• Title/Summary/Keyword: silica gel column chromatography

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Isolation and Purification of Resveratrol from a Grape Twig (포도 송이가지에서 레스베라트롤의 분리 정제)

  • Shin, Hyun-Jae;Kang, Byung-Sun;Ahn, Jun-Bae;Kim, Bok-Hee
    • KSBB Journal
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    • v.22 no.5
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    • pp.351-355
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    • 2007
  • Resveratrol, a polyphenolic compound with antioxidative property, was purified from the grape's twig to be used as functional additives of food and/or cosmetics. Extraction of the grape's twig was performed using 80% ethanol in ultrasonic extractor for 60 min. The crude extract was purified up to 99% after elution through silica gel open column chromatography. The stability of the purified resveratrol was as follows: a half life of 90 days at 40$^{\circ}C$ and 60 days at 25$^{\circ}C$. A sensory test of the commercial grape juice including the 1-10 ppm of purified resveratrol showed better preference than the grape juice without purified resveratrol additive. Color and smell test showed no difference between the samples. The grape twig can be used as a valuable resource for the extraction of resveratrol, which would be added to nutraceutical and cosmetic products.

Isolation and Characterization of an Antibacterial Substance from Rheum palmatum for Treatment of Bacterial Vaginosis (대황으로부터 세균성 질염 치료를 위한 항균성 물질의 분리 및 특성)

  • Jang, Jieun;Kang, Dong-Hee;Yoon, Jaewoo;Kim, Hyun-Soo
    • KSBB Journal
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    • v.32 no.2
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    • pp.133-139
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    • 2017
  • Rheum palmatum has traditionally been used as a preventive agent and medication against fever and infection. The aim of this study was to isolate and characterize an antibacterial substance from R. palmatum that is effective against bacterial vaginosis. A methanol extract from R. palmatum showed antibacterial activity against Lactobacillus vaginalis KC TC 3515, Chryseobacterium gleum KCTC 2904, and Sphingomonas paucimobilis KCTC 2834, which cause bacterial vaginosis. After extraction and pH control of the methanol extract from R. palmatum, we found that acidic and alkaline extracts did not show antibacterial activity. A neutral extract (50 mg/mL) displayed an inhibitory zone of 18 mm on a nutrient agar plate with C. gleum KCTC 2904. Fractions No. 11 and 12 among 41 fractions obtained by silica gel column chromatography produced inhibitory zones of 10 mm on nutrient agar plates with C. gleum KCTC 2904. $R_f0.15$ and $R_f0.17$ spots produced by TLC of fraction No. 11 showed antibacterial activity against C. gleum KCTC 2904. Isolation and purification of the peak at a retention time (Rt) of 9.427 min was achieved by HPLC of $R_f0.29spots$. The peak at Rt 9.427 min showed antibacterial activity against C. gleum KCTC 2904.

Screening of ${\beta}$-Catenin/TCF Transcription Factor Inhibitors in Medicinal Herb Extracts

  • Choe, Ye-Dang;Na, Byung-Jo;Park, Se-Yeon
    • The Journal of Korean Medicine
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    • v.32 no.3
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    • pp.35-43
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    • 2011
  • Objectives: This study was performed to screen target-specific inhibitors of ${\beta}$-catenin/TCF signaling whose functional activation plays an important role in early events in carcinogenesis. Methods: To investigate the activation or suppression of ${\beta}$-catenin/TCF transcription, we established a transiently transfected cell line with a constitutively active ${\beta}$-catenin mutant gene whose product is not degraded. This cell line was also co-transfected with luciferase reporter gene constructs containing either an optimized (TOPflash) or mutant (FOPflash) TCF-binding element. We investigated cytotoxic effects using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium salt (MTS) assay. To find effective inhibitors of ${\beta}$-catenin/TCF signaling from medicinal herbs, the crude extracts of 99 types of medicinal herbs were screened using a luciferase assay system in HEK-293 and SH-SY5y cells. Results: At a concentration of $50{\mu}g$/ml, extracts of Angelica koreanae radix, Cannabis sativa semen, Ephedrae intermedia Schrenk radix, and Vitis rotundifolia fruit showed the following inhibitory effects on ${\beta}$-catenin/TCF signaling: $40{\pm}5.6%$, $23{\pm}6.1%$, $8{\pm}5.1%$, and $22{\pm}9.8%$ in ${\beta}$-catenin-activated HEK-293 cells and $9{\pm}4.7%$, $39{\pm}8.1%$, $39{\pm}6.4%$, and $42{\pm}10.1%$ in ${\beta}$-catenin-activated SH-SY5y cells, respectively. Crude extracts of E. radix were isolated by silica gel column chromatography, and two non-polar fractions of these extracts showed inhibitory effects on ${\beta}$-catenin/TCF signaling. Conclusions: In this study, we established a transiently transfected cell line as a screening system and found that various medicinal herb extracts had inhibitory effects on ${\beta}$signaling.

