A simple, rapid and highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for determination of 2'-deoxy-2'-fluoro-5-azacytidine (2'F-5-AC), a novel promising antileukemic analog of azacitidine and decitabine, in the rat plasma using clofarabine as the internal standard (IS). Plasma samples were prepared by a one-step protein precipitation with methanol. Chromatographic analysis was carried out on a Zorbax Eclipse Plus C18 column (50 × 2.1 mm; 1.8 μm) with a gradient mobile phase consisting of 5 mM ammonium formate in methanol and in water. In multiple reaction monitoring (MRM) modes, 2'F-5-AC and IS were quantified using precursor-to-product ion transitions of m/z 247.0→113.1 and m/z 304.0→170.0, respectively. The analytical approach developed underwent validation in terms of selectivity, LLOQ 5 ng/mL, linearity (5.0 - 4000 ng/mL, (r) > 0.99) and accuracy (from 87.3% to 89.5%). This method was successfully employed in the pharmacokinetic study on male rats after the intravenous administration of 2'F-5-AC at a dose of 2 mg/kg. The pharmacokinetic parameters show that the maximum concentration of the compound (Cmax = 2036.8 ± 118.7 ng/mL) was observed immediately after administration, while the corresponding time to reach Cmax (Tmax) was 0.08 h, and the half-time (T1/2) was 1.5 h. Clearance (CL) and apparent volume of distribution (Vd) were 620 mL/h/kg and 1118 mL/kg. The present results would be valuable for further research and preclinical development of 2'F-5-AC.
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