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Development and validation of a sensitive assay for the quantification of imatinib using LC/LC-MS/MS in human whole blood and cell culture

Authors

  • Jelena Klawitter,

    1. Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Aurora, CO 80045-7503, USA
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  • Yan Ling Zhang,

    1. Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Aurora, CO 80045-7503, USA
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  • Jost Klawitter,

    Corresponding author
    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
    • Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Bioscience East, Suite 100, 1999 N. Fitzsimons Parkway, Aurora, CO 80045-7503, USA.
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  • Nora Anderson,

    1. Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Aurora, CO 80045-7503, USA
    2. Fraunhofer Institute for Toxicology and Experimental Medicine, Center of Drug Research and Medical Biotechnology, Hannover, Germany
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  • Natalie J. Serkova,

    1. Clinical Research and Development, Department of Anesthesiology, University of Colorado, Denver, Aurora, CO 80045-7503, USA
    2. University of Colorado Cancer Center, University of Colorado, Denver, Aurora, CO 80045-7503, USA
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  • Uwe Christians

    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
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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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