• Title/Summary/Keyword: Bio-column

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Determination of itraconazole in human plasma by high performance liquid chromatography (HPLC/UV를 이용한 혈장 중 이트라코나졸의 분석)

  • Jang, Hae Jong;Lee, Ye Rie;Lee, Kyung Ryul;Han, Sang-Beom;Kang, Seung Woo;Lee, Hee Joo
    • Analytical Science and Technology
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    • v.19 no.3
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    • pp.239-243
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    • 2006
  • This method is used for the determination of itraconazole in human plasma by liquid-liquid extraction and high performance liquid chromatography. Felodipine was used as an internal standard. After extraction of the plasma with diethyl ether, the centrifuged upper layer was then transferred. The supernantant was evaporated and then reconsituted with mobile phase. The mobile phase was composed of 10 mM ammonium acetate adjusted to pH 7 by phosphoric acid with a flow rate of 0.2 mL/min. A C18 reversed-phase column with a pre-column was used as the analytial column. Linear detection responses were obtained for itraconzole concentration range for 2~1,000 ng/mL. The correlation coefficient of linear regression($R^2$) was 0.9991, limit of quantification (LOQ) was 2 ng/mL, reproducibility was less than 10.8 %, and accuracy was 97.2~108.2%. This method has been successfully applied to the pharmacokinetic study of itraconazole in human plasma.

Evaluation of NH3 emissions in accordance with the pH of biochar

  • Yun-Gu, Kang;Jae-Han, Lee;Jin-Hyuk, Chun;Yeo-Uk, Yun;Taek-Keun, Oh;Jwa-Kyung, Sung
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.787-796
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    • 2021
  • Nitrogen (N) is the most important element during the process of plant growth, and the quality of crops varies depending on the amount of nitrogen present. Most of the nitrogen is used for plant growth, but approximately 10 - 20% of Nitrogen is carried away by the wind in the form of NH3. This volatilized NH3 reacts with various oxides in the atmosphere to generate secondary particulate matter. To address this, the present study attempts to reduce NH3 occurring in the soil using biochar at a specific pH. Biochar was used as a treatment with 1% (w·w-1) of the soil, and urea was applied at different levels of 160, 320, and 640 kg·N·ha-1. NH3 generated in the soil was collected using a dynamic column and analyzed using the indophenol blue method. NH3 showed the maximum emission within 4 - 7 days after the fertilizer treatment, decreasing sharply afterward. NH3 emission levels were reduced with the biochar treatment in all cases. Among them, the best reduction efficiency was found to be approximately 25% for the 320 kg·ha-1 + pH 6.7 biochar treatment. Consequently, in order to reduce the amount of NH3 generated in the soil, it is most effective to use pH 6.7 biochar and a standard amount (320 kg·N·ha-1) of urea.

Formation of Flavone Di-O-Glucosides Using a Glycosyltransferase from Bacillus cereus

  • Ahn, Byoung-Chan;Kim, Bong-Gyu;Jeon, Young-Min;Lee, Eun-Jeong;Lim, Yoong-Ho;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.387-390
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    • 2009
  • Microbial UDP-glycosyltransferases can convert many small lipophilic compounds into glycons using uridine-diphosphate-activated sugars. The glycosylation of flavonoids affects solubility, stability, and bioavailability. The gene encoding the UDP-glycosyltransferase from Bacillus cereus, BcGT-3, was cloned by PCR and sequenced. BcGT-3 was expressed in Escherichia coli BL21(DE3) with a glutathione S-transferase tag and purified using a glutathione S-transferase affinity column. BcGT-3 was tested for activity on several substrates including genistein, kaempferol, luteolin, naringenin, and quercetin. Flavonols were the best substrates for BcGT-3. The enzyme dominantly glycosylated the 3-hydroxyl group, but the 7-hydroxyl group was glycosylated when the 3-hydroxyl group was not available. The kaempferol reaction products were identified as kaempferol-3-O-glucoside and kaempferol-3,7-O-diglucoside. Kaempferol was the most effective substrate tested. Based on HPLC, LC/MS, and NMR analyses of the reaction products, we conclude that BcGT-3 can be used for the synthesis of kaempferol 3,7-O-diglucose.

