• Title/Summary/Keyword: Silica Gel Chromatography

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Characteristics of Histamine Forming Bacteria from Tuna Fish Waste in Korea (국내 참치 부산물 내 히스타민 생성 주요 세균의 특성 구명)

  • Bang, Min-Woo;Chung, Chang-Dae;Kim, Seon-Ho;Chang, Moon-Baek;Lee, Sung-Sil;Lee, Sang-Suk
    • Journal of Life Science
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    • v.19 no.2
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    • pp.277-283
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    • 2009
  • Biogenic amines are generally formed through the decarboxylation of specific free amino acids by exogenous decarboxylases released by microbial species associated with the fish products and fermented feeds. This study was conducted to investigate the properties of e tuna waste regarding the control of degradation of biogenic amines (histamine, tyramine, tryptamine, putrescine, and cadaverine) that might be related with the anti-nutritional factor of the tuna waste that is used for manufacturing domestic fish meal. The values of pH and the salt content were 6.51, 3.35% in tuna waste and 5.58 and 5.83% in tuna fish meal, respectively. The strains and dominant bacteria tested in the tuna waste sample were 9.20, 9.29, 5.67, 7.82 and 7.58 log CFU/g of total bacteria, aerobic plate count (APC), total coliform (TC), Lactobacillus spp. and Bacillus spp., respectively. The main histamine forming-bacteria (HFB) in tuna waste were detected by silica gel thin-layer chromatography (TLC) and 7 histamine-forming bacterial species were isolated among microbes grown in selective medium. The histamine concentration was determined by detection of fluorescence of ο-phthaldialdehyde (OPA) derivatives using HPLC and the date were used to reconfirm the identities of the amine-producing bacteria. The 15 histamine- forming bacteria strains grown in trypicase soy broth (TSB) supplemented with 1% L-histidine (TSBH) were identified as Lactococcus(L.) lactis subsp. lactis, Klebsiella pneummonlae, L. garvieae 36, Vibrio olivaceus, Hafnia alvei and L. garvieae which were main dominant amine - producing strains, and Morganella morganii identified by 16S ribosomal RNA (rRNA) sequencing with PCR amplification. A Phylogenetic tree generated from the 16S rRNA sequencing data showed different phyletic lines that could be readily classified as biogenic amine forming gram-positive and negative bacteria.

Identification of Growth Inhibitory Substance on Food-borne Microorganisms from Commiphora molmol Engl. and Its Application to Food Products (몰약(Commiphora molmol Engl.)의 식중독 미생물 증식 억제 물질의 구조동정 및 식품적용)

  • Han, Ji-Sook;Shin, Dong-Hwa;Baek, Nam-In
    • Korean Journal of Food Science and Technology
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    • v.33 no.4
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    • pp.401-408
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    • 2001
  • The ethanol extract and n-hexane fraction of Commiphora molmol Engl. showed minimum inhibitory concentration of 50 ppm and 25 ppm, respectively, on 5 strains of Listeria monocytogenes at $32^{\circ}C$. The purified substance, C3-3-2 fraction, was isolated by silica gel column and preparative thin layer chromatography from n-hexane fraction of Commiphora molmol Engl. The C3-3-2 fraction showed a strong bactericidal activity on 5 strains of L. monocytogenes at the concentration of 10 ppm in tryptic soy broth medium. At that concentration, the viable count was reduced $5{\sim}6$ log cycle from initial cell number. The n-hexane fraction of Commiphora molmol Engl. showed strong growth inhibition at the concentration of 25 ppm on Bacillus cereus and Staphylococcus aureus, at 50 ppm in broth on Salmonella enteritidis, and at 500 ppm on Vibrio parahaemolyticus. The purified antimicrobial substance, the C3-3-2 fraction, was identified as m-nonylphenol by on the basis of the $^1H-,\;^{13}C-NMR$ and EI/MS data. For the application test, the C3-3-2 fraction which was purely isolated from Commiphora molmol Engl. at 100 ppm were applied to minced Alaska pollack and ground beef at $32^{\circ}C$ and $5^{\circ}C$. The antimicrobial substances did not reduce L. monocytogenes ATCC 19113 at $32^{\circ}C$, while they reduced L. monocytogenes ATCC 19113 in viable number at $5^{\circ}C$. However, the antimicrobial effect of C3-3-2 fraction in food system was lower than that of broth condition.

