高效液相色谱法同时测定人尿液中布洛芬和吲哚美辛的浓度

Simultaneous Determination of Ibuprofen and Indomethacin in Human Urine by HPLC

目的:建立同时测定人尿液中布洛芬和吲哚美辛浓度的方法。方法:尿液样品经乙腈沉淀蛋白后,采用高效液相色谱法测定。色谱柱为Discovery C_(18),流动相为乙腈-20 mmol/L乙酸铵溶液(85∶15,V/V,用冰醋酸调节pH至3.5),紫外检测波长为220nm,流速为1.0 mL/min,柱温为室温,进样量为80μL。结果:布洛芬和吲哚美辛的尿药浓度均在0.1~50.0μg/mL范围内线性关系良好(r分别为0.999 6、0.999 5,n=3),定量下限均为0.1μg/mL,最低检测限均为0.03μg/mL;日内、日间RSD均小于10%(n=5),准确度为94.7%~97.2%;布洛芬和吲哚美辛的提取回收率分别为89.5%~91.8%、90.2%~92.4%(RSD均小于10%,n=15)。结论:该方法简便、快速,且选择性、精密度、准确度良好,适用于人尿液中布洛芬和吲哚美辛浓度的同时测定。

OBJECTIVE： To establish a method for simultaneous determination of ibuprofen and indomethacin concentration in human urine. METHODS： The urine samples were precipitated by acetonitrile. HPLC method was adopted. The determination was performed on Discovery C18 colunm with mobile phase consisted of acetonitrile-20 mmol/L ammonium acetate solution （85 ： 15, V/V, pH value adjusted to 3.5 with glacial acetic acid） at the flow rate of 1.0 mL/min. UV detection wavelength was set at 220 nm. The column temperature was room temperature, and sample size was 80 laL. RESULTS： The linear range of ibuprofen and indometha- cin were both 0.1-50.0 μg/mL （r=0.999 6, 0.999 5,n=3）. The limits of quantitation were both 0.1 μg/mL, and the limits of detec- tion were both 0.03 μg/mL. RSDs of inter-day and intra-day were all lower than 10% （n=5）, and accuracy ranged 94.7%-97.2%. The extraction recoveries of ibuprofen and indomethacin were 89.5 %-91.8 % and 90.2 %-92.4 % （all RSDs 〈 10 %, n = 15）, respec- tively. CONCLUSIONS： The method is simple and rapid with high selectivity, sensitivity and accuracy. It is suitable for simultane- ous determination of ibuprofen and indomethacin concentration in human urine.