• Title/Summary/Keyword: silica-gel column chromatography

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Flavonoids from the arial parts of Artemisia agryi and their antioxidant capacity through GSH recovery effect (황해쑥(Artemisia agryi)으로부터 flavonoid 화합물들의 분리 동정과 세포 내 GSH 회복능을 통한 항산화 활성 평가)

  • Hyeon Seon Na;Dahye Yoon;Hyeong-Ju Jeon;Dae Young Lee;Hyoung-Geun Kim
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.247-252
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    • 2022
  • The arial parts of Artemisia argyi were extracted with methanol : water (70:30), and the concentrates was partitioned into EtOAc (ethyl acetate), n-BuOH (normal butanol), and H2O (water) fractions. The repeated silica gel and ODS (octadecyl silica gel) column chromatographies for EtOAc and n-BuOH fractions led to isolation of four flavonoids without any ambiguity based on intensive interpretation of several spectroscopic data including nuclear magnetic resonance, and mass spectrometry. The chemical structure of the isolated compounds revealed to (2S)-naringenin (1), 3-methylkaempferol (2), 3,3'-dimethylquercetin (3), and 3,3',4'-trimethylquercetin (4). These four compounds were first isolated from A. argyi through this study. In this study, four compounds isolated from A. argyi showed an increase in glutathione mean and a decrease in glutathione heterogeneity so that the compounds uniformly raised the intracellular glutathione (GSH) level. Based on these results, it is considered that it can be used as a functional pharmacological material.

Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract (Eisenia bicyclis 에탄올 추출물로부터 분리한 Dieckol의 Lipase 저해 Mode)

  • Jung, Seul-A;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;Cho, Ji-Young;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.112-118
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    • 2013
  • This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

Adipocyte Differentiation Inhibitor Isolated from the Barks of Phellodendron amurense (황백(Phellodendri Cortex)으로부터 분리한 지방세포 분화 저해물질)

  • Kim, Kyung-Hee;Ahn, Soon-Cheol;Lee, Myung-Sun;Kweon, Oh-Song;Oh, Won-Keun;Kim, Min-Soo;Sohn, Cheon-Bae;Ahn, Jong-Seog
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.503-509
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    • 2003
  • For the development of the anti-obesity natural drug, the inhibitor of adipocyte differentiation was screened from Korean traditional medicinal plants. Phellodendri Cortex was selected as a candidate of adipocyte differentiation inhibitor. An inhibitory compound PC-4 was purified from the methanol (MeOH) extract of Phellodendri Cortex using silica gel and ODS RP-18 column chromatography and HPLC. PC-4 was obtained as yellow powder; UV ${\lambda}_{max}$ (MeOH): 230, 260, 340 and 430 nm. The chemical structure of PC-4 was determined as an isoquionoline alkaloid, berberine, on the basis of various NMR experiments including $^1H-\;and\;^{13}C-NMR$. The PC-4 inhibited the differentiation of preadipocyte NIH-3T3 L1 cells at a concentration of $1\;{\mu}g/mL$.

Separation of Antioxidant Compounds from Edible Marine Algae (식용 해조류에서 항산화 물질의 분리)

  • Park, Jae-Han;Kang, Kyoo-Chan;Baek, Sang-Bong;Lee, Yoon-Hyung;Rhee, Kyu-Soon
    • Korean Journal of Food Science and Technology
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    • v.23 no.3
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    • pp.256-261
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    • 1991
  • To isolate new antioxidants from twelve edible seaweeds, mixed methanol and chloroform extract of marine algae was fractionated into several subfractions and their antioxidant activities were measured by using AOM and 1, 1-diphenyl-2-picrylhydrazyl (DPPH). Especially the aqueous-methanol soluble fractions of brown and red algae showed a considerable antioxidant effect. Their antioxidant activities were stronger than synthetic antioxidants such as BHA, BHT, under the same concentration. Further fractionation of the aqueous-methanol soluble fractions using silica gel column chromatography yielded five subfractions. Among them methanol fraction exhibited high DPPH quenching activities. Also, it was confirmed to be benzene-derivative substances of two compounds by UV, HPLC, GC/MS analysis. Its each molecular weight was about 181, 238. These results suggested the existence of two effective natural antioxidant compounds in three edible marine algae.

