A novel chemiluminescence(CL)reaction was based on the oxidizing reaction of luminol by the trivalent copper-periodate complex(K5[Cu(HIO6)2],DPC)in alkaline medium.The CL intensity could be enhanced in the presence of amikacin sulfate(AKS).A new CL method was developed for the determination of AKS by coupling with flow injection(FI)technology.Because of the distinctive oxidative effect of DPC,the luminol-based CL reaction could occur at a low concentration of 107M.The relative CL intensity was proportional to the concentration of AKS in the range of 4.0 109–4.0 106g/mL with the detection limit of 1.2 109g/mL.The relative standard deviation was 2.1%for 8.0 109g/mL AKS(n 9).The proposed method was successfully applied to the direct determination of AKS at the level of ng/mL in serum samples.The recovery varied from 97.0%to 106.3%.A possible mechanism of the CL reaction was discussed in detail by relating to the CL kinetic characteristics and electrochemical activities of the oxidant DPC. A novel chemiluminescence (CL) reaction was based on the oxidizing reaction of luminol by the trivalent copper-periodate complex (K5 [Cu (HIO6) 2], DPC) in alkaline medium. The CL intensity could be enhanced in the presence of amikacin sulfate (AKS). A new CL method was developed for the determination of AKS by coupling with flow injection (FI) technology. Abstract of the distinctive oxidative effect of DPC, the luminol-based CL reaction could occur at a low concentration of 107M. relative CL intensity was proportional to the concentration of AKS in the range of 4.0 109-4.0 106 g / mL with the detection limit of 1.2 109 g / mL. The relative standard deviation was 2.1% for 8.0 109 g / mL AKS (n 9). proposed method was successfully applied to the direct determination of AKS at the level of ng / mL in serum samples. The recovery varied from 97.0% to 106.3%. A possible mechanism of the CL reaction was discussed in detail by relating to the CL kinetic characteristics and electrochemical activities of the oxidant DP C.