反相离子对高效液相色谱法测定人血浆中羟苯磺酸

RP-HPLC determination of dobesilic acid in human plasma

目的:建立测定羟苯磺酸血浆浓度的反相离子对高效液相色谱法,并用于临床药物动力学研究。方法:0.5 mL 血浆样品经乙腈沉淀蛋白处理后,以0.05 mol·L^(-1)磷酸二氢钾缓冲液(pH 3.0)-乙腈(87:13,v/v)为流动相(流动相中含0.1%四丁基氢氧化铵),Diamonsil C_(18)柱分离,检测波长为300 nm,流速为1.0 mL·min^(-1)。结果:测定血浆中羟苯磺酸方法的线性范围为0.364～36.4 mg·L^(-1),定量下限为0.364 mg·L^(-1)。日内、日间精密度(RSD)均小于8.7%,准确度(RE)在±4.5%以内。羟苯磺酸和内标对氨基苯甲酸的平均血浆样品处理回收率分别为74.8%和94.8%。结论:该方法选择性强,操作简单,适用于羟苯磺酸钙的临床药物动力学研究。

Objective：To develop and validate a sensitive and simple reversed -phase ion -pair HPLC method for the determination of dobesilic acid in human plasma. Methods：Mter a simple protein precipitation with acetonitrile, the analyte and the internal standard ,p -aminobenzoic acid ,were chromatographied on a Diamonsil C18 column with the mobile phase consisting of 0. 05 mol ·L^-1, potassium dihydrogen phosphate buffer （ pH = 3.0） - acetonitrile （87： 13, v/v） , containing 0. 1% tetra - n - butylammonium hydroxide, at a flow rate of 1.0 mL ·min^ - 1. The UV wavelength was set at 300 nm. Results：The linear calibration curves were obtained in the range of 0. 364 -36.4 mg · L^-1. The lower limit of quantification was 0. 364 mg·L^-1. The inter -and intra -day RSD acrossed the three validation runs were both below 8.7% ,and the accuracy （RE） was within 4. 5% for the analyte. The mean recoveries of dobesilic acid and internal standard p- aminobenzoic acid were 74.8% and 94. 8% ,respectively. Conclusion：The method is proved to be special, convenient, and suitable for clinical investigation and pharmacokinetics of calcium dobesilate.