The assessment of the functional status of the salivary glands has been used in the scintigraphic evaluation of xerostomia. Several quantitative methods derived from standard dynamic scintigraphy have been suggested. However, the indices proposed are quite variable and unlikely to be useful in clinical practice. The objectives of this study were to obtain reference values of major salivary glands uptake and excretion fraction in healthy subjects and to obtain normal ratios of Tc-pertechnetate uptake by the major salivary glands in comparison to the thyroid gland uptake. The standardization of these values has the purpose of making this evaluation faster and more objective. Fifty volunteers without clinical evidence of xerostomia or thyroid disease underwent static salivary glands scintigraphy with Tc-pertechnetate. Static images were obtained at 20 minutes and then at 3 minutes after oral stimulation with lemon juice. Percent uptake, excretion fraction and salivary gland to thyroid ratio rates were calculated for the parotid and the submandibular glands. The mean of the uptake values at 20 minutes for the right and left parotid glands were respectively 0.31% and 0.26%, and for the submandibular glands 0.15%. The excretion fraction of the tracer after the lemon juice stimulation was 70% for the parotids glands, 50% for the right and 49% for the left submandibular glands. The mean+/-SD salivary gland to thyroid count ratio was 0.79+/-0.45 for the right parotid, 0.78+/-0.5 for the left parotid, 0.67+/-0.33 and 0.66+/-0.34 for the right and left submandibular glands, respectively. Salivary glands scintigraphy with uptake and excretion fraction calculation is an easy to perform, non-invasive and objective method to investigate salivary glands function. These findings help the nuclear physician to interpret salivary gland scintigraphy more objectively, even in patients with thyroid gland dysfunction in whom Tc-pertechnetate thyroid uptake may be abnormal.