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The Experimentation on a New Saponin for Its Extraction Separation from Homonoia Riparia Lour and the Effects on Hyperuricemia

Author: CheRenYu
Tutor: ZhaoJing
School: Third Military Medical University
Course: Pharmacy
Keywords: Water willow Water willow saponin Hyperuricemia Xanthine oxidase RP-HPLC RT-PCR
CLC: R965
Type: Master's thesis
Year: 2009
Downloads: 93
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Abstract


Objective: To establish the method of determination of water willow saponin content to optimize the the water willow saponin extraction conditions in the water willow and study water willow saponins in the body of hyperuricemia animal. Method: the use of high performance liquid chromatography to establish a method for determination of the new natural compounds in water willow saponin content; uniform design method is preferable to use three factors affect the rate of the total water willow extract extract: extraction time, ethanol concentration and solid-liquid ratio good conditions; Orthogonal Design to affect the the total steroidal saponin extraction rate in the total extract 4 factors: extraction time, solvent type, extract the optimum conditions of temperature and solvent usage; mice administered adenine intraperitoneal hyperuricemia animal model, injection of uricase inhibitor potassium oxonate give high, medium, low doses of water willow saponins, as a positive control, allopurinol model animal serum levels of uric acid, xanthine oxidase enzyme activity influence total protein content of the liver, liver esterase mRNA expression levels, cholesterol and blood sugar levels and test drugs and laboratory animal pathology analysis, to observe the morphological changes. Results: A high performance liquid chromatography determination of the water willow saponins water willow extract: by RP-HPLC column was HYPERSILBDS-C18 column (200mm × 4.6mm, 5μm); mobile phase of acetonitrile: water = 75:25; flow rate of 1.0 ml / min; detection wavelength of 203nm; column temperature was 30 ° C; column pressure of 10.0 Mpa, the linear range of the method to 500μg/ml (r = 0.9998); average sample The recovery rate was 99.44%, RSD 0.52% (n = 15). The uniform design method, the the water willow total extract extract optimum conditions: percolation method with a concentration of 12 times the amount of 75% ethanol extract seven hours per 1g of water willow can get 0.13g water willow extract. Orthogonal design method, the total extract total steroidal saponins extracted the optimum: extracted four hours to 16 times the volume of n-butanol as a solvent refluxing extraction temperature of 70 ° C per 1g of water willow can be 0.138mg The water willow saponin. Modeling the first seven days, the animals of the model group, blood levels of uric acid and xanthine oxidase activity was significantly higher than the normal control group (p lt; 0.01), and pathological examination revealed the kidney model group with varying degrees of injury; administration 21 days, the animals of the model group, blood uric acid levels and xanthine oxidase enzyme activity was significantly higher than that in the normal control group, the water willow saponins high, in the low-dose drug group and the positive drug group (p lt; 0.01), and between the different dose groups The relationship between the presence of dose effect, high-dose group and low dose group difference (p-lt; 0.01); higher liver fat enzyme's mRNA expression analysis prompted the model group, the expression of the amount of than other each experimental group; lipids content analysis found the model group lipids significantly with high lipids in the normal control group, high, medium and low-dose group, and the positive control group (p lt; 0.01), and in addition to the low-dose group compared with normal controls (p lt; 0.01), no difference in the other group and the normal group ( p gt; 0.05); 21 days of the experiment, blood glucose levels of the mice in each group the difference was not statistically significant (p gt; 0.05); liver total protein content model group was significantly higher than the normal control group, high, medium and low-dose group, and positive drug the control group (p lt; 0.01). Conclusion: The use of high performance liquid extract water willow saponin content was measured to reproduce good, high precision, and quality control of medicinal herbs and their preparations for water willow. Establish water willow saponin extraction process safety, convenience, feasible extraction rate. The the water willow saponins intervention hyperuricemia animal model study confirmed that the compounds in the body can significantly inhibit the activity of xanthine oxidase, reduce animal model for the blood uric acid level and blood triglyceride levels, and the presence of a significant dose-response relationship , the compounds can be used as lead compounds for the development of anti-gout drug.

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