Phenolic content and DPPH radical scavenging activity of the flowers and leaves of Trifolium repens.

In order to facilitate the quality control of Trifolium repens flowers and leaves, a RP-HPLC method with UV detection was developed for the simultaneous quantitative determination of flavonols and isoflavones. The total flavonoid and phenolic (TPC) contents were determined spectrophotometrically in a visible part of the light spectrum at 425 and 760 nm, respectively. Additionally, evaluation of the antioxidant properties of the plant materials was performed using the DPPH in vitro test. The results showed that the flowers are the richest source of phenolics ranging from 28.7 to 38.8 mg GAE/g, and flavonoids, calculated for hyperoside, up to 20 mg HP/g, which hydrolyzed mainly to flavonols (the quercetin level greater than 6 mg/g). T. repens is poor in isoflavones; similar quantities ofca. 0.2 mg/g were detected in the flowers and leaves. The flower and leaf extracts showed antioxidant activity towards DPPH with EC50 values ranging from 72.3 to 179.3 microg/mL. Significant linear correlations were found between antioxidant potentials of the studied plant materials and total phenolic and flavonoid contents determined by HPLC and spectrophotometric methods (R2 in the range of 0.97 - 0.99).