Incrocio Bruni 54 is a little-known white grape variety developed in the Marche region (Italy) from a cross between Verdicchio and Sauvignon Blanc to combine aromatic freshness with structure. In light of the growing interest in minor and autochthonous cultivars, this study provides the first comprehensive chemical characterization of the aroma profile of Incrocio Bruni 54 wines. Seventeen commercial wines were analyzed for varietal compounds, such as terpenes, norisoprenoids, volatile thiols, methyl salicylate and its glycosides, and fermentative compounds, including esters, alcohols, acids, phenols, aldehydes, and ketones, using GC-MS/MS and LC-MS/MS. Odor activity value (OAV) calculations revealed an aroma profile dominated by ethyl esters, such as ethyl caproate and isopentyl acetate, β-damascenone, 4-vinylguaiacol, TDN, and the volatile thiols 3MH and 4MMP, imparting fruity, floral, spicy, and tropical notes. Comparison with datasets of 246 Italian monovarietal white wines and related sub-datasets composed of Verdicchio and Lugana showed significantly higher concentrations of 3MH and free methyl salicylate in Incrocio Bruni 54, but markedly lower levels of glycosylated methyl salicylate forms, suggesting a greater expression of this odorant in young wines balanced by a lower potential over aging. These findings highlight the distinctive aromatic fingerprint of Incrocio Bruni 54, combining parental traits with unique sensory potential, and support its knowledge and valorization in wine production.