Simple SummaryBleeding disorders can cause life-threatening illness in dogs. The need for fast recognition and diagnosis of these conditions is therefore of the utmost importance to have a positive impact on the patients’ survival. In the past decade, the use of viscoelastic testing for rapid assessment of global haemostasis has gained popularity. However, the most reliable time for testing after blood collection has not been determined. For this reason, blood samples were taken from healthy client-/staff-owned dogs and repeated measurements were performed at three different time points (10 min, 30 min, and 70 min after blood collection). Additionally, a group of currently ill patients was included and Ex-TEM S measurements were performed at the same three timepoints. We found that there was a significant change of results over time, suggesting the need for time-specific reference intervals. Which of these time points reflects the “true” coagulation status of our patients currently remains unknown.Viscoelastic testing as a bedside test to assess global haemostasis has gained popularity in the past decade, with rotational thromboelastometry (ROTEM) and thromboelastography (TEG) being the two commonly used devices. TEG studies suggest analysis 30 min after blood sampling. However, the reproducibility of results over time for ROTEM analysis using lyophilized samples in dogs has not been established. In this study, we investigated the influence of time on viscoelastic testing, using 33 healthy staff-/client-owned dogs for blood sampling and repeated measurements of ROTEM tracings at three different time points after blood collection. Additionally, a group of 21 hospitalized patients with suspected coagulation disorders were included to investigate whether stability over time was comparable between healthy and ill dogs. We demonstrated a significant difference of ROTEM tracings over time, with a tendency towards hypocoagulability over time. These changes do have a clinical relevance as they exceed reference intervals and could therefore lead to erroneous conclusions about a patient’s coagulation status. Therefore, time-specific reference intervals are proposed and presented in this publication.