억새뿌리·갯기름나물의 생리활성물질 동정 및 표준화연구

Abstract

The Roots of Miscanthus sinensis var. purpurascens have been used for acidemia and respiratory disease. The roots of Peucedanum japonicum Thunberg have been used for hypertension, diuresis, suppression and menorrhalgia. In this Study, I performed measurment of antioxidative activity, Whitening effect, anti-inflamatory activity and quantitive standardization from the Roots of Miscanthus sinensis var. purpurascens and Peucedanum japonicum Thunberg. 70% MeOH extracts of the Roots of Miscanthus sinensis var. purpurascens and Peucedanum japonicum Thunberg was fractionated by n-Hexane, Methylenchloride (MC), Ethyl acetate (EA) n-Butanol and Water, with five fractions, scavenging activity of DPPH radicals, test of inhibition on tyrosinase, test of inhibition on melanion synthesis by using B16F10 melanoma cell and test of inhbition on NO production by using 264.7 RAW cell were screened to determine antioxidant activity, whitening activity and anti-inflammatory activity. As a result, 70% MeOH extracts, MC and EA fractions of the Roots of Miscanthus sinensis var. purpurascens showed to have good qualified activities. And 70% MeOH extracts and EA fraction of Peucedanum japonicum Thunberg showed to have good qualified activities. Based on this result, components were isolated by activity guided fractionation. In MC fraction of the Roots of Miscanthus sinensis var. purpurascens, two compounds were isolated and elucidated as Methyl 4-hydroxy cinnamate (compound 1) and 4-hydroxybenzaldehyde (compound 2) by physicochemical data spectroscopic methods. In EA fraction of Peucedanum japonicum Thunberg, two compounds were 4-(2-hydroxy-1-methoxyethyl)phenol (compound 3) and 7-hydroxy-6-(2,3-dihy droxy-3-methylbutyl)-2H-chromen-2one(Peucedanol, compound 4) by physicochemical data spectroscopic methods. To investigate the melanogenesis inhibitory effects of these compounds, the melanin level and tyrosinase activity were examined in B16F10 melanoma cell. Compound 1 inhibited both tyrosinase activity and melanin synthesis compared with positive control arbutin. Quantitative analysis methods were developed by HPLC. Validation for verification of HPLC methods was carried out to check to specificity, linearity, intra-day precision, inter-day precision and accuracy by ICH guideline. At the same time, quantitative HPLC methods were established to measure content of each standard compound on extracts. These result suggest that methyl 4-hydroxycinnamate from he Roots of Miscanthus sinensis var. purpurascens might be a potent inhibitor against melanogenesis and these compounds could be further developed as skin whitening cosmeceuticals.