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Bioanalytical method validation for determination of arsenic speciation in dog plasma using HPLC-ICP/MS (Dog 혈장 중 HPLC-ICP/MS를 이용한 비소 화학종 분석법 검증)

  • Kim, Jong-Hwan;Kwon, Young Sang;Shin, Min-Chul;Kim, Su Jong;Seo, Jong-Su
    • Analytical Science and Technology
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    • v.29 no.5
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    • pp.234-241
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    • 2016
  • The approach presented in this article refers to the bioanalytical method validation for the detection and quantitative determination of arsenic species including arsenite (As(III)), arsenate (As(V)), dimethylarsinic acid (DMA) and monomethylarsonic acid (MMA) in dog plasma by high-performance liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICP/MS). The arsenic species were separated using an agilent As speciation column by a mobile phase of 2 mM sodium phosphate monobasic, 0.2 mM ethylenediaminetetraacetic acid disodium salt dehydrate, 10 mM sodium acetate, 3 mM sodium nitrate and 1 % ethyl alcohol at pH 11 (adjusted with 1M NaOH). The method validation experiment was obtained selectivity, linearity, accuracy, precision, matrix effect, recovery, system suitability, dilution integrity and various stabilities. All calibration curves showed good linearity (R2>0.999) within test ranges. The lower limit of quantitation (LLOQ) was 5 ng/mL for As(III), As(V) and DMA, and 20 ng/mL for MMA. The system suitability and dilution values were within 6.5 % and 7.7 %. Subsequently, the developed and validated HPLC-ICP/MS method was also successfully applied to determine the arsenic speciation in dog plasma samples, and the recoveries for the spiked samples were in the range of 91.5–102.2 %. Therefore, this method could be applied to the evaluation of arsenic exposure, health effect assessment and other bio-monitoring studies in biological samples.

Quantitative Cell Count of Vibrio vulnificus Cells Based on MPN-PCR Method (MPN-PCR 방법을 이용한 Vibrio vulnificus 균수 정량분석)

  • Jang, Yu-Mi;Park, Seul-Ki;Jeong, Hee-Jin;Lee, Jang-Won;Yoon, Yohan;Park, Kwon-Sam;Shin, Il-Shik;Kim, Young-Mog
    • Journal of Food Hygiene and Safety
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    • v.33 no.5
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    • pp.412-415
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    • 2018
  • The objective of this study was to establish a quantitative count method of Vibrio vulnificus cells. Plate count method is often used to count the number of V. vulnificus cells using thiosulfate citrate bile salts sucrose (TCBS) agar plate. However, this method is unsuitable for counting V. vulnificus cells due to growth inhibition and cell injuries in TCBS medium. In this study, we suggested a most probable number-polymerase chain reaction (MPN-PCR) method using alkaline peptone water medium for the quantification of V. vulnificus. This MPN-PCR method showed 2 log higher cell number than TCBS agar plate method. Similar results were also found in the control using, Luria-Bertani agar containing 2% NaCl. Thus, this MPN-PCR method can be used a sensitive method for quantitative count of viable V. vulnificus cells in fish and shellfish samples.

The Formation Characteristics of THMs and HAAs in Chlorination of Raw Water of Different Organic Matter Characteristics (상수원수의 유기물 특성에 따른 염소처리시 THMs 및 HAAs의 생성특성)

  • Oh, Sun-Mi;Kim, Seung-Hyun;Lee, Min-Gyu;Xu, Mei-Lan;Kam, Sang-Kyu
    • Journal of Environmental Science International
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    • v.15 no.8
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    • pp.785-797
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    • 2006
  • The formation characteristics of trihalomethanes (THMs) and haloacetic acids (HAAs) were investigated in chlorination of raw water of different organic mallet characteristics. The samples used in this study were hydrophobic (N-HPO) and hydrophilic fraction (N-HPI) (which were concentrated and separated from Nakdong river water), and humic acid (HA) (which is known as a strong hydrophobic acid) as a reference organic matter, the specific UV absorbance (SUVA) of which was 2.19, 1.15 and 7.92, respectively. With increasing chlorine contact time, THMFP and HAAFP (the formation potential of THMs and HAAs) increased, but their increase was different depending on the organic mallet characteristics (i.e., for N-HPI, THMFP was higher than HAAFP, but the inverse result was obtained for N-HPO and HA and the ratio between them was greater for HA), and the mainly formed chemical species were CHCI$_3$ in case of THMs and dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) in case of HAAs for N-HPO and HA (and the ratios of CHCI$_3$ to total THMs and DCAA and TCAA to total HAAs for HA were higher than those for N-HPO), but for N-HPI, the ratio of brominated THMs was a little higher than that of CHCI$_3$ and the ratio of DCAA and TCAA to total HAAs was lower than that of N-HPO, although they are main chemical species in case of HAAs. Comparing THMFP and HAAFP with the increase in bromide concentration added with those in not adding it, the former increased greatly and its increase was higher for the organic mallet with stronger hydrophobicity, but the latter was lower for N-HPO and N-HPI and was similar for HA. The main chemical species with increasing bromide concentration were CHBt$_3$ in case of THMs regardless of organic matter characteristics, and dibromoacetic acid (DBAA) for N-HPO and N-HPI, DBAA and tribromoacetic acid (TBAA) for HA in case of HAAs. With increasing reaction temperature and pH, THMFP and HAAFP increased for the former, but for the latter, THMFP increased and HAAFP decreased, although the rate of increase or decrease was different with organic mallet characteristics.

