Research has highlighted the nutritional benefits of ancient grains, especially emmer (Triticum dicoccum) and einkorn (Triticum monococcum), compared to modern wheat varieties, focusing on their higher levels of antioxidants and phytochemicals [1-4]. In this study, grains from traditional old Greek wheat varieties were evaluated and compared to commercial cultivars, like Simeto and Mexicali 81, in an effort to assess differences and eventually valorize local products. The samples studied in this research included emmer and einkorn as well as common wheat and durum wheat grains. The Internal Transcribed Spacer 2 (ITS2) nuclear region was amplified and sequenced as a barcode for species identification, allowing einkorn discrimination. The total content of bound and free polyphenols and flavonoids, as well as the antioxidant activity of bound and free compounds was also determined. Additionally, the level of variation of fatty acids was determined in all samples by gas chromatography. The results showed that local old landraces of emmer and einkorn had the highest polyphenol content, 2.2 and 3.1 times higher than the average value of durum wheat, respectively. Regarding the total flavonoid content einkorn and emmer had higher content than common wheat. The antioxidant activity of free or bound compounds was of the same level, at about 20-25% higher in both einkorn and emmer compared to common wheat. The results of this study demonstrate the high nutritional value of old local landraces underlining the importance of maintaining genetic material able to perform in a low input and environmental sustainable agriculture.