A simple and sensitive analytical method for quantification of carvedilol in pig serum was developed and validated. Carvedilol and internal standard (IS) were extracted into n-hexane-dichloromethane solvent system and separated using an isocratic mobile phase on a Phenomenex C(18) column. The eluent was monitored by spectroflourimetric detector at a flow rate of 1.0 mL/min. The linearity range of proposed method was 1-1000 ng/mL. The intra-day and inter-day coefficient of variation and percent error values of the assay method were less than 15%, and mean recovery was more than 89.95 and 94.27 for carvedilol and IS, respectively. The method is applicable for use in the pharmacokinetic characterization of carvedilol after administration of buccal patch (6.25 mg) in pigs.