A HPLC-MS/MS method for the determination of Nadolol in rat plasma: Development, validation, and application to pharmacokinetic study

Abstract

A sensitive validated method has been developed for the quantification of Nadolol in rat plasma by high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) using deuterated Nadolol (Nadolol D9) as internal standard (IS). The liquid–liquid extraction method using ethyl acetate was employed for the sample pretreatment. The separation was achieved on the Agilent Zorbax XDB C18 column (150 mm × 4.6 mm ID., 3.5 μm). The column temperature was controlled at 30°C. The components were eluted by using mobile phase A (10 mM ammonium formate) and mobile phase B (acetonitrile) in the ratio of 20:80 v/v with a flow rate of 0.5 mL/min. And 15 μL aliquot was injected in an isocratic elution mode with a total run time of 2.5 min. The multiple reactions monitoring transitions, m/z 310.20/254.10 for Nadolol and IS 319.20/255.00 were selected to achieve high selective analysis. The method exhibited great selectivity and linearity over the concentration range of 6 to 3000 ng/mL. The lower limit of quantification was found to be 6 ng/mL. The developed method proved acceptable results on selectivity, sensitivity, precision, accuracy, and stability studies as per Food and Drug Administration guidelines. This HPLC-MS/MS assay was successfully applied to get the pharmacokinetics parameters in rat plasma.