Ameliorating Effect of Gardenia jasminoides Extract on Amyloid Beta Peptide-induced Neuronal Cell Deficit

  • Choi, Soo Jung;Kim, Mi-Jeong;Heo, Ho Jin;Hong, Bumshik;Cho, Hong Yon;Kim, Young Jun;Kim, Hye Kyung;Lim, Seung-Taik;Jun, Woo Jin;Kim, Eun-Ki;Shin, Dong-Hoon
    • Molecules and Cells
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    • v.24 no.1
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    • pp.113-118
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    • 2007
  • The brains of Alzheimer's disease (AD) patients are characterized by large deposits of amyloid beta peptide ($A{\beta}$). $A{\beta}$ is known to increase free radical production in nerve cells, leading to cell death that is characterized by lipid peroxidation, free radical formation, protein oxidation, and DNA/RNA oxidation. In this study, we selected an extract of Gardenia jasminoides by screening, and investigated its ameliorating effects on $A{\beta}$-induced oxidative stress using PC12 cells. The effects of the extract were evaluated using the 2',7'-dichlorofluorescein diacetate (DCF-DA) assay and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. To find the active component, the ethanol extract was partitioned with hexane, chloroform, and ethyl acetate, respectively, and the active component was purified by silica-gel column chromatography and HPLC. The results suggested that Gardenia jasminoides extract can reduce the cytotoxicity of $A{\beta}$ in PC 12 cells, possibly by reducing oxidative stress.

Antioxidant Phenolic Components from the Whole Plant Extract of Cyperus amuricus Max. (방동사니 전초의 항산화 페놀성 성분)

  • Lee, Sa-Im;Choi, Hoon;Jeon, Hoon;Baek, Nam-In;Kim, Sung-Hoon;Kim, Hee-Ja;Cho, Chong-Hyeon;Ahn, Hyo-Cho;Yang, Jae-Heon;Chae, Byeong-Suk;Lim, Jong-Pil;Eun, Jae-Soon;Kim, Dae-Keun
    • Korean Journal of Pharmacognosy
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    • v.39 no.3
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    • pp.233-236
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    • 2008
  • In order to find the antioxidative components, fractionation of Cyperus amuricus (Cyperaceae) methanol extract was performed by measuring the DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging effect. Three compounds, 3,4-dimethoxy benzoic acid (1), 4-hydroxybenzoic acid (2), and piceatannol (3) were isolated from the active ethylacetate soluble fraction of C. amuricus through repeated silica gel and Sephadex LH-20 column chromatography. Among them, compound 3 showed the significant antioxidative effect on DPPH free radical scavenging test. These compounds are reported for the first time from this plant.

A Caspase Inducing Inhibitor Isolated from Forsythiae fructus (연교(Forsythiae fructus)로부터 분리한 caspase 유도 저해물질)

