• Title/Summary/Keyword: Preparative Chromatography

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Calcium Solubilization Ability and Anti-Inflammatory Effects of Hydrolyzed Casein

  • Kim, Da Young;Yoo, Jung Sik;Cho, Yoon Ah;Yoon, Ho Sik;Kim, Cheol-Hyun
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.687-700
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    • 2021
  • This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase®; Neutrase®; Protamex®; Flavourzyme®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex® and Neutrase® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.

Antiplatelet fraction from Ulmi cortex and its active components (유백피의 항혈전 활성 분획 및 유효성분에 관한 연구)

  • Kim, Dong-Seon;Yang, Won-Kyung;Sung, Yoon-Young;Lim, Sun Mi;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.39-44
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    • 2013
  • Objectives : The purpose of this study was to identify active fraction and components from antiplatelet Ulmi cortex extract. Methods : The 70% ethanol extract of Ulmi cortex was subjected to column chromatography over D101 resin and eluted with an 20% (W1), 30% (W2), 40% (W3), 50%(W4), 70% (W5), and 100% ethanol (W6) to yield 6 fractions. W6 was further fractioned and its active components were purified using semi-preparative HPLC. The isolated compounds were identified by MS and NMR, and their contents were simultaneously analyzed using HPLC/UV. Antiplatelet aggregation activities of the fractions and the compounds were evaluated using rat platelet-rich plasma in presence of collagen ($5{\mu}g/ml$), arachidonic acid (0.05 U/ml), or thrombin ($100{\mu}M$). Results : Among six fractions, W3 prominently inhibited platelet aggregation. At the concentration of $200{\mu}g/ml$, W3 strongly inhibited arachidonic acid- and collagen-induced platelet aggregations by 78.2% and 65.9%, respectivley, and weakly inhibited thrombin-inducded platelet aggregation by 32.6%. Catechin, epicatehin, and catechin-7-O-${\beta}$-D-glucopyranoside were isolated from W3 and their contents were revealed to be 15.1%, 0.87%, and 0.32%. Catechin and epicatechin at the concentrations of $100{\mu}M$ strongly inhibited collagen-induced platelet aggregation by 79.9% and 86.6%, respectively, but weakly inhibited arachidonic acid- and thrombin-induced platelet aggregations. Conclusions : A main active principle of anitplatelet Ulmi Cortex extract is W3 fraction, of which main active component is catechin considering its antiplatelet activity and content.

Studies on the Development of Natural Preservatives from Natural Products (전통식품 및 천연물에서 천연보존료 개발에 관한 연구)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Kwon, Yong-Kwan;Lee, Ju-Yeun;Kim, So-Hee;Ha, Sang-Chunl;Cho, Hong-Yon;Chang, Ih-Seop;Lee, Chul-Won;Kim, Kil-Saeng
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1667-1678
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    • 1999
  • Certain parts of 190 kinds of medicinal herbs and 171 kinds of original materials of food were extracted by methanol. The extracts were tested their microbial inhibition activities against several food spoilage microorganisms, Micrococcus luteus, Bacillus subtilis, Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli, Saccharomyces cerevisiae, Candida albicans, Penicillium citrinum, Aspergillus flavus and Aspergillus niger. The methanol extracts of Cornus officinalis, Evodia officinalis, Glycyrrhiza glabra, Salvia miltiorrhiza. Schizandrae fructus, Coptidis rhizoma, aroma hop and bitter hop were shown inhibitory effect on certain species of gram(+) bacteria. Aroma hop and bitter hop were shown inhibitory effect on certain species of gram(-) bacteria. The methanol extract of Salvia miltiorrhiza exhibited a strong antibacterial activities. It was purified by solvent fractionation, silicagel column chromatography, prep. TLC, prep. HPLC. The purified active substance was identified as cryptotanshinone by EIMS, $1^H-NMR,\;{13}^C-NMR$ and DEPT. Cryptotanshinone showed a strong antibacterial activity against gram positive bacteria $(MIC\;:\;3.91{\sim}62.50\;{\mu}g/mL)$. Especially, this compound was the most strong activity against Bacillus subtilis $(MIC\;:\;3.91\;{\mu}g/mL)$.

