title: Rapid and sensitive LC-MS/MS method for determination of megestrol acetate in human plasma: application to a human pharmacokinetic study.
abstract: 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 C18 column with an isocratic mobile phase, which consisted of 10 mm 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.
authors.affiliation: Department of Pharmaceutical Biochemistry, College of Pharmacy, Kyung Hee University, 1 Hoegi-Dong Dongdaemun-gu, Seoul, Republic of Korea.
authors.forename: Ji-Hyung
authors.initials: JH
authors.lastname: Seo
authors.forename: Ji-Sun
authors.initials: JS
authors.lastname: Park
authors.forename: Min-Ho
authors.initials: MH
authors.lastname: Jo
authors.forename: Mi-Sun
authors.initials: MS
authors.lastname: Park
authors.forename: Ju-Hee
authors.initials: JH
authors.lastname: Ryu
authors.forename: Young-Wuk
authors.initials: YW
authors.lastname: Cho
authors.forename: Wang-Sup
authors.initials: WS
authors.lastname: Shim
authors.forename: Gyu-Jeong
authors.initials: GJ
authors.lastname: Noh
authors.forename: Kyung-Tae
authors.initials: KT
authors.lastname: Lee
mesh_terms: D018931:Antineoplastic Agents, Hormonal / Q000097:blood*; D019540:Area Under Curve; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008297:Male; D019290:Megestrol Acetate / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000191:economics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000379:methods*; D013997:Time Factors
chemical_list: D018931:Antineoplastic Agents, Hormonal; D019290:Megestrol Acetate