A simple and fast spectrophotometric procedure has been developed for the determination of amikacin sulphate (AMK). The method is based on the interaction of ninhydrin in N,N′-dimethylformamide (DMF) medium, with primary amines present in amikacin. This reaction produced a blue-purple product which absorbed maximally at 590 nm. The effects of variables such as reagent concentration and reaction time were investigated to optimize the experimental conditions. Beer's law is obeyed in the concentration range of 1.0 - 21 μg/ml with RSD of 0.463% and molar absorptivity of 3.33 x 104 l/mol.cm. The proposed method has been applied successfully to the analysis of the bulk drug and its dosage forms and spiked human plasma. No interference was observed from common pharmaceutical adjuvant. Statistical comparison of the results with the reference method shows excellent agreement and indicates no significant difference in accuracy and precision.