Strawberry fruits are highly susceptible to cold burning, resulting in low storage periods at low temperatures. Plant extracts or essential oils (EOs) can potentially be used as preservatives in fruits throughout the refrigerated period. In the present study, the biochemicals, antioxidant characteristics, and shelf life of treated strawberries with Aloysia citrodora essential oil (ACEOs) were evaluated during keeping time. The treatments were produced as follows: T1, control; T2, 250 ppm ACEOs; T3, 500 ppm ACEOs; and T4, 750 ppm ACEOs. Total soluble solids (TSS), weight loss, titratable acidity (TA), antioxidant activity (DPPH assay), total phenolic (TPC), flavonoid and anthocyanin contents (TFC), and enzymes activity (peroxidase and ascorbate peroxidase) were evaluated during the refrigerated period (5 °C with relative humidity of 85–90% for 20 days). The results revealed that weight loss and TA were reduced in all treatments during storage, being that the rates were lower in samples treated with ACEOs. TPC, TFC, TSS, antioxidant, and enzymes activity were higher in treated fruits than control.