Biochemical Characterization of Recombinant L-Asparaginase (AnsA) from Rhizobium etli, a Member of an Increasing Rhizobial-Type Family of L-Asparaginases

  • Moreno-Enriquez, Angelica;Evangelista-Martinez, Zahaed;Gonzalez-Mondragon, Edith G.;Calderon-Flores, Arturo;Arreguin, Roberto;Perez-Rueda, Ernesto;Huerta-Saquero, Alejandro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.292-300
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    • 2012
  • We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

Heat-Induced Reaction of Bovine Whey Proteins (열처리로 야기되는 우유 유청 단백질의 반응)

  • 이유라;홍윤호
    • Food Science of Animal Resources
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    • v.22 no.2
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    • pp.179-182
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    • 2002
  • Using differential scanning calorimetry (DSC), changes underwent by a mixture of $\alpha$-lactalbumin ($\alpha$-La) and $\beta$-lactoglobulin ($\beta$-Lg) during heat treatment were studied, yielding useful information for the dairy industry. Results of the DSC showed that the heat denaturation temperature of the hobo-$\alpha$-La was higher than that of apo-$\alpha$-La, suggesting hole-$\alpha$-La‘s greater stability. The denaturation temperature of a mixture of holo-$\alpha$-La and $\beta$-Lg was also slightly lower than that of holo-$\alpha$-La alone. The denaturation temperature of an apo-$\alpha$-La and $\beta$-Lg mixture was higher than that of holo-$\alpha$-La and $\beta$-Lg, suggesting that the heat stability of apo-$\alpha$-La was increased by $\beta$-Lg. Based on these results, it is possible to conclude that a mixture of holo-$\alpha$-La and $\beta$-Lg is more intensively affected by an increase in temperature than other samples, and that free sulphydryl groups seem to take part in this heat-induced denaturation.

Fabrication of Chemical Sensors for the Detection of Acidic Gas using 1,3-bisdicyanovinylindane (1,3-bisdicyanovinylindane을 이용한 산성가스 감지용 화학 센서 제작)

  • Song, Hwan-Moon;Park, Young-Min;Son, Young-A;Lee, Chang-Soo
    • Korean Chemical Engineering Research
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    • v.46 no.1
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    • pp.184-188
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    • 2008
  • This study presented simple and efficient fabrication of chemical sensors for the detection of acidic gas using 1,3-bisdicyanovinylindane as an indicator because it can be promising materials having property of the rapid color change according to the variation of pH. The dissociation of proton and dye in acidic condition as changing of ion pairs give rise to dramatically change the absorbance intensity of 1,3-bisdicyanovinylindane, which can be easily applied to the development of chemical sensors. In addition, indicator dyes having negatively charge in aqueous phase can be easily fabricated using layer-by-layer (LBL) methods by way of electrostatic interaction. For the proof of concept, we demonstrated the abrupt presentation of skeleton symbol on the chemical sensor, which could be resulted from the reaction of 1,3-bisdicyanovinylindane as background color with acidic gas. Thus, the rapid appearance of symbol will induce user's caution under the emergency condition. The presented chemical gas sensor using 1,3-bisdicyanovinylindane have strong advantages. First, the fabrication process of gas sensor was very simple and low-cost. Secondly, sensors reacted by acidic gas could be reused for several times. Finally, the chemical gas sensor would be environmentally friend, which can be a basic tool for the realization of eco-organic sensor device.

