• Title/Summary/Keyword: silica gel column chromatography

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Isolation and Evaluation of Anti-oxidative Constituents from the Extracts of Ficus erecta var. sieboldii King Leaves (좁은잎천선과 잎 추출물 유래 항산화 활성 성분의 동정 및 효능 확인)

  • Park, Sung Hwan;Kim, Jung Eun;Yeum, Hyun Sook;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.321-328
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    • 2016
  • In this study, we investigated identification of anti-oxidative constituents from Ficus erecta var. sieboldii King (F. erecta) leaves. DPPH and $ABTS^+$ radical scavenging activities were screened for the ethanol extract and solvent fractions, and ethyl acetate fraction showed the most potent scavenging activities. Five constituents were isolated from the ethyl acetate fraction of F. erecta leaves; monoolein (1), oleic acid (2), lutein (3), afzelechin (4), catechin (5). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including NMR spectra, as well as comparison of the data to the literature values. As far as we know, all of the compounds 1 ~ 5 were isolated for the first time from this plant. Studies on DPPH and $ABTS^+$ radical scavenging activities were conducted for the isolated compounds. Among them, afzelechin (4) and catechin (5) showed strong DPPH and $ABTS^+$ radical scavenging activities, whose activities were comparable to a positive control vitamin C. Also, the content of catechin isolated from this plant was determined by HPLC and it was about 3.8 mg/g for the 70% ethanol extract and 20.8 mg/g for the ethyl acetate fraction. From these results, F. erecta leaves extract could be potentially applicable as anti-oxidant ingredients in cosmetic industries.

Biological Control of Phytophthora Blight of Red-pepper Caused by Phytophthora capsici.;II. Isolation and Antifungal Activity of the Substances (고추역병균(疫病菌)(Phytophthora capsici)의 생물학적(生物學的) 방제(防除);II. 항균물질(抗菌物質)의 분리(分離) 정제(精製) 및 항균활성(抗菌活性))

  • Chang, Yoon-Hee;Chang, Sang-Moon;Choi, Jyung;Lee, Dong-Hoon
    • Korean Journal of Environmental Agriculture
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    • v.15 no.4
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    • pp.399-405
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    • 1996
  • In the culture medium, the three antifungal fractions against P. capsici were separated by Sephadex G-25 column chromatography and Silica-gel chromatography. The substance A in white powder and the substance B in sticky oil were isolated by ethyl acetate : acetone mixture(7 : 3), and the substance C in yellow powder was isolated by chloroform : ethyl acetate mixture(95 : 5). The crude extract by ethyl acetate from the culture medium acidified to pH 2 was known to inhibit completely the growth of P. capsici at the level of $50mgkg^{-1}$. The substance A and B were known to be effective above the level of $5mgkg^{-1}$, and the substance C was effective above the level of $1mgkg^{-1}$. However, at the level of $20mgkg^{-1}$, the efficiency was in the order of A>C>B. It is apparent on a pot-experiment scale that the three substances effectively control Phytophthora blight of the red-pepper plant grown in the soil inoculated with P. capsici.

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Propolis from the Stingless Bee Trigona incisa from East Kalimantan, Indonesia, Induces In Vitro Cytotoxicity and Apoptosis in Cancer Cell lines

  • Kustiawan, Paula M;Phuwapraisirisan, Preecha;Puthong, Songchan;Palaga, Tanapat;Arung, Enos T;Chanchao, Chanpen
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6581-6589
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    • 2015
  • Background: Previously, stingless bee (Trigona spp.) products from East Kalimantan, Indonesia, were successfully screened for in vitro antiproliferative activity against human cancer derived cell lines. It was established that propolis from T. incisa presented the highest in vitro cytotoxicity against the SW620 colon cancer cell line (6% cell survival in $20{\mu}g/mL$). Materials and Methods: Propolis from T. incisa was extracted with methanol and further partitioned with n-hexane, ethyl acetate and methanol. The in vitro cytotoxicity of the extracts was assessed by the MTT assay against human colon (SW620), liver (Hep-G2), gastric (KATO-III), lung (Chago) and breast (BT474) cancer derived cell lines. The active fractions were further enriched by silica gel quick column, absorption and size exclusion chromatography. The purity of each fraction was checked by thin layer chromatography. Cytotoxicity in BT-474 cells induced by cardanol compared to doxorubicin were evaluated by MTT assay, induction of cell cycle arrest and cell death by flow cytometric analysis of propidium iodide and annexin-V stained cells. Results: A cardol isomer was found to be the major compound in one active fraction (F45) of T. incisa propolis, with a cytotoxicity against the SW620 ($IC_{50}$ of $4.51{\pm}0.76{\mu}g/mL$), KATO-III (IC50 of $6.06{\pm}0.39{\mu}g/mL$), Hep-G2 ($IC_{50}$ of $0.71{\pm}0.22{\mu}g/mL$), Chago I ($IC_{50}$ of $0.81{\pm}0.18{\mu}g/mL$) and BT474 (IC50 of $4.28{\pm}0.14{\mu}g/mL$) cell lines. Early apoptosis (programmed cell death) of SW620 cells was induced by the cardol containing F45 fraction at the $IC_{50}$ and $IC_{80}$ concentrations, respectively, within 2-6 h of incubation. In addition, the F45 fraction induced cell cycle arrest at the G1 subphase. Conclusions: Indonesian stingless bee (T. incisa) propolis had moderately potent in vitro anticancer activity on human cancer derived cell lines. Cardol or 5-pentadecyl resorcinol was identified as a major active compound and induced apoptosis in SW620 cells in an early period (${\leq}6h$) and cell cycle arrest at the G1 subphase. Thus, cardol is a potential candidate for cancer chemotherapy.

