The demand for natural Vanilla has increased rapidly, creating the need for more potential sources of high-quality Vanilla essence. Understanding the geographical influences on the aroma profile of Vanilla is essential. This study demonstrates the first comparative analysis of odorant compositions in the three most important Vanilla varieties: Vanilla planifolia, Vanilla pompona, and Vanilla tahitensis from different origins. Following the screening for odor-active molecules through gas chromatography-olfactometry and aroma extract dilution analysis (GC-O and AEDA), selected compounds were quantified using stable isotope dilution assays (SIDA) and their dose over threshold values (DoTs) were calculated. Vanillin was confirmed as the most important odor-active compound due to its highest DoT value, especially in the V. planifolia sample. Meanwhile, 4-methoxybenzyl alcohol and 4-methoxybenzaldehyde showed higher DoT factors than vanillin in V. pompona and partially in V. tahitensis samples. This indicates their role as discriminative odorants for these varieties. The heightened DoT values of 3-hydroxy-4,5-dimethyl-2(5H)-furanone in Uganda Vanilla samples unveil geographical influences on the odorant profile within V. planifolia species. Additionally, 2-methyl-3-(methyldithio)furan was identified for the first time in Vanilla samples with diverse DoT values from different species and origins.