AIM: To examine the predictive performance of the PPK software JPKD for the steady-state concentrations of amikacin and recommend the applicable conditions under fixed daily dosage of 400 mg and 600 mg. METHODS: Inpatients using amikacin in the First Affiliated Hospital of Bengbu Medical University from July 2022 to February 2024 were enrolled, and the measured concentrations of amikacin were detected by LC-MS/MS; Verified the predictive performance of JPKD software for peak and trough concentrations of amikacin; JPKD software was applied to predict the steady-state concentrations of amikacin in the patients at the infusion time of 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 h, and then compared the variability of steady-state concentrations with different levels of renal function at optimal infusion time, then the Cmax/MIC values were measured. RESULTS: A total of 69 patients were enrolled, including 18 patients with steady state trough concentrations and 17 patients with steady state peak concentrations. It was found that JPKD had a poor predictive ability for steady state trough concentrations but a good predictive ability for peak concentrations, the WRES<10% between predictive and measured concentrations, and a strong correlation existed between them (r = 0.806). With the shortening infusion time, the higher peak concentrations. The predicted peak concentrations at 0.5 h and 1.0 h infusion time groups were (34.81±6.87) μg/mL and (32.51±6.07) μg/mL, respectively. With the decline of the renal function, the peak concentrations showed a increasing trend. On the same level of renal function, the peak concentrations in the 600 mg group was higher than that of the 400 mg group. When MIC ≤ 2 μg/mL, 400 mg daily dose was chosen; when MIC=4 μg/mL, 400 mg daily dose could be used for CKD3b stage patients, and 600 mg daily dose could be used for CKD1, CKD2, and CKD3a stage patients; when MIC=8 μg/mL, it was predicted that a higher dose was needed to achieve the expected target. CONCLUSION: Amikacin is preferably administered intravenously for 0.5 to 1.0 h, fixed daily doses of 400 mg and 600 mg are indicated for some patients according to the target bacterial MIC and renal function.