• Title/Summary/Keyword: plasma column

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A Simple and Sensitive High Performance Liquid Chromatography-Electrospray Ionization/Mass Spectrometry Method for the Quantification of Ethyl Pyruvate in Rat Plasma

  • Kim, Hyun-Ji;Kim, Seung-Woo;Lee, Ja-Kyeong;Yoon, Sung-Hwa
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1221-1227
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    • 2011
  • Ethyl pyruvate (EP) is known as a scavenger of reactive oxygen species (ROS) in the body through its role in the donation of diketone groups to metals to form an EP-metal complex. In order to develop a method for the quantification of EP in biological media, a sensitive and specific, high-performance liquid chromatographyelectrospray ionization-mass spectrometry (HPLC-ESI/MS) method is used to determine the EP-alkali metal ion binding species. The analyte was separated on a ZORBOX SB-C8 ($3.5{\mu}m$, $30mm{\times}2.1mm$ I.D.) column and analyzed in selected ion monitoring (SIM) mode with a positive ESI interface using the m/z 255 $[2M + Na]^+$ ion. The method was validated over the concentration range of $0.5-60.0\;{\mu}g$/mL under 1/9 (v/v) of acetonitrile/methanol solvent system with flow rate 0.05 mL/min. The limit of quantification (LOQ) was $0.5{\mu}g$/mL.

A Study on the Control of Luminous Color in Gas Discharge Tubes

  • Lee, Jong-Chan;Her, In-Sung;Park, Yong-Sung;Masaharu Aono;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • v.4C no.1
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    • pp.5-9
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    • 2004
  • In this paper, pulsed discharge is used to control the luminous color in gas discharge tubes. The luminous color of the positive column in gas discharge tubes filled with Hg-Ar-Ne (1: 9, 60[Torr]) and having no phosphor material, varies from red to blue emitted by the Ne and Hg from the pulsed discharge. With changing of pulse-width and frequency, the electron temperature in the transient period affects changes to the residual ion and metastable atom densities. The first metastable atoms containing energy levels of about 16.6 [eV]have a very high probability that a collision will result in the ionization potential of Ar being 15.8 [eV]. The change of locus in the CIE chromaticity diagram with increasing pulse-width and frequency approves the variation of luminous color.

The Anticoagulant Fraction from the Leaves of Diospyros Kaki L. Has an Antithrombotic Activity

  • Sa You Seon;Kim Soo-Jin;Choi Hye-Seon
    • Archives of Pharmacal Research
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    • v.28 no.6
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    • pp.667-674
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    • 2005
  • The leaves of Persimmon (Diospyros kaki L.) has long been used for tea in Korea since it was thought to be effective against hypertension. An anticoagulant fraction was purified through gel filtration G-100, hydrophobic, gel filtration G-150, and FPLC, Phenyl superpose column chromatographies. The purified fraction was homogenous and its Mr was estimated 10,000 Da by gel filtration and SDS-PAGE. The purified fraction was sensitive to treatment of subtilisin B, but not to heat and its activity was not changed after periodate oxidation, indicating that the activity was not due to carbohydrates. It delayed thrombin time (TT), activated partial thromboplastin time (APTT), and prothrombin time (PT) using human plasma. TT was more sensitive than APTT and PT, suggesting that the anticoagulant activity may be caused by a degradation or a defect of fibrin or thrombin. It did not cause the hydrolysis of fibrin after incubation. However, it inhibited thrombin-catalyzed fibrin formation with a competitive inhibition pattern. These results indicate that it may be an antithrombotic agent and that it is bound to fibrinogen binding sites of thrombin.

