Objective: The objective of the study was to evaluate the pharmacokinetic parameters of abacavir sulphate mucoadhesive buccal films in vivo. Methods: Abacavir sulphate mucoadhesive buccal films were developed using the solvent casting method and the prepared buccal films were evaluated for qualitative and quantitative parameters. Pharmacokinetic parameters (maximum plasma concentration [Cmax], maximum plasma concentration [Tmax], area under the curve [AUC], and biological half-life [t1/2]) were evaluated in vivo using healthy albino white rabbits. The blood samples were collected evaluated and the results were compared with Ziagen a reference standard. The Modern Version 6 software and the pharmacokinetic function (Microsoft Excel add-in) applications were used to conduct the statistical study. Results: The abacavir sulphate mucoadhesive buccal films were prepared successfully and the evaluated qualitative and quantitative parameters were within in the acceptable range. The results of the study stated that Cmax, Tmax, AUC0-t, AUC0-α, and t1/2 of abacavir sulphate mucoadhesive buccal film were found to be 93.86 ng/ml, 8 h, 1652.21 ng/ml×h, 2939.76 ng/ml×h, and 17.96 h, respectively. These results were comparable with the reference standard. Conclusion: The overall absorption of abacavir sulphate was more in the test formulation with respect to the reference product at the same dose. Hence the study concludes that abacavir sulphate mucoadhesive buccal films achieved prolonged muchoadhesion and improved bioavailability compared to the conventional formulation.