The recent increase in demand for Okinawan pineapples has necessitated the development of new varieties with attractive aromas. This study aimed to evaluate the volatile characteristics of five Okinawan pineapple breeding lines, i.e., ‘No. 22’, ‘No. 25’, ‘No. 26’, ‘No. 27’, and ‘No. 28’. The total volatiles in the cryopulverized fruit flesh were examined using headspace gas-chromatography–mass-spectrometry-based electronic nose analysis. The total ion masses of the volatiles were visualized using principal component analysis, and three replicates of each line with comparable volatile characteristics were selected. Furthermore, the composition of the volatile components in these replicates was assessed, and the odor activity values (OAVs) were calculated. The breeding lines varied in the quantity and composition of their volatile compounds, which were predominantly esters, ketones, terpenes, and alcohols. The ‘No. 22’ fruit contained a greater content of volatiles than the other lines. Moreover, 14 volatiles with OAV > 1 were accounted as aroma-active compounds, and their variations were distinguished as follows: the highest OAV (786.96) was recorded for methyl 2-methylbutanoate of the ‘No. 26’ line; 2,5-dimethyl-4-methoxy-3(2H)-furanone was superior in the ‘No. 26’ and ‘No. 27’ lines; and δ-decalactone was only present in the ‘No. 22’ and ‘No. 27’ fruits, suggesting different potent practical uses for these new breeding lines.
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