• Title/Summary/Keyword: HPLC Chromatography

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Effects of an Antimicrobial Substance from Bombycis corpus on Antibiotic Resistant Microbes (백강잠으로부터 분리한 항균물질의 항생제 내성균에 대한 효과)

  • Lee, Hyeon-Woo;Um, Jeong-Sun;Ko, Mi-Kyung;Kim, Mi-Kyung;Eun, Jae-Soon;Jeon, Hoon;Leem, Jae-Yoon
    • YAKHAK HOEJI
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    • v.51 no.4
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    • pp.253-258
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    • 2007
  • Bombycis corpus, a batryticated silkworm and white-stiff silkworm, is an oriental drug consisting of the dried larva of silkworm, dead and stiffened due to the infection of Beauveria. An peptidyl antimicrobial molecule was purified from B. corpus by reverse phase-column chromatography and HPLC. Its molecular weight was determined to be 2295.45 by using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Its antimicrobial activity was diminished by trypsin digestion. It exhibited a broad antimicrobial spectrum against not only Gram-negative, but also Gram- positive bacteria. Furthermore, it was found to have an antimicrobial activity against vancomycin-resistant enterococci (VRE), methicillin-resistant S. arureus (MRSA), and vancomycin-intermediate resistant S. arureus (VISA). It may be a useful molecule for a new antibiotic development, especially against antibiotic resistant microbe. This substance may play a role in the defense system of this animal against Beauveria bassiana. This is the first report of a peptidyl antimicrobial substance from B. corpus.

Characterization of Norepinephrine Release in Rat Posterior Hypothalamus Using in vivo Brain Microdialysis

  • Sung, Ki-Wug;Kim, Seong-Yun;Kim, Ok-Nyu;Lee, Sang-Bok
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.1
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    • pp.9-14
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    • 2002
  • In the present study, we used the microdialysis technique combined with high performance liquid chromatography (HPLC) and electrochemical detection to measure the extracellular levels of norepinephrine (NE) in the posterior hypothalamus in vivo, and to examine the effects of various drugs, affecting central noradrenergic transmission, on the extracellular concentration of NE in the posterior hypothalamus. Microdialysis probes were implanted stereotaxically into the posterior hypothalamus (coordinates: posterior 4.3 mm, lateral 0.5 mm, ventral 8 mm, relative to bregma and the brain surface, respectively) of rats, and dialysate collection began 2 hr after the implantation. The baseline level of monoamines in the dialysates were determined to be: NE $0.17{\pm}0.01,$ 3,4-dihydroxyphenylacetic acid (DOPAC) $0.94{\pm}0.07,$ homovanillic acid (HVA) $0.57{\pm}0.05$ pmol/sample (n=8). When the posterior hypothalamus was perfused with 90 mM potassium, maximum 555% increase of NE output was observed. Concomitantly, this treatment significantly decreased the output of DOPAC and HVA by 35% and 28%, respectively. Local application of imipramine $(50\;{\mu}M)$ enhanced the level of NE in the posterior hypothalamus (maximum 200%) compared to preperfusion control values. But, DOPAC and HVA outputs remained unchanged. Pargyline, an irreversible monoamine oxidase inhibitor, i.p. administered at a dose of 75 mg/kg, increased NE output (maximum 165%), while decreased DOPAC and HVA outputs (maximum 13 and 12%, respectively). These results indicate that NE in dialysate from the rat posterior hypothalamus were neuronal origin, and that manipulations which profoundly affected the levels of extracellular neurotransmitter had also effects on metabolite levels.

Stability of Domoic Acid at Different Temperature, pH and Light (온도, pH 및 빛에 대한 Domoic Acid의 안정성)

  • Mok, Jong-Soo;Lee, Tae-Seek;Oh, Eun-Gyoung;Son, Kwang-Tae;Hwang, Hye-Jin;Kim, Ji-Hoe
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.1
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    • pp.8-14
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    • 2009
  • To prevent the food poisoning originated by consumption of shellfish contaminated with domoic acid, the quantitative analysis of domoic acid is to be very important. The stability of domoic acid at different temperature, pH and light was investigated using high performance liquid chromatography (HPLC). The mean recoveries of domoic acid in the methanol extracts from oyster (Crassostrea gigas), blue mussel (Mytilus edulis), short neck clam (Ruditapes philippinarum) and ark shell (Scapharca broughtonii) were 85.4-104.5%, 94.8-101.2%, 91.0-104.6%, and 95.7-109.6%, respectively. The working solutions of domoic acid standard were very stable for one month at $-18^{\circ}C$, $4^{\circ}C$, and room temperature. And domoic acid in the methanol extract from oyster was stable for a day at $4^{\circ}C$ and room temperature, and for a week at $-18^{\circ}C$. Therefore, this implies that quantitative analysis for domoic acid must consider the storage conditions of the standard solutions and the methanol extracts from shellfish. The standard solutions adjusted to pH 3-9 were also stable after heating at $121^{\circ}C$ for 30 min. The effect of light exposure on domoic acid was tested by exposing the methanol extracts to light. Domoic acid degraded slowly when the samples were kept in the dark (brown vial). However, following the light exposure the photodegradation became more rapid; no detectable domoic acid remained in $1.0{\mu}g/mL$ of methanol extract after 5 hours.