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Antioxidant Properties of 7 Domestic Essential Oils and Identification of Physiologically Active Components of Essential Oils against Candida albicans (식물정유 7종의 항산화능 분석 및 Candida albicans 생장 억제 정유의 생리활성 성분 구명)

  • LEE, Sang-Youn;LEE, Da-Song;CHO, Seong-Min;KIM, Jong-Chan;PARK, Mi-Jin;CHOI, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.49 no.1
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    • pp.23-43
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    • 2021
  • In this study, we selected two essential oils, Citrus unshiu and Cinnamomum cassia with superior antioxidant effects from the essential oils of 7 wild plants in South Korea and examined their antimicrobial activity against Candida albicans, which causes dermatitis to identify the antimicrobial components in the essential oils. As a result of measuring DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, SC50 value of the Citrus unshiu essential oil was 0.010 mg/mL, while for the Cinnamomum cassia essential oil, SC50 value was 0.09 mg/mL. In addition, when ABTS (2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity was measured, SC50 value of the Citrus unshiu essential oil was 0.09 mg/mL, while for the Cinnamomum cassia essential oil, it was 0.06 mg/mL, exhibiting high antioxidant activity. For the minimum inhibitory concentration (MIC), the essential oil of Cinnamomum cassia was 1.25 mg/mL and that of Citrus unshiu was 5 mg/mL, demonstrating a high antimicrobial activity of the Cinnamomum cassia essential oil. Through the thin layer chromatography (TLC) bioassay, we assessed the antimicrobial activity against C. albicans according to the fraction components of the two essential oils. Also, by using preparative TLC (prep. TLC), we obtained the active fractions, and by performing GC/MS analysis of the components with the same Rf value, we identified the antimicrobial-active components. As a result, the main components having antioxidant and antimicrobial activities were cinnamyl acetate, eucalyptol, linalool, and citral of the Cinnamomum cassia essential oil and linalool from the Citrus unshiu essential oil. Also, based on the analysis of the fractional components that showed antioxidant and antimicrobial activities in both of the two essential oils, it was found that linalool has antioxidant activity, while cinnamyl acetate, eucalyptol, citral, and geranyl acetate have antioxidant and antimicrobial activities.

Isolation and Identification of a Sterol and Three Glucosides from the Peel of Pear (Pyrus pyrifolia Nakai cv. Chuhwangbae) (추황배(Pyrus pyrifolia Nakai cv. Chuhwangbae) 과피로부터 1종의 Sterol과 3종의 배당체 화합물의 단리 · 동정)

  • Lee, Yu Geon;Cho, Jeong-Yong;Lee, Hyun Joo;Lee, Yong Hyun;Lee, Sang-Hyun;Han, Tae-Ho;Kim, Wol-Soo;Park, Keun-Hyung;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.557-564
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    • 2013
  • We isolated and identified antioxidants from acidic and neutral ethyl acetate fractions of the peel of pear (Pyrus pyrifolia N. cv. Chuhwangbae). We isolated 4 compounds from the methanol extract, by using 3 different types of column chromatography (Sephadex LH-20, silica gel, and octadecylsilane) and preparative HPLC. We identified the isolated compounds as (S)-(+)-2-cis-abscisic acid O-${\beta}$-D-glucopyranosyl ester (compound 1), 1-[4-O-${\beta}$-D-glucopyranosyl]phenyl ethanone (picroside, compound 2), ${\beta}$-sitosterol (compound 3), and ${\beta}$-sitosteryl 3-O-${\beta}$-D-glucopyranoside (compound 4) by nuclear magnetic resonance analysis. We are the first to report the identification of compounds 1, 2, and 4 from pear.