Induction of B Lymphocyte Differentiation by a Colostral Immunomodulatory Protein MIEF (초유에 함유되어 있는 면역조절물질인 MIEF가 B 세포의 분화에 미치는 영향)

  • Lee, Jong-Ho;Lee, Chong-Kil;Han, Seong-Sun
    • YAKHAK HOEJI
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    • v.38 no.3
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    • pp.351-357
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    • 1994
  • The levels of maternal immunity enhancing factor(MIEF), which is an immunomodulatory protein identified from bovine colostrum, were determined by indirect competitive enzyme-linked immunosorbent assay(ELISA) for the colostrum and normal milk collected during the first two weeks of lactation. The mean concentration of MIEF in the colostrum of the first day of lactation was $109\;{\mu}g/ml$, and fell from the third day of lactation to $3{\sim}4\;{\mu}g/ml$. The molecular weight of the purified MIEF determined by reducing SDS-PAGE and TSK G2000SW column chromatography was 22,000 and 24,000 daltons, respectively, showing that MIEF is a monomeric peptide in its native form. To examine the capacity of MIEF to induce differentiation of B Lymphocytes, human tonsillar Iymphocytes were cultured in the presence of different concentrations of MIEF, and then antibody secreting cells were enumerated by enzyme-linked immunospot(ELISPOT) assay. When added to cultures of human tonsillar Lymphocytes, MIEF induced differentiation of resting B Iymphocyte to antibody secreting plasma cells as efficiently as LPS.

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Anticoagulant 1,2,3,4,6-Pentagalloyl-$\beta$-D-Glucopyranose Isolated from Geranium (Pelargonium inquinans Ait)

  • Ji Myeong-Sim;Piao Xiang-Lan;Jin Yu-Lan;Park Ro Dong
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1037-1041
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    • 2005
  • Geranium (Pelargonium inquinans Ait) leaves were extracted with $80\%$ MeOH, and partitioned into n-hexane, ethyl acetate, BuOH and $H_2O$ to isolate the anticoagulant principles. The EtOAc fraction was found to be the most active, and was further purified using silica and octadecylisilane column chromatography employing a bioassay-guided fractionation method. The active compound was isolated and identified as $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$(PGG) (compound I). The isolated anticoagulant significantly prolonged the activated partial thrombin time (APTT) and thrombin time (TT) using normal human plasma. One microgram of $1,2,3,4,6-pentagalloyl-\beta-D-glucopyranose$ showed 0.063 heparin units in the APTT and 2.73 heparin units in the TT for anti-thrombosis. This is the first report of the isolation of PGG from geranium plants.

Evaluation of Recent Data Processing Strategies on Q-TOF LC/MS Based Untargeted Metabolomics

  • Kaplan, Ozan;Celebier, Mustafa
    • Mass Spectrometry Letters
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    • v.11 no.1
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    • pp.1-5
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    • 2020
  • In this study, some of the recently reported data processing strategies were evaluated and modified based on their capabilities and a brief workflow for data mining was redefined for Q-TOF LC-MS based untargeted metabolomics. Commercial pooled human plasma samples were used for this purpose. An ultrafiltration procedure was applied on sample preparation. Sample set was analyzed through Q-TOF LC/MS. A C18 column (Agilent Zorbax 1.8 µM, 50 × 2.1 mm) was used for chromatographic separation. Raw chromatograms were processed using XCMS - R programming language edition and Isotopologue Parameter Optimization (IPO) was used to optimize XCMS parameters. The raw XCMS table was processed using MS Excel to find reliable and reproducible peaks. Totally 1650 reliable and reproducible potential metabolite peaks were found based on the data processing procedures given in this paper. The redefined dataset was upload into MetaboAnalyst platform and the identified metabolites were matched with 86 metabolic pathways. Thus, two list were obtained and presented in this study as supplement files. The first list is to present the retention times and m/z values of detected metabolite peaks. The second list is the metabolic pathways related with the identified metabolites. The briefly described data processing strategies and dataset presented in this study could be beneficial for the researchers working on untargeted metabolomics for processing their data and validating their results.

High-performance liquid chromatographic assay and oral pharmacokinetics of new anti-HIV uracil derivatives, KR-V analogues, in rats

  • Lee, Youngmi;Heeok Shim;Seoungryong Yu;Hochul Shin;Moonkoo Chung;Jungkoo Roh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.123-123
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    • 1998
  • A number of uracil derivatives have been developed as anti-AIDS drugs having a mechanism of inhibiting cellular reverse transcriptase. A simple and rapid assay technique for recently synthesized KR-V analogues was developed using a high-performance liquid chromatography, and oral pharmacokinetics was examined for assessing their oral bioavailabilites. Plasma samples were analyzed by reversed-phase HPLC using an ODS column with an ultraviolet detection system. All the analogues were eluted within 12 min and the LOQ was 15-30 ng/$m\ell$. The extraction recoveries were higher than 85%, except KR-V1039, 1068 and 1720 having ester group. This chromatic method was well applied to the kinetic studies for KR-V analogues. Among 16 analogues tested in the present work, the 6 compounds including KR-V1123, 1122, 1784, 1783, 1736 and 1700 were found to be bioavailable for oral administration to rats.

