Objective: We aimed to investigate the potential of assessing thiol/disulfide homeostasis as novel oxidative stress markers to improve the challenging diagnosis of acute appendicitis in children. Material and Methods: A total of 60 pediatric patients (0-18 years) were enrolled in the study, 30 of which were in the control group and 30 in the acute appendicitis group. Native thiol, total thiol, dynamic disulfide, dynamic thiol, ischemia modified albümin, albumin, White blood cell, hemoglobin, hematocrit, and platelet counts of both groups were measured. The results of both groups were compared using the SPSS (Statistical Package for Social Sciences) version 17 (Chicago, USA) program. For all variables, p <0.05 was considered significant. Results: Total thiol (p<0.001), native thiol (p<0.001), and albümin (p<0.001) levels were significantly decreased while dynamic disulfide, dynamic thiol (p=0.003), and ischemia modified albümin (p<0.001) levels those indicating oxidant side were increased in acute appendicitis group compared to control. White blood cell counts in acute appendicitis group were higher (p<0.001), and platelet counts were lower (p=0.03) than the control group. Conclusion: In the differential diagnosis of acute appendicitis, especially in case that are difficult to diagnose, besides a physical examination, imaging, and current laboratory tests, quantification of thiol/disulfide homeostasis may be helpful in diagnosing. In addition, evaluating albumin and IMA levels may increase the specificity of the test. This test can be more helpful in cases diagnosis is difficult such as children small in their ages and mental retardation.