• Title/Summary/Keyword: Sephadex LH-20

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Antidiabetic Activities of Korean Red Pine (Pinus densiflora) Inner Bark Extracts

  • Min, Hee-Jeong;Kim, Eun-Ji;Shinn, Seong-whan;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.4
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    • pp.498-508
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    • 2019
  • This study was carried out to investigate the potential of Korean red pine (Pinus densiflora) inner bark extracts as an antidiabetic agent. The ethyl acetate soluble fraction of the bark extracts was chromatographed on a Sephadex LH-20 column to yield five compounds, which structures were elucidated by NMR spectroscopy. The isolated compounds were (+)-catehin, (-)-epicatechin, taxifolin, taxifolin-3'-O-${\beta}$-D-(+)-glucose and $\tilde{n}$-courmaric acid. The antidiabetic activity of the different fractions, including the crude extracts and isolated compounds, was evaluated by ${\beta}$-cells insulin secretion and glucose uptake in skeletal muscle cells. The insulin secretion was 128% for taxifolin at $25{\mu}g/mL$. However, the other samples had no effect on this test. For the glucose uptake activity assay, $1{\mu}M$ insulin and 2 mM metformin were used as controls. Both the crude extract and taxifolin showed relatively low activity values, but the other samples yielded glucose uptake values over 260%. ${\rho}$-courmaric acid showed the highest uptake (270%). The results confirmed that Korean red pine extracts may be used as a hypoglycemic agent.

Bacterial neuraminidase inhibitory linarin from Dendranthema zawadskii

  • Ju Yeon Kim;Jae Yeon Park;Yun Gon Son;Kyu Lim Kim;Jeong Yoon Kim
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.1-6
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    • 2023
  • Dendranthema zawadskii is a one of the popular plants as native in South Korea. In this study, linarin was isolated and purified using silica-gel, Diaion, and Sephadex LH-20 from the aerial parts of D. zawadskii. The chemical structure was completely identified through spectroscopic data including 1D, 2D nucleic magnetic resonance, and HRFABMS. Furthermore, linarin inhibited the bacterial neuraminidase (BNA) activity with 13.5 μM of IC50 dose-dependently. Through the enzyme kinetic experiments, linarin as BNA inhibitor exhibited a typical noncompetitive inhibition mode which Km was contestant and Vmax decreased as the concentration of the inhibitor increased. It was further identified that the inhibition constant was 16.0 μM. Linarin was the most abundance metabolite in the aerial part of D. zawadskii extract by UHPLC-TOF/MS analysis. Therefore, D. zawadskii and its main component are expected that it can be effectively used for the infection and inflammation caused by bacteria.

Purification and Identification of Inhibitory Compounds on Helicobacter pylori from Cheongmoknosang Callus for Biomass (Biomass를 위해 제조한 청목노상 callus로부터 Helicobacter pylori 억제물질의 분리 및 동정)

  • Yun, Dong-Hyuck;Cha, Won-Seup;Lee, Seon-Hon;An, Bong-Jeun;Kim, Jeung-Hoan;Chun, Sung-Sook;Bae, Jong-Ho;Cho, Young-Je
    • Journal of Life Science
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    • v.20 no.3
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    • pp.374-380
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    • 2010
  • The objective of this research was to evaluate the inhibitory activities of phenolic compounds isolated from Cheomoknosang callus on Helicobacter pylori. Total phenolic compounds of 80% ethanol extracts from callus were 15.3 mg/g. The activity of H. pylori inhibition at 80% ethanol extracts from Cheongmoknosang callus was determined as 14 mm clear zone. Isolation of inhibitory compounds was carried out on Sephadex LH-20 and MCI-gel CHP-20 column chromatography using a gradient elution procedure of increasing MeOH in $H_2O$. The chemical structure of the inhibitory compound against Helicobacter pylori was confirmed as protocatechuic acid, chlorogenic acid, caffeic acid and rosemarinic acid by spectroscopic analysis of FAB-MS, NMR and IR spectrum.

Hypoglycemic Effect of Cordyceps militaris (큰번데기동충하초(Cordyceps militaris)의 혈당강하효과)

