ABSTRACT
Objective To establish an efficient HPLC method for the determination of uric acid in plasma of hyperuricemia model mice,and the evaluation of uric acid lowering effect of Lesinurad.Methods The Laballiance Series Ⅲ HPLC system was adopted with Kromasil C18 column (100 mm × 4.6 mm,5 μm).The mobile phase consisted of methanol-0.5％ acetic acid (10:90) for isocratic elution with a flow rate of 0.4 mL/min.The detection wavelength was set at 283 nm.The established HPLC method was used to detect the plasma uric acid level of mice at 0.5,1.0,and 2.0 h time points after which being ip injected with 250 and 500 mg/kg uric acid.Lesinurad of 250 and 500 mg/kg was ig given to mice,0.5 h later,mice were ip injected with 500 mg/kg uric acid to establish hyperuricemia model,and 1 h later,the established HPLC method was used to detect the plasma uric acid level of mice.Results There was a good linear relationship between peak area and the concentration of plasma uric acid in the range of 7.5-150 μg/mL (r =0.997).The specificity,repeatability,precision,stability,and recovery of the established HPLC method was in accordance with the guiding rules of biological sample determination.Compared with the endogenous serum uric acid concentration of control group mice,serum uric acid concentration of 250 mg/kg dose group was significantly increased 0.5 h after ip administration with uric acid (P ＜ 0.01),and serum uric acid concentration of 500 mg/kg dose group was significantly increased 0.5,1.0,and 2.0 h after ip administration with uric acid.Compared with model group,the concentration of uric acid in plasma decreased significantly in low dosage group administered with Lesinurad (P ＜ 0.05),while decreased more significantly in high dosage group (P ＜ 0.01).Conclusion This convenient,rapid,and accurate method can be applied to the determination of uric acid in mouse plasma and the evaluation of relative drugs,which provide an efficient analysis way for establishing hyperuricemia model and screening relative drugs.