A simple HPLC-UV method for quantification of Atovaquone in rabbit plasma

A new simple, sensitive and specific high performance liquid chromatography with UV detection (HPLC-UV) method was developed and validated for determination of Atovaquone in rabbit plasma. The method utilized 250 mu L of plasma, and Atovaquone was extracted from the plasma by using liquid-liquid extraction method. Buparvaquone was used as the internal standard and the method was validated on a C-18 column using acetonitrile-ammonium acetate (pH 3; 0.02 M) (85 : 15, v/v) with UV detection at 254 nm. The assay was conducted at 45 degrees C and a flow rate of 1 ml/min. The calibration curve was linear between 80-4000 ng/ml (r(2) = 0.9993) with a limit of quantification (LOQ) of 80 ng/ml. The extraction recovery ranged from 84.5 to 97.9% and plasma samples containing Atovaquone were found to be stable in -20 degrees C for 4 weeks. Finally, the validated HPLC-UV method was successfully Applied in an in viva pharmacokinetic study involving New Zealand rabbits.