Study On Extraction Method And Process Of Gentianopsis Paludosa One Of The Tibetan Medicines That Combines Chemistry Combines Combines Chemistry And Pharmacodoynamics

Objective: To optimize the extraction process of Gentianopsis paludosa of Tibetan medicine by chemical means,and combine the study of protective effect of different solvent extracts of Gentianopsis paludosa on acute liver injury induced by CCL4 in mice and rats to provide technical support for the research,development and production Gentianopsis paludosa of Tibetan medicine in the future.Methods: The HPLC method was used to establish the determination method of concentration of swertisin,swertianolin and luteolin,three active ingredients in Gentianopsis paludosa.Swertisin,swertianolin and luteolin were used as indicators respectively,and the optimal extraction conditions for Gentianopsis paludosa were selected by single factor and orthogonal experiments.The acute liver injury model induced by CCL4 in mice and rats was established.The protective effect of different solvent extracts of Gentianopsis paludosa on acute liver injury of mice and rats was preliminarily verified.The optimal extraction process for the Gentianopsis paludosa was selected through the combination of chemistry and pharmacology.Results: Through experiment,the optimal extraction process was selected: 8 times of 70% ethanol are extracted for twice,90 minutes for each time.DHPLC conditions are: chromatographic column: AgilentSB-C18(250mm×4.6mm,5?m);moving phase: within 0 to 20 min,methanol: water(containing 0.04% H3PO4)is eluted by 40?60~63?37 linear gradient;Within 20 to 50 min,methanol: water(containing 0.04% H3PO4)= 63: 37 isocratic elution;flow velocity: 0.8mL/min;detection wavelength: 254 nm,column temperature: 30 °C.Under these conditions,the contents of the swertisin,swertianolin and luteolin,three active ingredients in Gentianopsis paludosa are 4.99 mg/g,1.89 mg/g,and 1.46 mg/g,respectively.Combined with the animal experiment results,it can be shown that under the extraction process conditions,the extracted solvent has the best protective effect on the CCL4-induced acute liver injury in mice and rats.Conclusion: This study realized the synchronous extraction of swertisin,swertianolin and luteolin,three active ingredients in Gentianopsis paludosa,and it verified the protective effect of Gentianopsis paludosa on the acute liver injury in mice and rats through the combination of pharmacology and chemistry,improving the comprehensive utilization rate of Gentianopsis paludosa,expanding the application range Gentianopsis paludosa and providing technical support for the clinical use and the preparation dosage form of Gentianopsis paludosa in the future.