This work presents a rapid and sensitive high performance liquid chromatography method for the determination of amikacin in water samples with solid phase extraction and pre-column derivatization. Amikacin residue was extracted from water samples with solid phase extraction cartridge. Then the extraction solution was derivatized with 4-chloro-3,5-dinitrobenzotrifluoride in the presence of triethylamine at 70°C in 20 min. The amikacin derivative was separated on a C18 column and detected by application of UV detection at 238 nm. The limit of detection is 0.2 μg/L with a signal-to-noise ratio of 3 and linearity is established over the concentration range from 0 to 500.0 μg/L. Recoveries of the amikacin in three types of water samples are from 87.5 % to 99.6 % and RSDs are 2.1 %-4.5 %. This method can be used for the quantification of amikacin residues in water samples.