HPLC法测定人血浆中瑞格列奈浓度

Determination of Repaglinide Concentration in Human Plasma by HPLC

目的：以往测定人血浆中瑞格列奈多采用LC-MS法和LC-MS/MS法,方法灵敏度高,但是由于试验成本较高无法普及,故研究建立人血浆中瑞格列奈浓度测定的HPLC方法。方法：采用高效液相色谱法,色谱柱为Agilent Zorbax C18柱（150mm×4.6mm,5μm）,流动相为乙腈-醋酸铵缓冲液（10mmol/mL,pH4.0）（78：22,V/V）;流速1mL/min,检测波长245nm,柱温34℃,进样量20μL。结果：血浆中杂质不干扰样品的测定;瑞格列奈血药浓度在2.0～100ng/mL范围内线性关系良好,定量下限为2.0ng/mL;高、中、低3种浓度的日内、日间RSD均〈10%;绝对回收率为83.8%~93.2%,相对回收率92.7%~105.3%。结论：本方法快速、简便、重现性好,可用于瑞格列奈的临床药动学和药效学研究。

Objective： Separation of repaglinide by LC-MS or LC-MS/MS has the advantage of high sensitivity, but its utility is limited for its expensive cost. The method for determination of repaglinide concentration in human plasma by HPLC was established. Method： Separation was performed on Agilent Zorbax C 18（150mm × 4.6 mm, 5mm） column with a mobile phase consisted of acetonitrile and ammonium acetate buffer （10mmol/L, pH4.0） （78：22, V/V） at the conditions of below： flow rate 1 mL/min, column temperature 34℃, detection wavelength 245 nm and injection volume 20 μ L. Results： The linear range of repaglinide was 2.0-100.0ng/mL, and the lower determination limit was 2.0ng/mL. RSDs of intra-day and inter-day were all less than 10%. Absolute recovery was 83.8%-93.2% and relative recovery was 92.7%-105.3%. Conclusion： Our method is rapid, convenient, sensitive and reproducible for clinical pharmacokinetic and pharmacodynamic study of repaglinide.