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Purpose: To evaluate thiol-disulfide homeostasis (TDH) and its relationship with clinical findings in patients with Graves' ophthalmopathy (GO). Methods: This study included 52 patients with GO and 34 healthy controls. Tests of TDH were conducted using the novel automated spectrophotometric method. ⋯ The mean disulfide levels and disulfide/native thiol % ratios of active GO patients were significantly higher than that of patients with inactive GO (p = 0.034, p = 0.001; respectively). There was a negative correlation between native thiol and CAS (r = -0.532, p = 0.001) and a positive correlation between disulfide/native thiol % ratio and CAS (r = 0.601, p < 0.001). Conclusions: The impairment of TDH indicates the presence of oxidative stress in moderate-to-severe GO, particularly in active GO and smokers.