Natural Antioxidant Activity of Ethanol Extracted from Bovine Bile ; Biological Effects and Characterization (초식동물 쓸개즙 추출물의 천연항산화 성분; 생물학적인 기능 및 특성규명)

  • Shim, Jae-Han;Park, Myung-Woo;Lim, Kye-Taek
    • Korean Journal of Environmental Agriculture
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    • v.18 no.3
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    • pp.221-228
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    • 1999
  • This study was carried out to extract the natural antioxidants from Bovine bile and to investigate their effects on various antioxidant activities. It also characterized the patterns of antioxidants by GC/FID and GC/MS. The antioxidative activities and chemical structure of the antioxidant were elucidated by examining the effects of biological activity and the analysis of GC/MS. The antioxidant materials extracted from bovine bile were isolated and purified by silica gel column chromatography and TLC. It was confirmed that there were effects of antioxidants such as Xanthine Oxidase(XO) and Glutathione-S-Transferase(GSH-T) on antioxidative activities. When they were compared with BHT, bile extracts showed the relative effects of 51.2% on the antioxidant activity, the inhibition effects of 48.3% on XO activity, and the synergism effects of 85.7% on the GSH-T activity. According to the results of investigation at neuron cell of mouse, the rate of cell activity in the treatment of 6mM glutathione was 96%, While it in the treatment of 140mg of bile extract was 78%. Based on the TLC analysis of EtOAc extracts from the Bovine bile, the antioxidant activity appeared at $R_f$ value, 0.72. These results suggested that the antioxidant may be coprostan 3-ol.

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A Study on the Antitumor Activity of Panax ginseng (고려인삼의 항암효과에 관한 연구)

  • Hwang, Woo-lk
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.52-60
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    • 1993
  • Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

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Isolation and Identification of Antioxidant Compound from the Lythrum Salicaria L. Roots (털부처꽃(Lythrum Salicaria L.) 뿌리로부터 항산화 물질의 분리 및 구조동정)

  • Lee, Kyeong-Hee;Lee, Dae-Young;Lee, Seung-Eun;Noh, Hyung-Jun;Lee, Jeong-Hoon;Choi, Jehun;Park, Chun-Geun;Kim, Seung-Yu;Lee, Jun-Su;Kim, Geum-Soog
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.359-363
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    • 2014
  • The roots of Lythrum salicaria L. were extracted in 80% aqueous MeOH and the concentrated extract was fractionated with EtOAc, n-BuOH, and $H_2O$, successively. The repeated silicagel and octadecyl $SiO_2$ column chromatographies of the EtOAc fractions led to isolation of an antioxidant compound and two major compounds. From the results of spectral data and the chemical characteristics including nuclear magnetic resonance, MS, and IR, the structures of compounds were determind as myricetin-3-O-${\beta}$-D-glucopyranoside (1), oleanolic acid (2), betulinic acid (3). This is the first reported isolation of compounds (1, 2) from L. salicaria. Compound 1 as well as EtOAc, n-BuOH, and $H_2O$ solvent fractions were evaluated for 2,2-dipicryl-1-phenylhydrazyl radical scavenging activity.

