ABSTRACT
Objective: To explore the relationship between chemical constituents of Isatidis Radix (IR)and the inhibition of bioactivity of neuraminidase (NA)on influenza virus in vitro. Methods: To establish the chemical fingerprints of IR extraction from different producing areas and determine the contents of common peaks by UPLC. To determine the activity of NA on influenza virus tested with IR by Kit. To analyze the correlation of the chemical information and biological effect by mathematical statistics methods such as grey correlation analysis and one-factor analysis. Results: The chemical contents and biological effect detection varied markedly in the products from different regions and by extracting methods. The result of comprehensive analysis showed that uridine and (R, S)-goitrin of common components in UPLC-fingerprints related with the inhibition of bioactivity of NA on influenza virus, and the content of (R, S)-goitrin was correlated significantly to the activity of NA. Conclusion: Based on the biological assay of NA on influenza virus, the correlation of the content of chemical components and biological effect is verified preliminarily, with a view to provide a more reliable data and reference for the selection and extraction of antiviral active ingredients fromIR.
ABSTRACT
Objective: To investigate the key factors in the process of IsatidisRadix (IR)preparation. The content changes of its main chemical ingredients (epigoitrin, uridine, guanosine, adenosine, and indirubin) and the correlation with its pharmacological activity had been studied, which would providetheoretical foundation for further optimization of process. Methods: The process of IR preparation (extracting-concentrating-alcohol sinking-drying-granulation) was simulated. HPLC was employed to determine the contents of the chemical ingredients in each step of IR preparation, and the clearance rate of OH•was used as the model to investigate the antioxidant activity. Cluster analysis, partial least sqμares (PLS), and multiple linear regression (MLR) were used to analyze the correlation between its chromatogram and pharmacological activity. Results: The content changes of chemical ingredients from IR preparation were generally decreased on the whole, and the highest loss rate of chemical ingredients occurred in the concentrating-drying process; Two metrological methods had been used toanalyze the correlationand the chromatogram ofindirubin, guanosine, and adenosine, which was positively related with the clearance rate of OH•.Conclusion: Each step in the process of IR preparation had the significant effect on the content of its chemical ingredients and pharmacological activity. The concentrating and drying process may be the key process which would finally influence the quality of preparation. Indirubin, guanosine, and adenosine may be the effective substances of IR with antioxidant activity.
ABSTRACT
Objective: To isolate and purify the polysaccharides (glycoproteins) from IsatidisRadix (Banlangen) systematically and to study the composition of them. Methods: Crude polysaccharides precipitated by 80% ethanol from thewater extract of IsatidisRadix, which has theanti-viral activity, were fractionated by DEAE-Sepharose Fast Flow, Sephacryl-200, and high gel chromatography system sequentially. The composition of monosaccharides and amino acids of polysaccharides (glycoproteins) was then determined by HPLC with pre-column derivatization using 1-phenyl-3-methyl-5-pyrazolone (PMP) and o-phthalaldehyde (OPA), respectively. Results: Two of homogeneous polysaccharides named IRPS1A and IRPS1B and two of homogeneous glycoproteins named IRPS2A and IRPS3A were obtained from IsatidisRadix by systematical separation and purification. The monosaccharide composition of IRPS1A and IRPS1B was of arabinose, mannose, galactose, and glucose. IRPS2A contained galacturonic acid, glucose, galactose, and arabinose. While IRPS3A contained mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose. The amino acid compositions of IRPS2A and IRPS3A were 14 kinds of amino acid residues including Asp, Glu, Ser, His, Gly, Thr, Arg, Ala, Cys, Val, Phe, Iso, Leu, and Lys. Besides all, IRPS2A also contained Tyr. Conclusion: This strategy can be used for the isolation and purification of homogeneouspolysaccharides/glycoproteins from IsatidisRadixwhich provides a possible support for the elucidation of the structure and the pharmacologic action of IsatidisRadixpolysaccharides (glycoproteins).