Disturbances in emotion regulation among opioid users receiving methadone maintenance treatment (MMT) was reported in a few small studies. We aimed to evaluate the prevalence and characteristics of patients in MMT with impaired emotion regulation. We studied a cross-sectional sample of 121 patients by means of the Difficulties in Emotion Regulation Scale (DERS). Data on clinical, sociodemographic and addiction variables were reviewed, including results of random observed urine testing for opioids, alcohol, cocaine, cannabis, and benzodiazepine (BDZ). The prevalence of a high DERS score (> 90) was 39%. Characteristics associated with high DERS scores (logistic regression model) were benzodiazepine misuse and 20 years of opioid use before admission to MMT. Among the subscales, IMPULSE and STRATEGIES were independently related to substance use (logistic regression). Greater cocaine usage was also more prevalent only in univariate analyses. The DERS score correlated inversely with years of education among patients with negative urine test. The DERS subscale score for AWARENESS was lower in females than in males as among patients with suicide attempts or involvement in physical violence. Since the IMPULSE and STRATEGIES scores predict drug use, these aspects of emotional regulation should be targeted first in third-wave cognitive-behavioral intervention in MMT.