Flavonoid content and composition in 124 accessions of forage and grain legumes was assessed by means of HPLC-DAD and HPLC-ESI-MS analyses of seed extracts. Excluding soybean, only 7 out of 77 grain legume accessions exhibited a total flavonoid content higher than 0.1 mg/g fresh seed weight (mean content 0.33 +/- 0.31 mg/ g fresh seed weight); while in 38 out of 47 forage legume, the mean flavonoid content was 1.49 +/- 0.89 mg/g fresh seed weight. On the basis of their higher seed flavonoid content and agronomic relevance, two grain and five forage legume accessions were selected for further HPLC-ESI-MS analyses aimed to identify bioactive compounds. Chemical analysis of the selected accessions permitted the identification of several flavonoids with pharmaceutical properties such as vitexin in Vigna angularis and Trigonella foenum-graecum, quercetin in all Trifolium spp and Medicago sativa, taxifolin in Trifolium pratense and Trifolium alexandrinum, myricetin in Trifolium repens, kaempferol in Phaseolus vulgaris and T. repens and luteolin in M. sativa and T. alexandrinum. Since much of literature information is regarding flavonoid content and composition of the green part of legume species, the obtained results contributed to fill the gap in knowledge about flavonoid content at the seed level.