Effect of temperature on fractional precipitation for paclitaxel purification

  • Jeon, Soon-Yim;Kim, Jin-Hyun
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.355-359
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    • 2005
  • Fractional precipitation is a simple, efficient method for pre-purifying paclitaxel from plant cell cultures. The storage temperature of factional precipitation was optimized in terms of the yield and purity of paclitaxel with a fixed methanol concentration in water (61.5%, v/v), paclitaxel content in the crude extract (0.5%, w/v), and storage time (3 days). The greatest yield $({\sim}84%)$ was obtained with storage at a constant temperature $(0^{\circ}C)$ for 3 days. Conversely, the highest purity $({\sim}79%)$ was obtained with stepwise reduction in temperature over 3 days. For a constant storage temperature, the maximum purity $({\sim}67%)$ was obtained at $0^{\circ}C$. There was little difference in the yield of paclitaxel between -20 and $12^{\circ}C$. This pre-purification process serves to minimize solvent usage, and the size and complexity of the high-performance liquid chromatography (HPLC) operation required for paclitaxel purification.

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Effect of Nitrogen Source on the Change of UDP-glucose and ${\beta}$-1,3-glucan Concentration

  • Park, Yang-Ho;Lee, Jung-Heon
    • KSBB Journal
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    • v.21 no.5
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    • pp.366-370
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    • 2006
  • In this research, analysis of UDP-glucose a precursor of ${\beta}$-1,3-glucan by high performance liquid chromatography(HPLC) was established using a reversed phase system. One of key metabolite UDP-glucose was selected and its concentration changes was measured with the change of fermentation conditions. The effects of fermentation conditions with/without nitrogen source for cell growth on ${\beta}$-1,3-glucan production were dependent on the UDP-glucose concentration. The UDP-glucose was synthesized rapidly during cell exponential growth period and maintained high during ${\beta}$-1,3-glucan production period. The UDP-glucose concentration was higher for ${\beta}$-1.3-glucan production fermentor than that for cell growth fermentor. The ${\beta}$-1,3-glucan production was optimal at pH 5.5 and synthesis of ${\beta}$-1,3-glucan was greatest at pH 5.5.

Determination of Loxoprofen Adsorption Isotherms by Frontal Analysis and Pulse Input Method (Frontal Analysis와 Pulse Input Method를 이용한 Loxoprofen의 등온흡착식 결정)

  • Lee, Eun;Park, Joon-Sub;Kim, In-Ho
    • KSBB Journal
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    • v.21 no.5
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    • pp.371-375
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    • 2006
  • Frontal analysis(FA) and Pulsed input method(PIM) have been frequently utilized to measure isotherm of single solute, as well as non-competitive isotherms of two solutes in chromatography(1). FA and PIM were used in this study as complementary methods to measure adsorption isotherms of loxoprofen racemate in HPLC. Prior to FA and PIM experiments, measurements of loxoprofen solubility were made at hexane/ethanol=50/50, 80/20, 95/5(v/v) with acetic acid(0.5%) for adjusting pH. The last composition(95/5) of hexane/ethanol allows us to separate loxoprofen racemate into two forms(retentate, extract). PIM and FA were used to determine the isotherms of re-and ex-loxoprofen.

High Performance Liquid Chromatographic Analysis of Free Amino Acids in Various Ginseng Products (고속액체(高速液體)크로마토그래피에 의한 각종(各種) 인삼제품(人蔘製品)중의 유리아미노산 조성의 분석(分析))

  • Lee, Sung-Woo;Kurozaki, Toshiharu;Woo, Sang-Kyu;Yoon, Tai-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.3
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    • pp.37-40
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    • 1982
  • Fifteen free amino acids except tryptophan, proline and cystine were identified from Korean red ginseng and dried ginsengs from Korea, America and Canada using by high performance liquid chromatography (HPLC). Arginine was 72.6% of total free amino acids in the red ginseng and 48.2 to 68.7% in the dried ginsengs. The content of each free amino acid was lower in the red ginseng than in Korean dried ginseng. Most free amino acids in Korean dried ginseng showed higher content than those in American and Canadian ones. Tryptophan, proline cystine, methionine and phenylalanine were not detected in the extracts of red ginseng and of Korean white ginseng. Arginine was highest in these extracts and all free amino acids were higher in the white ginseng extract.

