Previously, when selecting for flavor in the University of Florida southern highbush blueberry (SHB, Vaccinium corymbosum L. hybrids) breeding program, sugar/acid ratios and breeder preference were the only factors considered. A more precise method of evaluating flavor would include volatile compounds that may also contribute to the flavor experience. Therefore, volatile profiles of five SHB cultivars (Farthing, FL01-173, Scintilla, Star, and Sweetcrisp) were compared using gas chromatography–mass spectrometry. All cultivars were harvested on four separate dates within the harvest season, and fruit from each cultivar were also harvested at four developmental stages on the first harvest date. Among the cultivars, soluble solids content and volatile production tended to increase with fruit maturity, whereas titratable acidity decreased. All volatile components were more variable than measures of sugars and acids during the harvest season. Many of the volatiles present varied significantly between harvest dates, resulting in significant genotype × environment interactions during the harvest season. A closer examination of linalool, trans-2-hexenol, trans-2-hexenal, hexanal, and 1-penten-3-ol, five volatile compounds commonly associated with blueberry flavor, showed cultivar, developmental stage, and harvest date differences for each volatile. ‘Star’ experienced the least variation through the harvest period.