Journal
JOURNAL OF THE KOREAN SOCIETY FOR APPLIED BIOLOGICAL CHEMISTRY
Volume 58, Issue 5, Pages 715-722Publisher
KOREAN SOC APPLIED BIOLOGICAL CHEMISTRY
DOI: 10.1007/s13765-015-0097-8
Keywords
Antioxidant activity; Dermal fibroblast cells; Ethyl acetate; Rosmarinus officinalis L
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Funding
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - Ministry of Education [2014047495]
- Ministry of Knowledge Economy (MKE), Korea Institute for Advancement of Technology (KIAT)
- Jeju leading industry office through the Leading Industry Development for Economic Region
- National Research Foundation of Korea [2009-0094059] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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The solvent fractions of rosemary methanol extract were obtained by successive extraction with n-hexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction (EAF) contained a remarkable amount of polyphenol and flavonoid as well as high levels of alkyl and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity. The activity guided fractionation and repeated chromatographic separations over silica gel, RP C18, and Sephadex LH-20 led to isolation of six compounds from the EAF. H-1 NMR, C-13 NMR, 2D NMR, MS, and IR spectroscopies determined the compounds to be caffeic acid (1), rosmarinic acid (2), rosmarinic acid methyl ester (3), luteolin (4), apigenin (5), and hispidulin (6), and high-performance liquid chromatography quantification was used to determine concentrations in EAF. Among the six isolated compounds, rosmarinic acid methyl ester showed the highest scavenging activities against di(phenyl)-(2,4,6-trinitrophenyl) iminoazanium, alkyl and ABTS radicals. The EAF mixture, but not individual isolated compounds, shielded dermal fibroblast cells from H2O2-induced cytotoxicity at concentrations that encompass the SC50 of alkyl and ABTS radical. Therefore, our findings suggested for the first time that antioxidant capacity of the EAF mixture result in a synergistic effect on the antioxidant action.
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