The impact of harvest time on the physicochemical properties and antioxidant activity of Seolhyang strawberry varieties was evaluated. The Hunter color value, which indicates redness on the surface of strawberries, was significantly higher for strawberries with a third flower cluster. Our results also revealed that the strawberry pH tended to increase significantly with extension of harvest time. Soluble solids in strawberries from the fifth flower cluster were significantly lower throughout the harvest period. Strawberries from the first flower clusters exhibited the highest acidity, while strawberries from the third flower clusters held the lowest acidity. Strawberry antioxidant properties assessment revealed that total polyphenol and anthocyanin content tended to increase with delay of harvest time. Vitamin C content tended to significantly increase with increasing flower cluster number until cluster 4. Fifth flower cluster strawberries showed decreased vitamin C. Sensory acceptability scores for second-flower cluster strawberries were significantly higher than those of the others for appearance, taste, and overall acceptability. This study confirmed important impacts of harvest time on the physicochemical and sensory properties of strawberries. We expect that this data can be used to guide the development of processed strawberry products.