Antioxidative Properties of Asparagus cochinchinensis Root (천문동 건근의 항산화 효과에 관한 연구)

  • Koo, Kyoung Yoon;Kim, Won Baek;Park, So Hae;Kim, Minji;Kim, Bo Ram;Hwang, Jihoe;Kim, Min Jung;Son, Hong Joo;Hwang, Dae Youn;Kim, Dong Seob;Lee, Chung Yeoul;Lee, Heeseob
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.524-532
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    • 2016
  • This study was performed to compare the antioxidative activities of methanol extracts from Asparagus cochinchinensis with whole root (W-AC), flesh (F-AC), and root bark (B-AC). To evaluate the antioxidative properties of their methanol extracts, 1,1-diphenyl-2-picrylhydrazyl radical, nitrite, hydroxyl radical, 2,2'-azino-bis(3-ethylbenz thiazoline-6-sulfonate) radical scavenging activities, and contents of total flavonoid and polyphenol contents were measured. B-AC extract showed the highest antioxidative activity, whereas F-AC extract showed the lowest. For B-AC extract, caffeic acid was isolated by preparative high-performance liquid chromatography and confirmed by liquid chromatography-mass spectrometry and absorption spectroscopy, which showed 1.6% of total polyphenol contents among all methanol extracts.

Analysis of Residual Triflumizole, an Imidazole Fungicide, in Apples, Pears and Cucumbers Using High Performance Liquid Chromatography

  • Khay, Sathya;EI-Aty, A.M. Abd;Choi, Jeong-Heui;Shim, Jae-Han
    • Toxicological Research
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    • v.24 no.1
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    • pp.87-91
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    • 2008
  • The present study was conducted to monitor the level of triflumizole residues in fruits (apple and pear) and vegetable (cucumber) samples in order to assess risk posed by the presence of such residues to the consumer. Triflumizole was applied at a recommended dose rate to apple and pear pulps and to a cucumber sample. The samples were collected at harvesting time following several treatments (three and/or four treatments). Triflumizole was extracted with methanol and re-extracted into dichloromethane. The presence of triflumizole was determined by HPLC with UV detection at 238 nm following the cleanup of the extract by open preparative chromatographic column with Florisil. The versatility of this method was evidenced by its excellent linearity (> 0.999) in the concentration range between 0.2 and 4.0 mg/kg. The mean recoveries evaluated from the untreated samples spiked at two different fortification levels. 0.1 and 0.4 mg/kg, and ranged from 87.5${\pm}$0.0 to 93.3${\pm}$2.6 for the tested fruits and vegetable, respectively, and the repeatability (as relative standard deviation) from three repetitive determinations of recoveries were no larger than 6%. The calculated limit of detection was 0.02 mg/kg and the minimum detectable level of 4 ng for triflumizole was easily detected. When triflumizole was sprayed onto the apple trees three times at 50-40-30 and 40-30-21 days prior to harvesting and four times onto the pear trees at 40-30-21-14 days prior to harvesting, the mean residual amounts of 0.05 and 0.06 mg/kg for apples and pears, respectively, were not detected in all of the treatments. When the cucumber sample was fumigated four times at 7, 5, 3 and 1 day prior to harvesting, the mean residual amount was not detectable. Triflumizole can be used safely when sprayed (wettable powder, 30% active ingredient) and fumigated (10%) 4 times at 14 and 1 day prior to harvesting to protect the fruits and vegetable, respectively.

Studies on the Constituents of Korean Plant Edible Oils and Fats -Part 1. Composition of the sterol fraction of rape, sesame and perilla oils- (한국산(韓國産) 식물식용유지(植物食用油脂)의 성분(成分)에 관(關)한 연구(硏究) -제(第) 1 보(報) 채종(菜種), 호마(胡麻) 및 소마유(蘇麻油)의 sterol에 대하여-)