  • Kim, Jin-Hee;Kho, Yung-Hee;Kim, Mee-Ree;Kim, Hyun-A;Lee, Sang-Myung;Lee, Choong-Hwan
    • Korean Journal of Food Science and Technology
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    • v.34 no.1
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    • pp.114-117
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    • 2002
  • During the screening of inhibitors of caspase-3 induction in U937 human monocytic leukemia cells from natural sources, Forsythiae fructus, which showed a high level of inhibition, was selected. And then, the compound was purified from the methanol extract using silica gel column chromatography and HPLC. The inhibitor was identified as rengyolone, by spectroscophic methods of ESI-MS, $^1H-NMR$, $^{13}C-NMR$, DEPT, and HMBC. Rengyolone showed inhibitory activity of caspase-3 induction, a major protease of apoptosis cascade, with an $IC_{50}$ value of $6.25\;{\mu}g/mL$ after 7 h of treatment in U937 cells. It also showed inhibitory activity of caspace-1 induction, with an $IC_{50}$ value of $7.50\;{\mu}g/mL$ after 40 h of treatment in D10S cells. In addition, it showed protective effect against cell death with an $IC_{50}$ value of $11\;{\mu}g/mL$ on U937 cells induced by etoposide after 24 h of treatment, but did not show any cytotoxicity at the same condition without etoposide, a caspase 3 inducing agent.

In Vitro Peroxynitrite Scavenging Activity of 6-Hydroxykynurenic Acid and Other Flavonoids from Gingko biloba Yellow Leaves

  • Hyun, Sook-Kyung;Jung, Hyun-Ah;Chung, Hae-Young;Choi, Jae-Sue
    • Archives of Pharmacal Research
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    • v.29 no.12
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    • pp.1074-1079
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    • 2006
  • As part of our research on phytochemicals that exert protective effects against diseases related to reactive nitrogen species, we have evaluated the scavenging activity of the yellow leaves of Ginkgo biloba on $ONOO^{-}$. The methanol extract and ethyl acetate fraction obtained from yellow leaves of G. biloba evidenced a marked scavenging activity on authentic $ONOO^{-}$. Repeated column chromatography of the active ethyl acetate soluble fraction on silica gel, Sephadex LH-20, and RP-18, resulted in the purification of 15 known compounds, including sciadopitysin (1), ginkgolide B (2), bilobalide (3), isoginkgetin (4), kaempferol (5), luteolin (6), protocatechuic acid (7), bilobetin (8), amentoflavone (9), ${\beta}-sitosterol$ glucopyranoside (10), kaempferol 3-O-rhamnopyranoside (11), kaempferol 3-O-glucopyranoside (12), kaempferol $3-O-[{6^{'}-O-p-coumaroyl-{\beta}-D-glucopyranosyl(1{\rightarrow}2)-{\alpha}-L-rhamnopyranoside]$ (13), kaempferol 3-O-rutinoside (14), and 6-hydroxykynurenic acid (15). Among the compounds isolated, flavonoids (5, 6 and 11-14), protocatechuic acid (7), and 6-hydroxykynurenic acid (15) all exhibited marked scavenging activities on authentic $ONOO^{-}$. The $IC_{50}$ values of 5-7, 11-14 and 15 were as follows: $2.86{\pm}0.70,\;2.30{\pm}0.04,\;2.85{\pm}0.10,\;5.60{\pm}0.47,\;4.16{\pm}1.65,\;2.47{\pm}0.15,\;3.02{\pm}0.48,\;and\;6.24{\pm}0.27\;{\mu}M$, respectively. DL-Penicillamine ($IC_{50}=4.98{\pm}0.27\;{\mu}M$) was utilized as a positive control. However, the other compounds (1-4, 8-10) exerted no effects against $ONOO^{-}$.

Alkaloids from the Tuber of Aconitum chiisanense (지리바꽃 괴경의 알카로이드)

  • Lee, Moo-Taek;Sung, Hwan-Kil;Whang, Wan-Kyunn;Kim, Il-Hyuk
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.161-173
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    • 1997
  • Tuber of Aconitum chiisanense(Ranunculaceae) a specific medicinal plant in Korea, which is known to have the activity to recover reduced metabolism of feeble patients and has been used to symptoms such as pain, paralysis, atonia and coldness of extremities, etc. were studied. The powdered tubers of the plant were extracted with 10% EtOH 3 times and the combined extract was dissolved in 1N HCl solution and washed with ethyl acetate. The aqueous layer was basified with solid $Na_2CO_3$ and extracted with $CHCl_3$ to obtain an alkaloidal fraction. The alkaloidal fraction was subjected to column chromatography using silica gel, alumina and Sephdex LH 20, etc. From the alkaloidal fraction, five diterpene alkaloids, mesaconitine, aconitine, hypaconitine, 8-O-ethyl 14-benzoylmesaconine and talatizamine, were isolated and identified on the basis of their physico-chemical properties and spectroscopic evidences($^1H$-, $^{13}C$-NMR, EI-MS, IR, 2D-NMR) respectively. Especially the Compound IV, 8-O-ethyl 14-benzoylmesaconine, was assumed to be an artifact resulting from mesaconitine during extraction procedures. The contents of mesaconitine, aconitine and hypaconitine in the mother tuber of this plant were 0.300%, 0.024%, and 0.068%. And that of the attached tuber(new one) of this plant were 0.336%, 0.034% and 0.240% respectively.

