The isomers of 2,3-butanediol [R,R; S,S; R,S (meso-form)] and of acetoin (R,S) were determined in laboratory wine fermentations carried out by 50 yeast strains, 10 for each of the following species, Saccharomyces cerevisiae, Kloeckera apiculata, Candida stellata, Metschnikowia pulcherrima and Brettanomyces bruxellensis, in order to evaluate if such parameters might be used to differentiate wines obtained with different yeast species. According to analysis of variance (ANOVA), the strains of the same species behaved similarly, whereas the five yeast species behaved differently so that species-specific profiles were recognized. Moreover, the discriminant analysis grouped the wines into five groups, each including the 10 wines obtained by the 10 yeast strains of the same species. Trials were also included where musts partially fermented by non-Saccharomyces species were inoculated with a selected strain of S. cerevisiae to complete fermentation, and the content in 2,3-butanediol and acetoin isomers was again determined and statistical analysis was performed. Although the final values of these parameters resembled those obtained in pure fermentation with S. cerevisiae, statistical analysis discriminated wines according to the yeast species performing the first fermentation phase.