Purpose: This study aimed to identify the effect of work interruption that influenced workload and perception of patient safety culture on ward nurses.Methods: Participants were 184 ward nurses, with more than 12 months of work experience, from two tertiary hospitals in S city. A descriptive correlational study design was used. Collected data were analyzed using descriptive statistics, t-test, ANOVA, Mann-Whitney U test, Kruskal-Wallis test, Pearson's correlation, and hierarchical multiple

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... ssion analysis via SPSS version 27.0.Results: The results showed that general wards nurses had a higher degree of work interruption (t=5.632, <i>p</i><.001) and workload (t=3.603, <i>p</i><.001) compared to comprehensive nursing care service wards nurses. More interruption in nursing work caused more burden on work (γ=.440, <i>p</i><.001) and led to lesser perception of patient safety culture (γ=-.199, <i>p</i>=.007). Finally, the regression analysis showed that work interruption had a statistically significant relevance on nurses' workload (F=20.582, <i>p</i><.001) and perception of patient safety culture (F=8.792, <i>p</i><.001).Conclusion: To alleviate ward nurses' work interruption and decrease the negative effect on workload and perception of patient safety culture, it is necessary to mediate nurse staffing level and the number of assistants and utilize the environment.