Seroprevalence of Brucellosis and Isolation of Yersinia enterocolitica O:9 in Pigs (돼지에서 브루셀라병 항체조사 및 Yersinia enterocolitica O:9의 분리)

  • Jung, Byeong-Yeal;Byun, Jae-Won;Kim, Ha-Young;Shin, Dong-Ho;Park, Choi-Kyu;Jung, Suk-Chan
    • Journal of Life Science
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    • v.20 no.5
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    • pp.697-702
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    • 2010
  • Ten farrow-finish farms participated in this seromonitoring that was conducted to investigate the porcine brucellosis situation in Korea. In total, eight (80.0%) of the 10 farms and 139 (24.0%) of 578 pigs tested showed a positive response in the Rose Bengal test (RBT). Seroprevalence levels were determined using RBT according to age; 35 (14.6%) of 239 piglets, 36 (31.3%) of 115 growing pigs, and 68 (30.4%) of 224 finishing pigs and sows were positive, respectively. All positive samples in RBT were tested with the tube agglutination test (TAT) and competitive ELISA (C-ELISA), simultaneously. Although 48 samples came up positive in the TAT, all samples tested with C-ELISA were negative. Among 26 rectal swab samples from the TAT positive-pigs, Yersinia enterocolitica O:9 was isolated from seven samples (26.9%). Therefore, we speculated that the positive reaction of RBT and TAT in this study might be induced by the serologically cross-reacting bacteria with Brucella abortus.

Polydiacetylene-Based Chemo-/Biosensor of Label Free System with Various Sensing Tools (다양한 감지 방법을 갖고 있는 폴리디아세틸렌 기반 비표지 화학/바이오센서)

  • Park, Hyun-Kyu;Park, Hyun-Gyu;Chung, Bong-Hyun
    • KSBB Journal
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    • v.22 no.6
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    • pp.409-413
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    • 2007
  • Polydiacetylene(PDA)-based sensors possess a number of properties that can be successfully applied for label-free detection system. PDA is one of the most attractive color-generating materials, with growing applications as sensors. Here we introduce various PDA-based devices, used as biosensor, chemosensor, thermosensor, and optoelectronics sensor. In general, PDA liposomes and films are closely packed and properly designed for polymerization via 1,4-addition reaction to form an ene-yne alternating polymer chain. PDA-based two/three dimensional structures have been used for colorimetric or fluorescent devices, sensing biological as well as chemical components. This color-generating material also present a very high charge carrier mobility, allowing its application as field-effect transistor (FET). The immobilized PDA structures or films have distinct advantages for the detection of low concentration target molecules over the aqueous solution-based detection systems. In the present review, reported detection methods by using various PDA structures are summarized with updated references.

Effects of Heat Treatment Conditions on the Interfacial Reactions and Crack Propagation Behaviors in Electroless Ni/electroplated Cr Coatings (열처리 조건에 따른 무전해 Ni/전해 Cr 이중도금의 계면반응 및 균열성장거동 분석)

  • Son, Kirak;Choi, Myung-Hee;Lee, Kyu Hawn;Byon, Eungsun;Rhee, Byong-Ho;Park, Young-Bae
    • Journal of the Microelectronics and Packaging Society
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    • v.23 no.3
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    • pp.69-75
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    • 2016
  • This study investigated the effect of heat treatment conditions not only on the Cr surface crack propagation behaviors but also on the Ni/Cr interfacial reaction characteristics in electroless Ni/electroplated Cr double coating layers on Cu substrate. Clear band layer of Ni-Cr solid solutions were developed at Ni/Cr interface after heat treatment at $750^{\circ}C$ for 6 h. Channeling cracks formed in Cr layer after 1 step heat treatment, that is, heat treatment after Ni/Cr plating, while little channeling cracks formed after 2 step heat treatment, that is, same heat treatments after Ni and Cr plating, respectively, due to residual stress relaxation due to crystallization of Ni layer before Cr plating.

A study on the derivatization technique for tamoxifen metabolites in human urine by gas chromatography/mass spectrometry (기체크로마토그래피/질량분석기를 이용한 인체 내 뇨시료에서의 Tamoxifen 대사체 검출을 위한 유도체화 연구)

  • Kim, Yunje;Lee, Yoonjung
    • Analytical Science and Technology
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    • v.17 no.4
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    • pp.322-336
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    • 2004
  • The improved derivatization technique of tamoxifen metabolite in human urine is described for the acylation method that they are substituted by derivatization reagent like acyl anhydride for use of gas chromatography/mass spectrometry. The hydroxyl group of tamoxifen metabolite was derivatized by trifluoroacetic anhydride (TFAA), pentafluoroacetic anhydride (PFPA) and heptaflorobutylic anhydride (HFBA). It was investigated to the gas chromatography/mass spectrometry (GC/MS) technique use negative ion chemical ionization (NCI), positive ion chemical ionization (PCI) and electron impact (EI). In acylation of the metabolites of tamoxifen, the effective reaction temperature and time were shown to be at $50^{\circ}C$ for 30 min. The 4-hydroxytamoxifen, which is known to major metabolite of tamoxifen, was not detected in human urine, whileas the hydroxymethoxytamoxifen was detected. We thought that this result was from the single dose of tamoxifen.