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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.


Schematic of on-line screening HPLC-ABTS+ system.
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Related In: Results  -  Collection


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fig3: Schematic of on-line screening HPLC-ABTS+ system.

Mentions: A. tegmentosum extract was injected into a Dionex Ultimate 3000 HPLC system (Thermo Scientific). The chromatographic columns used in this experiment were commercially available and were purchased from RS-tech (0.46 × 25 cm, 5 μm, C18, Daejeon, Korea). The injection volume was 10 μL, and the flow rate of the mobile phase was 1.0 mL/min. The wavelength of the UV detector was fixed at 210, 254, 280, and 320 nm. The composition of the mobile phases was A: 99.9% water/trifluoroacetic acid (99.9/0.1 vol%) and B: 100% acetonitrile. The run time was 60 min, and the solvent program was the linear gradient method (90 : 10–60 : 40 A : B vol%, 70 min: initial condition) (Table 2). Figure 3 shows a schematic diagram of the on-line coupling of a HPLC to a DAD (diode array detector) and the continuous flow ABTS+ assay. Then, on-line HPLC was connected to a “T” piece where ABTS+ was added. The ABTS+ at a flow rate of 0.5 mL/min was delivered using a Dionex ultimate 3000 pump. After mixing through a 1 mL loop, maintained at 40°C, the absorbance was measured using a multiple wavelength detector (MWD) at 734 nm. Data were analyzed using the Chromeleon 7 software.


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)

Schematic of on-line screening HPLC-ABTS+ system.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: Schematic of on-line screening HPLC-ABTS+ system.
Mentions: A. tegmentosum extract was injected into a Dionex Ultimate 3000 HPLC system (Thermo Scientific). The chromatographic columns used in this experiment were commercially available and were purchased from RS-tech (0.46 × 25 cm, 5 μm, C18, Daejeon, Korea). The injection volume was 10 μL, and the flow rate of the mobile phase was 1.0 mL/min. The wavelength of the UV detector was fixed at 210, 254, 280, and 320 nm. The composition of the mobile phases was A: 99.9% water/trifluoroacetic acid (99.9/0.1 vol%) and B: 100% acetonitrile. The run time was 60 min, and the solvent program was the linear gradient method (90 : 10–60 : 40 A : B vol%, 70 min: initial condition) (Table 2). Figure 3 shows a schematic diagram of the on-line coupling of a HPLC to a DAD (diode array detector) and the continuous flow ABTS+ assay. Then, on-line HPLC was connected to a “T” piece where ABTS+ was added. The ABTS+ at a flow rate of 0.5 mL/min was delivered using a Dionex ultimate 3000 pump. After mixing through a 1 mL loop, maintained at 40°C, the absorbance was measured using a multiple wavelength detector (MWD) at 734 nm. Data were analyzed using the Chromeleon 7 software.

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.