• Title/Summary/Keyword: Thin Layer Chromatography(TLC)

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Immunochemical Studies of Starfish Gangliosides: Production of Monoclonal Antibody against AG-2, the Major Ganglioside of Starfish Acanthaster planci, and Detecting Its Distribution in Tissues by TLC Immunostaining

  • Miyamoto, Tomofumi;Yamamoto, Atsushi;Sakai, Maki;Tanaka, Hiroyuki;Shoyama, Yukihiro;Higuchi, Ryuichi
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.298-304
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    • 2006
  • In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

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Purifications of Phenoxyethanol Galactoside and Chlorphenesin Galactoside using Solvent Extraction followed by Gel Chromatography (Solvent Extraction과 Gel Chromatography를 이용한 Phenoxyethanol Galactoside와 Chlorphenesin Galactoside의 정제)

  • Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.954-961
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    • 2017
  • We investigated the purifications of PE-gal and CPN-gal, synthesized by transgalactosylation reaction using recombinant ${\beta}$-gal. The reaction mixture containing PE and PE-gal was first mixed with EA, and thereafter PE and PE-gal were distributed in two-phase (EA/water) system. In this system, PE and PE-gal was selectively moved into EA and water phase, respectively. Then, the water phase was collected, and silica gel chromatography was carried out using the collected water phase. Finally, we compared two purified PE-gal samples using HPLC and TLC analysis, in which the one was purified only by silica gel chromatography and the other was purified by EA extraction followed by silica gel chromatography. In the latter case, the residual PE was almost completely removed, whereas, in the former case, the residual PE was remained remarkably. Additionally, the purification yield of PE-gal was about 21% on the basis of mole. In the same purification protocol, CPN-gal was able to be purified using EA extraction followed by silica gel chromatography, in which the residual CPN was almost removed when CPN-gal was purified by EA extraction followed by silica gel chromatography.

Isolation and Identification of Antimicrobial Compound from Jakyak(Paeonia japonica var. pilosa N$_{AKAI}$ (백작약으로부터 식품부패 미생물에 대한 항균성 물질의 분리 및 동정)

  • 황재선;전희정;한영실
    • Korean journal of food and cookery science
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    • v.16 no.5
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    • pp.445-452
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    • 2000
  • Antimicrobial activity of Jakyak(Paeonia japonica var. pilosa $N_{AKAI}$) was investigated. Methanol extract of dried Jakyak was fractionated to hexane, chloroform, ethylacetate, butanol and aqueous fraction. Ethylacetate fraction among these fractions showed the highest inhibitory effect on the microorganisms such as B. subtilis, L. monocytogenes, and V. parahaemolyticus at 500 $\mu\textrm{g}$/disc. Ethylacetate fraction was further fractionated into 11 fractions by silica gel column and thin layer chromatography(TLC). The results showed that ethylacetate fractions No. 3, 4 and 5 had the highest antimicrobial activity. They were mixed again, re-separated, and 5 fractions were obtained. Among them, the highest inhibitory effect was obtained in No. 3 fraction, which was identified as cetyl alcohol by HPLC and GC-MS.S.

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Studies on the Indoles in Common Reed. -[Part 1] Indole Compounds Occuring in the Shoot of Common Reed [Phragmites Communis Trin.]- (갈대의 INDOLE 화합물(化合物) 연구(硏究) -[제1보](第一報) 갈대 유아(幼芽)의 Indole 화합물(化合物) 검색(檢索)-)

  • Kim, Y.H.;Lee, C.Y.;Kim, I.S.
    • Applied Biological Chemistry
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    • v.19 no.1
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    • pp.24-30
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    • 1976
  • Rhizomes of the common reed (Phragmites communis Trin.) were incubated for three days in the dark. Methanol extract of the shoots was thin layer chromatographed with several solvent systems and visualized with five reagents. The results may be summarized as follows: 1. Serotonin, tryptophan, and tryptamine were identified by cochromatography with the respective authentic compounds. Bufotenine, N-methylserotonin, and N,N-dimethyltryptamine were tentatively identified by their Rf values and colour reactions. The presence of skatole and gramine was suggested. 2. It was esteemed that the common reed might have an active methylation/hydroxylation system of indole compounds at least for a period of time. 3. The presently devised 'overlap' thin layer chromatographic technique may be a useful tool for the identification of a compound whose Rf value was diverse from that of the authentic one by the interferance containing in a sample material.

