Technical Abstract: Volatile composition of four southern highbush blueberry cultivars (‘Primadonna’, ‘Jewel’, ‘Snowchaser’, and Kestrel™) grown in two locations (Gainesville and Haines City, Florida) and harvested multiple times was investigated. Volatiles were extracted for chromatographic analysis using a 2-cm, tri-phase SPME fiber, and quantified using internal and external standards. A total of 42 volatiles were identified, including 9 esters, 11 terpenoids, 11 aldehydes, 7 alcohols, and 4 ketones. Twelve of these volatiles are reported for the first time in blueberries. Among them, 10 volatiles were positively identified, including (Z)-3-hexenal, (E, E)-2, 4-hexadienal, (E, Z)-2, 6-nonadienal, (E, E)-2, 4-nonedienal, methyl 2-methylbutanoate, butyl acetate, 2-methylbutyl acetate, and geranyl acetate. Another two, (Z)-dihydrolinalool oxide and (E)-dihydrolinalool oxide, were tentatively identified. The dominant volatiles in these cultivars were aldehydes, accounting for 73-91% of total volatiles. The second most abundant volatiles were terpenoids and esters, accounted for 3.2-18.5% and 0.4-15.5% of total volatiles, respectively. The volatile content in each blueberry cultivar varied in quality and quantity, and resulted in a distinct volatile profile. There were location and harvest date effects on some compounds for ‘Primadonna’, but ‘Jewel’ volatile production was stable regardless of location and harvest.