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Antidepressant Effect of the Subchronic Administration of the Methanolic Extract of Wild-ginseng and Cultivated-ginseng in Mice Tail Suspension Test (산삼과 인삼 메탄올 추출물 아만성 복용의 Mice Tail Suspension Test에서의 항우울 효과에 대한 비교연구)

  • Kwon, Sun-Oh;Choi, Soo-Min;Kim, Myung-Hwan;Lee, Bom-Bi;Park, Moo-Won;Lee, Hye-Jung;Park, Hi-Joon;Hahm, Dae-Hyun
    • Journal of Acupuncture Research
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    • v.26 no.4
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    • pp.99-106
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    • 2009
  • Objectives : The antidepressant effect of the subchronic administration of the methanolic extract of wild ginseng(WG) was investigated compared with that of cultivated ginseng(CG, panax ginseng) extract. Methods : To assess the antidepressant effect of the ginseng extracts, tail suspension test(TST) was executed in mice after daily administration of WG or CG extract for five consecutive days. Results : The WG extract at daily dose of 600mg/kg significantly reduced the total duration of immobility in the TST, whereas there was no significant reduction at daily dose of 300mg/kg WG and 600mg/kg CG. There were no individual differences between experimental groups in open field test (OFT) to evaluate psychostimulant effects of WG or CG extract. In the high performance liquid chromatography(HPLC) analysis of the extracts, it was found that WG included four times more ginsenoside Rg1 and Re, three times more Rf, and six times more Rb1 and Rc than CG. Conclusions : It is suggested that WG extract has stronger antidepressant effect than CG extract, which means it includes more antidepressant compounds than CG.

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Inhibitors of Tyrosinase and Melanogenesis from Galla rhois

  • Kim, Hyo-Jin;Jang, Dong-Il;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.2 no.4
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    • pp.285-290
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    • 1997
  • Previously, a 50% aqueous methanol extract of Galla rhois was shown to be the most potent tyrosinase inhibition activity with an {TEX}$IC_{50}${/TEX}(the concentration causing 50% inhibition of tyrosinase activity) of 0.2mg/ml of 205 crude drug extracts. To isolate tyrosinase inhibitors, the methanol extract was evaporated to a small volume in vacuo, and then partitioned stepwise with benzene and ethyl acetate(EtOAc). the EtOAc fraction was solubilized in 10% MeOH solution, and then fractionated successively by Diaion HP-20 and Sephadex LH-20 column chromatography, and preparative HPLC. Three phenolic compounds were isolated, and characterized as gallic acid(GA), methyl gallate(MG) and 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose(PGG) by UV, IR, {TEX}${1}^H${/TEX}-&{TEX}${13}^C${/TEX}-NMR, and FAB-MS spectroscopy, PGG({TEX}$IC_{50}${/TEX}=50$\mu\textrm{g}$/ml) showed a considerable inhibitory effect against mushroom tyrosinase, while GA({TEX}$IC_{50}${/TEX}=1.6mg/ml) and MG({TEX}$IC_{50}${/TEX}=234$\mu\textrm{g}$/ml) did not show an appreciable effect. Meanwhile, MG inhibited greatly melanogenesis in a murine melanocyte cell line, Mel-Ab. MG and PGG showed typical noncompetitive inhibition patterns against mushroom tyrosinase. These results suggest that PGG and MG may be potentially useful as either anti-browning or anti-melanogenic agents in foods and cosmetics.

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DPPH Radical Scavenging Effect and in vitro Lipid Peroxidation Inhibition Activity of Portulaca oleracea and Its Active Principles (쇠비름(Portulaca oleracea) 추출물의 DPPH radical 소거능과 in vitro 지질과산화 억제 효과와 활성성분)

  • Lee, Hee-Jung;Lee, Burm-Jong;Lee, Dong-Seok;Seo, Young-Wan
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.668-672
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    • 2003
  • An antioxidative activity of Portulaca oleracea was tested by in vitro experimental models. The antioxidative activities were determined by evaluation the DPPH radical scavenging activity and by measuring lipid peroxide using 2-thiobarbituric acid (TBA). The crude extract was sequentially partitioned with n-hexane, 15% aq. MeOH, EtOAc, n-BuOH, $H_2O$. Among them, a remarkable antioxidative effect was observed in the fractions of EtOAc and n-BuOH. The DPPH radical scavenging effect $(IC_{50}=17.90{\mu}g/ml)$ of the n-BuOH soluble fraction was comparable with that of natural antioxidant, ${\alpha}-tocopherol(IC_{50}=\;6.99{\mu}g/ml)$ and the inhibitory effect of lipid peroxidation in mouse liver homogenate was similar to that of natural antioxidant, L-ascorbic acid at a concentration of 0.1mg/ml to 5mg/ml. From the BuOH soluble fraction yielded two biophenolic glycosides, 3-hydroxy-1-(2-hydroxyethyl)phenyl-4-O-${\beta}$-D-glucopyranoside(1) and 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside(2) using column chromatography and revered-phase HPLC. In particular, the DPPH radical scavenging activity of 2 was comparable to that tocopherol$(IC_{50}=6.59{\mu}g/ml)$.

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