BACKGROUND ROTEM(®) test results can be affected by the citrate to blood ratio in the tested sample. Overfilling or underfilling specimen tubes can change this ratio. OBJECTIVES The aim of this study was to determine the impact of under- and overfilling citrate test tubes on ROTEM(®) EXTEM measurements. RESULTS Overall repeated measures ANOVA demonstrated significant differences of CT (p = 0.004), CFT (p = 0.005), A5 (p = 0.001), A10 (p <0.001), and MCF (p < 0.001). Pairwise comparison revealed that underfilling significantly altered the results of those parameters reflecting functional clot firmness (A5, A10, and MCF), while overfilling led to a prolongation of the CT results only. No differences were observed for alpha angle and maximum lysis. CONCLUSION Both underfilling and overfilling specimen tubes have significant influence on the results of the ROTEM(®) EXTEM test, although the small observed bias is likely of limited clinical relevance. However, it seems prudent to limit the maximum allowed difference in filling to less than ± 10%. All ROTEM(®) operators should be aware of this pre-analytical variable.