The influence of four day/night growing temperature combinations (18/12, 25/12, 25/22, and 30/22 C) on phenolic acid, flavonol, and anthocyanin content and their antioxidant activities against peroxyl radicals (ROO), superoxide radicals (O2-), hydrogen peroxide (H2O2), hydroxyl radicals (OH), and singlet oxygen (1O2) in fruit juice of Earliglow and Kent strawberry (Fragaria ananassa Duch.) cultivars was studied. Pelargonidin-based anthocyanins such as pelargonidin 3-glucoside (291.3-945.1 g/g fresh wt.), pelargonidin 3-rutinoside (24.7-50.9 g/g fresh wt.), and pelargonidin 3-glucoside-succinate (62.2-244.0 g/g fresh wt.) were the predominant anthocyanins in strawberry fruit juice. The content of cyanidin-based anthocyanins, cyanidin 3-glucoside and cyanidin 3-glucoside-succinate, was much lower than that of pelargonidin-based anthocyanins. Strawberry growth in high temperature conditions significantly enhanced the content of p-coumaroylglucose, dihydroflavonol, quercetin 3-glucoside, quercetin 3-glucuronide, kaempferol 3-glucoside, kaempferol 3-glucuronide, cyanidin 3-glucoside, pelargonidin 3-glucoside, pelargonidin 3-rutinoside, cyanidin 3-glucoside-succinate, and pelargonidin 3-glucoside-succinate in strawberry juice. Plants grown in the cool day and cool night temperature (18/12 C) generally had the lowest phenolic acid, flavonols, and anthocyanins. An increase in night temperature from 12 to 22 C, with the day temperature kept constant at 25 C, resulted in a significant increase in phenolic acid, flavonols, and anthocyanins. These conditions also resulted in a significant increase in antioxidant capacity. The highest day/night temperature (30/22 C) yielded fruit with the most phenolic content as well as ROO, O2-, H2O2, OH, and 1O2 radical absorbance capacity. Fruit of Kent cv. strawberry had higher values of phenolic acid, flavonols, anthocyanins, and antioxidant capacities than fruit of Earliglow cv. strawberry under all temperature regimes.