反相HPLC法测定比格犬血浆中乐福昔布的含量

RP-HPLC determination of lefucoxib in Beagle Dog plasma

目的:建立比格犬血浆中创新药物乐福昔布的测定方法。方法:以塞来昔布为内标。采用Agilent 1100 HPLC仪,以Kromasil C_(18)(250mm×4.6mm,5μm)色谱柱为分析柱,5mmol·L^(-1)醋酸铵缓冲液(用36%醋酸调pH至5)-乙腈(34.8:65.2)为流动相,流速为1.0mL·min^(-1),柱温为25℃,检测器波长为254nm,进样量为50μL。结果:在5～500ng·mL^(-1)浓度范围内,测定方法具有良好的线性关系,线性方程为:Y=0.00729X-0.00141,r=0.9984。准确度、精密度均符合生物样品的测定要求,最低定量浓度为5ng·mL^(-1)。结论:该检测方法简便、准确、专属性强,能够满足乐福昔布在比格犬体内药代动力学研究的需要。

Objective： To develop a method for determination of lefucoxib in plasma of Beagle Dog. Method： Celecoxib was used as the internal standard. Lefucoxib was chromatographed on Agilent 1100 HPLC with a C18column （Kromasil, 250 mm × 4. 6 mm,5 μm） , and detected at 254 nm. The mobile phase was 5 mmol· L^-1 ammonium acetate solution（adjusting pH to 5 with 36% acetic acid） - acetonitrile（34. 8：65.2） ,the flow rate was 1.0 mL·min^-1 ,the column temperature was 25 ℃ ,the injection volume was 50 μL. Result： A good linear relationship was obtained between the peak area ratio of lefucoxib/celecoxib and the concentration of lefucoxib over the range of 5 to 500 ng· mL^ - 1. The linear equation was Y = 0. 00729X - 0. 00141, r = 0. 9984. The limit of quantitation was 5 ng ·mL^-1. Conclusion ：The method is simple, fast, and accurate. It can be employed to the study of pharmacokinetics in Beagle dog.