© 2020, Springer Nature B.V.Purpose: To evaluate the ability of thiol-disulphide homeostasis to predict renal injury in ureteral obstruction by matching renal scintigraphy and pathological findings in an experimental rat model. Methods: 24 rats were randomized and divided into 3 groups as sham, partial and complete. Blood samples for biochemical evaluations and dimercaptosuccinic acid (DMSA) renal scintigraphy were performed at the beginning and on the 7th day postoperatively. Only a midline abdominal incision was done in sham group. Left ureter of partial group was dissected and gently buried into the psoas muscle. In complete group, left ureter was dissected and ligated. Results: Statistically significant association was found between left kidney function loss percentage, native thiol, total thiol and creatinine alterations and pathological EGTI (Endothelial, Glomerular, Tubular, Interstitial) scores (r = − 0.867, r = − 0.815, r = 0.745, r = − 0.911; p = 0.000). Statistically significant difference was detected between groups in terms of postoperative native thiol and total thiol values (p = 0.000, p = 0.001). There was also a significant difference in preoperative and postoperative comparisons of these parameters in partial and complete obstruction groups (p = 0.012, p = 0.018). There was a significant difference in terms of postoperative disulphide/native thiol and native thiol/total thiol ratios in complete obstruction group compared to other groups (p1 = 0.011, p2 = 0.040; p1 = 0.015, p2 = 0.028). Conclusions: Association with scintigraphic and pathological results empowers the value of this parameter. Disulphide/native thiol and native thiol/total thiol ratios indicate complete obstruction. These findings may shed light to more comprehensive studies.