WFRF:(Rosing J)
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- Kip, A E (author)
Quantification of miltefosine in peripheral blood mononuclear cells by high-performance liquid chromatography-tandem mass spectrometry.
- Article/chapterEnglish2015
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- Elsevier BV,2015
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- LIBRIS-ID:oai:DiVA.org:uu-494592
- https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-494592URI
- https://doi.org/10.1016/j.jchromb.2015.06.017DOI
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- Language:English
- Summary in:English
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- Phagocytes, the physiological compartment in which Leishmania parasites reside, are the main site of action of the drug miltefosine, but the intracellular pharmacokinetics of miltefosine remain unexplored. We developed a bioanalytical method to quantify miltefosine in human peripheral blood mononuclear cells (PBMCs), expanding from an existing high performance liquid chromatography-tandem mass spectrometry method for the quantification of miltefosine in plasma. The method introduced deuterated miltefosine as an internal standard. Miltefosine was extracted from PBMC pellets by addition of 62.5% methanol. Supernatant was collected, evaporated and reconstituted in plasma. Chromatographic separation was performed on a reversed phase C18 column and detection with a triple-quadrupole mass spectrometer. Miltefosine was quantified using plasma calibration standards ranging from 4 to 1000ng/mL. This method was validated with respect to its PBMC matrix effect, selectivity, recovery and stability. No matrix effect could be observed from the PBMC content (ranging from 0.17 to 26.3×10(6)PBMCs) reconstituted in plasma, as quality control samples were within 3.0% of the nominal concentration (precision less than 7.7%). At the lower limit of quantitation of 4 ng/mL plasma, corresponding to 0.12ng/10(6) PBMCs in a typical clinical sample, measured concentrations were within 8.6% of the nominal value. Recovery showed to be reproducible as adding additional pre-treatment steps did not increase the recovery with more than 9%. This method was successfully applied to measure intracellular miltefosine concentrations in PBMC samples from six cutaneous leishmaniasis patients up to one month post-treatment.
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- Rosing, H (author)
- Hillebrand, M J X (author)
- Castro, M M (author)
- Gomez, M A (author)
- Schellens, J H M (author)
- Beijnen, J H (author)
- Dorlo, T P C (author)
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- In:Journal of chromatography. B: Elsevier BV998-999, s. 57-621570-02321873-376X
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- NATURAL SCIENCES
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- Uppsala University
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