title: Simultaneous determination of AMN107 and Imatinib (Gleevec, Glivec, STI571) in cultured tumour cells using an isocratic high-performance liquid chromatography procedure with UV detection.
abstract: A reversed phase high-performance liquid chromatographic (HPLC) method with UV detection was developed for the simultaneous determination of imatinib (Gleevec, Glivec, STI571) and AMN107 in cultured tumour cells, using clozapine as an internal standard. The compounds of interest were extracted by liquid-liquid extraction using TOXI-TUBES((R)) A extraction tubes. Chromatographic separation was performed on a Phenomenex Gemini C18 reversed phase column (150 mm x 2.0 mm, 5 microm particle size), using a mixture of 65% CH(3)OH (methanol) and 35% NH(4)Ac (Ammonium acetate) buffer (20mM, pH 10). Separation was achieved under isocratic conditions at a flow rate of 0.5 ml/min. Imatinib, clozapine and AMN107 are detected by UV detection at 260 nm. Calibration curves were linear from 50 to 7500 ng/ml with correlation coefficients (r(2)) better than 0.998. The limit of quantitation (LOD) was 50 ng/ml. The method has been successfully applied to a cellular kinetics study.
authors.affiliation: Lab of Experimental Oncology, KU Leuven/UZ Gasthuisberg, Department of General Medical Oncology, Leuven, Belgium. Gunther.guetens@ua.ac.be
authors.forename: Gunther
authors.initials: G
authors.lastname: Guetens
authors.forename: Hans
authors.initials: H
authors.lastname: Prenen
authors.forename: Gert
authors.initials: G
authors.lastname: De Boeck
authors.forename: Allan
authors.initials: A
authors.lastname: van Oosterom
authors.forename: Patrick
authors.initials: P
authors.lastname: Schöffski
authors.forename: Martin
authors.initials: M
authors.lastname: Highley
authors.forename: Ernst A
authors.initials: EA
authors.lastname: de Bruijn
mesh_terms: D000970:Antineoplastic Agents / Q000032:analysis*; D001549:Benzamides; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D000068877:Imatinib Mesylate; D010879:Piperazines / Q000032:analysis*; D011743:Pyrimidines / Q000032:analysis*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate; C498826:nilotinib