• Title/Summary/Keyword: High performance liquid chromatography-UV

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Antimicrobial Substance against Escherichia coli O157:H7 Produced by Lactobacillus amylovorus ME1

  • Jung, Byung-Moon;Woo, Suk-Gyu;Chung, Kun-Sub
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.679-682
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    • 2008
  • A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

Characteristics of pre-extracted hemicelluloses from Korean mixed wood by hot water and alkali solution and its effect on handsheet properties (열수 및 알칼리 용액을 이용하여 국산 목재 칩으로부터 선추출한 헤미셀룰로오스의 특성과 이에 따른 수초지 물성 변화)

  • Seo, Dong-Il;Lee, Sang-Hoon;Sim, Kyu-Jeong;Lee, Hak-Lae;Youn, Hye-Jung
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.43 no.5
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    • pp.60-67
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    • 2011
  • Hemicelluloses pre-extracted from Korean mixed wood chip were investigated as a wet-end additive. Hemicelluloses dissolved in hot water and alkali solution were isolated by ethyl alcohol precipitation from pre-extractives. They showed molecular weight of 9,000 ~ 27,000 g/mol as revealed by size exclusion chromatography. The reduction of molecular weight through hot water extraction was caused by autohydrolysis. Chemical composition of the hemicelluloses were analyzed with high-performance liquid chromatography and UV-Vis spectroscopy. As the surface charge of isolated hemicelluloses were negative, the adsorption of hemicelluloses onto softwood unbleached kraft pulp fiber was promoted by poly-DADMAC. The physical properties of handsheets increased as the molecular weight of hemicellulose increased. On the other hands, the optical property decreased with hemicellulose adsorption.

Characterization of novel EGCG (Epigallocatechin gallate) analogues synthesized by glucansucrase from Leuconostoc mesenteroides B-1299CB

  • Moon, Young-Hwan;Lee, Jin-Ha;Nam, Seung-Hee;Kim, Do-Won;Oh, Deok-Kun;Kim, Do-Man
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.502-506
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    • 2005
  • Three EGCG analogues were synthesized by the acceptor reaction of a glucansucrase from Leuconostoc mesenteroides B-1299CB with EGCG and sucrose. The transfer products was purified using Sephadex LG-20 column chromatography and high performance liquid chromatography (HPLC). EGCG-G1 and EGCG-G2 were novel compounds for the first time reported in this paper. EGCG glycosides showed similar or slower antioxidative effects according to their structures $(EGCG{\geq}EGCG-G1>EGCG-G1'>EGCG-G2)$. However, the water solubilities of the EGCG-G1, EGCG-G1' and EGCG-G2 were 52, 76 and 140 times higher than that of EGCG. Furthermore, they showed more browning resistance against UV irradiation than EGCG.

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Molecular Cloning and Characterization of Bacillus cereus O-Methyltransferase

  • Lee Hyo-Jung;Kim Bong-Gyu;Ahn Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.619-622
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    • 2006
  • Biotransformation is a good tool to synthesize regioselective compounds. It could be performed with diverse sources of genes, and microorganisms provide a myriad of gene sources for biotransformation. We were interested in modification of flavonoids, and therefore, we cloned a putative O-methyltransferase from Bacillus cereus, BcOMT-2. It has a 668-bp open reading frame that encodes a 24.6-kDa protein. In order to investigate the modification reaction mediated by BcOMT-2, it was expressed in E. coli as a His-tag fusion protein and purified to homogeneity. Several substrates such as naringenin, luteolin, kaempferol, and quercetin were tested and reaction products were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). BcOMT-2 could transfer a methyl group to substrates that have a 3' functional hydroxyl group, such as luteolin and quercetin. Comparison of the HPLC retention time and UV spectrum of the quercetin reaction product with corresponding authentic 3'-methylated and 4'-methylated compounds showed that the methylation position was at either the 3'-hydroxyl or 4'-hydroxyl group. Thus, BcOMT-2 transfers a methyl group either to the 3'-hydroxyl or 4'-hydroxyl group of flavonoids when both hydroxyl groups are available. Among several flavonoids that contain a 3'- and 4'-hydroxyl group, fisetin was the best substrate for the BcOMT-2.