Validation of Method Determining Coixol in Coix lachryma-jobi var. ma-yuen Roots Extract (율무근 추출물의 Coixol 성분 분석법 검증)

  • Kwon, Jin Gwan;Seo, Changon;Choi, Yun-Hyeok;Choi, Chun Whan;Kim, Jin Kyu;Jeong, Wonsik;Lee, Ji Eun;O, Kyeong Hee;Hong, Seong Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.952-956
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    • 2017
  • An high performance liquid chromatography (HPLC) analysis method was developed for standard determination of coixol as a functional cosmetic material in Coix lachryma-jobi var. ma-yuen roots extract. HPLC was performed on a $C_{18}$ Unison US column ($4.6{\times}250mm$, $5{\mu}m$ column) using a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analyte was detected at 290 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limit of detection and quantitation were 0.07 and 0.25 mg/mL, respectively. Calibration curves showed good linearity ($R^2$>0.9995), and the precision of analysis was satisfied (less than 0.29%). Recoveries of quantified compounds ranged from 98.36 to 100.30%. This result indicates that the established HPLC method is very useful for the determination of a marker compound in C. lachryma-jobi var. ma-yuen roots extracts.

Dermal Papilla Cells Proliferation Constituent of Schisandra chinensis Fruits and Optimization Using Response Surface Methodology (오미자의 모유두세포 증식 활성성분과 반응표면분석을 이용한 추출조건의 최적화)

  • Cho, Hyun Dae;Jeong, JiYeon;Ryu, Hwa Sun;Lee, JungNo;Park, Sung-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.4
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    • pp.415-424
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    • 2020
  • In the present study, we have refined gomisin N, which represents activity in the proliferation of dermal papilla cells (HFDPCs) from the fruit of Schisandra chinensis (S. chinensis), and have identified optimal extraction conditions for obtaining extracts with high content of gomisin N. The activity of the extracts and fractions was evaluated, and the results indicated approximately 29% proliferation activity in the group treated with 1 ㎍/mL of n-hexane fraction. Column chromatography was used to assess the active ingredient in the n-hexane fraction, and two compounds, namely gomisin N(1) and schisandrin(2), were isolated and identified. When the HFDPCs proliferation activity was tested for the isolated compounds, gomisin N exhibited ≥ 20% proliferation activity. Thus, via response surface methodology (RSM), the optimum extraction conditions to obtain the maximum level of gomisin N from the fruit of S. chinensis were determined, where ethanol proportion, extraction time, and extraction temperature were used as the independent variables. The results revealed coefficient of determination ≥ 0.95 and p-value ≤ 0.05, which confirmed the fit of the model. The optimum extraction conditions to achieve the maximum content of gomisin N were as follows: ethanol proportion 83.8%, extraction temperature 80 ℃, and extraction time 8.7 h. The content of gomisin N using these conditions was predicted as 378,300 ppm, and a mean value close to the predicted value (376,884 ppm) was obtained while validating the aforementioned conditions.

Development of Analytical Method of Polycyclic Aromatic Hydrocarbons Deposited on Tree Leaves by GC/MS (GC-MS 에 의한 나뭇잎에 침착된 다환방향족 탄화수소의 분석)

  • Chun, Man-Young;Lim, Ceoel-Soo;Kim, Tae-Wook
    • Korean Journal of Environmental Agriculture
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    • v.18 no.2
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    • pp.135-139
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    • 1999
  • A new effective and economic method was developed, removing interferences such as chlorophyll and lipid from leaves with small amounts of reagents and solvents in order to analyse PAHs(Polycyclic Aromatic Hydrocarbons). The extract from a soxhlet containing $4{\sim}5g$ of leaves and 100ml of dichlormethane and refluxed for 20 hrs was concentrated and eluted with 60ml of a hexane:dichloromethane (1:1) mixture through a column of 9mm wide inner diameter and 130mm long, packed from the bottom with 2.5g of $Al_2O_3$, 1.5g of $SiO_2$and 2g of anhydrous $Na_2SO_4$. The eluent was concentrated and loaded on a GPC column of 20mm wide inner diameter and 280mm long, packed with 12g of Bio-beads. The column was washed with 37ml of the hexane:dichloromethane(1:1) mixture. Another 43ml of the mixture was eluted as a PAH fraction and collected. This eluent was concentrated under gentle nitrogen to $50{\mu}l$ and analysed using GC-MS. The recoveries, obtained by comparing with the amounts of the internal standards of deuterated PAHs were $43.3{\sim}107.5%$(RSD $2.2{\sim}9.5%$).

