• Bulletin of the Korean Chemical Society
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• v.28 no.5
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• pp.737-744
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• 2007

An isotope dilution-liquid chromatography/tandem mass spectrometric method was developed as a candidate reference method for the accurate determination of acrylamide in potato chips, starch-rich foodstuff cooked at high temperature. Sample was spiked with 13C3-acrylamide and then extracted with water. The extract was further cleaned up with an Oasis HLB solid-phase extraction (SPE) cartridge and an Oasis mixed-phase cation exchange (MCX) SPE cartridge. The extract was analyzed by using LC/ESI/Tandem MS in positive ion mode. LC with a medium reversed-phase (C4) column was optimized to obtain adequate chromatographic retention and separation of acrylamide. MS was operated to selectively monitor [M+H]+ ions of the analyte and its isotope analogue at m/z 72 and m/z 75, respectively. Sample was also analyzed by the LC/MS with selectively monitoring the collisionally induced dissociation channels of m/z 72 → m/z 55 and m/z 75 → 58. Compared to the LC/MS chromatograms, the LC/MS/MS chromatograms showed substantially reduced background chemical noises coming from solvent clusters formed during ESI spray processes and interferences from sample matrix. Repeatability and reproducibility studies showed that the LC/MS/MS method is a reliable and reproducible method which can provide a typical method precision of 1.0% while the LC/MS results are influenced by chemical interferences.

• YAKHAK HOEJI
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• v.57 no.6
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• pp.420-425
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• 2013

Phentermine (PT) and phenmetrazine (PM) have been widely used as anti-obesity drugs. These drugs should be used with caution due to its close relation to amphetamine in its structure and toxicity. PT and PM, amphetamine-type anorectics, have recently been considered as alternatives for methamphetamine abuse in Korea. In addition, the misuse and abuse of PT and PM obtained by illegal sources such as the internet become a serious social problem. In the present study, a simultaneous detection and quantification method for determining PT and PM in human urine was developed and validated according to the international guidelines. The urine samples were screened using a fluorescence polarization immunooassay and analyzed by gas chromatography mass spectrometry (GC-MS) after extraction using automatic solid phase extraction (SPE) with a mixed-mode cation exchange cartridge and derivatization with pentafluoropropionic anhydride (PFPA). The validation results for selectivity, linearity, limits of detection (LOD) and quantification (LOQ), intra- and inter-assay precision and accuracy and recovery were satisfactory. The validated method was successfully applied to authentic urine samples collected from 38 drug abuse suspects. PT and/or PM were identified with or without methamphetamine in urine samples. Abuse of PT and PM have increased continuously in Korea, therefore, closer supervision of the inappropriate use of anoretics is necessary.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.379-387
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• 1996

The objects of the present study were to establish the method of purification, subunit dissociation of verotoxin-2 (VT2) produced by Escherichia coli O157:H7, and to investigate the characteristics of purified verotoxin-2 such as molecular weight and composition of amino acid. The results were summerized as follows; Verotoxin-2 was extracted by addition of polymyxin B sulfate into bacterial cell lysate prepared from Escherichia coli O157:H7(KSC109). As an initial step, the bacterial cell lysate was precipitated with 30% saturated ammonium sulfate. The precipitated crude toxin was then subjected to anion-exchange, chromatofocusing and cation-exchange chromatography. Using this scheme, we obtained highly purified toxin with a specific activity of $1.1{\times}10^9$ $CD_{50}/mg$. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) for purified VT2 showed two protein bands. The upper band, approximately 32 Kd, was supposed as A subunit and the lower band, approximately 7.7 Kd, was supposed as B subunit. When the toxin was separated in the subunit-dissociating solution, two peaks emerged with retention times of 15 and 28 min by HPLC. These peaks represented A subunit and B subunit, respectively. The amino acid composition of purified VT2 were made up in order of glutamic acid, histamine, asparaginic acid, histidine, lysine, alanine and leucine etc. The largest amount among the amino acid composing VT2 was methionine.

• Analytical Science and Technology
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• v.21 no.2
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• pp.143-147
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• 2008

U(IV) ion, the valance state of uranium presumed at in a deep-depth disposal of a spent fuel, was prepared and separated from U(VI) ion. In order to prepare U(IV) ion, tests were performed by adding several reducing agents into a uranyl solution or by dissolution of uranium oxide in a mixed acid added with a reducing agent. The valance states of the uranium in the prepared solutions were identified by separating two ions with a Dowex AG 50W-X8 cation exchange resins and measuring the solutions using a laser-induced fluorescence spectroscopy. However, U(IV) and U(VI) were not separated by a Lichroprep Si60 exchange resin in the same separation condition of Pu(IV) and Pu(VI).

• Journal of the Korean Wood Science and Technology
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• v.12 no.3
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• pp.25-34
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• 1984

