Purpose: The aim of this study was to evaluate any possible effects of brain motion. CSF pulsations and other possible sources of physiological motion in electrocardiographic (ECG) triggered and non-triggered single-shot echo-planar imaging (EPI) measurements of diffusion. Material and Methods: Three different triggering protocols were evaluated in 6 healthy volunteers: 1) ECG triggering with time delay (TD) 100 ms; 2) ECG triggering with TD 400 ms; and 3) no triggering at all. Results: The results obtained showed that white matter mean apparent diffusion coefficient (ADC) values were similar for all triggering protocols and that the reproducibility in ADC measurements using diffusion-weighted (DW) EPI was good. The average standard deviation of the ADC values was, however, higher for ADC maps obtained without ECG triggering. Conclusion: The use of prospective diastolic ECG triggering significantly improves the accuracy of quantitative diffusion measurements but for routine clinical diffusion imaging, where quantitative data is of less importance, the accuracy obtained without ECG triggering can be considered adequate.