Free radical scavenging actions of three Trifolium species in the protection of blood plasma antioxidant capacity in vitro

Context: Three clover [Trifolium L. (Leguminosae)] species were selected on the basis of data from traditional medicine, phytochemical profiles, and agricultural significance. Objective: The in vitro evaluations of free radical scavenging properties, ferric reducing abilities, and antioxidant effects of extracts from T. pratense L. (crude extract and phenolic fraction), T. pallidum L., and T. scabrum L. (phenolic fractions) were performed. Materials and methods: Activities of the Trifolium extracts were determined at their final concentrations of 1.5-50 mu g/ml. Free radical scavenging properties of methanol extract solutions were estimated by the reduction of DPPH center dot and ABTS(center dot) radicals. Measurements of the total antioxidant capacity (TAC) were carried out to assess the antioxidant activities of the extracts in human blood plasma under conditions of oxidative stress, induced by 200 mu M peroxynitrite. Results: The phenolic fraction of T. pratense displayed the strongest ABTS(center dot) and DPPH center dot radical scavenging effects (EC50 value of 21.69 and 12.27 mu g/ml, respectively). The EC50 value for T. pallidum extract attained 29.77 and 30.06 mu g/ml. The two remaining extracts were less potent scavengers (EC50 value higher than 50 mu g/ml). Similar differences were obtained during evaluation of the ferric reducing abilities. Analysis of antioxidant properties of the extracts in blood plasma did not provide such evident differences in their actions, however, it indicated that the T. pratense phenolic fraction displayed the strongest effect. Conclusions: The examined Trifolium extracts partly protected blood plasma and enhanced its non-enzymatic antioxidant defense against harmful action of peroxynitrite in vitro.