Background: The objective of this study was to determine whether manually performed calibration and quality control (QC) processes could be replaced with an automated laboratory system when installed analyzers fail to provide automated calibration and QC functions.Methods: Alanine aminotransferase (ALT), total cholesterol (TC), creatinine (Cr), direct bilirubin (DB), and lipase (Lip) items were used as analytes. We prepared pooled serum samples at 10 levels for each test item and divided them into two groups; five for the analytical measurement range (AMR) group and five for the medical decision point (MDP) group. Calibration and QC processes were performed for five consecutive days, and ALT, TC, Cr, DB, and Lip levels were measured in the two groups using automated and manual methods. Precision and the mean difference between the calibration and QC methods were evaluated using the reported values of the test items in each group.Results: Repeatability and within-laboratory coefficients of variation (CVs) between the automated system and the conventional manual system in the AMR group were similar. However, the mean reported values for test items were significantly different between the two systems. In the MDP group, repeatability and within-laboratory CVs were better with the automation system. All calibration and QC processes were successfully implemented with the Aptio total laboratory automation system.Conclusion: The Aptio total laboratory automation system could be applied to routine practice to improve precision and efficiency.