A Study on Discriminative Criteria of 6 Kinds of Achyranthis Radix Using HPLC/DAD;Isolation and Identification of 20-hydroxyecdysone from Aclryranthes japonica $N-{AKAI}$ and Comparison of Patterns of Achyranthis Radix from Different Locations by HPLC (HPLC/DAD를 이용한 6종(種) 우슬(牛膝)의 분류기준 연구;우슬(牛膝)(쇠무릎, Achyranthes japonica $N_{AKAI}$)로부터 20-hydroxyecdysone 분리.동정 및 산지별 우슬의 HPLC 패턴 비교)

  • Kim, Jeong-Hi;Kim, Jong-Mun;Kang, Dae-Hoon
    • The Korea Journal of Herbology
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    • v.23 no.1
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    • pp.109-116
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    • 2008
  • Objectives : This study was performed to investigate the discriminative criteria of 6 kinds of Achyranthis Radix by HPLC/DAD. Methods : 20-hydroxyecdysone is isolated by silica gel column chromatography ($CHCl_3$:MeOH, 7:1-1:1 v/v) and identified by nuclear magnetic resonance, A high-performance liquid chromatographic method with diode array detection was used to identify 20-hydroxyecdysone in A. japonica. The analysis was performed using $C_{18}$ column with isocratic elution consisted of 18% acetonitrile and 82% water and the detection was carried out by DAD at 254 nm. 6 kinds of Achyranthis Radix from different locations were extracted in MeOH. Each extracts was analyzed by HPLC in same condition as used in analysis of 20-hydroxyecdysone. The identities of each extracts were determined by comparing the retention time and UV spectrum with that of reference compound. Results : 1. A. japonica and A. bidentata showed the similar patterns of HPLC chromatogram and 20-hydroxycedysone was present in both of them because the peaks having the same retention time and UV spectrum as 20-hydroxyecdysone were shown in the HPLC chromatograms of A. japonica and A. bidentata 2. Cyathula officinalis and C. capitata showed the similar patterns of HPLC chromatogram. The peak having the same retention time and UV spectrum as 20-hydroxyecdysone was shown in the HPLC chromatogram of C. capitata but not shown in the HPLC chromatogram of C. officinalis. 3. Two species of medicinal drugs from Sacheon province showed similar patterns of HPLC chromatogram. Achyranthis Radix from Sacheon(wild) did not have 20-hydroxycedysone but Achyranthis Radix from Sacheon(cultivated) showed the peak having the same retention time as 20-hydroxyecdysone but UV spectrum of the peak was different from that of 20-hydroxyecdysone. Conclusions : These results suggested that 20-hydroxyecdysone could be the discriminative criteria for Achyranthis Radix contain 20-hydroxyecdysone though they belong to different genus and species. And the patterns of HPLC chromatogram also could be the discriminative criteria as the different species of Achyranthis Radix belonging to the same genus showed similar patterns of HPLC chromatogram.

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In vitro and In vivo Antimicrobial Activities of Medicinal Plants against Crown Gall in Grapevine (포도나무 줄기혹병균에 대한 약용식물의 항균활성 및 병발생억제)

  • Kim, Eun Su;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.34 no.4
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    • pp.537-548
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    • 2016
  • The objective of this study was to evaluate the antimicrobial activities of 9 kinds of medicinal plants against crown gall in grapevine. The medicinal plants extracted with several solvent systems were screened for in vitro antibacterial activity by the disc diffusion method. The ethanol and ethyl acetate extracts from magic lily flowers, tachys roots, asian plantain flowers and seeds, sweet wormwood leaves, stems and flowers, immature bitter melon fruits, cockscomb flowers, and peach tree resin showed in vitro antimicrobial activities against Rhizobium vitis with growth inhibition zones ranging from 10 to 27 mm in diameter. The minimum inhibitory concentration values of extracts against R.vitis ranged from 10,000 in Asian plantain flower and 50,000 fold diluted extracts in sweet wormwood flowers, stems, leaves, cockscomb leaves and immature bitter melon fruits. The active fractions of ethyl acetate and ethanol extracts from the medicinal plants were partially separated through silica gel column chromatography and thin layer chromatography (TLC). The active fractions were separated at Rf 0.36, 0.69, 0.75, 0.84, and 0.94 in sweet wormwood extracts, Rf 0.96 and 0.99 in cockscomb flower extracts, Rf 0.92 and 0.97 in cockscomb leaf extracts, and Rf 0.85 in immature bitter melon fruit extracts in TLC analysis developed with hexane:ethyl acetate (20:80, v/v) and methanol:chloroform (20:80, v/v). Among extracts from plants with in vitro antimicrobial activities, sweet wormwood, cockscomb leaves, and immature bitter melon fruits showed in vivo antimicrobial activities with inhibition activity of 100, 67, and 83.3%, respectively, in 'Kyoho' grapevine inoculated with R. vitis compared with the untreated control. These findings indicate that extracts of medicinal plants could be used as sustainable candidates to control crown gall disease caused by R. vitis in grapevines.

