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A simple, specific, sensitive and rapid LC-ESI-MS/MS method has been developed and validated for the quantification of GSK-199 in mice plasma using warfarin as an internal standard (IS). Sample preparation was accomplished through a liquid-liquid extraction process. Chromatographic separation of GSK-199 and IS was performed on a Chromolith column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (25:75, v/v) at a flow rate of 1.0 ml/min. Elution of GSK-199 and IS occurred at ~1.32 and 1.81 min, respectively. The total chromatographic run time was 2.50 min. A linear response function was established in the concentration range of 2.52-1009 ng/ml. Method validation was performed as per regulatory guidelines and the results met the acceptance criteria. The intra- and inter-day accuracy and precision were in the range of 1.75-12.3 and 4.99-12.3%, respectively. GSK-199 was found to be stable under various stability conditions. This novel method has been applied to a pharmacokinetic study in mice.