ABSTRACT
A rapid LC coupled with electrospray ionization (ESI) MS/MS method was developed and validated for the quantification of paroxetine in heparinized human plasma. The plasma samples were prepared by the solid-phase extraction method without drying or reconstitution. Elution was done with 0.5 mL 0.2% (v/v) formic acid in methanol-acetonitrile (65 + 35, v/v). The analyte and the internal standard (IS; imipramine hydrochloride) were chromatographed on a BDS Hypersil C18 column. The analyte was analyzed by LC/MS/MS with only 1.7 min run time. An ESI interface was chosen for ionization of the analyte from the sample matrix. Selected reaction monitoring mode for detection of paroxetine and the IS were achieved by using m/z 330.17/192.10 and 281.13/86.14, respectively. The LC retention times for paroxetine and imipramine were 0.94 and 1.05 min, respectively. The method was linear in the concentration range of 0.5-80.0 ng/mL with r > or = 0.9995. Recovery of paroxetine and imipramine ranged from 90 to 95%. The assay has been successfully applied to bioequivalence study samples for estimation of paroxetine in healthy human subjects.
Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Reverse-Phase/methods , Paroxetine/blood , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Blood Chemical Analysis/methods , Chromatography, High Pressure Liquid/statistics & numerical data , Chromatography, Reverse-Phase/statistics & numerical data , Drug Stability , Humans , Imipramine/blood , Imipramine/standards , Reference Standards , Selective Serotonin Reuptake Inhibitors/blood , Spectrometry, Mass, Electrospray Ionization/statistics & numerical data , Tandem Mass Spectrometry/statistics & numerical dataABSTRACT
A selective, rapid and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is described for assay of donepezil in human plasma using escitalopram as an internal standard. Chromatographic separation was achieved on a Betabasic-C(8), 5 microm, 100 x 4.6 mm column using methanol:water:formic acid (90:9.97:0.03, v/v/v) as mobile phase. Detection of donepezil and internal standard was achieved by ESI MS/MS in positive ion mode using 380.20/91.10 and 325.13/262.00 transitions, respectively. The linearity over the concentration range of 0.15-50 ng/mL for donepezil was obtained and the lower limit of quantification was 0.15 ng/mL. For each level of quality control samples, inter-day and intra-day precisions (RSD) were < or =8.92 and 10.35% and accuracy (%RE) were < or =7.33% and 9.33%, respectively. The recovery was more than 88.50% for both donepezil and internal standard by solid-phase extraction, eliminating evaporation and reconstitution steps.