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Isolation and Identification of Terpenoids from the Flower of Rhododendron yedoense var. poukhanense (산철쭉(Rhododendron yedoense var. poukhanense) 꽃으로부터 Terpenoid의 분리.동정)

  • Hong, Yoon-Hee;Song, Myoung-Chong;Han, Jae-Taek;Jang, Tae-O;Lee, Youn-Hyung;Kim, Sung-Hoon;Kim, Seung-Ae;Park, Mi-Hyun;Baek, Nam-Ln
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.144-149
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    • 2003
  • Extracts were obtained from the flower of Rhododendro yedoense var. poukhanense. (7 kg) in 80% aqueous MeOH and successively fractionated with solvent of EtOAc, n-BuOH and $H_2O$, successively. Silica gel and ODS column chromatographies of the EtOAc and n-BuOH fractions were repeatedly carried out by using the various solvent systems to give five terpenoids. Chemical structures of the isolated terpenoids were determined as $2{\alpha},3{\beta}-dihydroxylolean-12-ene$ (1), ursolic acid (2), grayanotoxin IV (3), grayanotoxin I (4) and grayanotoxin III (5) based on the interpretation of several spectral data including 2D-NMR such as $^1H-^1H\;COSY$, HMQC and HMBC.

A Study on Dietary Supplement to Reduce Obesity by the Mechanism of Decreasing Lipid and Carbohydrate Absorption (지방 및 탄수화물 흡수억제 메커니즘을 활용한 비만 개선 식이 연구)

  • Kim, Seok-Gi;An, Guk-Hwan;Yoon, Seung-Won;Lee, Young-Chun;Ha, Sang-Do
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.519-526
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    • 2003
  • Pancreatic lipase and ${\alpha}$-amylase inhibitory activities in purified extracts of pumpkin and job's tear were studied. Pancreatic lipase inhibitory activity was determined by measuring the rate of releasing oleic acid from triolein, and ${\alpha}$-amylase inhibitory activity was determined by iodometric method. The extracts of pumpkin and job's tear were purified using silica gel and C-18 gel column chromatographies. Treatment of pumpkin extract $(120\;{\mu}g/mL)$ in 3T3-L1 preadipocyte decreased differentiation about 95% and blocked accumulation of lipid. Body weights of rats fed high-fat diet containing dietary supplement decreased about 13% as compared with those fed only highfat diet. These results revealed dietary supplement is a good obesity-reducing material for decreasing lipid and carbohydrate absorptions.

Anti-inflammatory Activity of Opuntia ficus-indica (손바닥선인장의 항염증 활성)

  • Park, Eun-Hee;Hwang, Sung-Eun;Kahng, Ja-Hoon
    • YAKHAK HOEJI
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    • v.42 no.6
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    • pp.621-626
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    • 1998
  • Cactus (Opuntia ficus-indica var saboten Makino) is a tropical or subtropical plant, which is widely used as folk medicine for burned wound, edema and indigestion. We previously found that the ethanol extract of cactus stem showed anti-inflanunatory action. This investigation was designed to isolate the active fraction of anti-inflanimatory action from cactus stem by solvent extraction and colunm chromatography. Carrageenan-induced paw edema in rats and acetic acid-induced writhing test in mice were used as animal models to search anti-inflammatory and analgesic activities. respectively. The ethanol extract of cactus stem was consecutively extracted with hexane, ethyl acetate, and n-butanol. The hexane fraction was the most effective in carrageenan-induced paw edema, and then was separated in colunm chromatography of silica gel by the elution with hexane/ethyl acetate mixture. The most effective fraction 1 was separated in a second colunm chromatography by eluting with hexane/diethyl ether mixture. The most effective fraction 1-5 was obtained, and separated in a third column chromatography by eluting with hexane/chloroform mixture. It produced the most effective fraction 1-5-1. Moreover, fraction 1-5-1 showed an inhibitory effect on acetic acid-induced writhing in the doses of 30mg/kg and 60mg/kg,p.o.,indicating that it also contained analgesic activity.

