A new chemiluminescence (CL) reaction between luminol and diperiodatoargentate [K(2) [Ag (H(2)IO(6)) (OH)(2)]] was observed in alkaline medium. The CL intensity could be greatly enhanced by amikacin sulfate. Therefore a new CL method for the determination of amikacin sulfate was built by combining with flow injection technology. A possible mechanism of the CL reaction was proposed via the investigation of the CL kinetic characteristics, the CL spectrum and the UV absorption spectra of some related substance. The concentration range of linear response was 5.1 x 10(-8) to 5.1 x 10(-6)mol L(-1) with a detection limit of 1.9 x 10(-8) mol L(-1) (3sigma). The proposed method had good reproducibility with a relative standard deviation of 2.8% (n = 7) for 5.1 x 10(-7) mol L(-1) of amikacin sulfate. It was successfully applied to determine amikacin sulfate in serum.