Women with substance use disorder (SUD) often have experienced adverse childhood experiences (ACEs). The intergenerational nature of ACEs also put their children at risk for experiencing ACEs. However, no research has explored the prevalence of ACEs in children whose mothers have SUD. This study assessed ACE scores in mothers with SUD and their children and compared them with non-SUD participants. Females with SUD were recruited from a treatment center (n = 50) and compared to females without SUD from the same area (n = 50). The ACE scores of the participants and their children were measured as well as sociodemographic variables. ANOVA and Fisher’s Exact tests were used to examine univariate differences. Multivariate regression models assessed the difference in ACE scores between the groups and their children and the relationship between maternal and child ACE scores while including sociodemographic confounders. The mean ACE score was significantly higher in SUD participants (4.9, SD = 2.9) when compared to non-SUD participants (1.9, SD = 2.0) after controlling for sociodemographic variables (p < .01). Children of treatment participants also had significantly higher mean ACE scores (3.9, SD = 2.3) than children of comparison participants (1.3, SD = 2.0, p < .01). Maternal ACE score was positively related to children’s ACE score after controlling for sociodemographic variables. Given the intergenerational nature of ACEs and their high burden in both mothers and children in substance use treatment, these preliminary findings suggest that mother–child trauma-informed interventions may be appropriate for this population.