Development and validation of a simple and sensitive LC-MS/MS method for the quantification of cefazolin in human plasma and its application to a clinical pharmacokinetic study

Cefazolin is widely used during surgery to prevent surgical site infections (SSIs). Although cefazolin re dosing is often needed due to its short half-life, the appropriate redosing schedule remains controversial and there is limited information on cefazolin disposition following repeated doses during surgery. In parallel with an ongoing cefazolin redosing clinical study, we have developed and fully validated a simple and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of cefazolin in human plasma. A simple protein precipitation was used for sample preparation. MS/MS analysis was performed using multiple reaction monitoring (MRM) under a positive ionization mode. The lower limit of quantification (LLOQ) for cefazolin was evaluated at 0.25 mu g/mL and a linearity ranging from 0.25 to 300 mu g/mL. Accuracy was <_ 114.3% for quality controls and <_ 118.2% for LLOQ; intra-day and inter-day precision ranging from 1.9% to 14.2% for all quality controls and LLOQ. Matrix effect, extraction recovery, stability testing, dilution integrity, hemolysis effects and whole blood stability have all been investigated. total of 17 parameters were validated and passed their validation criteria. The method was applied in the quantification of cefazolin in clinical plasma samples and was able to successfully determine the concentrations in patients undergoing various surgeries. In comparison with other prior published methods, our method has a simple sample preparation combined with a short analysis run time, a wide dynamic range and low limit of quantification, and is a fully validated assay that abides by FDA guidance. (c) 2021 Elsevier B.V. All rights reserved.

Automated summary

Cefazolin is commonly used to prevent surgical site infections during surgery, but the appropriate redosing schedule is controversial. A new LC-MS/MS method for quantifying cefazolin in human plasma was developed, with accurate results and validation criteria met. This method offers a simple sample preparation, short analysis time, wide dynamic range, and low limit of quantification, making it a reliable tool for determining cefazolin concentrations in clinical settings.