Preanalytical variability is a common source of errors in coagulation testing, as clotting assays are particularly susceptible to poor standardization of the whole analytical process. To investigate the effect of a short-term venous stasis on routine coagulation testing, we measured activated partial thromboplastin time, prothrombin time, fibrinogen and D-dimer in plasma specimens collected either without venous stasis or following the application of a 60 mmHg constant, standardized external pressure by a sphygmomanometer, for 1 (1-min stasis) and 3 min (3-min stasis). When compared with blood specimens collected without stasis, the Pearson's correlation coefficients and the corresponding slopes of the Passing and Bablok regression line of samples collected following 1 and 3-min stasis were acceptable. However, statistically significant differences by paired Student's t-test could be observed for all parameters tests following 3-min stasis, and for all but the activated partial thromboplastin time after 1-min stasis. Significant difference between specimens collected after 1- and 3-min stasis was also achieved for prothrombin time (P < 0.01), fibrinogen (P < 0.01) and D-dimer (P < 0.05). The agreement between measurements was yet acceptable after 1-min stasis, but achieved clinical significance for prothrombin time, fibrinogen and D-dimer after 3-min stasis. Taken together, results of the present investigation confirm that the effects of venous stasis during venipuncture are clinically meaningful. As hematocrit values and activities of clotting factors VII, VIII and XII significantly increased, whereas that of activated factor VII remained unchanged, we hypothesize that a short-term venous stasis, as induced by up to 3-min tourniquet placing, might not be sufficient to produce additional procoagulant responses besides hemoconcentration.