• Title/Summary/Keyword: Solvent fraction

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Identification of Anticoagulant Components in Korean Red Ginseng

  • Wee, Jae-Joon;Kim, Young-Sook;Kyung, Jong-Soo;Song, Yong-Bum;Do, Jae-Ho;Kim, Dong-Chung;Lee, Sung-Dong
    • Journal of Ginseng Research
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    • v.34 no.4
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    • pp.355-362
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    • 2010
  • In this study, the anticoagulant compounds in Korean red ginseng (KRG) were investigated. KRG powder was extracted using hot methanol, and the methanol extract was fractionated into n-hexane, ethylacetate, n-butanol, and aqueous fractions by solvent partitioning. The remains from the methanol extraction were further extracted with water and then dialyzed to obtain low and high molecular weight fractions. The anticoagulant activities of the seven fractions were evaluated in terms of thrombin time, prothrombin time, and activated partial thromboplastin time. Among these fractions, the ethylacetate fraction showed the most potent anticoagulant activity. The active components in the ethylacetate fraction were identified as the phenolic compounds vanillic, caffeic, ferulic, and p-coumaric acid via TLC and HPLC. These findings suggest that the anticoagulant activities of phenolic compounds contribute to the cardiovascular effects of KRG.

Study on the Antitumor Substance Isolated from Oldenlandiae diffusae Herba (백화사활초(白花蛇活草)로부터 분리(分離)된 항암성물질(抗癌性物質)에 관(關)한 연구(硏究))

  • kim, Sung-Hoon
    • Journal of Haehwa Medicine
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    • v.4 no.2
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    • pp.273-297
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    • 1996
  • Hexane fraction of Oldenlandiae diffusae Herba(ODH) which was being used for the treatment of caner in oriental medicine showed the best cytotoxicity against L1210 and A549 in the solvent fractions. Antitumor substance isolated from hexane fraction of ODH was identified as ursolic acid(UA) by photometric analysis. IC50 of UA against cancer cells as SNU-1, HCT15, XF498, SK-MEL2 and A549 was $13{\mu}g/m{\ell}$, $15{\mu}g/m{\ell}$, $12{\mu}g/m{\ell}$, $9{\mu}g/m{\ell}$ and $11{\mu}g/m{\ell}$ respectively. It significantly inhibited the metastasis to lungs and kidneys from pulmonary colonization assay and study on histological changes of organs and showed the enhancing effect on B cell dosage-dependently by FACS analysis. T/C % of UA against S-180 cells was 171 % and its cytotoxicity against SNU-1 iant was confirmed from the morphological changes by elctronic microscopes such as SEM and TEM that it induced undulated membrane 4 hr after UA treatment, and the breakdown of cell membrane and nucleus 24 hr after UA treatment.

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The Effective Preparation of Flavonoids from Scutellaria baicalensis GEORGI by Diaion HP-20 Resin

  • Yu, Young-Beob
    • Korean Journal of Plant Resources
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    • v.27 no.6
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    • pp.635-641
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    • 2014
  • Scutellaria baicalensis $G_{EORGI}$ (Scutellariae Radix) has been used to clear heat and to dry dampness in the stomach or intestines, which manifests as diarrhea or dysenteric disorder. In this study, we investigated the effective preparation of active components in Scutellariae Radix using the methods of solvent extraction and absorption fractionation for the development of new functional food or pharmaceuticals. The marker substances, baicalin, baicalein, wogonoside, and wogonin were directly isolated from the Scutellariae Radix. There chemical structures were elucidated by spectroscopic analysis. The Scutellariae Radix was extracted with hot water. To enhance yield of flavonoids in Scutellariae Radix, the hot water extract was dissolved in ethanol with concentration dependent manner. The precipitates were separated using centrifugal techniques at 10,000 rpm. Supernatant liquid was applied to the HPLC for quantification of major compounds. Separately, the hot water extract was absorbed on Diaion HP-20 resin. And then, the absorbed fraction was eluted with methanol for HPLC. The contents of baicalin, baicalein, wogonoside and wogonin in different treatment methods were analyzed by HPLC. Total amount of four major components were 16.9% in 50% ethanol extract, 21.7% in 70% ethanol extract, 20.5% in 90% ethanol extract, and 39.3% in absorbed fraction of Diaion HP-20 resin. In these results, we found that resin absorption method is suitable for the extraction of enriched flavonoids from Scutellariae Radix.

