Simple and robust analysis of cefuroxime in human plasma and bone tissues by LC-MS/MS

In end-stage osteoarthritis, total joint arthroplasty (TJA) represents the definitive therapeutic intervention. Cefuroxime, a second-generation cephalosporin, exhibits a broad spectrum of activity against both Gram-negative and Gram-positive microorganisms, making it a cornerstone of surgical antimicrobial prophylaxis (SAP) to mitigate prosthetic joint infection (PJI) risk. However, the escalating demand for revision arthroplasties has paralleled rising implant-associated infections, necessitating target-site pharmacokinetic optimization to ensure effective antibiotic exposure at the bone-implant interface. Therefore, we developed a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for simultaneous quantification of cefuroxime in human plasma and bone tissues. The separation was completed in 7.5 min on a BEH C₁₈ column (2.1 × 50 mm, 3.5 μm), and the gradient elution was performed in a mobile phase consisting of 0.1% formic acid in acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL min^-1. The correlation coefficients of calibration curves were all greater than 0.99. The detection accuracy of plasma ranged from 93.11% to 98.60% (89.15-106.2% for bone). The intra- and inter-assay precision for both plasma and bone measurements were within 15% (20% at the lower limit of quantitation, LLOQ). The matrix effects were 2.34% to 2.91% in plasma and 3.13-5.17% in bone, while extraction recoveries ranged from 99.8% to 102.0% for plasma and 105.0-107.0% for bone. Upon stability assessment under varying storage conditions, all samples exhibited a difference of less than 15.0%. The method was successfully applied to the determination of cefuroxime in plasma and bone tissues of actual patients.