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Identification of Sugar from Korean Ginseng Saponins by Acid-hydrolysis (인삼(人蔘) Saponin 산가수분해물(酸加水分解物)의 당류(糖類)에 관(關)한 연구(硏究))

  • Yim, Kook-Yi
    • Journal of Nutrition and Health
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    • v.10 no.1
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    • pp.26-33
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    • 1977
  • In order to identify the sugars of saponin originated from Korean ginseng, experimentations were made on the four kinds of ginseng. The results are summarized as follows. 1. The starch content of Ginseng-tail, White-ginseng 3 and 6 years old-whole ginseng were 10.4, 31.5, 8.2, 25.6% and total sugar of its were 37.0, 61.5, 64.5, 62.5% and free sugar were 7.6, 10.5, 11.3, and 10.7% respectively. 2. Saponins were separated from Ginseng-tail, White-ginseng, 3 and 6 years old-whole ginseng by modified SHIBATA method. looms of crude saponin was used for the Thin layer Chromatography (TLC) and thirteen to twelve spots of saponin were isolated by double development $(Solvent:\;CHCL_3\;:\;MeOH\;:\;H_2O=65\;:\;35\;:\;10)$ and by two dimensional development. $(Solvent:\;nBuOH\;:\;HOAC\;:\;H_2O=4\;:\;1\;:\;5)$ The Pattern of spot was not significantly different according to Ginseng sample. 3. Glucose was identified from the acid-hydrolyzate of Ginsen-tail saponin by paper chromatography and isolated the unknown chromatogram seems to be pentose.

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Triglyceride Composition of Kidney Bean (Phaseolus vulgaris L.) (강남콩(Phaseolus vulgaris L.)의 Triglyceride 조성)

  • Kwon, Yong-Ju;Uhm, Tai-Boong;Kim, Choong-Ki;Kim, Sang-Phil;Ko, Seuk-Beum;Lee, Tae-Kyoo;Yang, Hee-Cheon
    • Korean Journal of Food Science and Technology
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    • v.19 no.6
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    • pp.533-536
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    • 1987
  • Lipids of Kidney bean were extracted by the mixture of chloroform-methanol-water (1:2:0.8, v/v) and from these lipids, triglycerides were separated by thin-layer chromatography(TLC) and then reseparated into different fractions by high performance liquid chromatography (HPLC) depending on the polarity of the triglycerides. The triglyceride and fatty acid composition of these fractions were determined by gas liquid chromatography (GLC). From these results, the major triglycerides of Kidney bean lipids were estimated to be $C_{18:2}C_{18:3}C_{18:3}$ (26.6%), $C_{18:2}C_{18:2}C_{18:3}$ (18.5%), $C_{18:3}C_{18:3}C_{18:3}$ (14.9%).

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Antioxidant and α-Glucosidase Inhibitory Activities of the Extract from Sparganium stoloniferum Buch.-Ham. Root and Its Constituent Compounds

  • Xu, Ming Lu;Wang, Lan;Hu, Jian He;Wang, Myeong-Hyeon
    • Preventive Nutrition and Food Science
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    • v.14 no.4
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    • pp.354-357
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    • 2009
  • Three compounds, vanillic acid, p-hydroxylcinnamic acid, p-hydroxybenzaldehyde have been isolated from the ethylacetate extract of Sparganium stoloniferum Buch.-Ham roots using silica gel open column chromatography, preparative thin-layer chromatography (pTLC) and reverse phase high performance liquid chromatography. The structures of the compounds were established on the basis of IR, extensive 1D NMR, and MS analyses. The ethylacetate (EtOAc) extract, vanillic acid, and p-hydroxybenzaldehyde showed $\alpha$-glucosidase inhibition activity of 72.71%, 20.13%, and 30.42%, at the concentration of 10 ${\mu}g/mL$, respectively. The EtOAc extract exhibited strong antioxidant activity with an $IC_50$ value of 24.37 ${\mu}g/mL$ against DPPH radical scavenging activity, the vanillic acid, p-hydroxylcinnamic acid, and p-hydroxybenzaldehyde with an $IC_50$ value of 2.10 ${\mu}M$, 1.59 ${\mu}M$, and 2.72 ${\mu}M$ against DPPH, respectively.

Isolation and Identification of Antimicrobial Compounds from Licorice Extracts (감초 추출물로부터 항균성 물질의 분리 및 동정)

  • Lee, Jin-Man;Lee, Yoon-Won;Hur, Sang-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.2
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    • pp.255-263
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    • 2016
  • Antimicrobial properties of Licorice(Glycyrrhizae radix L.) against food spoilage microorganism, Bacillus subtilis KCTC 1021 was investigated. Antibacterial activity of the essential oil was as equivalent as Potassium metabisulfite and myconazole. The licorice extracts was fractionated to hexane, chloroform, ethyl acetate, butanol and water fraction. Chloroform fraction showed the highest inhibitory effect on the Bacillus subtilis KCTC 1021. Chloroform fraction was further fractionated by silica gel column chromatography and thin layer chromatography(TLC). The antibacterial compound was isolated from their fractions and its chemical structures was identified as (R)-glabridin by ESI-MS, $^1H$-NMR and $^{13}C$-NMR.

Sterol Compositions in Three Solanaceous Seed Oils (3종(種)의 가지과식물종자유중(料植物種子油中)의 Sterol 조성(組成))

  • Jeong, Tae-Myoung;Yang, Min-Suk;Nah, Hyo-Hwan
    • Applied Biological Chemistry
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    • v.21 no.1
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    • pp.51-57
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    • 1978
  • Sterol compositions of three solanaceous plant seed oils i.e., tobacco(Nicotiana tabacum L.), datura (Datura stramonium L.) and Chinese lycium (Lycium chinense Mill.) were analyzed by thin layer chromatography, gas liquid chromatography and gas liquid chromatography-mass spectrometry. Cholesterol, cholest-7-enol, campesterol, 24-methylenecholesterol, stigmasterol, sitosterol, and 28-isofucosterot were identified in 4-des-methylsterol fraction. 31-Norlanost-8-enol, $4{\alpha}$-methylcholest-8-enol, lophenol, 31-norlanosterol, obtusifoliol, 31-norcycloartenol, cyloeucalenol, gramisterol (24-methylenelophenol), and citrostadienol in 4-monomethylsterol fraction, and lanost-8-enol, cycloartanol, lanosterol, ${\beta}$-amyrin, 24-methylenelanost-8-enol, cycloartenol, lupeol and 24-methylenecycloartanol in 4,4-dimethylsterol fraction were identified respectively.

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Comparison of Triglyceride Composition between Red Pepper Seed Oils Harvested from the Chungsong and Youngyang Areas (청송.영양산 고추씨 기름의 Triglyceride 조성의 비교)

  • 김동수;전선미;정동윤;김현대;박영호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.225-232
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    • 1991
  • A Study was carried out to elucidate the triglyceride compositions of the red pepper seed oils harvested from two different areas. The oil was extracted from the red pepper seed with nhexane. Each triglyceride of the oil was separated by thin layer chromatography (TLC) and fractonated by reverse phase high performance liquid chromatography (HPLC) on the basis of acyl carbon numbers, and partition number group(PN) and fatty acid composition of triglyceride were analyzed by gas liquid chromatography (GLC). From the results, it was found that the red pepper seed oils of the Chungsong and Youngyang areas consisted of 14 and 18 kinds of triglycerides, respectively. The red pepper seed oil of the Chungsong area consisted of (C18:2, C18:2, C18:2=41.0%), (C16:0, C18:2, C18:2=37.1%), and that of the Youngyang area consisted of (C18:2, C18:2, C18:2=41.0%), (C16:0, C18:2, C18:2=36.3%) and (C16:0, C16:2, C18:2=8.4%), as the major triglycerides.

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