Antagonism of Bacterial Extracellular Metabolites to Freshwater-Fouling Invertebrate Zebra Mussels, Dreissena polymopha

  • Gu, Ji-Dong;Ralph Mitchell
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.133-138
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    • 2001
  • We investigated the antagonism of indigenous bacteria isolated from stressed mussels and their extracellular metabolites on the adult zebra mussel, Dreissena polymorpha. Selective bacterial isolates including Aeromonas media, A. salmonicida, A. veronii, and Shewanella putrefaciens, showed strong lethality against adult mussels and 100% mortality was observed within 5 days of incubation. Bacterial metabolites, fractionated and concentrated from stationary-phase culture supernatants of these bacterial isolates, displayed varying degrees of antagonistic effects on zebra mussels. Among the three size fractions examined, <5, 5-10, and >10 kDa, the mast lethal fraction seems to be >10 kDa for three of the four isolates tested. Further chemical analyses of these size fractions revealed that the predominant constituents were polysaccharides and proteins. No 2-keto-3-deoxyoctanoic acid (2-KDO), deoxyribonucleic acids (DNA) or uranic acid were detectable. Extraction of supernatants of two antagonistic isolates with polar solvent suggested that polar molecules are present in the active fraction. Our data suggest that extracellular metabolites produced by antagonistic bacteria are also involved in disease development in zebra mussels and elucidation of the mechanisms involved may offer a novel strategy for control of biofouling invertebrates.

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Inhibitory effect of Fucofuroeckol-A from Eisenia bicyclis on tyrosinase activity and melanin biosynthesis in murine melanoma B16F10 cells

  • Shim, Kil Bo;Yoon, Na Young
    • Fisheries and Aquatic Sciences
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    • v.21 no.11
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    • pp.35.1-35.7
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    • 2018
  • Background: The aim of this study was to investigate the in vitro inhibitory effects of Fucofuroeckol-A isolated from Eisenia bicyclis against tyrosinase activity and 3-isobutyl-1-methylxanthine (IBMX)-induced melanin biosynthesis in B16F10 melanoma cells. Result: Among the ethanolic (EtOH) extract of E. bicyclis and its organic solvent fractions, the ethyl acetate (EtOAc) soluble fraction showed a noticeable inhibitory effect on mushroom tyrosinase with an $IC_{50}$ value of $37.6{\pm}0.1{\mu}g/mL$. Repeated column chromatography of the active EtOAc fraction resulted in the isolation of Fucofuroeckol-A. It evidenced more potent tyrosinase inhibitory effect with an $IC_{50}$ value of $11.4{\pm}1.4{\mu}M$ than arbutin ($IC_{50}=1076.6{\pm}44.3{\mu}M$), which was used as a positive control. Lineweaver-Burk plots suggest that Fucofuroeckol-A plays as a noncompetitive inhibitor against tyrosinase. Furthermore, we have evaluated the inhibitory effects of Fucofuroeckol-A on IBMX-induced melanin formation in B16F10 melanoma cells. Fucofuroeckol-A ($12.5-100{\mu}M$) exhibited a significant inhibition of melanin production in the melanoma cells. Conclusion: In the present study, we suggested that Fucofuroeckol-A might prove possibility as a novel inhibitor of melanin biosynthesis in cosmetic applications.

