Isolation and Bioactivity Analysis of Ethyl Acetate Extract from Acer tegmentosum Using In Vitro Assay and On-Line Screening HPLC-ABTS(+) System.

Kwang Jin Lee,Won-Kyung Cho,Na-Young Song,You Chang Oh,Jin Yeul Ma

doi:10.1155/2014/150509

Abstract

The Acer tegmentosum (3 kg) was extracted using hot water, and the freeze-dried extract powder was partitioned successively using dichloromethane (DCM), ethyl acetate (EA), butyl alcohol (n-BuOH), and water. From the EA extract fraction (1.24 g), five phenolic compounds were isolated by the silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatography. Based on spectroscopic methods such as 1H-NMR, 13C-NMR, and LC/MS the chemical structures of the compounds were confirmed as feniculin (1), avicularin (2), (+)-catechin (3), (−)-epicatechin (4), and 6′-O-galloyl salidroside (5). Moreover, a rapid on-line screening HPLC-ABTS+ system for individual bioactivity of the EA-soluble fraction (five phenolic compounds) was developed. The results indicated that compounds 1 and 2 were first isolated from the A. tegmentosum. The anti-inflammatory activities and on-line screening HPLC-ABTS+ assay method of these compounds in LPS-stimulated murine macrophages were rapid and efficient for the investigation of bioactivity of A. tegmentosum.

Highlights

Traditional remedies based on natural products could be traced back over five millennia to written documents of the early civilizations [1]
This study showed that among the soluble fractions from the hot water extract of A. tegmentosum, the ethyl acetate (EA)-soluble fraction
Compounds in the dried twigs of A. tegmentosum were extracted with hot water and partitioned successively using DCM, EA, normal butyl alcohol (n-BuOH), and water

Summary

Introduction

Traditional remedies based on natural products could be traced back over five millennia to written documents of the early civilizations [1]. DPPH (ABTS) radical is another simple, rapid on-line method for the detection of antioxidants from crude plant extracts [12] It combines HPLC with an assay involving a stable radical species [1,1-diphenyl2-picrylhydrazyl radical (DPPH) and 2,2󸀠-azinobis-(3ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS)] in the HPLC-DPPH (ABTS) method [13, 14]. This method was successfully applied for the screening and identification of natural bioactive compounds from complex mixtures, for the extracts of OMHs [15,16,17]. We investigated the applications of on-line screening HPLC-ABTS+ assays for bioactivity screening to find a more practical approach toward the use of on-line screening HPLC-ABTS+ assays for the rapid pinpointing of peaks in chromatograms corresponding to bioactive compounds

Experimental

Results and Discussion

Anti-Inflammatory Activities Screening

Conclusions