Abstract
A sensitive, acetonitrile-free, high performance liquid chromatography method was developed for the quantitation of alibendol concentrations in plasma. To date, only one other method has been reported for the determination of alibendol in plasma. However, that method may lack the sensitivity needed to determine the low plasma levels encountered in pharmacokinetic studies. Following simple extraction, sample components were separated by isocratic reverse-phase chromatography and quantified using UV detection at 315 nm. The mobile phase consisted of a mixture of 0.02 M phosphate buffer and methanol (50:50, v/v), adjusted to pH 3.0 with phosphoric acid. The assay’s lower limit of quantitation (LLOQ) was determined to be 20 ng/ml in a plasma sample volume of 0.5 ml. Sildenafil was used as an internal standard. Observed retention times were approximately 5.0 and 11.5 min for alibendol and sildenafil, respectively. The within-run precision showed RSD values between 5.83 and 16.96 %. The between-run RSD values varied from 6.73 to 17.99 % at the LLOQ. This method was successfully employed to track the concentration of alibendol in beagle dogs following oral administration of the drug. This assay is advantageous as it does not use expensive acetonitrile and is more sensitive than previously reported method.
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