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

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。

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

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 C18 column(250 mm × 4. 6 mm,4 μm),with a mobile phase consisting of acetonitrile 50 mmol/L,citric acid solution 1 mmol/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.