Determination of Cefalothin and Cefazolin in Human Plasma, Urine and Peritoneal Dialysate by UHPLC‐MS/MS: application to a pilot pharmacokinetic study in humans

Abstract

An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the analysis of cefazolin and cefalothin in human plasma (total and unbound), urine and peritoneal dialysate has been developed and validated. Total plasma concentrations are measured following protein precipitation and are suitable for the concentration range of 1-500 µg/mL. Unbound concentrations are measured from ultra-filtered plasma acquired using Centrifree(®) devices and are suitable for the concentration range of 0.1-500 µg/mL for cefazolin and 1-500 µg/mL for cefalothin. The urine method is suitable for a concentration range of 0.1-20 mg/mL for cefazolin and 0.2-20 mg/mL for cefalothin. Peritoneal dialysate concentrations are measured using direct injection, and are suitable for the concentration range of 0.2-100 µg/mL for both cefazolin and cefalothin. The cefazolin and cefalothin plasma (total and unbound), urine and peritoneal dialysate results are reported for recovery, inter-assay precision and accuracy, and the lower limit of quantification, linearity, stability and matrix effects, with all results meeting acceptance criteria. The method was used successfully in a pilot pharmacokinetic study with patients with peritoneal dialysis-associated peritonitis, receiving either intraperitoneal cefazolin or cefalothin. Copyright © 2015 John Wiley & Sons, Ltd.

Comparative total and unbound pharmacokinetics of cefazolin administered by bolus versus continuous infusion in patients undergoing major surgery: a randomized controlled trial

“…Cefazolin concentrations in plasma were measured by a validated Ultra-high pressure liquid chromatography-tandem mass spectrometer (UHPLC-MS/MS) method from 1 to 500 lg ml À1 (total cefazolin, Columbia, MD, USA) and from 0.1 to 500 lg ml À1 (unbound cefazolin, Columbia, MD, USA) on a Shimadzu Nexera UHPLC connected to a Shimadzu 8030þ triple quadrupole mass spectrometer. 12 Clinical samples were assayed alongside plasma calibrators and quality controls and met batch acceptance criteria (www.fda.gov/downloads/Drugs/Guidances/ ucm070107.pdf, accessed 25 July 2016).…”

Simultaneous quantification of fentanyl, sufentanil, cefazolin, doxapram and keto‐doxapram in plasma using liquid chromatography–tandem mass spectrometry

“…In general, for all analytes, our assay performed better than or comparable to prior reported assays, even in comparison with assays measuring only a single analyte, which makes it easier to achieve good performance on run time, required sample volume and matrix effects (Aranda et al, ; Carlier et al, ; Clavijo et al, ; Fernandez Mdel et al, ; Hisada et al, ; Kiriazopoulos et al, ; LeGatt et al, ; Lillico et al, ; Mahlke et al, ; Nichol et al, ; Nosseir et al, ; Palleschi et al, ; Parker et al, ; Robson & Prescott, ; Saari et al, ; Suzuki et al, ). Furthermore, most assays use a different drug as an internal standard, whereas we used a deuterated form of fentanyl, which shows better comparable behavior to the analytes that are measured than using a different drug.…”

“…On the other hand, Suzuki et al needed a 12 min run time and used propranolol as an internal standard, where we needed a 5 min run time and used deuterated fentanyl, and our sample preparation required fewer operational steps. Regarding cefazolin, multiple assays have been reported with comparable performance (Carlier et al, ; Kiriazopoulos et al, ; Lillico et al, ; Parker et al, ). The reported fentanyl assays required larger sample volumes except for Hisada et al (), who needed only 20 μL and reached a lower LLOQ of 0.05 μg L −1 compared with our 0.1 μg L −1 .…”

“…Stability during three cycles of freezing and thawing, and freezer stability studies of the analytes in the matrix are not presented because they were already carried out in previous published papers and do not depend on the analytical method. No relevant effects of freezing and thawing were found for all analytes, together with good stability at −20°C for 4 weeks (Clavijo et al, ; Kiriazopoulos et al, ; Mahlke et al, ; Nosseir et al, ; Palleschi et al, ; Parker et al, ; Saari et al, ; Suzuki et al, ).…”

Pharmacokinetics of Intraperitoneal Cefalothin and Cefazolin in Patients Being Treated for Peritoneal Dialysis-Associated Peritonitis

“…Cefazolin and cefalothin were measured by UHPLC-MS/ MS (17). Depending on the matrix measured, the lower limit of quantitation for cefazolin in plasma and PD effluent was 0.1 -1.0 mg/L and cefalothin was 0.2 -1 mg/L.…”