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Estimation of polycyclic aromatic hydrocarbons emission from sewage sludge of sewerage treatment plants in Korea

  • Kim, Dong-Hwan;Hwang, Jong-Seob;Lee, Min-Hee;Ok, Gon
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11a
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    • pp.85-88
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    • 2003
  • Polycyclic aromatic hydrocarbons (PARs) are an important group of organic contaminants present in sewage sludge, due to their persistence, toxic, bioaccumulative and long range transfer. These characters make themselves as Persistent Organic Pollutants(POPs) in Long Range Transboundary Air Pollutions convention(LRT AP) of Europe. A method of the gas chromatographic-mass spectrometric (GC-MS) determination of PARs present in sewage sludge was developed and applied to analyzed samples from five sewerage treatment plants (SWTPs), having different treatment types. PARs were extracted from freeze-dried samples by toluene 16 hours in a soxhlet extraction system. The sludge extracts were cleaned-up by an activated silica gel column chromatography. The sum of the 16 US Environmental Protection Agency PARs sewage sludge samples varied from 2.44 to 4.82 ${\mu}g$/g. Concentration of emission carcinogen PARs(PARcarc), such as Benzo(a)anthracene, Benzo(b)fluoranthene, Benzo(k)fluoranthene, Benzo(a)pyrene, Dibenzo(a, h)anthracene and Indeno(1, 2, 3-cd)pyrene ranged from 0.62 to 1.03 ${\mu}g$/g. The total amount of PAHs emission from sewage sludge in Korea was calculated as a top-down approach. PARs and $\sum$PAHcarc from sewage treatment plants had several pathway each by-products. In the ocean dumping, PAHs and $\sum$PAHcarc emissions were 1155.95 kg/year and 5040.32 kg/year. In recycle, PAHs and $\sum$PAHcarc emissions were 98.36 kg/year and 428.87 kg/year. In the landfill, PAHs and $\sum$PAHcarc emissions were 190.40 kg/year and 830.21 kg/year. In the incineration, PAHs and $\sum$PAHcarc emission were 33.10 kg/year and 830.21 kg/year. (In case of incineration, the whole provisions of PARs and $\sum$PAHcarc contained to flowed in sludge was supposed to be exhausted to environment through exhaust after incineration.)

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Antioxidant Activity of Daidzin and Puerarin toward Oxidation of Human Low Density Lipoprotein (갈근에서 분리한 Daidzin 및 Puerarin의 사람 Low Density Lipoprotein 대한 항산화 효과)

  • 박종옥;김경순;지영애;류병호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.25-31
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    • 1997
  • Antioxidative activity of daidzin and puerarin isolated from Puerariae radix against oxidation of low density lipoprotein(LDL) was investigated. The concentration of daidzin at 100$\mu\textrm{g}$/$m\ell$ and puerarin at 60$\mu\textrm{g}$/$m\ell$ inhibited Cu$^{2+}$-mediated oxidation of LDL almost completely. The electrophoretic mobility of oxidized LDL by addition of daidzin(100$\mu\textrm{g}$/$m\ell$) and puerarin(60$\mu\textrm{g}$/$m\ell$) was faster than that of native LDL, but slower than that of oxidized LDL. The oxidized LDL induced by J774 or macrophage was inhibited strongly in the presence of 100$\mu\textrm{g}$/$m\ell$ daidzin and 60$\mu\textrm{g}$/$m\ell$ Puerarin. The formation of conjugated dienes in the oxidized LDL was strongly inhibited by 100$\mu\textrm{g}$/$m\ell$ daidzin and 60$\mu\textrm{g}$/$m\ell$ puerarin.n.

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