Pretreatment Effect on the Analysis of Insoluble Hexavalent Chromium (전처리방법에 따른 불용성 6가 크롬 분석에 관한 연구)

  • Lee, Eun Jung;Roh, Jae Hoon;Won, Jong Uk;Chun, Mi Ryonng;Cho, Myung Hwa;Kim, Chi Nyon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.6 no.2
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    • pp.292-300
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    • 1996
  • Hexavalent chromium($Cr^{+6}$) compounds are considered to be particularly hazardous, primarily because of the associated risk of allergic reaction and cancer. The analytic method of hexavalent chromium such as the s-diphenylcarba-zide(DPC) method and all ether previously used methods are often made uncertain due to significant interferences from organic components. This report can provide a technique for the more rapid and simple determination of total hexavalent chromium. than other currently using methods. The s-diphenylcarbazide method proposed by the U.S. National Institute for Occupational Safety and Health has low recovery rate(15.67 - 48.20%) due to interference, iron chloride and nickel chloride. A microwave oven technique has high recovery rate(about 70%) of insoluble hexavalent chromium. For the difference of ionic charges of $Cr^{+3}$-ethylenediamine tetraacetic acid(EDTA) chelate and $CrO_4{^{-2}}$, we could detect them simultaneously by ion exchanged high performance liquid chromatography. The confirmation of $Cr^{+3}$ and $Cr^{+6}$ were checked by fraction collector and flameless atomic absorption spectrometer. We observed that the small amount of hexavalent chromium is converted to trivalent chromium due to enhancement of chromium reduction by $Fe^{+3}$ or $Ni^{+2}$. As a result of this study, on the analysis of insoluble hexavalent chromium with microwave oven was used for, it may be better and more precise analysis after pretreatment by 2% NaOH-3% $Na_2CO_3$ and then analysis UV-spectrophotometer. It should be done for various studies on insoluble hexavalent chromium on the basis work environmental monitoring so called welding, painting etc.

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Determination of thyroid hormones in plasma samples by high performance liquid chromatograph/diode array detector/electrospray ionization mass spectrometer (HPLC/DAD/ESI-MS를 이용한 혈장시료 중 갑상선 호르몬 분석)

  • Kwak, Sun Young;Moon, Myeong Hee;Pyo, Heesoo
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.424-433
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    • 2007
  • An analytical method for the determination of thyroid hormones in plasma samples has been studied by solid-phase extraction and high-performance liquid chromatography/diode array detector (DAD)/electrospray ionization (ESI)-mass spectrometer. Seven thyroid hormones were successfully separated by gradient elution on the reverse phase Hypersil ODS column (4.6 mm I.D., 250 mm length, particle size $5{\mu}m$) with ammonium formate buffer and acetonitrile. In addition, these compounds were confirmed by UV spectra and ESI-mass Spectra. The extraction recoveries of thyroid hormones in the plasma sample (at pH 3) were in the range of 74.5-115.7 % with solid-phase extraction by C18, followed by elution with 4 mL of methanol. The calibration curves showed good linearity with the correlation coefficients ($r^2$) varying from 0.9939 to 0.9978 and the detection limits of all analytes were obtained in the range of 20-50 ng/mL (38.1-162.8 pmol/mL). As a result, thyroxine was found in the range of 50.98-112.97 ng/mL in normal plasma samples.

Determination of Acethylsalicylic acid (Aspirin) and Salicylic acid in Eel (Anguilla japonica) Plasma by High-performance Liquid Chromatography (HPLC에 의한 뱀장어 (Anguilla japonica) 혈장중의 acethylsalicylic acid (aspirin)과 salicylic acid의 측정)

  • Kim, Dong-Wan;Koo, Jae-Geun;Park, Sung-Woo
    • Journal of fish pathology
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    • v.20 no.2
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    • pp.139-145
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    • 2007
  • To decrease stress in eel (Anguilla japonica) during its culture or transportation, aspirin (ASA) known as analgesic, antiinflammatory and antithrombic agent was administrated by dipping or oral routes. Concentrations of aspirin (ASA) and salicylic acid (SA) in eel plasma were simultaneously measured by a high performance liquid chromatography (HPLC). The plasma was acidified with 0.2 M HCl and 0.2 M orthophosphoric acid, and mixed with acetonitrile. ASA and SA extracted with acetonitrile were analyzed by the HPLC equipped with reversed phase Novapak C18 column (4 ㎛ silica, 150×4 mm) and UV detector(237 nm). The mobile phase consisted of 740 ㎖ water, 900 ㎕ orthophosphoric acid (85%) and 180 ㎖ acetonitrile. The retention times of ASA, SA and 2-methylbenzoic acid(MBA) were 4.8 min, 8.4 min and 11.5 min, respectively. The limit of quantification was 0.01 ㎍/㎖ for SA and 0.05 ㎍/㎖ for ASA. The mean recovery from eel plasma was 70.8~99.6% for ASA and 95.2~100.3% for SA. This HPLC method was applied to analyze ASA and SA of eel plasma after either dipping in a concentration of 20 ppm or feeding the feed supplemented with 50 ㎎/kg BW. Only SA was detected in eel plasma after the administration of ASA by dipping or oral routes because the drug was quickly decomposed into SA in eel plasma. The amount of SA in eel plasma reached the highest value at 3hr in dipping and 7 days in oral administration. When the ASA-administrated eel were kept in ASA free aquaria, 0.02-0.03 ㎍/㎖ of SA were detected 48 hr after the administration in both routes.

