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

Lee KJ, Song NY, Oh YC, Cho WK, Ma JY - J Anal Methods Chem (2014)

Bottom Line: Based on spectroscopic methods such as (1)H-NMR, (13)C-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).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.

View Article: PubMed Central - PubMed

Affiliation: Korean Institute of Oriental Medicine (KIOM), KM-Based Herbal Drug Development Group, 1672 Yuseongdae-ro, Yuseong-gu, Daejeon 305-811, Republic of Korea.

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 (1)H-NMR, (13)C-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.

No MeSH data available.


Effect of five compounds on the production of (a) TNF-α, (b) IL-6, and (c) IL-1β cytokines in macrophages. Cells were pretreated with five compounds for 30 min before being incubated with LPS for 24 h. Production of cytokines was measured by ELISA. Data shows mean ± SE values of duplicate determinations from three independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.
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fig9: Effect of five compounds on the production of (a) TNF-α, (b) IL-6, and (c) IL-1β cytokines in macrophages. Cells were pretreated with five compounds for 30 min before being incubated with LPS for 24 h. Production of cytokines was measured by ELISA. Data shows mean ± SE values of duplicate determinations from three independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.

Mentions: The inhibitory effect of the five compounds on the production of inflammatory cytokines, another parameter of inflammation, was investigated. In this study, we examined the effect of the five compounds on TNF-α, IL-6, and IL-1β expressions. Figure 9(a) shows that feniculin slightly repressed TNF-α production at a concentration of 10 μM (Figure 9(a)). However, avicularin, (+)-catechin, (−)-epicatechin, and 6′-O-galloyl salidroside did not show any inhibitory effect on LPS-induced TNF-α production. As shown in Figure 9(b), consistent with TNF-α results, feniculin slightly repressed IL-6 production at a concentration of 10 μM. Moreover, avicularin inhibited IL-6 production at concentrations of 30 and 50 μM. (+)-Catechin inhibited IL-6 production at 30 μM or more. (−)-Epicatechin and 6′-O-galloyl salidroside did not show any inhibitory effect on LPS-induced IL-6 secretion. However, all the compounds except 6′-O-galloyl salidroside did not inhibit IL-1β (Figure 9(c)).


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

Lee KJ, Song NY, Oh YC, Cho WK, Ma JY - J Anal Methods Chem (2014)

Effect of five compounds on the production of (a) TNF-α, (b) IL-6, and (c) IL-1β cytokines in macrophages. Cells were pretreated with five compounds for 30 min before being incubated with LPS for 24 h. Production of cytokines was measured by ELISA. Data shows mean ± SE values of duplicate determinations from three independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4216704&req=5

fig9: Effect of five compounds on the production of (a) TNF-α, (b) IL-6, and (c) IL-1β cytokines in macrophages. Cells were pretreated with five compounds for 30 min before being incubated with LPS for 24 h. Production of cytokines was measured by ELISA. Data shows mean ± SE values of duplicate determinations from three independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.
Mentions: The inhibitory effect of the five compounds on the production of inflammatory cytokines, another parameter of inflammation, was investigated. In this study, we examined the effect of the five compounds on TNF-α, IL-6, and IL-1β expressions. Figure 9(a) shows that feniculin slightly repressed TNF-α production at a concentration of 10 μM (Figure 9(a)). However, avicularin, (+)-catechin, (−)-epicatechin, and 6′-O-galloyl salidroside did not show any inhibitory effect on LPS-induced TNF-α production. As shown in Figure 9(b), consistent with TNF-α results, feniculin slightly repressed IL-6 production at a concentration of 10 μM. Moreover, avicularin inhibited IL-6 production at concentrations of 30 and 50 μM. (+)-Catechin inhibited IL-6 production at 30 μM or more. (−)-Epicatechin and 6′-O-galloyl salidroside did not show any inhibitory effect on LPS-induced IL-6 secretion. However, all the compounds except 6′-O-galloyl salidroside did not inhibit IL-1β (Figure 9(c)).

Bottom Line: Based on spectroscopic methods such as (1)H-NMR, (13)C-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).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.

View Article: PubMed Central - PubMed

Affiliation: Korean Institute of Oriental Medicine (KIOM), KM-Based Herbal Drug Development Group, 1672 Yuseongdae-ro, Yuseong-gu, Daejeon 305-811, Republic of Korea.

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 (1)H-NMR, (13)C-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.

No MeSH data available.