This experiment was carried out to investigate the effects of monosaccharides extracted by saturated portland cement solution on the cement setting in comparision with the inhibitory index (I) of each lignocellulosic-cement system. The wood species which have been widely reforested in Korea, Populus alba-grandulosa, Larix leptolepis, Abies holophylla, Pinus koraiensis, Pinus rigida, Pinus densiflora and agricultural wastes of rice husk and rice stalk were used at this study. The wood meal, 0.50g on dry weight basis, through 0.83 mm(20 mesh) and retained on 0.35mm (40 mesh) screen was extracted by 25 ml saturated portland cement solution and the pH of saturated portland cement solution Was 12.7. To eliminate cation exsisting in the extracted solution, the cation exchange column was used (Fig. 4). Afterwards the extracted monosaccharides were reduced into alditols with sodium borohydride and analyzed by the gas-liquid chromatography for xylan, mannan, arabinan, galactan, gluean. The heat of cement hydration for lignocellulosic-cement system was measured in Dewar flask (Fig. 2). And then the inhibitory indices were calculated from maximum hydration temperature, time and maximum slops of hydration curve of ligno cellulosic-cement systems. The results obtained were as follows; (1) The inhibitory index of pines-Pinus rigida (I=29.33) and Pinus densiflora (I=35.76), were lower than that of poplar-Populus alba-glandulosa (I=41.48), and the index of Larix ieptoiepis (I=73.00) was the highest among eight lignocellulosic-cement systems, and accordingly both Pinus rigida and Pinus des(flora were seemed to be good wood species for wood-cement composite manufacture. (2) In case of Pinus rigida, the inhibitory index was 29.33 and the ratio of the hexoses to the pemoses was 6.04 and in case of Larix leptolepis, the index and the ratio were 73.00 and 35.19, respectively. Therefore the inhibitory index increased with increasing the ratios of the hexoses to the pentoses. (3) The richer amount of xylose and mannose in species caused decreasing the slops of the hydration curve of the lignocellulosic-cement system, prohahly due to the chemical adsorption of the acetyl groups in the hemicellulose on the surface of cement grains. (4) The amoun of xylose and mannose were significant to the inhibitory index of each lignocellulosic-cement system but any specific relation between the amount of glucose and inhibitory index was not found.

• Proceedings of the Korean Radioactive Waste Society Conference
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• 2005.06a
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• pp.121-128
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• 2005

A study on the selective separation of $^{99}Tc,\;^{94}Nb,\;^{55}Fe,\;^{90}Sr$ and $^{59}Ni(^{63}Ni)$ from various radioactive wastes discharged from the nuclear power plants in Korea is being performed for use in their quantifications which are indispensible for the evaluation of the radionuclide inventory. Separation behaviour of Ce, Ca, Mg, Al, Cr, Ti, Mn and Cu recovered along with Ni during the separation of Re (as a surrogate of $^{99}Tc$), Nb, Fe and Sr by anion exchange and Sr-Spec extraction chromatography was investigated by cation exchange and Ni-Spec extraction chromatography using synthetic radioactive waste dissolved solutions containing matrix elements such as Re, Nb, Fe, Sr, Ni, B, Na, K, Ce, Co, Ca, Mg, Al, Zn, Cr, Pb, Cd, Mo, Mn, Cu, Zr, Ti and U. To purify the Ni fraction recovered and prepare a radionuclide source available for gas proportional counting, an application of the Ni precipitation procedure with dimethylglyoxime in the medium of ammonium citrate and tartaric acid solutions as a masking agent for co-existing metal ions was described in detail.

• Journal of Life Science
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• v.23 no.12
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• pp.1486-1494
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• 2013

A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

• KSBB Journal
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• v.14 no.1
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• pp.9-16
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• 1999

Transforming growth factor $\beta$1(TGF-$\beta$1) has potentials to be used as a new therapeutic agent. However, studies with TGF-$\beta$ were hindered by its high cost. In this study, we developed an improved method to purify TGF-$\beta$1 from human platelets, for which four purification steps were used: platelet extraction, gel filtration, cation exchange chromatography, and reverse phase high performance liquid chromatography. After a final step of purification, a pure protein with a molecular weight of 25,000 corresponding to the commercially available TGF-$\beta$1 was obtained, which were confirmed by silver staining and Western blotting after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). It was confirmed by the inhibitory effects of TGF-$\beta$1 on a mink lung epithelial cell line that the purified TGF-$\beta$1 had its biological activity, whose activity is slightly higher than that of the commercially available TGF-$\beta$1. About 3.7$\mug of the purified TGF-$\beta$1 was obtained from 10 units of concentrated human platelets, the final yield of which is about 21%.

• Applied Biological Chemistry
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• v.42 no.2
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• pp.99-104
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• 1999

Soil actinomycetes of 703 strains were isolated from 25 sampling points in Cheju Island using 4 different media. Arginine glycerol salts agar containing soil extract was found to be the best medium for the isolation of soil actinomycetes. Soil samples from pasture land showed higher population and diversity of the actinomycetes than those from citrus field, forest, island, hill or valley. When the antibacterial activity of the 526 isolates was tested against three bacterial strains, Escherichia coli, Staphylococcus aureus and Pseudomonas solanacearum the frequency of the isolates with antibacterial activity varied much depending upon the media used for isolation and cultivation. BL106Ba, one of the 10 isolates that showed antibacterial activity against all the above 3 test strains, was chosen based upon the pH and heat stability of its antibacterial metabolites, and was identified as Streptomyces sp. based upon its cultural, morphological and physiological characteristics. The partially purified white crystalline substance obtained from the culture supematant of BL1063a through cation exchange chromatography(AG MP-50) and three times consecutive gel filtration(Sephadex G-10) showed high antimicrobial activity against gram positive and negative bacteria, but low activity against yeasts. The partially purified substance was found to contain at least four different compounds with antibacterial activity by both thin layer chromatography and high performance liquid chromatography.

• Analytical Science and Technology
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• v.15 no.1
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• pp.20-25
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• 2002

An accurate and simple analytical technique for $^{89}Sr$ and $^{90}Sr$, overcoming the demerits of the conventional method, has been developed with extraction chromatography and liquid scintillation counting. The Sr fraction was separated from hindrance elements with oxalate coprecipitation or cation exchange resin and purified with Sr-Spec column. With liquid scintillation counter, $^{89}Sr$ was measured by Cerenkov radiation method, and $^{90}Sr$ was measured by spectrum unfolding method. The developed radioactive strontium separation method was validated by application to the IAEA-reference material (IAEA-375, Soil) and radioactive waste samples.