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Radiolysis Assessment of $^{18}F$-FDG According to Automatic Synthesis Module (자동합성장치에 따른 $^{18}F$-FDG의 방사선분해 평가)

  • Kim, Si-Hwal;Kim, Dong-Il;Chi, Yong-Gi;Choi, Sung-Wook;Choi, Choon-Ki;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.1
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    • pp.8-11
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    • 2012
  • Purpose : Among quality control items, the radiochemical impurity must be below 10% of total radioactivity. In this regard, as the recently commercialized automatic synthesis module produces a large amount of 18F-FDG, radiolysis of radiopharmaceuticals is very likely to occur. Thus, this study compared the changes in radiochemical purity regarding radiolysis of $^{18}F$-FDG according to automatic synthesis module. Materials and methods : Cyclotron (PETtrace, GE Healthcare) was used to produce $^{18}F$ and automatic synthesis module (FASTlab, Tracerlab MX, GE Healthcare) was used to achieve synthesis into FDG. For radiochemical purity, Radio-TLC Scanner (AR 2000, Bioscan), GC (Gas Chromatograph, Agilent 7890A) was used to measure the content of ethanol included in $^{18}F$-FDG. Glass board applied with silica gel ($1{\times}10cm$) was used for stationary phase while a mixed liquid formed of acetonitrile and water (ratio 19:1) was used for mobile phase. High-concentration and low-concentration $^{18}F$-FDG were produced in each synthesis module and the radiochemical purity was measured every 2 hours. Results : The purity in low-concentration (below 2.59 GBq/mL) was measured as 99.26%, 98.69%, 98.25%, 98.09% in Tracerlab MX and as 99.09%, 97.83%, 96.89%, 96.62% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. The purity in high-concentration (above 3.7 GBq/mL) was measured as 99.54%, 96.08%, 93.77%, 92.54% in Tracerlab MX and as 99.53%, 95.65%, 92.39%, 89.82% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. Also, ethanol was not detected in GC of $^{18}F$-FDG produced in FASTlab, while 100~300 ppm ethanol was detected in Tracerlab MX. Conclusion : Whereas the change of radiochemical purity was only 3% in low-concentration $^{18}F$-FDG, the change was rapidly increased to 10% in high-concentration. Also, higher radiolysis were observed in $^{18}F$-FDG produced in FASTlab than Tracerlab MX. This is because ethanol is included in the synthesis stage of Tracerlab MX but not in the synthesis stage of FASTlab. Thus, radiolysis is influenced by radioactivity concentration than the inclusion of ethanol, which is the radioprotector. Therefore, after producing high-concentration $^{18}F$-FDG, the content must be diluted through saline to lower concentration.

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Effects of 3,5-di-O-Caffeoylquinic acid from Artemisia scoparia Waldstein et Kitamura on the Function of HPV 16 Oncoproteins (인진에서 분리한 3,5-di-O-Caffeoylquinic acid가 자궁경부암 바이러스 발암단백질의 기능에 미치는 영향)

  • Baek, Tae-Woong;Lee, Kyung-Ae;Ahn, Min-Jung;Joo, Hae-Kyung;Cho, Min-Chul;Kang, Jung-Woo;Kim, Hee-Seo;Shim, Jung-Hyun;Lee, Hee-Gu;Oh, Hyun-Cheol;Ahn, Jong-Seok;Cho, Yong-Kwen;Myung, Pyung-Keun;Yoon, Do-Young
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.368-374
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    • 2004
  • Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human papillomavirus) type 16. The HPV is cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Artemisia scoparia Waldstein et Kitamura has inhibitory effects on the binding between E6 oncoprotein and tumor suppressor p53, or the binding between E6 and E6 associated protein (E6AP), an E3 ubiquitin-protein ligase. HPV oncoprotein inhibitors from Artemisia scoparia W. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and the inhibitory compounds were finally purified by HPLC using an ELISA screening system based on the binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on the functions of E6 oncoprotein. We investigated whether 3,5-di-O-caffeoylquinic acid (DCQA) extracted from Artemisia scoparia W. Could inhibit the function of E6 oncoprutein. DCQA inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that DCQA inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

Potent Anticarcinogenic Action of Moutan radix for Mouse Ascites Cancer Induced by Mouse Sarcoma 180 Cells (Moutan radix의 mouse sarcoma 180 cell로 유발한 mouse ascites cancer에 대한 항암효과)