  • Kwon, Young-Min;Cho, Su-Min;Kim, Jee-Hun;Lee, Jae-Hee;Lee, Yeon-Ah;Lee, Seung-Jung;Lee, Min-Won
    • Korean Journal of Pharmacognosy
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    • v.32 no.4 s.127
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    • pp.327-329
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    • 2001
  • Cordyceps militaris (CM) has been used as a tonics in the traditional medicine. To investigate the anti-diabetic principle of CM, activity guided fractionation was conducted. Hot water extract of CM was fractionated into 3 parts: above 100,000(A), $100,000{\sim}20,000(B)$, below 20,000(C) in molecular weight using in membrane filter system. All fractions showed mild hypoglycemic activity in streptozotocin (STZ)-induced diabetic rats by oral administration (300 mg/kg). The fraction C which was most active among them was fractionated again into two parts, C-1 and C-2 by Sephadex LH 20 column chromatography. The fraction C-1 showed hypoglycemic activity but C-2 did not show activity compared with control in STZ mice. In glucose-fed hyperglycemic mice, fraction C, C-1 and C-2 also showed significant glucose lowering activity. Their decreasing rates of plasma glucose level after 1 hours administrations of fraction C, C-1 and C-2 were 24.5%, 29.3% and 22.0%, respectively (Tolbutamide: 48.4%). These results suggested that CM has both insulin like and insulin release promoting activity and could be developed as an antidiabetic agent.

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Antioxidantive, Phospholipase $A_2$ Inhibiting, and Anticancer Effect of Polyphenol Rich Fractions from Panax ginseng C. A. Meyer (한국산 인삼의 Polyphenol 분획물의 항산화, Phospholipase $A_2$ 및 암세포증식 억제효과)

  • Choi, Hee-Jin;Han, Ho-Suk;Park, Jung-Hye;Son, Jun-Ho;Bae, Jong-Ho;Seung, Tae-Su;Choi, Cheong
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.251-256
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    • 2003
  • The polyphenol fractions of Korean ginseng were purified using Sephadex LH-20, MCI gel, Bondapak $C_{18}$ TLC, and HPLC from the 60% acetone soluble fraction. Fraction I showed 48.16%, 79.71% and 43.55% inhibition at 150 ppm against lipid oxidation in the presence of copper ion, superoxide and hydrogen peroxidation. Electron donating abilities of fraction II showed 35.17% inhibition at 200 ppm. Fraction III showed 48.49% and 25% inhibition at 150 ppm against lipid oxidation in the presence of ferrous ion and hydroxy radical ion. The phospholipase $A_2$ inhibitory effect of fraction III was 48.9% at the concentration of $60\;{\mu}g/ml$. The cytotoxic effects of fraction II was the highest (73.29% at 0.25 mg/ml) among the tested polyphenol fractions.

Biological activity and analysis of α-glucosidase inhibitor from mulberry (Morus alba L.) wine (오디와인의 생리활성 및 α-glucosidase 저해제의 분석)

  • Son, Woo-Rim;Choi, Sang-Won
    • Food Science and Preservation
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    • v.20 no.6
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    • pp.877-885
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    • 2013
  • Wine extracts of four different berry fruits, such as mulberry, blueberry, strawberry, and raspberry, were investigated for antioxidant, anti-tyrosinase, and ${\alpha}$-glucosidase activities by using in vitro assays. Additionally, quantitative changes of ${\alpha}$-glucosidase inhibitor in mulberry wine were determined by HPLC according to mulberry cultivars and fermentation process. Among four berry wines examined, mulberry wine showed the most potent ${\alpha}$-glucosidase inhibitory activity with 69.37% at 0.23 mg/mL, while blueberry and strawberry wines exhibited the strongest inhibition against DPPH radical and tyrosinase activity, respectively. Four compounds were isolated and purified from mulberry wine by a series of isolation procedures, such as solvent fractionation, and Diaion HP-20, ODS-A, and Sephadex LH-20 column chromatographies. Among them, Comp. 4 exerted the strongest ${\alpha}$-glucosidase inhibitory activity ($I_C_{50}=31.57{\mu}M$), and its chemical structure was identified as quercetin by UV and NMR spectral analysis. Finally, the "Daeseongppong" (16.83 ppm) muberry wine had larger amount of quercetin than the "Iksuppong" (14.85 ppm) and "Cheongilppong" (8.92 ppm) mulberry wines, but their contents of three mulberry wines decreased considerably with aging process. These results suggest that mulberry wine containing quercetin acted as ${\alpha}$-glucosidase inhibitor may be useful as a potential functional wine for improving diabetic disorder.

Anti-inflammatory Effect of Myricetin from Rhododendron mucronulatum Turcz. Flowers in Lipopolysaccharide-stimulated Raw 264.7 Cells (Lipopolysaccharide로 유도된 Raw264.7 cell에서 Rhododendron mucronulatum Turcz. Flower으로부터 분리한 myricetin에 의한 염증 억제효과)

