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目的:建立以反相高效液相色谱法测定人血浆中吡嗪酰胺血药浓度的方法。方法:采用Diamonsil C_(18)(250 mm×4.6mm,5μm)色谱柱,流动相为0.05 mol·L~(-1)乙酸铵-乙腈(91:9),流速为1.0 ml·min~(-1),检测波长为268 nm,柱温为25℃,采用蛋白沉淀法处理血浆样品。结果:吡嗪酰胺血浆浓度在1.645～105.28μg·ml~(-1)(r=0.999 5)范围内线形关系良好,低、中、高三种浓度的平均相对回收率分别为(85.70±0.75)%,(99.53±2.82)%,(98.93±0.62)%,日内、日间RSD均小于10%。结论:本方法简便、快速、准确、灵敏度高,可用于吡嗪酰胺血药浓度测定和人体药动学研究。
Objective: To establish a method for the determination of pyrazinamide in human plasma by reversed-phase high performance liquid chromatography. Methods: Diamonsil C 18 column (250 mm × 4.6 mm, 5 μm) was used. The mobile phase consisted of 0.05 mol·L -1 ammonium acetate and acetonitrile (91: 9) at a flow rate of 1.0 ml · min ~ -1). The detection wavelength was 268 nm and the column temperature was 25 ℃. Plasma samples were processed by protein precipitation. Results: The linear relationship of pyrazinamide plasma concentration in the range of 1.645-105.28μg · ml -1 (r = 0.999 5) was good, and the average relative recoveries of low, middle and high concentrations were (85.70 ± 0.75) %, (99.53 ± 2.82)%, (98.93 ± 0.62)% respectively. The intra-day and inter-day RSD were less than 10%. Conclusion: The method is simple, rapid, accurate and sensitive. It can be used in the determination of pyrazinamide plasma concentration and pharmacokinetics in humans.