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Development of Quantitative Analytical Method for Isoflavonoid Compounds from Fruits of Cudrania Tricuspidata (꾸지뽕나무 열매 isoflavonoid 화합물에 대한 정량분석법 개발)

  • Yoon, Sun Young;Kim, Seok Ju;Sim, Su Jin;Lee, Hak-Ju
    • Journal of the Korean Wood Science and Technology
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    • v.44 no.3
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    • pp.337-349
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    • 2016
  • In this study, an analytical method to evaluate the quality of isoflavonoid compounds purified and isolated from the fruits of Cudrania tricuspidata was developed and validated using Ultra Performance Liquid Chromatography (UPLC). The fruits of C. tricuspidata were extracted with methanol and further fractioned with n-hexane, ethyl acetate and water. The resulting ethyl acetate extract separated into four isoflavonoid compounds by a combination of silica gel and sephadex LH-20 column chromatography. The structures of the compounds were elucidated as alpinumisoflavone, 6,8-diprenyl genistein, 6,8-diprenyl orobol, 4'-O-methylalpinumisoflavone by various techniques such as UV-Vis, ESI-MS, $^1H\;NMR$ and $^{13}C\;NMR$ spectroscopy. Finally, a method to characterize the compounds was developed by using the UPLC equipped with a $C_{18}$ column and a gradient mobile phase system consisting of 2% acetic acid in water (solvent A) and 2% acetic acid in methanol (solvent B). The developed method was validated with the parameters such as selectivity, linearity, limit of detection, limit of quantitation, accuracy, and precision, which are defined by the ICH (International Conference on Harmonization). Using the validated method, the compounds in the fruits harvested in different months were also quantitatively analyzed. We propose this approach this approach can readily be utilized as an efficient evaluation method to quantify the extracts of C. tricuspidata.

Secondary Metabolites from the Mycelial Culture Broth of Phellinus linteus (상황(Phellinus linteus) 배양 균사체의 2차 대사산물에 대한 화학적 연구)

  • Song, K.S.;Cho, S.M.;Ko, K.S.;Han, M.W.;Yoo, I.D.
    • Applied Biological Chemistry
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    • v.37 no.2
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    • pp.100-104
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    • 1994
  • From the 48 hour-cultured mycelial broth of Phellinus linteus, six compounds were isolated by means of ethyl acetate extraction, silica gel column chromatography and preparative thin layer chromatography, consecutively. Compound 1 was identified as a succinic acid by the comparison of its spectral data with authentic sample. Compounds 2 and 3 were identified as p-hydroxyphenyl acetic acid methyl ester and p-hydroxybenzaldehyde by spectroscopic studies, respectively. NMR and MS studies of compound 6 revealed that it was 2,5-dihydroxymethyl furan. Compound 4, which showed similar NMR absorptions and MS fragmentation pattern with those of compound 6 was identified as 2-hydroxymethyl-5-methoxymethylfuran. These structures were verified by the spectral data of the acetate derivatives of the compounds. Compound 5 was supposed to be a N-acetyltyramine from its $^1H-NMR$ and EI-MS data, and its structure was confirmed by a synthesis starting from tyramine.

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Isolation and Identification of Growth Inhibition Substance on L. monocytogenes from Dystaenia takesimana Kitagawa (섬바디로부터 L. monocytogenes에 대한 생장억제 물질의 분리 및 구조동정)

  • Oh, Jin-Ah;Shin, Dong-Hwa;Baek, Nam-In
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.984-993
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    • 1999
  • The ethanol extracts and its n-hexane fraction of Dystaenia takesimana Kitagawa exhibited growth inhibition on Listeria monocytogenes ATCC 19111, ATCC 19112, ATCC 19113, ATCC 19114 and ATCC 15313. The minimum inhibitory concentration of the ethanol extract and its n-hexane fraction were 50 ppm and below 30 ppm on Listeria monocytogenes respectively. By silica gel column chromatography, the active fraction A8 was obtained from the ethanol extract of Dystaenia takesimana Kitagawa. After three times of column chromatography, the SBD-1 and SBD-2 were separated from the A8 fraction of the ethanol extract of Dystaenia takesimana Kitagawa. Antimicrobial activity of the SBD-l and SBD-2 was lower than that of the A8. And the A8 exhibited growth inhibition on five strains of Listeria monocytogenes at the level of $10{\sim}30$ ppm and the bactericidal effect was confirmed at same the level. The purified antimicrobial active compound was identified as (9z)-heptadeca-l,9-dien-4,6-diyn-3,8-diol, falcarindiol, by EI/MS, $^{1}H-NMR$ and $^{13}C-NMR$.

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