| OBJECTIVESThe aim of this study is to research the extraction and purification of aesculin and it’s effect on the level of uric acid in chronic hyperuricemia mice.METHODS1. Extraction of aesculin from cortex fraxiniThe extractin of aesculin from Cortex Fraxini was optimized by single factor and L9(3)4orthogonal design. The influences of alcohol concentration, solid-liquid ratio, extraction time, number of ectraction, and extraction temperature on the content of aesculin were investigated.2. Purification of aesculinCombining the method of macroporous resins and recrystallization to isolate and purify the aesculin. By researching the adsorption and desorption of aesculin on the resins of HPD-100,HPD-400,HPD-500, ADS-7,ADS-17,D101and AB-8to find the best resin and purification condition.3. Aesculin’s effect on the level of uric acid in chronic hyperuricemia miceHealthy male Kunming mouse of clean grade35±5g were selected and randomly divided into6groups and6mouse per group:control group, model group, low dose(50mg/kg) aesculin group, medium dose (100mg/kg) aesculin group, high dose(200mg/kg) aesculin group and positive group(50mg/kg). all the mouse were given xanthine600mg/kg (ip.) and the potassium oxonate100mg/kg (ig.) except the control group mouse given same dose normal saline.low dose group were given50mg/kg aesculin, medium group were given100mg/kg aesculin, high group were given200mg/kg aesculin, positive group were given50mg/kg allopurinol. Continue7days. Administration1hour later, remove all the mouse eyeballs to get blood at7th days. Draw serum to detect the BUA,BUN and CR with automatic biochemical analyzer.RESULTS1. Extraction of aesculin from Cortex FraxiniThe optimum extraction condition was as follows:solid-liquid ratio1:10. extracting2times at65℃,each time for2h with75%ethanol.2. Purification of aesculinThe experimental results show that the resin ADS-7is the best resin for the purification of Aesculin. The optimum isolation process is as follows:the concentration of cortex fraxini is0.192mg/mL and maximum loading quantity of aesculin is4.02mg/mL. First, the macroporous resin was eluted by distilled water to wipe off impurities. Second, the macroporous resin was eluted by30%alcohol.and the first half of eluting sovent was collected and placed it in the refrigerator for24hours at4-6℃. After recrystallization and filtered, dried at room temperature. The purity of aesculin is more than97%.3. Aesculin’s effect on the level of uric acid in chronic hyperuricemia miceThe results showed that the model group and the control group compared to mice BUA was significantly increased (P<0.01), suggesting that the model was successful. Compared with model group, The high, medium and low dose aesculin group can significantly reduce the BUA of chronic hyperuricemia mice. Medium and high dose aesculin group can reduced the BUA of chronic hyperuricemia mouse to below the normal level, the inhibition rates were63.45%and80.69%, respectively. The CR of model group is higher than the control group (P<0.01). The CR of high, medium and low dose aesculin group slightly higher than the control group, but no statistical significance. Compared with the control group, the BUN of model group, high, medium and low dose aesculin group was no statistical significance. The CR and BUN of positive group of mouse were significantly higher than the control group,model group and Aesculin groups(P<0.01)CONCLUSIONSThis study supply a efficiently method to prepare high purity aesculin. The pharmacodynamic test results indicated that Aesculin has strongly lowering uric acid effect and no damage of renal function, aesculin is more significantly safety than allopurinol in the treatment of hyperuricemia. |
