目的 建立一種快速測定高尿酸模型小鼠血漿中尿酸濃度的高效液相色譜（HPLC）法，并將其用于典型藥物L(fēng)esinurad的降尿酸作用評價。方法 采用Laballiance Series III HPLC系統(tǒng)，色譜柱為Kromasil C₁₈柱（100 mm×4.6 mm，5 μm）；流動相為甲醇/0.5%乙酸（10：90），等度洗脫，體積流量0.4 mL/min，檢測波長為283 nm；應(yīng)用建立的HPLC法測定小鼠ip給予250、500 mg/kg尿酸0.5、1.0、2.0 h后血漿中尿酸濃度；ig給予小鼠50、150 mg/kg Lesinurad 0.5 h后，ip 500 mg/kg尿酸制備模型，1 h后HPLC法測定血漿中尿酸濃度。結(jié)果 在建立的HPLC法中，血漿尿酸濃度在7.5～150 μg/mL與峰面積呈良好線性關(guān)系（r=0.997），方法專屬性、重復(fù)性、精密度、樣品穩(wěn)定性、回收率均符合生物樣品檢測指導(dǎo)原則規(guī)定。與對照組小鼠內(nèi)源性血尿酸濃度比較，250 mg/kg劑量組在0.5 h血漿尿酸濃度顯著升高（P<0.01），500 mg/kg組在0.5、1.0、2.0 h血漿尿酸濃度均顯著升高（P<0.01）。與模型組比較，50、150 mg/kg Lesinurad組血尿酸濃度均顯著下降（P<0.05、0.01），且呈劑量相關(guān)性。結(jié)論 本研究建立的HPLC法簡便、快速、準確，可用于小鼠血漿尿酸濃度測定及相關(guān)藥物的藥效評價，為小鼠高尿酸模型建立及后續(xù)降尿酸藥物的篩選提供了快速可靠的分析手段。

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.

“精準醫(yī)學(xué)研究”國家重點研發(fā)計劃（2016YFC0904903），天津市科技計劃項目（16YFZCSY00910）