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Comparison of Surface and Core Peptide Fraction from Apo B-100 of Human LDL (Low Density Lipoprotein)

  • Cho, Hyun-Mi;Shin, Seung-Uon;Kim, Tae-Woong
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.145-151
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    • 1999
  • Apolipoprotein B-100 (apo B-100) is an important component in plasma low density lipoproteins (LDL). It function as the ligand for the LDL receptor in peripheral cells. The LDLs are removed from the circulation by both high-affinity receptor-mediated and receptor-independant pathways. LDLs are heterogeneous in their lipid content, size and density and certain LDL subspecies increase risk of atherosclerosis due to differences in the conformation of apo B in the particle. In the present study , surface and core peptide fraction of Apo B-100 have been characterized by comparing peptide-mapping and fluorescence spectroscopy. Surface fragments of apo B-100 were generated by digestion of LDL with either trypsin , pronase, or pancreatin elastase. Surface fractions were fractionated on a Sephadex G-50 column. The remaining core fragments were delipidated and redigested with the above enzymes, and the resulting core peptides were compared with surface peptides. Results from peptide-mapping by HPLC showed pronase-digestion was more extensive than trypsin -digestion to remove surface peptide fraction from LDL. Fluorescence spectra showed that core fractions contained higher amount of tryptophan than surface fractions, and it indicated that core fraction wa smore hydrophobic than surface fractions. A comparison of the behavior of the core and surface provided informations about the regions of apo B-100 involved in LDL metabolism and also about the structural features concerning the formation of atherosclerosis.

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Purification and Characterization of Anti-Coagulant Activity Fraction from Persimmon Stem (감꼭지로부터 혈액응고저해물질의 정제와 특성)

  • 사유선;김경아;최혜선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1323-1327
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    • 2003
  • Persimmon has been considered to have therapeutic values for various diseases in Korea. Dried persimmon has been applied to wounded parts for anti-inflammatory and analgesic activities. Anti-coagulant fraction from Persimmon stem was purified through gel filtration, phenyl Sepharose, DEAE-Sephadex and additional gel filtration column chromatographies. Its molecular weight was estimated to be 130,000 ∼ 180,000. By element analysis, its main components were C, H, and O. The anti -coagulant was heat- stable and completely inhibited after periodate oxidation, indicating that it was a complex carbohydrate.

Identification of Higenamine nad its Metabolites in Rat by Gas Chromatography/Mass Spectrometry

  • Ryu, Jae-chun;Song, Yun-Seon;Kim, Myung-Soo;Cho, Jung-Hyuck;Yunchoi, Hye-Sook
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.213-218
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    • 1993
  • ($\pm$)-Higenamine is known as a cardiotonic principle of aconite root (root of Aconitum spp., Ranunculaceae). A simple and sensitive detection method for higenamine was developed by using gas chromatography-mass spectrometry (GC/MS). The recovery of higenamine after extraction and concentration with XAD-2 resin column was around 95% from rat biological fluids such as bile, plasma and urine. The limits of detection of higenamine in these biological fluids were approximately 0.1 ng/ml each. It has well been suggested that tetrahydroisoquinolines possessing catechol moiety such as higenamine should be subjected to the catechol-O-methyl transferase (COMT) activity in vivo. We detected two major peaks of presumed metabolites of higenamine in the total ion chromatogram obtained from the rat urine sample after the oral adminstration of ($\pm$)-higenamine. The scan mass spectrum of one of the metabolties coincided with those obtained from coclaurine $(C_6$-O-methyl higenamine) and those of the other metabolite are suggestive of isococlaurine $(C_7$-O-methyl higenamine).

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