A validated UPLC–MS/MS assay using negative ionization mode forhigh-throughput determination of pomalidomide in rat plasma
Shakeel, Muzaffar Iqbala, Essam Ezzeldin, Khalid A. Al-Rashood, Faiyaz . 2015
tIn this study, a sensitive UPLC–MS/MS assay was developed and validated for high-throughput determi-nation of pomalidomide in rat plasma using celecoxib as an internal standard (IS). Liquid liquid extractionusing dichloromethane was employed to extract pomalidomide and IS from 200 L of plasma. Chromato-graphic separation was carried on Acquity BEHTMC18column (50 mm × 2.1 mm, 1.7 m) using an isocraticmobile phase of acetonitrile: 10 mM ammonium acetate (80:20, v/v), at a flow rate of 0.250 mL/min.Both pomalidomide and IS were eluted at 0.66 ± 0.03 and 0.80 ± 0.03 min, respectively, with a totalrun time of 1.5 min only. A triple quadruple tandem mass spectrometer using electrospray ionizationin negative mode was employed for analyte detection. The precursor to product ion transitions of m/z272.01 → 160.89 for pomalidomide and m/z 380.08 → 316.01 for IS were used to quantify them respec-tively, multiple reaction monitoring mode. The developed method was validated according to regulatoryguideline for bioanalytical method validation. The linearity in plasma sample was achieved in the con-centration range of 0.47–400 ng/mL (r2≥ 0.997). The intra and inter-day precision values were ≤11.1%(RSD, %) whereas accuracy values ranged from −6.8 to 8.5% (RE, %). In addition, other validation resultswere within the acceptance criteria and the method was successfully applied in a pharmacokinetic studyof pomalidomide in rats.
