Simultaneous determination of aceclofenac and its three metabolites in plasma using liquid chromatography-tandem mass spectrometry.
A new LC/MS/MS-based method allows simultaneous determination of aceclofenac and its three metabolites (4'-OH-aceclofenac, diclofenac, and 4'-OH-diclofenac) in plasma. After acetonitrile-induced precipitation of proteins from the plasma samples, aceclofenac, 4'-OH-aceclofenac, diclofenac, 4'-OH-diclofenac, and flufenamic acid (an internal standard) were chromatographed on a reverse-phase C(18) analytical column. The isocratic mobile phase of acetonitrile/0.1% formic acid((aq)) [80:20 (v/v)] was eluted at 0.2 mL/min. Quantification was performed on a triple-quadrupole mass spectrometer employing electrospray ionization, and the ion transitions were monitored in multiple reaction-monitoring mode. The monitored transitions for aceclofenac, diclofenac, 4'-OH-diclofenac, 4'-OH-aceclofenac and flufenamic acid were m/z 352.9-->74.9, 296.1-->251.7, 311.8-->267.7, 368.9-->74.9, and 279.9-->235.9, respectively. The coefficient of variation of the assay precision was less than 6.5%, and the accuracy ranged from 93% to 103%. The limits of detection were 2 ng/mL for aceclofenac and 0.2 ng/mL for both diclofenac and 4'-OH-diclofenac. This method was used successfully to measure the concentrations of aceclofenac and its three metabolites in plasma from healthy subjects after administration of a single 100-mg oral dose of aceclofenac. This analytic method is a very simple, sensitive, and accurate way to determine the pharmacokinetics of aceclofenac and its metabolites.
Kang
Wonku
W
College of Pharmacy, Catholic University of Daegu, Kyoungbuk 712-702, South Korea.
Kim
Eun-Young
EY
D002853:Chromatography, Liquid / Q000379:methods*; D004008:Diclofenac / Q000031:analogs & derivatives* / Q000097:blood*; D006801:Humans; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
C108035:4'-hydroxyaceclofenac; D004008:Diclofenac; C108036:4'-hydroxydiclofenac; C056498:aceclofenac