There is marked variability in the cerebral blood flow (CBF) between the ictal and interictal state in epilepsy, and it would therefore be desirable to increase the reliability of ictal/interictal single-photon emission tomography (SPET) difference images. We aimed to improve the step of quantitative normalization of images by finding the best possible reference region. In 16 patients (11 with lateralization of the epileptogenic focus, five with bilateral foci) both ictal and inter-ictal SPET scans were performed after injection of technetium-99m labelled tracer. Then, each region among a selected set (brain+cerebellum, brain, cerebellum, hemispheres, and for patients with an expected lateralization, cortical lobe containing the focus and symmetrical contralateral lobe) was investigated by comparison of the regional ictal/inter-ictal variance in counts. Among patients with a suspected lateralized focus, the distribution of CBF in the contralateral cortical lobe appeared to vary less between ictal and inter-ictal states than in other investigated areas. As a consequence, this latter region constitutes the best choice as a reference region. For patients with bilateral foci, the cerebellum appears to be a good compromise even though it presents with significant CBF changes.