Folate (Vitamin B9) is critical for a range of biological functions in adults and children, including DNA, protein and neurotransmitter synthesis. It is also essential for the healthy development of the foetus in early pregnancy and for the prevention of neural tube defects, such as spina bifida. Strawberries are considered a tasty and healthy fruit consumed all over the world and may potentially be an important dietary source of natural folates. However, the relative importance of strawberry as a dietary source will depend on the total folate concentration, vitamer profile, storage stability and bioavailability to humans. Red Rhapsody, an important commercial strawberry cultivar in Australia, was screened for its folate content and storage stability by stable isotope dilution assay (SIDA). Total folate content ranged from 90-118 μg/100 g fresh weight (fw), which was well above the value in the Australian Food Composition Database (39 μg/100 g fw). 5-Methyltetrahydrofolate, the biologically active form in humans, was the principal vitamer present. Furthermore, folate remained relatively stable during refrigerated (4°C) storage (loss of only 28% after 14 days of storage). This information is relevant for consumers since the inherent perishability of strawberry fruit makes refrigerated storage common practice in Australian households.