Antimicrobial, Antioxidant and Anticoagulation Activities of Korean Radish (Raphanus sativus L.) Leaves (무청의 항균, 항산화 및 항혈전 활성)

  • Lee, Ye-Seul;Kwon, Kyung-Jin;Kim, Mi-Sun;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.228-235
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    • 2013
  • Radish (Raphanus sativus) is a common cruciferous vegetable, and its aerial parts, called Mu-chung in Korean, have plentiful nutritional components such as vitamins, minerals and dietary fibers. Mu-chung has been used as a kimchi, a traditional Korean fermented dish, and dried Mu-chung is an important component of soups commonly consumed during winter in Korea. Since the advent of the mass production of radish in Korea, with the segregation of farm areas and towns and changing diets, Mu-chung has mostly been discarded instead of utilized. In addition, studies concerning the efficient utilization and useful bioactivities of Mu-chung are still lacking worldwide. In this study, we prepared the ethanol extract of Mu-chung and its subsequent solvent fractions. Antimicrobial, antioxidation, and anticoagulation activities were then evaluated in the hopes of developing a functional biomaterial from Korean radishes' aerial parts. The ethanol extraction yield for hot-air dried Mu-chung was 5.6%, and the fraction yields of n-hexane (H), ethylacetate (EA), butanol (B) and water residue were 25.3, 3.6, 19.4, and 51.7%, respectively. Analysis of total polyphenol and total flavonoid contents showed that the EA fraction had the highest content (97.57 and 152.91 mg/g) amongst the fractions. In antimicrobial activity assays, the H and EA fractions were effective against gram positive bacteria (Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis), but not effective against gram negative bacteria (Escherichia coli and Pseudomonas aeruginosa). The B fraction also exhibited moderate antibacterial activity, suggesting that the extract of Mu-chung has various antibacterial components. In antioxidation activity assays, the EA fraction showed strong DPPH, ABTS and nitrite scavenging activities ($69-222{\mu}g/ml$ of $IC_{50}$), including reducing power. In anticoagulation activity assays, the EA fraction demonstrated strong inhibition activity against human thrombin and prothrombin. Prominent anticoagulation activity was found in aPTT assays; the aPTT of the EA fraction was extended 15-fold compared than that of the solvent control. Our results suggest that Mu-chung is an attractive nutritional food material possessing useful bioactivities, and the EA fraction of Mu-chung could be developed as a functional food ingredient.

Preparation of Active Fraction from Radish Water Extracts for Improving the Intestinal Functions and Constipation Activities (무(Raphanuse sativa var. nigra L.) 물 추출물로 부터 장기능 및 변비질환 개선을 위한 활성 분획의 제조)

  • Baik, Soon-Ok;Lee, Yoo-Hui;Kim, Young-Sook;Ryu, Myeong-Hyeon;Kim, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.47 no.3
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    • pp.315-320
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    • 2004
  • The aim of this study was to develop an activator, 3-10 kDa fraction from radish water extracts, that will improve the intestinal function and bowel movement in the colons. Radish water extracts were investigated for their intestinal function effects according to the charcoal meal transit method, employing Balb/c mice: also, their anti-constipation activities were compared utilizing the loperamide-induced constipation method, employing SD rats. The result suggested that the effects of the charcoal meal transit increased remarkably in radish water extract administrated rats in comparison to loperamide administrated rats. Futhermore, the effects of various solvent extracts of radish on charcoal meal transit in Balb/c mice increased remarkably in radish water fraction administrated rats than in different solvent fraction administrated rats. Radish extraction was tested and isolated into 4 groups: below 3 kDa, 3-10 kDa, 10-300 kDa, and over 300 kDa. 3-10 kDa was the most effective on the intestinal function and bowel movement in the colons; also, 3-10 kDa fraction of radish water extraction was found to be the most effective charcoal meal transit. The dry weight and moisture content of feces remarkedly increased in the 3-10 kDa administrated rats group than in the loperamide only group. Experimental results revealed that 3-10 kDa fraction of radish water extract was the most effective on the intestinal function and bowel movement was the crypt epithelial cells that contained more MUC2 in the 3-10 kDa administrated group than the loperamide only group: in addition, the thickness of mucus layer stained with alcian blue was significantly thicker in 3-10 kDa administrated rats than in loperamide administrated rats. Crypt epithelial cells secreted more MUC2 in the 3-10 kDa administrated group than the loperamide only group and the stained cells clearly showed the MUC2 with antibody Biogenex AM358.

