A simple, precise and sensitive UV method have been developed for the estimation of Abacavir sulphate in bulk drug and pharmaceutical dosage form by Difference Spectrophotometric method. Abacavir sulphate has exhibited maximum absorbance at about 248.38 nm and 283.79 nm in acidic and basic solution respectively. Beer's law was obeyed in the concentration range of 2-12mcg/ml in both the cases. The proposed method was successfully applied for the determination of Abacavir sulphate in commercial tablet preparation. As per ICH guidelines the results of the analysis were validated statistically and were found to be satisfactory.