高效液相色谱法测定家兔血浆中安妥沙星浓度及药代动力学参数

Determination of antofloxacin concentration in rabbits plasma by high performance liquid chromatography and its pharmacokinetics

目的:建立测定家兔血浆中安妥沙星浓度的高效液相色谱方法,探讨安妥沙星在家兔体内的药代动力学特征。方法:取安妥沙星片,按25 mg/kg剂量,研细,分散于0.5%羧甲基纤维素钠溶液中,给家兔灌胃,采血,血样经高氯酸处理后进行高效液相色谱法分析。色谱柱条件:Phenomenex C18柱(250 mm×4.6 mm,4μm);乙腈∶50 mmol/L枸橼酸溶液∶1 mol/L醋酸胺溶液(19∶80∶1,体积比)为流动相,流速1.0 ml/min;检测波长为295 nm,柱温40℃。结果:安妥沙星在家兔血浆中的线性范围为0.164~10.5μg/ml,最低定量限为0.164μg/ml。日内及日间变异均<10%,准确度相对误差<5%。血浆中回收率>80%。实验条件下安妥沙星在家兔体内的药代动力学参数:血药浓度时间曲线下面积为(46.17±13.99)mg·L^(-1)·h~(^(-1)),达峰时间为(0.9±0.14)h,峰浓度为(5.15±0.54)mg/L,生物半衰期为(11.98±4.02)h,清除率为(0.58±0.19)L·kg^(-1)·h^(-1)。结论:该方法经考察符合生物样品的测定要求,可用于测定家兔血浆安妥沙星浓度和药代动力学的研究。

Objective： To develop a high performance liquid chromatography method to assess the concentration of antofloxacin in rabbit plasma,and explore the drug metabolism of antofloxacin in rabbits Kinetic characteristics. Methods： Antofloxacin tablets were taken,powdered,added to 0. 5% sodium carboxymethyl cellulose solution and mixed. The solution was intragastric administrated to rabbits at a dose of 25 mg / kg. Blood samples were collected. Following protein precipitation,the analytes were separated on a Phenomenex C18column（ 250 mm × 4. 6 mm,4 μm）,with a mobile phase consisting of acetonitrile 50 mmol / L,citric acid solution 1mmol / L,ammonium acetate solution（ 19∶ 80∶ 1,volume / volume） at a flow rate of 1. 0 ml / min,and the eluent was detected at 295 nm,column temperature was 40 ℃. Results： Linear calibration was generated over antofloxacin＇s concentration range of 0. 164 to 10. 5 μg /ml for plasma samples. Intra-and inter-days precision was 10%,and the accuracy of the relative error was 5%,which were acceptable for all quality control. Samples with relative errors were within the prescribed scopes,including the lowest limit of quantification of 0. 164 μg / ml. The mean recovery of antofloxacin from plasma was 80%. Pharmacokinetic parameters of antofloxacin in rabbits were estimated as follows： the area under concentration-time curve was（ 46. 17 ± 13. 99） mg·L^-1·h^-1,the time peak was（ 0. 9 ± 0. 14） h,the peak concentration was（ 5. 15 ± 0. 54） mg/L,the half time was（ 11. 98 ± 4. 02） h,and the clearance rate was（ 0. 58 ± 0. 19） L·kg^-1·h^-1. Conclusions： This specific,sensitive and precise method is suitable for the pharmacokinetic study of antofloxacin in rabbits.