Purpose: To evaluate dynamic thiol/disulfide homeostasis in patients with chronic central serous chorioretinopathy (cCSC). Methods: This prospective study included 34 cCSC cases and 37 healthy individuals who were age- and sex-matched. A new colorimetric method for measuring thiol/disulfide homeostasis was used. Native thiol, total thiol/disulfide levels, disulfide/native thiol, disulfide/total thiol, and native thiol/total thiol ratios were measured. Results: The age and gender distributions were similar in both main groups. The mean duration of disease was 47.29 ± 24 months. Native and total thiol levels were significantly lower among the cCSC group relative to the control group (p < 0.001). There was not a statistically significant difference between the cCSC and the control group in terms of disulfide levels (p = 0.492). While disulfide/native thiol and disulfide/total thiol ratios were elevated, the native thiol/total thiol ratio was decreased in the cCSC group (p = 0.017, 0.021, 0.036, respectively). Ratios obtained using plasma native thiol, total thiol, and disulfide levels differed significantly between the both groups. Conclusion: Disulfide/thiol ratio was significantly greater in cCSC patients relative to healthy control subjects. Our results suggest that the oxidative process is involved in the pathogenesis of the cCSC.