| Objective: To optimize the Riparsaponin extraction procedure from Homonoia riparia Lour. A method for determining the content of Riparsaponin was developed for the evaluation. Moreover the effects of Riparsaponin on the hyperuricemia animal model in vivo were investigated,Methods: A HPLC method for determining the content of Riparsaponin from Homonoia riparia Lour have been established. A uniform probability design method was used to optimize the extraction system for the extractum from Homonoia riparia Lour which included the factors of concentration of alcohol, the time of extraction and the solid-to-liquid ratio in the system. An orthogonal experimental design was used to optimize the technologcial conditions for the total saponins extraction from the extractum which included the factors of time of extraction,types of solvent,temperature of extraction and quantity of solvent. hyperuricemia mouse model was established by combination use of adenine and postassium oxonate. Different evaluation targets included serum uric acid level activity of xanthine oxidase,concentration of total protein,mRNA expression quantity of hepaticlipase,serum triglyceride level,and serum glucose level had been detected after administration per os with three different dosages of Riparsaponin on the model comparing with allopurinol-treated group.also,some mouse kidneys and livers were collected for pathology analysis to evaluate the pathological morphology.Results: The Riparsaponin test was performed on the HYPERSILBDS-C18 (200mm* 4.6mm,5μm)with mobile phase consisted of acetonitrile-water (75:25) at a flow rate of 1.0 ml/min.The detection wavelength was set at 203nm,the column temperature was 30℃,and the column pressure was 10.0 MPa.The calibration curves of Riparsaponin was in a good linearity over the ranges of 5μg/ml~500μg/ml(r=0.9998),and the average recovery was 99.44%, RSD was 0.52%(n=15).The optimum extraction conditions of Homonoia riparia Lour extractum were:extracting with 60% alcohol in the ratio of 1:12 for 7hours,it could be got 0.13g Homonoia riparia Lour extractum per gram of Homonoia riparia Lour.The optimization extraction procedure of Riparsaponin from Homonoia riparia Lour was as follows:with the temperature of 70℃,hot refluxing w ith A cetic Ether in the ratio of 16:1 for 48h,it could be got 0.138mg Riparsaponin per gram of Homonoia riparia Lour. The serum Uric Acid level and activity of Xanthine Oxidase were significantly increased(p<0.01) in the model group compared with control group after administration for 7days,and the pathology analysis indicated that there were serious damages in model group compared with control group.The serum Uric Acid level and activity of Xanthine Oxidase were significantly increased(p<0.01) in the model group compared with control group,high-dosage group, Intermindiat-dosage group,low- dosage group and allopurinol-treated group.The effect of dose-respones relationship had been observed in the high-dosage group, Intermindiat-dosage group and low- dosage group,the effect between high-dosage group and low- dosage group was significantlyn different(p<0.01).Analysis on mRNA expression quantity of Hepaticlipase in liver was indicated that the mRNA expression quantity of model group was high than other groups.The serum Triglyceride level in the model group was significantly increased compared with other groups.Meanwhile,there were no differences(p>0.05) between control group and other groups except(p<0.01) low- dosage group.The serum glucose level and concentration of total protein were no differences between control group and other groups(p>0.05)after the administration for 21days.Conclusion: The HPLC method which established to detect the content of Riparsaponin has good reproducibility and attainable precision, It can be applied to quality control.The optimization extraction procedure of Riparsaponin showed safe, convenient,feasibility and high performance. It was indicated that the constituent of Riparsaponin has outstanding inhibition on the activity of Xanthine Oxidase, could reduce the serum Uric Acid level and serum Triglyceride accompanying with effect of dose-respones relationship in vivo from the intervention experiment on hyperuricemia animal model.We do anticipate that Riparsaponin can be used as lead compound to develop novel anti-gout drugs.
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