A small number of sweetening compounds, low in calories and with sensory profiles similar to sucrose, are commercially used in various food products. Among them, it is notorious the interest that in the last decade have generated natural sweeteners, particularly glycoside obtained from leaves of Stevia rebaudiana because of its high sweetness and synergism with other sweeteners. The objective of this study was the characterization of two stevia cultivars coming from Venezuela, Colombia and France to establish differences according to genotype and place of origin. The characterization included particle size, dry matter, matter insoluble in alcohol (MIA), ash, pH, acidity, fat, protein, polyphenols, total soluble solids (TSS), color and turbidity in dehydrated leaves, along with quantification of steviolglycosides (stevioside and rebaudioside A). Similar values were found in terms of ash, protein, pH, acidity and polyphenols in samples of different origin. The Morita II cultivar showed similarity between the Venezuelan and Colombian samples for most of the variables. The French clonal variety recorded higher values of dry matter, MIA, grease and turbidity, lower values of TSS and color, and no trend in grain size. With regard to the contents of steviolglycosides, the cultivar Morita II showed higher proportion of rebaudioside A, while in French clonal variety the stevioside showed the highest proportion. Both of them presented good thermal stability. Given the higher yield and better sensory profile of rebaudioside A, Morita II, both Venezuelan and Colombian, looks promising to be used as a substitute for sucrose.