Development and validation of a sensitive assay for the quantification of imatinib using LC/LC‐MS/MS in human whole blood and cell culture |
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| Authors: | Jelena Klawitter Yan Ling Zhang Jost Klawitter Nora Anderson Natalie J Serkova Uwe Christians |
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| Institution: | 1. Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Aurora, CO 80045‐7503, USA;2. Eurofins Medinet Denver, Aurora, CO 80045‐7503, USA;3. Fraunhofer Institute for Toxicology and Experimental Medicine, Center of Drug Research and Medical Biotechnology, Hannover, Germany;4. University of Colorado Cancer Center, University of Colorado, Denver, Aurora, CO 80045‐7503, USA |
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| Abstract: | We developed and validated a semi‐automated LC/LC‐MS/MS assay for the quantification of imatinib in human whole blood and leukemia cells. After protein precipitation, samples were injected into the HPLC system and trapped onto the enrichment column (flow 5 mL/min); extracts were back‐flushed onto the analytical column. Ion transitions M + H]+ of imatinib (m/z = 494.3 → 394.3) and its internal standard trazodone (372.5 → 176.3) were monitored. The range of reliable response was 0.03–75 ng/mL. The inter‐day precisions were: 8.4% (0.03 ng/mL), 7.2% (0.1 ng/mL), 6.5% (1 ng/mL), 8.2% (10 ng/mL) and 4.3% (75 ng/mL) with no interference from ion suppression. Autosampler stability was 24 hs and samples were stable over three freeze–thaw cycles. This semi‐automated method is simple with only one manual step, uses a commercially available internal standard, and has proven to be robust in larger studies. Copyright © 2009 John Wiley & Sons, Ltd. |
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| Keywords: | imatinib gleevec liquid chromatography sample enrichment tandem mass spectrometry LC/LC‐MS/MS |
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