title: High-throughput determination of nonsteroidal anti-inflammatory drugs in human plasma by HILIC-MS/MS.
abstract: A simple and sensitive method was developed and validated here for the analysis of thirteen nonsteroidal anti-inflammatory drugs (NSAIDs) in human plasma samples by hydrophilic interaction liquid chromatography (HILIC)-tandem mass spectrometry (MS/MS). A small volume of plasma (20μL) spiked with compounds was diluted with 80μL of 10-mM ammonium acetate followed by a simple protein precipitation with 400μL of acetonitrile. After centrifugation, the clear supernatant extract was directly injected into the HILIC-MS/MS, without any solvent evaporation and reconstitution steps. The chromatographic separation of the NSAIDs was achieved on a Unison UK-Amino HILIC column (50mm×3mm i.d., particle size 3μm) with a linear gradient elution system composed of 10mM ammonium acetate (pH 6.8) and acetonitrile at a flow rate of 0.4mL/min. The mass spectra obtained by HILIC-MS showed base peak ions due to [M+H](+) for indomethacin, oxaprozin, ketoprofen, alminoprofen, zaltoprofen, tiaprofenic acid, pranoprofen, and ketoprofen-d3 and due to [M-H](-) for etodolac, ibuprofen, diclofenac, fenoprofen, loxoprofen, naproxen, and ibuprofen-d3. Recoveries of these thirteen NSAIDs in plasma were 34.8-113% and the lower limits of quantitation were 0.125-1.25μg/mL. The intra- and interday coefficient of variations for all drugs in plasma were less than 14.6%. The data obtained from actual plasma determinations of zaltoprofen, ibuprofen, and diclofenac are also presented.
authors.affiliation: Department of Legal Medicine, Showa University School of Medicine, Tokyo 142-8555, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Nemoto
authors.forename: Xiao-Pen
authors.initials: XP
authors.lastname: Lee
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kumazawa
authors.forename: Chika
authors.initials: C
authors.lastname: Hasegawa
authors.forename: Masaya
authors.initials: M
authors.lastname: Fujishiro
authors.forename: Akemi
authors.initials: A
authors.lastname: Marumo
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Shouji
authors.forename: Katsunori
authors.initials: K
authors.lastname: Inagaki
authors.forename: Keizo
authors.initials: K
authors.lastname: Sato
mesh_terms: D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000284:Administration, Oral; D000328:Adult; D000893:Anti-Inflammatory Agents / Q000097:blood*; D000894:Anti-Inflammatory Agents, Non-Steroidal; D001774:Blood Chemical Analysis; D016903:Drug Monitoring; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008297:Male; D008875:Middle Aged; D011786:Quality Control; D012044:Regression Analysis; D015203:Reproducibility of Results; D012997:Solvents / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D000085:Acetates; D000097:Acetonitriles; D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D012997:Solvents; C018824:ammonium acetate; C032159:acetonitrile
keywords: Human plasma; Hydrophilic interaction liquid chromatography (HILIC); Nonsteroidal anti-inflammatory drugs (NSAIDs); Tandem mass spectrometry (MS/MS); UK-Amino column