Abstract：
BACKGROUND: The investigation of drug disposition in tissues is critical to improving dosing strategy and maximizing treatment effectiveness, yet developing a multi-tissue bioanalytical method could be challenging due to the differences among various matrices. Herein, we developed an LC-MS/MS method tailored for the quantitation of piperacillin (PIP), cefazolin (CFZ), and cefoxitin (CFX) in rat plasma and 12 tissues, accompanied by validation data for each matrix according to the FDA and EMA guidelines.
RESULTS: The method required only a small sample volume (5 μL plasma or 50-100 μL tissue homogenates) and a relatively simple protocol for simultaneous quantitation of PIP, CFZ, and CFX within different biological matrices. Mobile phase A was composed of 5 mM ammonium formate and 0.1 % formic acid in water, while mobile phase B contained 0.1 % formic acid in acetonitrile. The mobile phase was pumped through a Synergi Fusion-RP column equipped with a guard column with a gradient elution program at a 0.3 mL/min flow rate. The mass spectrometer was operated in positive ionization mode (ESI+) using multiple reaction monitoring.
CONCLUSIONS: The validated method has been successfully applied to quantify PIP, CFZ, and CFX from the plasma and tissue samples collected in a pilot rat study and will further be used in a large pharmacokinetic study. To our knowledge, this is also the first report presenting long-term, freeze-thaw, and autosampler stability data for PIP, CFZ, and CFX in rat plasma and multiple tissues.
RESULTS: The method required only a small sample volume (5 μL plasma or 50-100 μL tissue homogenates) and a relatively simple protocol for simultaneous quantitation of PIP, CFZ, and CFX within different biological matrices. Mobile phase A was composed of 5 mM ammonium formate and 0.1 % formic acid in water, while mobile phase B contained 0.1 % formic acid in acetonitrile. The mobile phase was pumped through a Synergi Fusion-RP column equipped with a guard column with a gradient elution program at a 0.3 mL/min flow rate. The mass spectrometer was operated in positive ionization mode (ESI+) using multiple reaction monitoring.
CONCLUSIONS: The validated method has been successfully applied to quantify PIP, CFZ, and CFX from the plasma and tissue samples collected in a pilot rat study and will further be used in a large pharmacokinetic study. To our knowledge, this is also the first report presenting long-term, freeze-thaw, and autosampler stability data for PIP, CFZ, and CFX in rat plasma and multiple tissues.
摘要：
背景：研究组织中的药物处置对于改善给药策略和最大化治疗效果至关重要，然而，由于各种基质之间的差异，开发多组织生物分析方法可能具有挑战性。在这里，我们开发了一种专门用于定量哌拉西林（PIP）的LC-MS/MS方法，头孢唑啉（CFZ），和头孢西丁（CFX）在大鼠血浆和12个组织中，根据FDA和EMA指南，附有每个矩阵的验证数据。
结果：该方法仅需要少量样品体积（5μL血浆或50-100μL组织匀浆）和相对简单的方案即可同时定量PIP，CFZ,和CFX在不同的生物基质中。流动相A由5mM甲酸铵和0.1%甲酸水溶液组成,流动相B在乙腈中含有0.1%甲酸。将流动相以0.3mL/min的流速泵送通过配备有具有梯度洗脱程序的保护柱的SynergiFusion-RP柱。使用多反应监测以正离子化模式(ESI+)操作质谱仪。
结论：经过验证的方法已成功用于量化PIP，CFZ,和CFX的血浆和组织样品收集在一个试验大鼠研究，并将进一步用于大型药代动力学研究。据我们所知,这也是第一份长期报告，冻融,和PIP的自动进样器稳定性数据，CFZ,和CFX在大鼠血浆和多个组织中。
结果：该方法仅需要少量样品体积（5μL血浆或50-100μL组织匀浆）和相对简单的方案即可同时定量PIP，CFZ,和CFX在不同的生物基质中。流动相A由5mM甲酸铵和0.1%甲酸水溶液组成,流动相B在乙腈中含有0.1%甲酸。将流动相以0.3mL/min的流速泵送通过配备有具有梯度洗脱程序的保护柱的SynergiFusion-RP柱。使用多反应监测以正离子化模式(ESI+)操作质谱仪。
结论：经过验证的方法已成功用于量化PIP，CFZ,和CFX的血浆和组织样品收集在一个试验大鼠研究，并将进一步用于大型药代动力学研究。据我们所知,这也是第一份长期报告，冻融,和PIP的自动进样器稳定性数据，CFZ,和CFX在大鼠血浆和多个组织中。