Antioxidant and Antiproliferative Activities of the Halophyte Angelica japonica Growing in Korean Coastal Area (한국 연안지역에 서식하는 갯강활의 항산화 및 암세포증식 억제 활성)

  • Jayapala, Priyanga S.;Oh, Jung Hwan;Kong, Chang-Suk;Sim, Hyun-Bo;Seo, Youngwan
    • Journal of Life Science
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    • v.32 no.10
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    • pp.749-761
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    • 2022
  • This study evaluated the antioxidizing and antiproliferative effects of Angelica japonica extract and its solvent-partitioned fractions. A dried sample of the halophyte A. japonica was extracted twice using methylene chloride (CH2Cl2) and extracted twice again using methanol (MeOH). The combined crude extracts were then fractionated by solvent polarity into distilled water (water), n-butanol (n-BuOH), 85% aqueous methanol (85% aq.MeOH), and n-hexane fractions. The antioxidant activities of the crude extracts and their solvent-partitioned fractions were assessed according to their DPPH radical and peroxynitrite scavenging abilities, formation of intracellular reactive oxygen species (ROS), DNA oxidation, NO production, and ferric reducing antioxidant power (FRAP). The crude extract showed significant antioxidant activity in the overall antioxidizing bioassay systems. Among solvent-partitioned fractions, good antioxidant activities were observed in n-BuOH and 85% aq.MeOH fractions and significantly correlated with the polyphenol and flavonoid contents of the samples. Furthermore, all samples tested, including the crude extract, not only showed cytotoxic effects against human cancer cells (AGS, HT-29, MCF-7, and HT-1080) but also prevented cell migration in a dose-dependent manner in the wound healing assay using HT 1080. Among the solvent-partitioned fractions, the 85% aq.MeOH fraction most effectively inhibited the invasion of HT-1080 cells. Therefore, these results suggest that A. japonica may be a potential antioxidizing and antiproliferative agent.

Anti-invasive Effect of Artemisia scoparia Halophyte Extract and its Solvent-partitioned Fractions in Human Fibrosarcoma Cells (인간 섬유육종세포에서 비쑥 추출물과 유기용매 분획물의 암전이 억제 효과)

  • Kim, Junse;Kong, Chang-Suk;Sim, Hyun-Bo;Seo, Youngwan
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1100-1109
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    • 2021
  • The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H2O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.

Antioxidant Activities of Solvent Extracts from Pomegranate Endocarp (석류 내피 용매별 추출물의 항산화 활성)

  • Jin, So-Yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1635-1641
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    • 2011
  • This study was designed to investigate antioxidant activities of pomegranate endocarp extracts. Pomegranate endocarp extract contains the highest antioxidant function compared to pomegranate extracts from other parts. Pomegranate endocarp extract was fractionated with hexane, dichloromethane, ethyl acetate, butanol extract, and water, followed by evaluation for antioxidant activity. During this experiment, various antioxidant tests such as nitrite scavenging activity, reducing power, superoxide anion scavenging activity, $ABTS^+$ scavenging activity, SOD like-activity, and DPPH radical scavenging activity were conducted on the $CH_2Cl_2$, EtOAc, BuOH, $H_2O$ fractions of pomegranate endocarp extract. Results showed that the ethyl acetate fraction contained the highest DPPH radical scavenging activity among the samples. For SOD like-activity, the dichloromethane fraction had the highest antioxidant activity. For superoxide anion scavenging activity, the ethyl acetate fraction had the highest antioxidant activity. In the $ABTS^+$ scavenging activity test, the ethyl acetate fraction $IC_{50}$ was 39.26 ${\mu}g$/mL, whereas that of the butanol fraction was 40.95 ${\mu}g$/mL. In testing reduction power at 0.1 mg/mL, the O.D. of the ethyl acetate fraction was highest at 1.404 and showed higher activity than ascorbic acid at 1.332. The results of the nitrite scavenging activity test were very similar to those of the SOD like-activity test. These results suggest that pomegranate endocarp extract may have value as a natural antioxidant.