Development and validation of an LC/MS/MS assay for mycophenolic acid in human peripheral blood mononuclear cells.
The aim was to develop a LC/MS/MS method able to quantify mycophenolic acid (MPA) in the peripheral blood mononuclear cells (PBMCs) of transplanted patients. PBMCs were isolated from blood by a density gradient separation. The chromatographic separation was carried out on a Zorbax Stable Bond CN, 150 mmx2.1 mm, and MS/MS detection was performed after positive electrospray ionisation of the protonated parent ion. The calibration range was from 0.25 to 100 ng/sample. Extraction from the cells and ionisation recoveries reached 73.5 and 37.9%, respectively. Inaccuracy was always <10% with CVs<15%. MPA was stable at room temperature in the autosampler over 48 h and at -20 degrees C over 1.5 months. Application to clinical samples taken from patients treated with mycophenolate mofetil indicated that the method is suitable for measuring intracellular MPA.
Bénech
Henri
H
CEA, iBiTecS, Service de Pharmacologie et d'Immunoanalyse, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France. henri.benech@cea.fr
Hascoët
Sophie
S
Furlan
Valérie
V
Pruvost
A
A
Durrbach
A
A
D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D009173:Mycophenolic Acid / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D009173:Mycophenolic Acid