  • Ko, Young-Su;Chang, You-Kyung;Lee, Hyo-Jee;Woo, Sang-Kyu;Yang, Cha-Buym
    • Journal of Nutrition and Health
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    • v.10 no.2
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    • pp.44-53
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    • 1977
  • The Korean origin edible oil sterol part of the rape, sesame and perilla oil can be separated from the other constituents of the non-saponifiable fraction, by the thin layer chromatography on the preparative Plates. The composition of sterols have been determined by gas liquid chromatography and thin layer chromatographic anlysis. Two sterols, ${\beta}$-sitosterol and campesterol were present in all of the oils. And brassicasterol were found in rapeseed oil in addition to the two sterols that were common to all of the oils studied. It was noted that ${\beta}$-sitosterol was the major sterol in the Korean original edible oils. The results showed that contents of sterols were campesterol 24.31%, ${\beta}$-sitosterol 58.90% ana Brassicasterol 11.54%, and $\Delta^7$-sterol 5.25% by method of triangulation and campesterol 26.16%, ${\beta}$-sitosterol 57.50%, brassicasterol 11.70% and ${\Delta}^7$-sterol 4.64% by method of Planimetry of rape seed oil. By sesame seed oil sterol compositions were campesterol 20.35%, stigmasterol 9.15%, ${\beta}$-sitosterol 43.49%, ${\Delta}^7$-sterol 11.25% and others 15.76% by method of triangulation and campesterol 16.79%, stigmasterol 8.69%, ${\beta}$-sitosterol 44.58%, ${\Delta}^7$-sterol 14.28% and others 15.56% by method of planimetry. Campesterol 12.45%, stigmasteriol 5.40%, ${\beta}$-sitosterol 72.32% and ${\Delta}^7$-Sterol 9.83% by method of triangulation-and campesterol 13.00%, stigmasterol 3.76%, ${\beta}$-sitosterol 74.57% and ${\Delta}^7$ sterols 8.67% by method of planimetry of perilla oil. Contents of totalsterol in Korean edible oils were 0.82% by rape, 0.58% by sesame and 0.45% by perilla, respectively.

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Isolation and Purification of Garlic Specific Organic Compounds (마늘 특이 유기화합물의 분리 및 정제)

  • Oh, Tae-Young;Kyung, Kyu-Hang
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.553-557
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    • 2011
  • Garlic specific organic compounds were separated and purified using a recycling preparative high-performance liquid chromatography (HPLC) from blanched garlic cloves. Identification of the compounds involved comparing the previously reported HPLC retention times as well as other identification methods including $^1H$- and $^{13}C$-nuclear magnetic resonance and liquid chromatography-mass spectrometry. The yields of garlic specific organic compounds were 12.2, 42.5, 1.6, 1.2, and 4.8% on wet weight basis of garlic for alliin(S-allyl-L-cysteine sulfoxide), isoalliin(S-1-propenyl-L-cysteine sulfoxide), ${\gamma}$-glutamyl-S-allylcysteine, ${\gamma}$-glutamyl-S-1-propenylcysteine and ${\gamma}$-glutamyl-phenylalanine, respectively. All the compounds, except for ${\gamma}$-glutamylphenylalanine, contained sulfur.

Isolation and Characterization of Cathepsin B inhilbitor Produced by Streptomyces luteogriseus KT-10 (Streptomyces luteogriseus KT-10 이 생산하는 Cathepsin B 저해물질의 분리 및 특성)

  • 한길환;김상달
    • Microbiology and Biotechnology Letters
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    • v.29 no.2
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    • pp.84-89
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    • 2001
  • Isolation and Characterization of Cathepsin B inhibitor Produced by Streptomyces luteogriseus KT-IO. Han, Kil~Hwan and Sang~Dal Kim*. Department of Applied Microbiology, Yeungnam Universit}/t Kyongsan 712749, Korea - The cathepsin B inhibitor produced by Streptomyces luteogriseus KT-IO was very stable in heat, acidic and alkaline conditions. The cathepsin B inhibitor was isolated from the extracted fraction of culture broth with butanol, methanol and chloroform subsequently, the inhibitor was purified with following several column chromatography sLlch as DEAE-Sephadex A-25, Sephadex G-15, silica gel 60, Sephadex LH-20, and preparative HPLC. The cathepsin B inhibitor showed positively to detective reaction of ninhydrine, 5% H2S04, iodine, but negatively to the reaction of Ehrlich's reagent, DNS, aniline. The molecular formular of cathepsin B inhibitor was elucidated by JR, lH and 13C-NMR, FAB mass and elemental analyzer. Consequently, it was identified as C4HlI04N6. The cathepsin B inhibitor had the mode of competitive inhibition with the reaction of cathepsin B.

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