Objective: The objective was to compare the biochemical changes of amikacin by autoinjector delivery and manual injection in rats. Materials and Methods: Amikacin drug cartridge (500 mg/2 mL) for autoinjectors was diluted to 63 mg/mL and rats were administered 1.2 mL, i.p. One group was given 3 and a second group 7 injection on consecutive days. 3 and 7 days manual injection of same dose of amikacin (about 500 mg/kg, i.p.) and a control group (saline) were also included (total 5 groups). On day 4 or 8 biochemical parameters were studied. Results: Significant increase in urea, creatinine and aspartate aminotransferase were observed in 7 days administration in both autoinjector and manual injection groups compared to control group. All other parameters viz., glucose, cholesterol, total triglycerides, bilirubin, uric acid, total protein, albumin, alanine aminotransferase and alkaline phosphatase did not show any significant change. No significant change was observed in 3 days administration groups. Conclusion: High dose of amikacin for longer duration is known for its nephrotoxicity which is evidenced by the increase in urea and creatinine in both autoinjector and manual injection groups. This study shows that autoinjector device for amikacin which is new can be considered for further research work. © 2016 Anitha Roy et al.