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Structural Safely Analysis of a Modified 1-2W Type Greenhouse Enhanced for Culturing Paprika (착색단고추 재배용 1-2W형 개조온실 구조의 안정성 검토)

  • Suh, Won-Myung;Choi, Man-Kwon;Bae, Yong-Han;Lee, Jong-Won;Yoon, Yong-Cheol
    • Journal of Bio-Environment Control
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    • v.17 no.3
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    • pp.197-203
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    • 2008
  • This study was performed to check the structural safety of modified 1-2W Greenhouses to be utilized fur growing Paprika. This type of greenhouse was derived from being remodeled by enhancing the column height of conventional 1-2W type greenhouses. According to the results of structural analysis performed by SAP-2000, there was not significant change in critical snow depth in spite of increasing the column height of 1.2 m by welding. But the critical wind velocities were shown to be $26.0\sim4l.0m/s$, which were $3\sim18%$ lower wind velocities compared with those critical velocities estimated for typical type of 1-2W greenhouse. Under the wind loads, those maximum section forces such as shear force, axial force, and bending moment, together with the deformed frame shape of strained greenhouse, were almost similar in both typical type and modified type. Maximum bending moment of column was found at eave's height of column on windward side. Under the snow loads, those maximum section forces such as shear farce, axial force, and bending moment, together with the deformed frame shape of strained greenhouse, were almost similar in both typical type and modified type. Maximum section forces except axial force was found at eave's height of column. Maximum axial force was found at inner column. Soil bearing capacity together with the total foundation resistance against wind upheaval was found to be consistently safe enough to resist to both wind load and snow load.

Determination of rebamipide in human plasma by column-switching high- performance liqiud chromatography.

  • Koung, Joung-Sun;Park, Chang-Hun;Kim, Ho-Hyun;Lee, Hee-Joo;Beom, Han-Sang
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.278.2-278.2
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    • 2003
  • A column-switching semi-micro HPLC method with fluorescence detection was developed for the direct analysis of rebamipide in human plasma. Plasma was filtered through a 0.45 $\mu\textrm{m}$ membrane filter and 5 ${\mu}\ell$ of the filtrate was directly injected onto the pre-column. After elution of the plasma proteins to waste, the retained rebamipide and internal standard(ofloxacin) were transferred to a C18 semi-microcolumn (5$\mu\textrm{m}$, 150 ${\times}$2.0mm) where they were separated using acetonitrile-1.4% acetic acid (40:60, v/v) as mobile phase. (omitted)

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Inhibitory Effects of Acinetobacter sp. KTB3 on Infection of Tobacco mosaic virus in Tobacco Plants

  • Kim, Young-Sook;Hwang, Eui-ll;O, Jeong-Hun;Kim, Kab-Sig;Ryu, Myong-Hyun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.293-296
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    • 2004
  • During the screening of antiviral substances having inhibitory effects on Tobacco mosaic virus (TMV) infection on tobacco plants, we found a bacterial isolate KTB3, and identified it as Acinetobacter sp. which strongly inhibited the infection of TMV When the culture filtrate from KTB3 was applied on the upper surface of the Xanthi-nc tobacco leaves at the same time, or 24 hours before TMV inoculation, almost complete inhibition was achieved. Likewise, 86% inhibition was achieved, when the culture filtrate was applied on the underside of the leaves. In field trials, transmission of TMV from diseased seedlings to healthy ones during transplanting work was reduced by 92%, when the culture filtrate was sprayed onto the tobacco seedlings, cv. NC82, 24 hours before transplanting. No toxic effect was observed on the tobacco plants. Antiviral substance from the culture filtrate was purified by ethanol precipitation, dialysis, DEAE-cellulose, and Sephadex G75 gel column chromatography. The partially purified active material which showed positive color reaction to sugar and protein inhibited TMV infection by 60% at 1 ${\mu}$g/ml.

Purification and Characterization of an Intracellular Inulinase from Bacillus sphaericus 188-1

  • Kim, Jae-Ho;Kwak, Yoon-Jin;Lee, Jong-Tae;Park, Shin-Yang;Lee, Jong-Soo
    • Preventive Nutrition and Food Science
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    • v.7 no.4
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    • pp.421-426
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    • 2002
  • In order to obtain basal data for industrial application of inulinase from Bacillus sphaeicus 188-1, its intracellular inulinase was purified by ammonium sulfate fractionation and column chromatography on DEAE-Sephadex A-50 and Sephadex G-100. The enzyme was homogeneous as judged by SDS-polyacrylamide gel electrophoresis, with an apparent molecular weight of 29 kDa. Inulinase activity was optimal at pH 6.5 and 4$0^{\circ}C$. The enzyme activity was significantly inhibited by Cu$^{2+}$, Cd$^{2+}$ and Hg$^{2+}$. The inulinase exhibited an apparent Km value of 0.014% for inulin.