  • Choi, Moo-Young;Hong, Shin-Hyup;Cho, Jun-Hyo;Park, Hye-Jin;Jo, Jae-Bum;Lee, Jae-Eun;Kim, Dong-Hee;Kim, Byung-Oh;Cho, Young-Je
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1245-1252
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    • 2016
  • As a research of inflammation inhibitory activity using natural resource, the inflammation inhibitory activity by purified active compound from Rhododendron mucronulatum flower was experimented. Rhododendron mucronulatum flower components were purified and separated with Sephadex LH-20 and MCI gel CHP-20 column chromatography, Purified compound was confirmed as myricetin by $^1H-NMR$, $^{13}C-NMR$ and Fast atom bombardment (FAB)-Mass spectrum to have inhibition activity on inflammatory factors secreted by Raw 264.7 cells in response to lipopolysaccharide stimulation. Myricetin inhibited nitric oxide (NO) expression in a concentration dependent manner, approximately 40% inhibition was observed at a concentration of $50{\mu}M$. The inhibition effect of myricetin on inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression was 20% and 80%, respectively, at a concentration of $25{\mu}M$. Myricetin also inhibited expression of the inflammatory cytokines, tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, IL-6 and prostaglandin $E_2(PGE_2)$ in a concentration dependent manner; a concentration of $50{\mu}M$, 70%, 80%, 80% and 95% inhibition was observed, respectively. Therefore myricetin isolated from Rhododendron mucronulatum flowers is expected to have an anti-inflammatory effect in Raw 264.7 cell induced by lipopolysaccharides. The results can be expected myricetin from Rhododendron mucronulatum flower to use as functional resource for anti-inflammatory activity.

Development of Biologically Active Compounds from Edible Plant Sources XXII. Triterpenoids from the Aerial Parts of Sajabalssuk (Artemisia princeps PAMPANINI) (식용식물자원으로부터 활성물질의 탐색-XXII. 사자발쑥(Artemisia princeps PAMPANINI)의 지상부로부터 Triterpenoid의 분리)

  • Bang, Myun-Ho;Cho, Jin-Gyeong;Song, Myoung-Chong;Lee, Dae-Young;Han, Min-Woo;Chung, Hae-Gon;Jeong, Tae-Sook;Lee, Kyung-Tae;Choi, Myung-Sook;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.223-227
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    • 2008
  • The aerial parts of Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk) was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and $H_2O$, successively. From the EtOAc fraction, three cycloartane-type triterpnoids and one ursane-type triterpenoid were isolated through the repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the results of physico-chemical data including NMR, MS and IR, the chemical structures of the triterpenoids were determined as wrightial (1), wrightial acetate (2), 27-norcycloart-20(21)-ene-25-al-3${\beta}$-ol acetate (3) and ursolic acid (4). No report has been found for isolation of compound 3 in the literature so far, and compounds 1, 2 and 3 were the first to be isolated from Sajabalssuk (Artemisia princeps PAMPANINI, Sajabalssuk). Also, compound 1 showed Acyl-CoA:Cholesterol acyltransferase (hACAT-1) and hACAT-2 inhibitory activity with the $IC_{50}$ values of 33.0 and 45.0 ${\mu}g/ml$, respectively. Compounds 2 and 3 inhibited hACAT-1 activity with the $IC_{50}$ values of 12.0 and 16.0 ${\mu}g/ml$, respectively.

Inhibitors of Tyrosinase and Melanogenesis from Galla rhois

  • Kim, Hyo-Jin;Jang, Dong-Il;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.2 no.4
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    • pp.285-290
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    • 1997
  • Previously, a 50% aqueous methanol extract of Galla rhois was shown to be the most potent tyrosinase inhibition activity with an {TEX}$IC_{50}${/TEX}(the concentration causing 50% inhibition of tyrosinase activity) of 0.2mg/ml of 205 crude drug extracts. To isolate tyrosinase inhibitors, the methanol extract was evaporated to a small volume in vacuo, and then partitioned stepwise with benzene and ethyl acetate(EtOAc). the EtOAc fraction was solubilized in 10% MeOH solution, and then fractionated successively by Diaion HP-20 and Sephadex LH-20 column chromatography, and preparative HPLC. Three phenolic compounds were isolated, and characterized as gallic acid(GA), methyl gallate(MG) and 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose(PGG) by UV, IR, {TEX}${1}^H${/TEX}-&{TEX}${13}^C${/TEX}-NMR, and FAB-MS spectroscopy, PGG({TEX}$IC_{50}${/TEX}=50$\mu\textrm{g}$/ml) showed a considerable inhibitory effect against mushroom tyrosinase, while GA({TEX}$IC_{50}${/TEX}=1.6mg/ml) and MG({TEX}$IC_{50}${/TEX}=234$\mu\textrm{g}$/ml) did not show an appreciable effect. Meanwhile, MG inhibited greatly melanogenesis in a murine melanocyte cell line, Mel-Ab. MG and PGG showed typical noncompetitive inhibition patterns against mushroom tyrosinase. These results suggest that PGG and MG may be potentially useful as either anti-browning or anti-melanogenic agents in foods and cosmetics.

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Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.