title: Optimization of the simultaneous determination of imatinib and its major metabolite, CGP74588, in human plasma by a rapid HPLC method using D-optimal experimental design.
abstract: A simple, rapid and specific HPLC method has been developed and validated for the simultaneous determination of imatinib, a tyrosine kinase inhibitor, and its major metabolite, CGP74588, in human plasma. The optimization of the HPLC procedure involved several variables, of which the influences of each was studied. After a series of preliminary-screening experiments, the composition of the mobile phase and the pH of the added buffer solution were set as the investigated variables, while the resolution between imatinib and CGP74588 peaks, the retention time and the imatinib peak width were chosen as the dependent variables. Applying D-optimal design, the optimal chromatographic conditions for the separation were defined. The method proved to show good agreement between the experimental data and predictive values throughout the studied parameter range. The optimum assay conditions were achieved with a Chromolith™ Performance RP-8e 100 mm × 4.6 mm column and a mixture of methanol/acetonitrile/triethylamine/diammonium hydrogen phosphate (pH 6.25, 0.048 mol L(-1)) (20:20:0.1:59.9, v/v/v/v) as the mobile phase at a flow rate of 2 mL min(-1) and detection wavelength of 261 nm. The run time was less than 5 min, which is much shorter than the previously optimized methods. The optimized method was validated according to FDA guidelines to confirm specificity, linearity, accuracy and precision.
authors.affiliation: Biopharmaceutics and Pharmacokinetic Division, Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran 14155-651, Iran.
authors.forename: Ali-Akbar
authors.initials: AA
authors.lastname: Golabchifar
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Rouini
authors.forename: Bijan
authors.initials: B
authors.lastname: Shafaghi
authors.forename: Saeed
authors.initials: S
authors.lastname: Rezaee
authors.forename: Alireza
authors.initials: A
authors.lastname: Foroumadi
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Khoshayand
mesh_terms: D000970:Antineoplastic Agents / Q000097:blood*; D001549:Benzamides; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D000068877:Imatinib Mesylate; D057230:Limit of Detection; D010879:Piperazines / Q000097:blood*; D011743:Pyrimidines / Q000097:blood*; D015203:Reproducibility of Results
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; C464561:CGP 74588; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate