• Title/Summary/Keyword: Ion-Exchange Chromatography

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Purification and Characterization of Apolipophorin-III from Haemolvmph of Fall Webworm Hvphantria cunea Drury (미국흰불나방(Hyphantria cunea Drury) 혈림프부터 apolipophorin-III의 순수정제 및 특성)

  • 윤화경;서신자김학열
    • The Korean Journal of Zoology
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    • v.37 no.4
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    • pp.488-494
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    • 1994
  • Apolipophorin-III (ApoLp-III) was purified from adult haemolynph of Hyphantriu cuneo and their molecular weight and synthetic place were investigated. ApoLp-III purification was performed by KBr-density gradient ultracentrifugation followed by gel permeation chromatographv (Sephadex G-1001 and ion-exchange chromatography (CM-52) and their purity was confirmed on 10% SDS-PAGE. ApoLp-III has the molecular weight of 18 ItDa and is synthesized by fat body.

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Studies on the Carbohydrates of Ginseng (part, 1) Free sugars in Ginseng (人蔘成分에 關한 硏究 (第1報) 遊離糖類에 對하여)

  • Lee, Tae-Young;Kwon, Tai-Wan
    • Journal of the Korean Chemical Society
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    • v.5 no.1
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    • pp.73-75
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    • 1961
  • The Compositions of the free sugars in Korean Ginseng were determined by use of the ion exchange chromatography and anthrone reaction. The free sugars were sucrose, fructose and glucose. The contents of each sugar in dried and wet Ginseng were Sucrose: 8.525 and 5.280, Fructose: 0.498 and 0.245, glucose: 0.433 and 0.975, respectively.

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A STUDY OF THE PHOTOTOXICITY OF ORANGE FLOWER ABSOLUTE

  • Yang, Hai-Joo;Lee, Neung-An
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.13 no.1
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    • pp.1-6
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    • 1987
  • We have confirmed the phototoxicity of orange flower absolute, studied the phototoxic ingredient and developed a method to eliminate it. To confirm the phototoxicity, we tested French orange flower absolute, Moroccan absolute and Egyptian absolute with Hartley strain guinea Pig under UV irradiation ranging from 320400 nm supplied with fluorescent lamps. Using a combination of isolation techniques including HPLC and IR, we were able to confirm that the phototoxic agent was bergapten. The development of a non-phototoxic orange flower absolute using ion exchange chromatography is described.

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Purification and Properties of Polygalacturonase from Ganoderma lucidum (Ganoderma lucidum이 생산하는 Polygalacturonase의 정제 및 특성)

  • Yoon, Sook;Kim, Myung-Kon;Hong, Jai-Sik;Kim, Myeong-Sook
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.298-308
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    • 1994
  • The properties of polygalacturonase by Ganoderma lucidum in liquid culture were investigated. The enzyme was composed of an endo- and an exo-polygalacturonase. The endo- and exo-polygalacturonase were purified approximately 56 and 9.2-fold, respectively, through ammonium sulfate fractionation, gel filtration on Biogel P-100, anion exchange chromatography on DEAE-cellulose, gel chromatography on Sephadex G-150 and re-gel chromatography on Sephadex G-150. The endo- and exo-polygalacturonase had higher affinity for apple pectin than for citrus pectin or pectic acid. The Km values of the endo- and exo-polygalacturonase for apple pectin, determined on the Lineweaver-Burk plot, were 1.44 and 10.6 mg $ml^{-1}$ for apple pectin, respectively. Purified endo-polygalacturonase was found to be homogeneous electrophoretically and had a molecular weight of 54,000 estimated on SDS polyacrylamide gel. The optimal pH for the activity of the enzymes was 4.0. The endo- and exo-polygalacturonase were stable in the pH range of 4.0 to 6.0 and 3.5 to 5.5, respectively. The optimal temperatures of the endo- and exo-polygalacturonase were 40 and $60^{\circ}C$, respectively. The exo-polygalacturonase was more resistant to heat than the endo-polygalacturonase, requiring heating for 40 min at $80^{\circ}C$ for complete inactivation. The activity of the endo-polygalacturonase was increased by $Ca^{++}$ and $Mn^{++}\;ions$, while that of the exo-polygalacturonase was increased by $Ca^{++}\;ion$ only, and was not affected by $Mn^{++}\;ion$.

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Studies on the Nonvolatile Organic Acids in Kimchis fermented at Different Temperatures (숙성온도(熟成溫度)에 따른 김치의 비휘발성(非揮發性) 유기산(有機酸)에 관(關)한 연구(硏究))

  • Kim, Hyun-Ock;Rhee, Hei-Soo
    • Korean Journal of Food Science and Technology
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    • v.7 no.2
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    • pp.74-81
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    • 1975
  • Nonvolatile organic acids were determined in raw Chinese cabbage, Kimchis fermented at $6^{\circ}{\sim}7^{\circ}C$ and at $22^{\circ}{\sim}23^{\circ}C$. Total acids were isolated by ion-exchange chromatography and quantitative determination of individual acids was performed by silicic acid partition column chromatography. Acids were identified qualitatively by paper chromatography. Palatability of Kimchis was evaluated by nine pannels. Results were as follows: Maleic, fumaric, lactic, succinic, malonic, oxalic, glycolic, malic, citric, tartaric, sulphuric and phosphoric acid were found in raw Chinese cabbage. Kimchis fermented at $6^{\circ}{\sim}7^{\circ}C$, and at $22^{\circ}{\sim}23^{\circ}C$. In raw Chinese cabbage, most of organic acids were in salt form and malic acid was highest in amount. In Kimchi fermented at $6^{\circ}{\sim}7^{\circ}C$, the amount of lactic acid and succinic acid was higher than that in Kimchi fermented at $22^{\circ}{\sim}23^{\circ}C$, and, tartaric, oxalic, malic, malonic, maleic, glycolic and fumaric acids were less than those in Kimchi fermented at $22^{\circ}{\sim}23^{\circ}C$. Kimchi fermented at $6^{\circ}{\sim}7^{\circ}C$ had a more flavorous sour taste than Kimchi fermented at $22^{\circ}{\sim}23^{\circ}C$. It is assumed that large amount of lactic acid and succinic acid in Kimchi fermented st $6^{\circ}{\sim}7^{\circ}C$ have a connection with flavorous sour taste.

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A Multidimensional System for Phosphopeptide Analysis Using TiO2 Enrichment and Ion-exchange Chromatography with Mass Spectrometry

  • Cho, Kun;Yoo, Ji-Sun;Kim, Eun-Min;Kim, Jin-Young;Kim, Young-Hwan;Oh, Han-Bin;Yoo, Jong-Shin
    • Bulletin of the Korean Chemical Society
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    • v.33 no.10
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    • pp.3298-3302
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    • 2012
  • Although offline enrichment of phosphorylated peptides is widely used, enrichment for phosphopeptides using $TiO_2$ is often performed manually, which is labor-intensive and can lead to irreproducible results. To address the problems associated with offline enrichment and to improve the effectiveness of phosphopeptide detection, we developed an automated online enrichment system for phosphopeptide analysis. A standard protein mixture comprising BSA, fetuin, crystalline, ${\alpha}$-casein and ${\beta}$-casein, and ovalbumin was assessed using our new system. Our multidimensional system has four main parts: a sample pump, a 20-mm $TiO_2$-based column, a weak anion-exchange, and a strong cation-exchange (2:1 WAX:SCX) separation column with LC/MS. Phosphorylated peptides were successfully detected using the $TiO_2$-based online system with little interference from nonphosphorylated peptides. Our results confirmed that our online enrichment system is a simple and efficient method for detecting phosphorylated peptides.

A Study on the Characteristics of Anion Exchange Membrane According to Aliphatic Alkyl Chain Spacer Length Introduced into Branched Poly (Arylene Ether Sulfone) (수지상 폴리(알릴렌 이써 설폰)에 도입된 지방족 알킬사슬 연결자길이에 따른 음이온교환막의 특성 연구)

  • KIM, HYUN JIN;YOO, DONG JIN
    • Transactions of the Korean hydrogen and new energy society
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    • v.33 no.3
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    • pp.209-218
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    • 2022
  • Recently, research on the development of anion exchange membranes (AEMs) has received considerable attention from the scientific community around the world. Here, we fabricated a series of AEMs with branched structures with different alkyl spacers and conducted comparative evaluations. The introduction of these branched structures is an attempt to overcome the low ionic conductivity and stability problems that AEMs are currently facing. To this end, branched polymers with different spacer lengths were synthesized and properties of each membrane prepared according to the branched structure were compared. The chemical structure of the polymer was investigated by proton nuclear magnetic resonance, Fourier transform infrared, and gel permeation chromatography, and the thermal properties were investigated using thermogravimetric analysis. The branched anion exchange membrane with (CH2)3 and (CH2)6 spacers exhibited ionic conductivities of 8.9 mS cm-1 and 22 mS cm-1 at 90℃, respectively. This means that the length of the spacer affects the ionic conductivity. Therefore, this study showing the effect of the spacer length on the ionic conductivity of the membrane in the polymer structure constituting the ion exchange membrane is judged to be very useful for future application studies of AEM fuel cells.

Effects of Extraction Method on Anserine, Protein, and Iron Contents of Salmon (Oncorhynchus keta) Extracts (연어(Oncorhynchus keta) 추출물 중의 Anserine, 단백질 및 철분 함량에 미치는 추출방법의 영향)

  • Min, Hye-Ok;Park, In-Myoung;Song, Ho-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.2
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    • pp.220-228
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    • 2017
  • Effects of extraction methods on reducing concentrations of pro-oxidants (total iron and protein) of salmon was determined. For development of the extraction process, the effectiveness of several extraction methods was determined and compared, including heat treatment (60, 80, and $100^{\circ}C$), ion exchange and carboxymethyl (CM)-cellulose column chromatography, and ultrafiltration (UF). Protein, total iron, and anserine contents of salmon extracts were 23.64 mg/mL, $16.20{\mu}g/mL$, and 5.47 mg/mL in non-heated extracts, 7.40 mg/mL, $2.32{\mu}g/mL$, and 5.20 mg/mL in heated extracts at $60^{\circ}C$, 7.64 mg/mL, $1.20{\mu}g/mL$, and 5.21 mg/mL at $80^{\circ}C$, and 7.04 mg/mL, $0.68{\mu}g/mL$, and 4.04 mg/mL at $100^{\circ}C$, respectively. Heating and UF decreased contents of protein and total iron, whereas only UF slightly decreased anserine content. Application of the primary ion exchange method increased the content of anserine up to 16%. Protein and total iron contents by the primary ion exchange method decreased by 70 and 98%, respectively. Secondary ion exchange (CM-cellulose) treatment after primary ion exchange and UF resulted in lower anserine content than the primary ion exchange method. However, the content of impurities (protein, total iron) was lower than in all other salmon extracts. Therefore, primary ion exchange, UF, and secondary ion exchange method were the best extraction processes in this study.

Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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Isolation, Purification and Characterization of Polysaccharides that induce in vitro Immuno-Stimulation of Macrophases derived from Liquid Culture of Cordyceps militaris

  • Kwon, Jeong-Seok;An, Hyo-Sil;Hong, Eock-Kee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.287-292
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    • 2005
  • The crude polysaccharides(C-CPF, C-CPM, C-CPB) derived from fruiting body, mycelia and mycelia free broth of cordyceps militaris were obtained by ethanol precipitation of hot water extracts. After a batch fermentation of C. militaris was carried out in a 5 L jar vessel, endo-polysaccharide and exo-polysaccharide were obtained. They were demonstrated as the hetero polysaccharides which were composed of glucsose, galactose and mannose by performed with HPAEC(high pH anion exchange chromatography) and conformation of random coil by its complex forming ability with congo red reagent. They were purified by ion exchange (DEAE-cellulose) and gel filtration chromatography. They were monitered by phenol-sulfuric acid method and Bradford method. The NO induction activities of crude polysaccharides and purified polysaccharides derived from mycelia free broth were enhanced rather than LPS(lipo polysaccharide) which was used as a general NO inducer. These effects presumably contibute to the antitumor activities. The homogenieties and molecular weights of polysaccharides were determined by using Sepharose CL-6B. The yield, molecular weights and NO induction activities of C-CPFN Fr.III, C-CPMN Fr.III, C-CPBN Fr.II were 0.387, 0.408 and 0.153, 127 K 210 K and 36 K, 40.79%, 88.72%, and 104.17%, respectively.

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