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Development and validation of a sensitive LC/MS/MS method for the determination of γ ‐tocotrienol in rat plasma: application to pharmacokinetic studies
Author(s) -
Liang Su,
Bian Xiaomei,
Ma Jing,
Arogunjo Motolani,
Deorukhkar Amit A.,
Krishnan Sunil,
Xie Huan
Publication year - 2013
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.2748
Subject(s) - chemistry , chromatography , electrospray ionization , pharmacokinetics , correlation coefficient , standard curve , calibration curve , selected reaction monitoring , analytical chemistry (journal) , coefficient of variation , mass spectrometry , detection limit , tandem mass spectrometry , pharmacology , mathematics , medicine , statistics
γ ‐Tocotrienol has attracted much attention owing to its multiple health benefits. This study developed and validated a simple, specific, sensitive and reliable LC/MS/MS method to analyze γ ‐tocotrienol in rat plasma. Plasma samples (50 μL) were extracted with internal standard solution (25 ng/mL of itraconazole) in acetonitrile (200 μL) with an average recovery of 44.7% and an average matrix effect of −2.9%. The separation of γ ‐tocotrienol and internal standard from the plasma components was achieved with a Waters XTerra® MS C 18 column with acetonitrile–water as mobile phase. Analysis was performed under positive ionization electrospray mass spectrometer via the multiple reaction monitoring. The standard curve was linear over a concentration range of 10–1000 ng/mL with correlation coefficient values >0.997. The method was validated with intra‐ and inter‐day accuracy (relative error) ranging from 1.79 to 9.17% and from 2.16 to 9.66%, respectively, and precision (coefficient of variation) ranged from 1.94 to 9.25% and from 2.37 to 10.08%, respectively. Short‐term stability, freeze–thaw stability and the processed sample stability tests were performed. This method was further applied to analyze γ ‐tocotrienol plasma concentrations in rats at various time points after administration of a 2 mg/kg single intravenous dose, and a pharmacokinetic profile was successfully obtained. Copyright © 2012 John Wiley & Sons, Ltd.