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Rapid and sensitive LC‐MS/MS method for determination of megestrol acetate in human plasma: application to a human pharmacokinetic study
Author(s) -
Seo JiHyung,
Park JiSun,
Jo MinHo,
Park MiSun,
Ryu JuHee,
Cho YoungWuk,
Shim WangSup,
Noh GyuJeong,
Lee KyungTae
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.2807
Subject(s) - chemistry , megestrol acetate , chromatography , ammonium acetate , ammonium formate , formic acid , pharmacokinetics , calibration curve , selected reaction monitoring , extraction (chemistry) , ethyl acetate , detection limit , mass spectrometry , high performance liquid chromatography , tandem mass spectrometry , pharmacology , medicine , cancer
A rapid, simple and fully validated LC‐MS/MS method was developed and validated for the determination of megestrol acetate in human plasma using tolbutamide as an internal standard (IS) after one‐step liquid–liquid extraction with methyl‐ tert ‐butyl‐ether. Detection was performed using electrospray ionization in positive ion multiple reaction monitoring mode by monitoring the transitions m / z 385.5 → 267.1 for megestrol acetate and m / z 271.4 → 155.1 for IS. Chromatographic separation was performed on a YMC Hydrosphere C 18 column with an isocratic mobile phase, which consisted of 10 m m ammonium formate buffer (adjusted to pH 5.0 with formic acid)–methanol (60:40, v/v) at a flow rate of 0.4 mL/min. The achieved lower limit of quantitation (LLOQ) was 1 ng/mL (signal‐to‐noise ratio > 10) and the standard calibration curve for megestrol acetate was linear ( r > 0.99) over the studied concentration range (1–2000 ng/mL). The proposed method was fully validated by determining its specificity, linearity, LLOQ, intra‐ and inter‐day precision and accuracy, recovery, matrix effect and stability. The validated LC‐MS/MS method was successfully applied for the evaluation of pharmacokinetic parameters of megestrol acetate after oral administration of a single dose 800 mg of megestrol acetate (Megace™) to five healthy Korean male volunteers under fed conditions. Copyright © 2012 John Wiley & Sons, Ltd.