Background Electronic prescribing (EP) systems are online technology platforms by which medicines can be prescribed, administered, and stock controlled. The actual impact of EP on patient safety is not truly understood. This study seeks to assess the impact of the implementation of an EP system on safety culture, as well as assessing differences between clinical respondent groups and considering their implications. Methods Staff completed a modified Safety Attitudes Questionnaire survey, 6 weeks following the introduction of EP across surgical services in a hospital in Dorset, England. Responses were assessed and differences between respondent groups compared. Rates of self-reported adverse events were compared before and after implementation. Results Overall response rate was 34.5%. There was no significant difference between usage patterns and previous experience with EP between user groups. Overall safety was felt to have been reduced by the introduction of EP. Significant differences between clinician and nonclinicians were seen in ability to discuss errors (3.23 ± 0.5 versus 2.8 ± 0.69, P = 0.004), drug chart access, and ease of medication prescribing. Regression analysis did not identify any confounding factors. Despite a significant reduction in the adverse event rate in other divisions of the hospital that did not implement EP at the same time, this same reduction was not seen in the surgical department. Conclusions This is the first study to assess the impact of EP on safety culture using a validated assessment tool (Safety Attitudes Questionnaire). Overall safety culture deteriorated following introduction of EP. Problems with system usability/intuitiveness, nonstandardized implementation, and competence assessment strategies may have all contributed to this result. Centers seeking to implement EP in future must consider these factors to ensure a positive impact on patient safety and outcomes.