title: A validated UPLC-MS/MS assay using negative ionization mode for high-throughput determination of pomalidomide in rat plasma.
abstract: In this study, a sensitive UPLC-MS/MS assay was developed and validated for high-throughput determination of pomalidomide in rat plasma using celecoxib as an internal standard (IS). Liquid liquid extraction using dichloromethane was employed to extract pomalidomide and IS from 200μL of plasma. Chromatographic separation was carried on Acquity BEH™ C18 column (50mm×2.1mm, 1.7μm) using an isocratic mobile phase of acetonitrile: 10mM ammonium acetate (80:20, v/v), at a flow rate of 0.250mL/min. Both pomalidomide and IS were eluted at 0.66±0.03 and 0.80±0.03min, respectively, with a total run time of 1.5min only. A triple quadruple tandem mass spectrometer using electrospray ionization in negative mode was employed for analyte detection. The precursor to product ion transitions of m/z 272.01→160.89 for pomalidomide and m/z 380.08→316.01 for IS were used to quantify them respectively, multiple reaction monitoring mode. The developed method was validated according to regulatory guideline for bioanalytical method validation. The linearity in plasma sample was achieved in the concentration range of 0.47-400ng/mL (r(2)≥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 results were within the acceptance criteria and the method was successfully applied in a pharmacokinetic study of pomalidomide in rats.
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia; Bioavailability Laboratory, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia. Electronic address: muziqbal@gmail.com.
authors.forename: Muzaffar
authors.initials: M
authors.lastname: Iqbal
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia; Bioavailability Laboratory, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
authors.forename: Essam
authors.initials: E
authors.lastname: Ezzeldin
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
authors.forename: Khalid A
authors.initials: KA
authors.lastname: Al-Rashood
authors.affiliation: Department of Pharmaceutics, College of Pharmacy, King Saud University, P. O. Box 2457, Riyadh 11451, Saudi Arabia.
authors.forename: Faiyaz
authors.initials: F
authors.lastname: Shakeel
mesh_terms: D000818:Animals; D000068579:Celecoxib; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D057166:High-Throughput Screening Assays / Q000379:methods*; D057230:Limit of Detection; D008297:Male; D011720:Pyrazoles / Q000737:chemistry; D011786:Quality Control; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D013449:Sulfonamides / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013792:Thalidomide / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry
chemical_list: D011720:Pyrazoles; D013449:Sulfonamides; D013792:Thalidomide; C467566:pomalidomide; D000068579:Celecoxib
keywords: Pharmacokinetics; Pomalidomide; Rat plasma; UPLC–MS/MS