A validated method for the quantification of fosfomycin in human plasma by liquid chromatography–tandem mass spectrometry

• Unambiguous detection and quantification of fosfomycin in human plasma by MS/MS.
• Fast and easy sample preparation by protein precipitation. No derivatization necessary.
• Chromatographic separation applying the HILIC mechanism on a silica column.
• The chemically close analog propylphosphonic acid used as I.S.
• Calibration range optimized for patients receiving intravenous fosfomycin therapy.

Fosfomycin is a small, hydrophilic antibiotic drug with activity against Gram-positive as well as Gram-negative pathogens. It is in increasing use in intensive care units as a last line antibiotic since it shares no cross-resistance with other antibiotics. It is not metabolized and plasma levels are dependent on renal excretion rate and renal replacement therapy such as hemofiltration or hemodialysis. Measurement of fosfomycin plasma concentrations is therefore highly desirable in order to optimize dosing. We have developed a method for the quantification of fosfomycin in human plasma using HILIC chromatography on a silica stationary phase and tandem mass spectrometric detection. Sample preparation consisted only of protein precipitation without derivatization. Propylphosphonic acid was used as internal standard. Two calibration ranges from 15 to 150 μg/ml and 100 to 750 μg/ml were necessary to cover the whole range of plasma concentrations expected from intensive care patients. Intraday precision ranged from 4.0% to 6.4%, depending on the concentration level, with accuracies ranging from −1.1% to 11.5%. The corresponding interday precisions and accuracies were 2.0–11.0% and 0.6–7.8%, respectively. Fosfomycin was stable in human plasma under all storing conditions relevant for clinical samples. First experiences with this method in clinical routine use confirmed the applicability and ruggedness of the analytical procedure.