Bioanalytical development and validation of liquid chromatographic–tandem mass spectrometric methods for the quantification of total and free cefazolin in human plasma and cord blood

Abstract

ObjectivesCefazolin is a commonly prescribed β-lactam antibiotic for prophylaxis against skin infections following surgery, including caesarean sections. Assessment of maternal and neonatal exposure is important for correlating drug concentrations to clinical outcomes. Thus, bioanalytical methods for the quantification of both total and free cefazolin in maternal plasma and cord blood can assist in the comprehensive evaluation of cefazolin exposure.Design and methodsSpecimen preparation for the measurement of total cefazolin was performed via protein precipitation with acetonitrile containing the internal standard cloxacillin. Ultrafiltration was used to isolate free cefazolin. Processed samples were analyzed on a Prelude SPLC system coupled to a TSQ triple quadrupole Vantage mass spectrometer. Methods were validated following FDA bioanalytical guidelines.ResultsThe analytical measuring ranges of these methods were 0.48–480 µg/mL and 0.048–48 µg/mL for total and free drug, respectively. Calibration curves were generated using 1/x2 weighted linear regression analysis. Total cefazolin demonstrated inter- and intra-assay precision of ≤20% at the LLOQ and ≤11.2% at other levels. Free cefazolin demonstrated inter- and intra-assay precision of ≤18.5% at the LLOQ and ≤12.6% at other levels, respectively. Accuracy (%DEV), carryover, matrix effects, recovery and stability studies were also acceptable based on FDA recommendations. Furthermore, it was demonstrated that samples prepared in cord blood can be accurately quantified from an adult plasma calibration curve, with recoveries ≤9.1% DIF and ≤11.9% DIF for total and free cefazolin, respectively.ConclusionsThe described LC–MS/MS methods allow for the measurement of total and free cefazolin in both plasma and cord blood.

Effect of Maternal Obesity on Maternal-Fetal Transfer of Preoperative Cefazolin at Cesarean Section

“…Total and free cefazolin concentrations were determined in maternal and umbilical cord plasma by using a previously described liquid chromatographic-tandem mass spectrometric (LC-MS/MS) assay. 16 The method was validated as per US Food and Drug Administration Bioanalytical Method Validation guidelines. Cefazolin-spiked plasma calibrators, quality control samples, and unknown specimens were prepared identically.…”

Preanalytical Stability of Flucloxacillin, Piperacillin, Tazobactam, Meropenem, Cefalexin, Cefazolin, and Ceftazidime in Therapeutic Drug Monitoring: A Structured Review

“…Seven published articles included data on the preanalytical stability of cefazolin. 14,28,36,49–52 These data are shown in Figure 1E. At room temperature, cefazolin has been reported to be stable for up to 24 hours, 49 48 hours, 14 and 72 hours, 52 with no instability reported in other studies investigating shorter times.…”

“…14,28,36,49–52 These data are shown in Figure 1E. At room temperature, cefazolin has been reported to be stable for up to 24 hours, 49 48 hours, 14 and 72 hours, 52 with no instability reported in other studies investigating shorter times. 28,36,50,51 As data beyond 48 hours are only based on 1 study, caution should be taken after 48 hours.…”

Development of Thin-Layer Chromatography–Densitometric Procedure for Qualitative and Quantitative Analyses and Stability Studies of Cefazolin

“…Baranowska et al developed the RP-HPLC-DAD method to measure the concentration of various drugs, including CFZ, in urine samples [18]. Due to the use of CFZ in the treatment of postoperative infections, especially after a cesarean section, an LC-MS/MS method was also developed for the quantification of total and free cefazolin in maternal plasma and umbilical cord blood after its isolation using an ultrafiltration technique [19]. The HILIC-ESI/MS methodology was used for the simultaneous quantification of antibiotics such as cefuroxime, cefoxitin, and CFZ in breast milk.…”