Everolimus quantification in peripheral blood mononuclear cells using ultra high performance liquid chromatography tandem mass spectrometry

A reliable ultra high performance liquid chromatography tandem mass spectrometry method has been developed for the determination of everolimus in human peripheral blood mononuclear cells (PBMCs). Protein precipitation was used for sample preparation. Analysis was performed on an UPLC Waters Acquity...

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Bibliographic Details
Published in:Journal of pharmaceutical and biomedical analysis 2012-07, Vol.66, p.278-281
Main Authors: Roullet-Renoleau, Francois, Lemaitre, Florian, Antignac, Marie, Zahr, Noel, Farinotti, Robert, Fernandez, Christine
Format: Article
Language:English
Subjects:
ammonium acetate
Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Everolimus
Follow-Up Studies
formic acid
Fundamental and applied biological sciences. Psychology
General pharmacology
Heart Transplantation - methods
Humans
Immunosuppressive Agents - analysis
Leukocytes, Mononuclear - metabolism
Liquid chromatography tandem mass spectrometry
Medical sciences
methanol
mononuclear leukocytes
Peripheral blood mononuclear cells
Pharmacology. Drug treatments
Quality Control
Reproducibility of Results
Sirolimus - analogs & derivatives
Sirolimus - analysis
tandem mass spectrometry
Tandem Mass Spectrometry - methods
Therapeutic drug monitoring
ultra-performance liquid chromatography
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Summary:A reliable ultra high performance liquid chromatography tandem mass spectrometry method has been developed for the determination of everolimus in human peripheral blood mononuclear cells (PBMCs). Protein precipitation was used for sample preparation. Analysis was performed on an UPLC Waters Acquity system. Chromatography was carried out using a cartridge column MassTrak TDM C18 (2.1×10mm) with 0.1% formic acid and 2mM of ammonium acetate in water and 0.1% formic acid and 2mM of ammonium acetate in methanol mixture as a mobile phase delivered at a flow rate of 0.4mL/min in gradient mode. The assay was linear over a range of 0–12.5ng/mL. The analysis of quality control samples at 2.5, 5.0 and 10.0ng/mL demonstrated good precision with relative standard deviation of less than 15%. Recoveries at concentrations of 2.5, 5.0 and 10.0ng/mL were all greater than 83%. The method was successfully applied to the analysis of everolimus in PBMCs from blood samples of transplant recipients.
ISSN:0731-7085
1873-264X
DOI:10.1016/j.jpba.2012.03.042