Highly sensitive and rapid determination of tacrolimus in peripheral blood mononuclear cells by liquid chromatography–tandem mass spectrometry

Autor: Soma Bahmany, Lucia E.A. de Wit, Birgit C. P. Koch, Teun van Gelder, Brenda C. M. de Winter, Nauras Shuker, Bart C. H. van der Nagel, Carla C. Baan, Dennis A. Hesselink
Přispěvatelé: Pharmacy, Internal Medicine
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Clinical Biochemistry
intracellular concentration
Ficoll
030226 pharmacology & pharmacy
01 natural sciences
Biochemistry
High-performance liquid chromatography
Peripheral blood mononuclear cell
Tacrolimus
Analytical Chemistry
03 medical and health sciences
0302 clinical medicine
Drug Stability
Liquid chromatography–mass spectrometry
Limit of Detection
Tandem Mass Spectrometry
Drug Discovery
Lysis buffer
medicine
Protein precipitation
Humans
Molecular Biology
Research Articles
Whole blood
Pharmacology
Chromatography
medicine.diagnostic_test
Chemistry
010401 analytical chemistry
UPLC–MS/MS
Reproducibility of Results
General Medicine
peripheral blood mononuclear cells
0104 chemical sciences
surgical procedures
operative
Therapeutic drug monitoring
Leukocytes
Mononuclear
Linear Models
Research Article
Chromatography
Liquid
Zdroj: Biomedical Chromatography
Biomedical Chromatography, 33(1):e4416. John Wiley & Sons Ltd.
ISSN: 1099-0801
0269-3879
Popis: After solid organ transplantation, tacrolimus is given to prevent rejection. Therapeutic drug monitoring is used to reach target concentrations of tacrolimus in whole blood. Because the site of action of tacrolimus is the lymphocyte, and tacrolimus binds ~80% to erythrocytes, the intracellular tacrolimus concentration in lymphocytes is possibly more relevant. For this purpose, we aimed to develop, improve and validate a UPLC–MS/MS method to measure tacrolimus concentrations in isolated peripheral blood mononuclear cells (PBMCs). PBMCs were isolated using a Ficoll separation technique, followed by a washing step using red blood cell lysis. A cell suspension of 50 μL containing 1 million PBMCs was used in combination with MagSiMUS‐TDMPREP. To each sample we added 30 μL lysis buffer, 20 μL reconstitution buffer containing 13C2H4‐tacrolimus as internal standard, 40 μL MagSiMUS‐TDMPREP Type I Particle Mix and 175 μL Organic Precipitation Reagent VI for methanol‐based protein precipitation. A 10 μL aliquot of the supernatant was injected into the UPLC–MS/MS system. The method was validated, resulting in high sensitivity and specificity. The method was linear (r 2 = 0.997) over the range 5.0–1250 pg/1 × 106 PBMCs. The inaccuracy was
Databáze: OpenAIRE
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