Despite the high risk of bacterial contamination, no studies have been found that evaluate the optimal hanging time of open-system enteral sets under room temperature and intensive care conditions. This study aims to determine the differences in microbial loads of solutions taken from open-system enteral feeding sets hanging for 24 h under room temperature and intensive care conditions. This study is a descriptive type of research. The study sample consisted of a total of 40 samples, with 20 samples under room temperature (25.2-23.3°C) and 20 samples under intensive care conditions (21.1-24.2°C). A simple random sampling method was used for sample selection. The assessment of microbial quality was based on the US Food and Drug Administration online Bacteriological Analytical Manual. In this study, it was determined that 5% of the samples taken from intensive care and 15% of the samples taken at room temperature exceeded the accepted limit for aerobic mesophilic bacteria count. The count of catalase and coagulase-negative Staphylococcus spp was 5% in intensive care samples and 40% in room temperature samples. The total coliform count was not detected in intensive care samples but was 35% in room temperature samples. Escherichia coli was not detected in either group. Differences in microbial loads were observed when commercial enteral solutions were administered using open-system enteral feeding sets under room and intensive care conditions for 24 h. Therefore, large-scale studies are needed to determine the safe use of open-system enteral feeding sets under room temperature conditions.