Structural characterization of phenolic antioxidants from purple perilla (Perilla frutescens var. acuta) leaves

The antioxidant activities of various extracts from purple perilla (Perilla frutescens var. acuta) leaves based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, 2,2-azino-bis-(3-ethylenebenzothiozoline-6-sulfonic acid) (ABTS) radical cation scavenging ability, and reducing power were investigated. Purple perilla leaves were initially extracted with 80% ethanol and then sequentially fractionated according to solvent polarity. Of all fractions, the ethyl acetate fraction had the highest antioxidant activity. This fraction was separated into four sub-fractions (sub-fractions 1-4) by Sephadex LH-20 chromatography. Of the four sub-fractions, sub-fraction 3 had the highest antioxidant activity. The EC₅₀ values of sub-fraction 3 for DPPH radical scavenging activity and reducing power were 2.4 and 1.7 times lower than those of butylated hydroxytoluene (BHT), respectively. Based on HPLC analysis, the most abundant phenolic acid in sub-fraction 3 of purple perilla leaves was rosmarinic acid, at 314.3 mg/g. The structure of rosmarinic acid was confirmed by ESI-IT-TOF MS and NMR analysis.