Identification of Biological Effective Constituents from the Potentilla supina

Potentilla supina (PS) is a member of the Rosaceae family that is native to China, Japan, Korea, India, and Malaysia. It produces one or more erect stems from a branching caudex and system of rhizomes. It grows 20 to 60 cm tall, and is slightly hairy to nearly hairless. The leaves are ternate, divided into three leaflets. The basal leaves are largest, borne on long petioles. Each has oval leaflets up to 3-cm long that are deeply cut into blunt teeth. Smaller leaves occur higher on the stem. The inflorescence is a cyme of one or more flowers. The flower has usually five yellow petals up to 1-cm long on a calyx of pointed sepals and narrower pointed bractlets. Potentilla supina is often used in Korean traditional systems of medicine as a remedy for hemostasis, dysentery cough, pertussis, sore throat, and external bleeding. Generally, drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. The purpose of this study was to determine whether the total polyphenol content, anti-oxidative and anti-inflammatory activities were examined on the extract from PS and their biologically active constituents were identified. The total polyphenol contents appeared to be highest in EtOAc fraction(316.4 ± 11.8 mg/g) of PS. The DPPH radical scavenging activity was highest in an EtOAc fraction of PS of 95.4 ± 0.3% at a concentration of 250 μg/mL (P < 0.05).The anti-inflammatory activities of EtOAc fraction of PS was evaluated for inhibitory activities against lipopolysacchride(LPS) induced nitric oxide(NO) and prostaglandin E₂(PGE₂) production in RAW264.7 celllines. The EtOAc fraction of PS was high inhibitory activity for both tests with IC₅₀ values showed in the ranges of 29.34~50.75 μg/mL. The biological effective compounds in the whole parts of PS were isolated by a bioassay guided purification using the anti-oxidative and anti-inflammatory activities. The repeated column chromatography of ethyl acetate-soluble layer in this ethanol extract led to isolation two constituents such as caffeic acid (1) and quercitrin (2). The structures of these compounds were identified by spectroscopic methods and by comparing their data to those in the literature. As far as we know, two compounds were isolated for the first time from this plant. Both compounds were already reported ingredients but the two compounds are considered to exhibit a high physiological activity. This result revealed that EtOAc fraction of PS is expected to be good candidate for development into source of prophylactic agent.