title: Everolimus quantification in peripheral blood mononuclear cells using ultra high performance liquid chromatography tandem mass spectrometry.
abstract: 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 (2.1×10 mm) with 0.1% formic acid and 2 mM 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 0.4 mL/min in gradient mode. The assay was linear over a range of 0-12.5 ng/mL. The analysis of quality control samples at 2.5, 5.0 and 10.0 ng/mL demonstrated good precision with relative standard deviation of less than 15%. Recoveries at concentrations of 2.5, 5.0 and 10.0 ng/mL were all greater than 83%. The method was successfully applied to the analysis of everolimus in PBMCs from blood samples of transplant recipients.
authors.affiliation: Clinical Pharmacy Department - EA 4123, College of Pharmacy, Paris 11 University, 5 rue Jean Baptiste Clement, 92296 Chatenay Malabry, France.
authors.forename: Francois
authors.initials: F
authors.lastname: Roullet-Renoleau
authors.forename: Florian
authors.initials: F
authors.lastname: Lemaitre
authors.forename: Marie
authors.initials: M
authors.lastname: Antignac
authors.forename: Noel
authors.initials: N
authors.lastname: Zahr
authors.forename: Robert
authors.initials: R
authors.lastname: Farinotti
authors.forename: Christine
authors.initials: C
authors.lastname: Fernandez
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000068338:Everolimus; D005500:Follow-Up Studies; D016027:Heart Transplantation / Q000379:methods; D006801:Humans; D007166:Immunosuppressive Agents / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D011786:Quality Control; D015203:Reproducibility of Results; D020123:Sirolimus / Q000031:analogs & derivatives* / Q000032:analysis; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007166:Immunosuppressive Agents; D000068338:Everolimus; D020123:Sirolimus