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

J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Apr 1;865(1-2):55-62. doi: 10.1016/j.jchromb.2008.02.005. Epub 2008 Feb 20.


A sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the quantification of miltefosine is presented. A 250 microL 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 x 2.0 mm I.D., 5 microm) 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 microL 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.

Publication types

  • Validation Study

MeSH terms

  • Antiprotozoal Agents / blood*
  • Antiprotozoal Agents / pharmacokinetics
  • Chromatography, Liquid / methods*
  • Humans
  • Phosphorylcholine / analogs & derivatives*
  • Phosphorylcholine / blood
  • Phosphorylcholine / pharmacokinetics
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry / methods*


  • Antiprotozoal Agents
  • Phosphorylcholine
  • miltefosine