 30 min, respectively. The ethanol concentration was 85%.Chapter four: CZE is used for the analysis method to investigate different degradation methods. The result showed that the monosaccharide types and content degaraded by H2O2 degaradation method and mixed acid degradation were not as good as the superacid degradation. Finally, we selected superacid degradationg method. The best degrading condition of Isatis polysaccharide was with 1 mol/L sulfate solution at 95℃for 5 h. Chapter five: The method that extracting, degradation of Isatis polysaccharide, and monosaccharide derivatives analysis by CZE was established. The results showed that the method is stable and reproducible. CZE can be applied to the component analysis of Isatis monosaccharide , and the method is accurate, reliable with satisfactory results.