  • Bahn, Kyeong-N.;Lee, Eun-J.;Yang, Min-S.;Kim, Jeong-O.;Ha, Yeong-L.
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.364-369
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    • 1995
  • Anticarcinogenic activity of Moutan radix for mouse ascites cancer induced by mouse Sarcoma 180 (S-180) cells was investigated. Methanol extract of Moutan radix including other folk medicinal plants (Taxus cuspidata, Curcuma longa, Artemisia capillaris, Ligrstri fructus, and Liriope platyphylla) used to remedy or cure many chronic human diseases like cancer was fractionated into hexane, chloroform ($CHCl_3$), ethylacetate (EtOAc), and butanol (BuOH) fractions. Anticarcinogenic activity of the fractions, exhibited a strong cytotoxicity for L1210 and S-180 cells, was examined for mouse ascites cancer induced by S-180 cells. Male ICR mice (7 mice/treatment, $5{\sim}6$ weeks of age, $23{\pm}1\;g$ were injected i.p. with S-180 cells ($1{\times}10^{7}\;cell/1\;ml$ PBS). One day later, each mouse was given 0.1 ml of 10% DMSO containing sample ($30\;{\mu}g/g$ body weight) every day for 10 consecutive days. Control mice were only given 0.1ml S-180 cells and 0.1 ml 10% DMSO. Mice treated with EtOAc fraction of Moutan radix showed 28.7 days of life, which is 167% of control mice's life. Based on the dose-dependant experiment mice treated with $30\;{\mu}g$ showed longer life relative to mice treated with ootherr doses (5, 15, $60\;{\mu}g$), and mice treated with $60\;{\mu}g$ exhibited toxic symptoms. Body weight of mice treated with Moutan radix was significantly reduced relative to that of control mice (p<0.05). GC-MS analysis in conjunction with silica-gel column chromatography revealed that the EtOAc fraction contained 2-methoxylphenol, benzoic acid, 1-(4-hydroxy-3-methoxyphenyl)ethanone, 8-methyl-2,4(1H,3H)pteridinedione and 2,5-furan-dicarboxylic dimethyl ester as regards to the anticarcinogenic property of the EtOAc fraction. These results suggest that Moutan radix might be included as an anticarcinogenic medicinal plant for treatment of ascites cancer.

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Effects of Flavonoid from Rhus verniciflua on Testosterone Secretion by Rat Leydig Cells In Vitro (옻나무 유래 Flavonoid 처리가 흰쥐 Leydig 세포의 체외배양에서 Testosterone 분비에 미치는 영향)

  • 성환후;최선호;장유민;민관식;우제현;장원경;정남철;나천수;정일정
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.125-130
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    • 2001
  • This study was performed to report a direct dose dependent stimulatory effect of the Flavonoid(F) on basal testosterone secretion and a dose dependent effect on LH induced testosterone production by Leydig cell of matured rats in vitro culture. F was obtained kom the Rhus vernicifua through aceton extraction and silica gel adsorption column chromatography. Leydig cells (1$\times$10$^{6}$ cells/well) from 12 weeks old rats were incubated with or without F(0, 20, 40, 80, 160 ng) or insulin-like growth factor-I(IGF-I) in the presence or absence of LH(10, 100ng). 1. The maximal stimulatory concentrations of testosterone in culture media were showed at 24hr of culture. but these testosterone level were decreased at 36 hr of culture. 2. Flavonoid(80ng) were significantly(P < 0.05) increased testosterone production compared with control groups for 12 hr culture. 3. Testosterone secretion by Leydig cells stimulated with LH(10, 100ng) for 6 hr and 12hr culture compared with 3 hr culture. 4. LH 10 ng augmented testosterone were increased by addition of F 40 ng for 12 hr culture. 5. F(0 and 40 ng) also enhanced LH 10 ng stimulated testosterone for 3 hr Leydig cells culture. 6. Addition of IGF-I 100 ng to the culture medium for 6 hr were increased the concentration of testosterone by Leydig cells stimulated with 100 ng LH. These results indicate that Flavonoid has a direct stimulatory effect on basal testosterone secretion in rat Leydig cells, and also modulates LH mediated testosterone. Therefore, Flavonoid may act as a modulator on gonadal development or gonadal steroidogenesis in direct or indirect.

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Anti-hepatotoxic Activity of Chrysanthemum coronarium L. var. spatiosum Extract (쑥갓의 간독성 보호작용)