Analysis of Bioconversed-components in Fermented Jaeumganghwa-tang by Lactobacillus (Lactobacillus로 발효한 자음강화탕의 생물전환 성분분석)

  • Lee, Kwang Jin;Song, Na-Young;Roh, Joo-Hwan;Liang, Chun;Ma, Jin Yeul
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.131-135
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    • 2013
  • Jaeumganghwa-tang (JGT) is a traditional herbal medicine used for chronic bronchitis and inflammatory diseases. The variation in the amount of bioactive components of JGT and its fermentation JGT with ten species of microorganism was investigated via high performance liquid chromatography coupled with diode array detection (HPLC-DAD). Simultaneous qualitative and quantitative analysis of eight bioactive compounds; 5- hydroxymethylfurfural (5-HMF), paeoniflorin, nodakenin, hesperidin, nodakenetin, palmatine, berberine and glycyrrhizin were achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard compounds. In the result, the paeoniflorin amount was 6.95 mg/g that as a main compound in JGT. The amount of nodakenetin was the highest in the fermented-JGT with Lactobacillus fermentum KFRI 145 ($0.47{\pm}0.01mg/g$), which was increased by 2,250% compared to that in non-fermented JGT ($0.02{\pm}0.00mg/g$). In the fermented JGT using Lactobacillus acidophilus KFRI 162, most components were increased than non-fermented JGT, except paeoniflorin and hesperidin.

Antioxidant, Anti-wrinkle and Whitening Effect of Fermented Extracts of Hwangryunhaedoktang (황련해독탕 발효물의 항산화, 항주름 및 미백 효과)

  • Um, Ji Na;Min, Jin Woo;Joo, Kwang Sik;Kang, Hee Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.1
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    • pp.69-78
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    • 2017
  • Hwangryunhaedoktang (HHT) has been traditionally used as a preventive and therapeutic medicine to treat enervation and diverse chronic diseases. This study was designed to compare the antioxidant, anti-wrinkle and whitening effects of HHT extract and its fermented extract by Leuconostoc mesenteroides (FHHT). FHHT was prepared by inoculation of L. mesenteroides after the extraction procedure with 70% ethanol. HHT and FHHT was investigated via high-performance liquid chromatography (HPLC). Simultaneous qualitative analysis of two bioacitive components, berberine and palmatine. was achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard components. Cell viability test results indicated that both HHT and FHHT were non-toxic. In DPPH radical scavenging ability, $SC_{50}$ values of the FHHT was $68.85{\mu}g/mL$, which is more effective than HHT. Moreover, FHHT showed higher expression in production of procollagen type I than HHT. In nontoxic concentration range, FHHT showed strong melanin production inhibitory effect in ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$)-stimulated B16F10 cell ($IC_{50}=9.82{\mu}g/mL$). These results suggested that fermented extracts of hwangryunhaedoktang had considerable potential as a cosmetics ingredient with an antioxidant and anti-wrinkle and whitening effects.

Enhancement of Antioxidant and Whitening Effect of Fermented Extracts of Scutellariae baicalensis (황금 발효물의 항산화 및 미백 효과 증진)

  • Um, Ji Na;Min, Jin Woo;Joo, Kwang Sik;Kang, Hee Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.3
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    • pp.201-210
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    • 2017
  • Scutellariae baicalensis (S. baicalensis) has been traditionally used for anti-inflammatory effect. This study was designed to compare the antioxidant and whitening effects of S. baicalensis extract and its fermented extract by Leuconostoc mesenteroides (L. mesenteroides). Fermented extract of S. baicalenins was prepared by inoculation of L. mesenteroides after the extraction procedure with 70% ethanol. S. baicalensis extract and its fermented extract was investigated via high-performance liquid chromatography (HPLC). Simultaneous qualitative analysis of two bioactive components; baicalin and baicalein was achieved by comparing their retention times ($t_R$) and UV spectra with those of the standard components. Cell viability test results indicated that both S. baicalensis extract and its fermented extract were non-toxicity. In DPPH radical scavenging ability, $SC_{50}$ values of the fermented extract was $34.43{\mu}g/mL$ as a result of more effective than S. baicalensis extract. In nontoxic concentration rage, fermented extract of S. baicalensis showed strong melanin production inhibitory effect in ${\alpha}$-melanocyte stimulating hormone (MSH)-stimulated B16F10 cell ($IC_{50}=68.17{\mu}g/mL$). These results suggested that fermented extracts of S. baicalensis has considerable potential as a cosmetics ingredient with an antioxidant and anti-wrinkle and whitening effects.