Background and Objectives According to our national standards, whole blood units are stored at 22±2°C prior to processing. Since there has been no study about the quality of RBCs prepared after the storage of whole blood units at 22±2oC in Iran, we decided to investigate the quality effect of whole blood stored for 8 and 24 hours at this temperature.Materials and Methods Twelve whole blood units in pediatric bags were collected and placed in the cooling plate box.After 8 and 24 hours, RBCs were produced from these stored units. Then, the bags were analyzed for hemolysis, 2, 3-diphosphoglycerate, lactate dehydrogenase, glucose, and sodium.Results Although a higher percentage of hemolysis, lactate dehydrogenase levels, and lower 2, 3-DPG, sodium and glucose levels were determined in RBCs prepared after the 24-hour WB storing time, no significant differences, except for 2, 3 DPG, were observed between 8-hour (205±12) and 24-hour (113±13) WB storing time periods.Conclusions Although storing whole blood at 22±2oC for 24 hours prior to RBCs production had an impact on its quality, the property of prepared components is defined to be within an acceptable range of quality control.