SummaryThe COVID‐19 pandemic has highlighted the importance of environmental ventilation in reducing airborne pathogen transmission. Carbon dioxide monitoring is recommended in the community to ensure adequate ventilation. Dynamic measurements of ventilation quantifying human exhaled waste gas accumulation are not conducted routinely in hospitals. Instead, environmental ventilation is allocated using static hourly air change rates. These vary according to the degree of perceived hazard, with the highest change rates reserved for locations where aerosol‐generating procedures are performed, where medical/anaesthetic gases are used and where a small number of high‐risk infective or immunocompromised patients may be isolated to reduce cross‐infection. We aimed to quantify the quality and distribution of ventilation in hospital by measuring carbon dioxide levels in a two‐phased prospective observational study. First, under controlled conditions, we validated our method and the relationship between human occupancy, ventilation and carbon dioxide levels using non‐dispersive infrared carbon dioxide monitors. We then assessed ventilation quality in patient‐occupied (clinical) and staff break and office (non‐clinical) areas across two hospitals in Scotland. We selected acute medical and respiratory wards in which patients with COVID‐19 are cared for routinely, as well as ICUs and operating theatres where aerosol‐generating procedures are performed routinely. Between November and December 2022, 127,680 carbon dioxide measurements were obtained across 32 areas over 8 weeks. Carbon dioxide levels breached the 800 ppm threshold for 14% of the time in non‐clinical areas vs. 7% in clinical areas (p < 0.001). In non‐clinical areas, carbon dioxide levels were > 800 ppm for 20% of the time in both ICUs and wards, vs. 1% in operating theatres (p < 0.001). In clinical areas, carbon dioxide was > 800 ppm for 16% of the time in wards, vs. 0% in ICUs and operating theatres (p < 0.001). We conclude that staff break, office and clinical areas on acute medical and respiratory wards frequently had inadequate ventilation, potentially increasing the risks of airborne pathogen transmission to staff and patients. Conversely, ventilation was consistently high in the ICU and operating theatre clinical environments. Carbon dioxide monitoring could be used to measure and guide improvements in hospital ventilation.