HPLC-MS/MS法检测兔离体角膜实验中妥布霉素及其药物动力学研究

HPLC-MS/MS Determination and Pharmacokinetic Study of Tobramycin through Isolated Cornea of Rabbits

目的:建立测定谷胱甘肽-碳酸氢钠林格液(GBR)溶液中妥布霉素的液质方法,并对其在家兔离体角膜实验中的药动学特征进行研究。方法:以GBR溶液为释放介质,用液质联用法测定妥布霉素在兔离体角膜中的累积释药量。色谱柱为Waters XTerra RP C18柱(150mm×4.6mm,3μm);流动相为2mM.L-1醋酸铵缓冲液(用甲酸调节pH至3.2)-甲醇(90:10);流速0.6mL.min-1;柱温40℃;采用选择反应监测方式,定量分析的离子分别为m/z468.2→163.0(妥布霉素)和m/z448.2→160.0(内标西索米星)。结果:对照组和冰片组中妥布霉素的渗透释药曲线均符合零级药代动力学方程,线性浓度范围为0.217μg.mL-1～108.5μg.mL-1,最低定量浓度为0.217μg.mL-1。天然冰片组及合成冰片组比对照组的累积渗透量增加近4倍,但冰片组之间没有显著差别。结论:该方法专属性强,灵敏度高,简便快捷,可用于妥布霉素离体角膜透过实验的监测及药动学研究。

Objective:Using LC/MS method to detect tobramycin in glutathione-sodium bicarbonate Ringer's solution and studying its pharmacokinetic in isolated cornea of rabbits.Methods:The cumulative osmotic drug releasing amount of tobramycin in the isolated cornea of rabbits was determined by HPLC/MS with GBR solution as releasing medium.The column is Waters XTerra RP C18(150mm×4.6mm,3μm);the mobile phase is 2mM·L-1 ammonium acetate buffer solution(adjust pH to 3.2 with formic acid)–methanol(90:10);flow rate was 0.6mL·min-1;the column temperature was 40℃.Quantification was based on selected reaction monitoring(SRM) using the precursor→product ion combinations of m/z468.2→163.0 and m/z448.2→160.0 for tobramycin and sisomicin(internal standard) respectively.Results:The osmotic drug releasing of tobramycin in the control group and borneol group all conformed to the zero-grade kinetics equation.The assay range of this method was 0.217μg·mL-1–108.5μg·mL-1,The limit of quantification was 0.217μg·mL-1.The cumulative infiltration of natural and synthetic borneol are nearly 4-fold than that of the control group,but there is no significant difference between these two borneol groups.Conclusion:This method is specificity,sensitive,accurate,simple and quick.It can be applied to detect the tobramycin in vitro transcorneal experiment,also study the pharmacokinetics for it.