  • Kang, Hyun-Jung;Lee, Eun-Ju;Sung, Sang-Hyun;Kim, Young-Choong;Song, Eun-Sook;Park, Mi-Jung;Lee, Heum-Sook
    • Korean Journal of Food Science and Technology
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    • v.35 no.1
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    • pp.138-143
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    • 2003
  • Total methanolic extract of Chrysanthemum coronarium L. var. spatiosum (Compositae) was revealed to have anti-hepatotoxic activity against galactosamine-induced toxicity on primary cultured rat hepatocytes. After successive partitioning with chloroform, n-butanol, and water, the chloroform fraction showed a significant inhibition activity of 51% at 50 ppm, compared with that of silybin, 45.9% at $100\;{\mu}M$. The chloroform fraction was subjected to silica gel column chromatography and yielded active CH-II, CH-V and CH-VI subfractions, and the anti-hepatotoxic activity of these subfractions were 47.6, 56.3, and 23.2%, respectively, at 50 ppm. Total glutathione contents of CH-II, CH-V, and CH-VI increased by 49.8, 43.9, and 47.5% of the control, respectively at 50 ppm, whereas that of silymarin was, 59.7% at $100\;{\mu}M$ after challenged with galactosamine. The ratio of (reduced glutathione) / (total glutathione) in CH-II, CH-V and CH-VI subfraction showed similar values of $0.86{\sim}0.87$ at 50 ppm, whereas that of silymarin was, 0.85 at $100\;{\mu}M$. The incorporation of $[^3H]-uridine$ uptake into RNA was not affected by these active subfractions.

Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.

Antimicrobial Activity of Ethanol Extract from Sargassum thunbergii (지충이(Sargassum thunbergii) 에탄올 추출물의 항균활성)

  • Lee, So-Young;Song, Eu-Jin;Kim, Koth-Bong-Woo-Ri;Yoon, So-Young;Kim, Seo-Jin;Lee, So-Jeong;Hong, Yong-Ki;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.502-508
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    • 2009
  • Antimicrobial activity of Sargassum thunbergii was determined by paper disc assay and minimum concentration inhibitor (MIC) test. A water extract of S. thunbergii did not show the antimicrobial activity, but an ethanol extract of S. thunbergii (SHE) inhibited Serratia liquefaciens, Salmonella Typhimurium, Pseudomonas aerogenosa and all of the tested gram-positive bacteria at 4 mg/mL. Especially, Bacillus subtilis, Clostridium perfringens and Listeria monocytogenes were susceptible to SHE. As the results of MIC test, SHE inhibited the growth of B. subtilis, Staphylococcus aureus and Listeria monocytogenes at concentration of $0.1{\sim}0.3%$, and inhibited C. perfringens at 0.01%. In the thermal and pH stability test for SHE, antibacterial activities of SHE were maintained when the SHE was treated at $121^{\circ}C$ for 15 minutes or under pH $2{\sim}8$. SHE was partitioned in the order of n-hexane, chloroform, ethyl acetate and butanol. As the results of the MIC test for each obtained fraction, no fraction exhibited higher antibacterial activity than that of the crude SHE. However, a mixture of chloroform, ethylacetate and ethanol fractions showed higher antibacterial activity than SHE.

Screening of Biologically Active Compound from Edible Plant Sources-IX. Isolation and Identification of Sesquiterpene Lactons Isolated from the Root of Ixeris dentata forma albiflora; Inhibition Effects on ACAT, DGAT and FPTase Activity (식용식물자원으로부터 활성물질의 탐색-IX. 흰씀바귀(Ixeris dentata forma albiflora)뿌리에서 Sesquiterpene Lactone 화합물의 분리 및 구조 동정; ACAT, DGAT 및 FPTase 효소 활성의 저해)

  • Bang, Myun-Ho;Jang, Tae-O;Song, Myoung-Chong;Kim, Dong-Hyun;Kwon, Byoung-Mog;Kim, Young-Kuk;Lee, Hyun-Sun;Chung, In-Sik;Kim, Dae-Keun;Kim, Sung-Hoon;Park, Mi-Hyun;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.251-257
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    • 2004
  • The root of lxeris dentata forma albiflora was extracted with 80% aqueous MeOH and solvent fractionated with EtOAc, n-BuOH and water, successively. From the EtOAc and n-BuOH fractions, four sesquiterpene compounds were isolated through the repeated silica gel and ODS column chromatographies. The chemical structures were determined as zaluzanin C (1), $9{\alpha}-hydroxyguaian-4(l5),10(14),11(13)-triene-6,12-olide$ (2), $3{\beta}-O-{\beta}-D-glucopyranosyl-8{\alpha}-hydroxyguaian-4(15),10(14 )-diene-6,12-olide$ (3), and $3{\beta}-O-{\beta}- D-glucopyranosyl-8{\beta}hydroxyguaian-10(14)-ene-6,12-olide$ (4) through the interpretation of several spectral data including 2D-NMR. Some showed the inhibitory effects on DGAT (Diacylglycerol acyltransferase), ($IC_{50}$ values of 1, 2: 0.13, 0.10 mM), the catalyzing enzymes of the intracellular esterification of diacylglycerol and FPTase (Famesyl-protein transferase), ($IC_{50}$ values of 1, 2: 0.15, 0.18 mM), the farnesylation enzyme for Ras protein charge of cancer promotion.