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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 (a) cell viability and LPS-induced (b) NO production in RAW 264.7 cells. RAW 264.7 cells were pretreated with five compounds for 30 min before incubation with LPS for 24 h. (a) Cytotoxicity was evaluated by an MTT assay. (b) The culture supernatant was analyzed for nitrite production. As a control, the cells were incubated with vehicle alone. Data shows mean ± SE values of triplicate determination from independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.
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fig8: Effect of five compounds on (a) cell viability and LPS-induced (b) NO production in RAW 264.7 cells. RAW 264.7 cells were pretreated with five compounds for 30 min before incubation with LPS for 24 h. (a) Cytotoxicity was evaluated by an MTT assay. (b) The culture supernatant was analyzed for nitrite production. As a control, the cells were incubated with vehicle alone. Data shows mean ± SE values of triplicate determination from independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.

Mentions: We evaluated the cytotoxicity of five compounds using the MTT assay to determine the optimal concentration effective for anti-inflammation with minimum toxicity. As shown in Figure 8(a), all the five compounds did not affect cell viability up to 100 μM, indicating that the compounds were not toxic to cells.


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 (a) cell viability and LPS-induced (b) NO production in RAW 264.7 cells. RAW 264.7 cells were pretreated with five compounds for 30 min before incubation with LPS for 24 h. (a) Cytotoxicity was evaluated by an MTT assay. (b) The culture supernatant was analyzed for nitrite production. As a control, the cells were incubated with vehicle alone. Data shows mean ± SE values of triplicate determination from 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

fig8: Effect of five compounds on (a) cell viability and LPS-induced (b) NO production in RAW 264.7 cells. RAW 264.7 cells were pretreated with five compounds for 30 min before incubation with LPS for 24 h. (a) Cytotoxicity was evaluated by an MTT assay. (b) The culture supernatant was analyzed for nitrite production. As a control, the cells were incubated with vehicle alone. Data shows mean ± SE values of triplicate determination from independent experiments. *P < 0.01 and **P < 0.001 were calculated from comparing with LPS-stimulation value.
Mentions: We evaluated the cytotoxicity of five compounds using the MTT assay to determine the optimal concentration effective for anti-inflammation with minimum toxicity. As shown in Figure 8(a), all the five compounds did not affect cell viability up to 100 μM, indicating that the compounds were not toxic to cells.

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.