Sunitinib is an inhibitor of several receptor tyrosine kinases and is used for treatment of gastrointestinal stromal tumors as well as advanced renal cell carcinoma. Considering the need for low-cost cancer treatments, it is essential to develop reliable and sensitive bioanalytical methods of sunitinib determination in human plasma for conducting pharmacokinetic studies of new generic drug products.
Currently available bioanalytical methods are fast and sensitive, predominantly based on protein precipitation and MS/MS detection, providing LLOQs (lower limit of quantification) at the level of 0.1 ng/mL [1]. Presented work describes validation of a method developed for sunitinib determination using LC/MS instrument, conducted according to respective FDA (U.S. Food and Drug Administration) [2] and EMA (European Medicines Agency) [3] guidelines. In order to achieve required LLOQ, liquid-liquid extraction of 500 µL plasma sample with hexane/isopropanol (90/10 v/v) mixture was used. Chromatographic analysis was carried out using Zorbax SB-C18 150 x 3 mm, 3.5 µm column and isocratic elution with 55/45 (v/v) mixture of 0,1% HCOOH and ACN/MeOH (80/20 v/v). The detector was a single quadrupole setup with ESI (+) probe, operating in Single Ion Monitoring (SIM) mode.
Applied sample preparation procedure as well as instrumental conditions allowed recoveries of approximately 70 % for both sunitinib and sunitinib-d10 (internal standard) and linear detection range of 0.1-150.0 ng/mL.
Presented study was supported by the European Union (European Regional Development Fund) in frame of the Innovative Economy Operational Programme 2007-2013 (project No. UDA-POIG.01.03.01-14-069/08)
[1] R. Honeywell, K. Yarzadah, E. Giovannetti, N. Losekoot, E. F. Smit, M. Walraven, J. S. W. Lind, C. Tibaldi, H. M. Verheul, G. J. Peters, J. Chromatogr. B 878 (2010) 1059-1068.
[2] Guidance for Industry. Bioanalytical Method Validation. U.S. Department of Health and Human Services. Food and Drug Administration. Center for Drug Evaluation and Research (CDER). Center for Veterinary Medicine (CVM). May 2001.
[3] Guideline on bioanalytical method validation. European Medicines Agency (EMEA/CHMP/EWP/192217/2009). London, 21 July 2011.
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