Development and validation of a quantitative assay for the measurement of miltefosine in human plasma by liquid chromatography–tandem mass spectrometry

A sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay for the quantification of miltefosine is presented. A 250 μL human EDTA plasma aliquot was spiked with miltefosine and extracted by a solid-phase extraction method. Separation was performed on a Gemini C18 column (150 mm × 2.0 mm I.D., 5 μm) using an alkaline eluent. Detection was performed by positive ion electrospray ionization followed by triple-quadrupole mass spectrometry. The assay has been validated for miltefosine from 4 to 2000 ng/mL using 250 μL human EDTA plasma samples. Results from the validation demonstrate that miltefosine can be accurately and precisely quantified in human plasma. At the lowest level, the intra-assay precision was lower than 10.7%, the inter-assay precision was 10.6% and accuracies were between 95.1 and 109%. This assay is successfully used in a clinical pharmacokinetic study with miltefosine.