Cytotoxic and Anti-inflammatory Activities of Lipids from the Nuruk (Rhizopus oryzae KSD-815) (누룩(Rhizopus oryzae KSD-815)으로부터 분리한 지질화합물의 세포독성 및 항염증 활성)

  • Kwak, Ho-Young;Lee, Sang-Jin;Lee, Dae-Young;Bae, Nark-Hyun;Jung, La-Koon;Hong, Sung-Youl;Kim, Gye-Won;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.51 no.2
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    • pp.142-147
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    • 2008
  • Nuruk is the Korean traditional Koji that contains various microorganisms and has been used to make the traditional fermented foods including alcoholic beverages. Rhizopus oryzae KSD-815 was isolated from the alcohol-fermenting Nuruk used for manufacturing traditional alcohol. In this study, the authors reported the isolation and identification of four lipids from the Nuruk (Rhizopus oryzae KSD-815) that inoculated wheat with Rhizopus oryzae KSD-815. The dried and powdered Nuruk (Rhizopus oryzae KSD-815) were extracted three times at room temperature with 80% aqueous MeOH. The extracts were partitioned with EtOAc, n-BuOH, and water, successively. The EtOAc extract was suspended in 80% MeOH and partitioned repeatedly with n-hexane. From the n-hexane fraction, four lipids were isolated through the repeated silica gel and ODS column chromatographies. According to the results of physico-chemical data including NMR, GC and MS, the chemical structures of the compounds were determined as linolenic acid methyl ester (1), palmitic acid methyl ester (2), linoleic acid (3), palmitic acid (4). Cytotoxicity was evaluated in huamn breast cancer cells, MDA-MB-231 and human hepatocarcinoma, SK-HEP-1 cells using MTT assay. Exposure of compounds 1 and 3 led to a dose-dependent inhibition of cell viability in both cancer cell lines. In addition, treatment of RAW264.7 cells with compound 3 caused inhibition of lipopolysaccharide/interferon-${\gamma}$-induced nitric oxide production.

Enzymatic Interesterification and Melting Characteristic for Asymmetric 1,2-Distearoyl-3-Oleoyl-rac-Glycerol Triacylglycerol Enriched Product (효소적 반응을 이용한 비대칭형 1,2-Distearoyl-3-Oleoyl-rac-Glycerol 혼합물의 생성 및 융점 특성)

  • Kim, Jin Young;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.93-101
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    • 2014
  • Asymmetric 1,2-distearoyl-3-oleoyl-rac-glycerol (SSO) triacylglycerol (TAG) is used as a cocoa butter replacer (CBR). In this study, it was produced by lipase-catalyzed interesterification of fully hydrogenated soybean oil (FHSBO) and oleic ethyl ester (OEE) in a batch type reactor at $75^{\circ}C$, 250 rpm. Different molar ratios (FHSBO : OEE=1:1, 1:2 and 1:3, w/w) and various reaction times (1, 2, 3, 4, and 5 hr) were also tested. The optimized condition for SSO was a FHSBO : OEE molar ratio of =1:1 at reaction times of 2, 3, 4, and 5 hr. Enzymatic synthesis generated SSO/SOS, as well as the other TAGs (e.g., PSO/POS, SOO/OSO, SSS), ethyl esters, monoacylglycerol (MAG), and diacylglycerol (DAG). After scale-up, fractionation by solvent (methanol and acetone) fractionation and column chromatography was applied. To reduce ethyl esters, high-melting TAGs (e.g., SSS), and SOO/OSO in reactants, solvent fractionation was applied. Using a silica gel column (sample : silica gel=2:1, wt%), MAG and DAG were removed at $25^{\circ}C$. The major fatty acid composition of the final products (with a high SSO/SOS content) was palmitic acid (C16:0, 10.9~12.9 area%), stearic acid (C18:0, 52.2~54.9 area%), and oleic acid (C18:1, 34.2~35.5 area%). In reversed-phase HPLC analysis, the major TAG species of the final product (FHSBO : OEE=1:1, 2 hr) were SSO/SOS (82.31 area%) and PSO/POS (14.51 area%). Based on the $[SS]^+$ : $[SO]^+$ ratio obtained by RP-HPLC/APCI-MS, the final product had a higher SSO (AAB type TAG) content than cocoa butter (CB). The solid fat index (SFI) of CB and the final product obtained were similar with a narrow melting point range around ~32 to $35^{\circ}C$.