A fast and sensitive LC–MS/MS method for the quantification of fosfomycin in human urine and plasma using one sample preparation method and HILIC chromatography

摘要

Highlights ? Fosfomycin is gaining popularity in antibiotic prescription. ? Analysis of urine and plasma concentrations is needed for PK/PD characterization. ? PK/PD of antibiotics is crucial for therapy optimization and resistance prevention. ? The method is validated over a specific clinically relevant concentration range. ? The method is unique because low LLOQ, fast runtime and uniform sample preparation. Abstract Fosfomycin is an old antibiotic that is increasingly prescribed because of emergence of the antibiotic resistance and the growing incidence of multi-drug resistant infections. Surprisingly, little is known about its pharmacokinetics (PK) and the pharmacodynamics (PD). Quantification of fosfomycin in both urine and plasma provides insight into the PK/PD characteristics of fosfomycin, which is crucial for the optimization of the therapy and the prevention of the emergence of resistance. An analytical method is therefore needed for the quantification of fosfomycin in both urine and plasma. A fast and sensitive tandem mass spectrometry method in combination with HILIC chromatography for the quantification of fosfomycin with a universal sample preparation method for urine and plasma was developed and validated according to FDA guidelines. The universal sample preparation method only requires 100μL of a sample, the addition of the internal standard fosfomycin-13C3 benzylamine and an ultrafiltration step. The method is applicable for the concentration range of 0.75–375mg/L (R2 of 0.9998 in both matrices) encompassing the clinically relevant concentration range based on the susceptibility of possible (uro)pathogens in the clinical setting. The validation results for urine and plasma for all QC levels, were 2.1% and 3.2% for accuracy, 1.5% and 1.7% for within day precision and 5.0% and 3.8% for between day precision, respectively. No matrix effects were encountered and the total recovery in urine and plasma was high (102.5% and 99.4%). Prepared samples were stable at 4°C and 15°C for at least 72h and stored samples at ?80°C were stable for at least 6 months. Selectivity and s