• Title/Summary/Keyword: Reaction monitoring

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Direct Stem Blot Immunoassay (DSBIA): A Rapid, Reliable and Economical Detection Technique Suitable for Testing Large Number of Barley Materials for Field Monitoring and Resistance Screening to Barley mild mosaic virus and Barley yellow mosaic virus

  • Jonson, Gilda;Park, Jong-Chul;Kim, Yang-Kil;Kim, Mi-Jung;Lee, Mi-Ja;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.260-265
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    • 2007
  • Testing a large number of samples from field monitoring and routine indexing is cumbersome and the available virus detection tools were labor intensive and expensive. To circumvent these problems we established tissue blot immunoassay (TBIA) method an alternative detection tool to detect Barley mild mosaic virus (BaMMV) and Barley yellow mosaic virus (BaYMV) infection in the field and greenhouse inoculated plants for monitoring and routine indexing applications, respectively. Initially, leaf and stem were tested to determine suitable plant tissue for direct blotting on nitrocellulose membrane. The dilutions of antibodies were optimized for more efficient and economical purposes. Results showed that stem tissue was more suitable for direct blotting for it had no background that interferes in the reaction. Therefore, this technique was referred as direct stem blot immunoassay or DSBIA, in this study. Re-used diluted (1:1000) antiserum and conjugate up to 3 times with the addition of half strength amount of concentrated antibodies was more effective in detecting the virus. The virus blotted on the nitrocellulose membrane from stem tissues kept at room temperature for 3 days were still detectable. The efficiency of DSBIA and RT-PCR in detecting BaMMV and BaYMV were relatively comparable. Results further proved that DSBIA is a rapid, reliable and economical detection method suitable for monitoring BaMMV and BaYMV infection in the field and practical method in indexing large scale of barley materials for virus resistance screening.

Review of Analytical and Assessment Techniques of Terminal Electron Accepting Processes (TEAPs) for Site Characterization and Natural Attenuation in Contaminated Subsurface Environments (오염 지중환경 특성화와 자연저감평가를 위한 말단전자수용과정(TEAPs) 분석 및 평가기술 소개)

  • Song, Yun Sun;Kim, Han-Suk;Kwon, Man Jae
    • Journal of Soil and Groundwater Environment
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    • v.25 no.2_spc
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    • pp.1-15
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    • 2020
  • Monitoring and assessing terminal electron accepting processes (TEAPs) are one of the most important steps to remediate contaminated sites via various in-situ techniques. TEAPs are a part of the microbial respiration reactions. Microorganisms gain energy from these reactions and reduces pollutants. Monitoring TEAPs enables us to predict degradability of contaminants and degradation rates. In many countries, TEAPs have been used for characterization of field sites and management of groundwater wells. For instance, US Environmental Protection Agency (EPA) provided strategies for groundwater quality and well management by applying TEAPs monitoring. Denmark has also constructed TEAPs map of local unit area to develop effective groundwater managing system, particularly to predict and assess nitrogen contamination. In case of Korea, although detailed soil survey and groundwater contamination assessment have been employed, site investigation guidelines using TEAPs have not been established yet. To better define TEAPs in subsurface environments, multiple indicators including ion concentrations, isotope compositions and contaminant degradation byproducts must be assessed. Furthermore, dissolved hydrogen concentrations are regarded as significant evidence of TEAPs occurring in subsurface environment. This review study introduces optimal sampling techniques of groundwater and dissolved hydrogen, and further discuss how to assess TEAPs in contaminated subsurface environments according to several contamination scenarios.

Analytical method development for residual metamizol in meat using LC/MS/MS (식육 중 메타미졸 잔류물의 LC/MS/MS 시험법 개발)

  • Kim, Tae-Wook;Yang, Yeung-Kyong;Gwoak, Soon-Chul;Kang, Dong-Young
    • Analytical Science and Technology
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    • v.24 no.4
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    • pp.237-242
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    • 2011
  • In this study, an analytical method was developed for residual metamizol in beef and pork using LC/MS/MS. 4-methylaminoantipyrin (MAA), the main metabolite of metamizol was targeted for analysis instead of its parent compound. MAA was simply extracted from meat by acetonitrile, purified and then analyzed by multiple reaction monitoring method (MRM). Standard addition method was used for calibration. The calibration curves showed the linearity of $r^2$ > 0.99 for both matrices included. The developed method was validated by six-time intra-lab tests and inter-lab tests with two other institutes. The validation of the whole procedure for beef showed the intra-lab accuracies of 78-102% (CV 5.5-9.1%) and the inter-lab accuracy of 98% (CV 14%); the intra-lab accuracies of 95-99% (CV 3.9-5.6%) and the inter-lab accuracy of 111% (CV 13%).

A Liquid-Based Colorimetric Assay of Lysine Decarboxylase and Its Application to Enzymatic Assay

  • Kim, Yong Hyun;Sathiyanarayanan, Ganesan;Kim, Hyun Joong;Bhatia, Shashi Kant;Seo, Hyung-Min;Kim, Jung-Ho;Song, Hun-Seok;Kim, Yun-Gon;Park, Kyungmoon;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2110-2115
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    • 2015
  • A liquid-based colorimetric assay using a pH indicator was introduced for high-throughput monitoring of lysine decarboxylase activity. The assay is based on the color change of bromocresol purple, measured at 595 nm in liquid reaction mixture, due to an increase of pH by the production of cadaverine. Bromocresol purple was selected as the indicator because it has higher sensitivity than bromothymol blue and pheonol red within a broad range and shows good linearity within the applied pH. We applied this for simple determination of lysine decarboxylase reusability using 96-well plates, and optimization of conditions for enzyme overexpression with different concentrations of IPTG on lysine decarboxylase. This assay is expected to be applied for monitoring and quantifying the liquid-based enzyme reaction in biotransformation of decarboxylase in a high-throughput way.

Implementation of a Black-Box Program Monitoring Abnormal Body Reactions (부정기적 발생 신체이상 모니터링 블랙박스 프로그램 구현)

  • Kim, Won-Jin;Yoon, Kwang-Yeol
    • The Journal of the Korea institute of electronic communication sciences
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    • v.7 no.3
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    • pp.671-677
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    • 2012
  • A black-box program was implemented in order to monitor abnormal symptoms of human body irregularly occurring during sleep. The system consists of sensor probing body signals, auxiliary devices such as the alarm, lamp, network camera, and signal monitoring computer. Various types of sensors, PPG, ECG, EEG, temperature, respiration sensor, G-sensor, and microphone were used to more exactly identify the causes of abnormal symptoms. If a symptom occurs, the system records the patient's condition to provide information being utilized in the treatment. The sensors are attached on some locations of body being proper to check a specific type of abnormal reaction. Based on the normal range and type of measurement data, criteria of signal levels were set to distinguish abnormal reaction. An abnormal signal being probed, the program starts to operate the lamp, alarm, and network camera at the same time and stores the signal and video data.

Monitoring of Maillard Reaction Characteristics under Various Roasting Conditions of Polygonatum odoratum Root (둥굴레 근경의 가열조건에 따른 갈변반응 특성의 모니터링)

  • 박난영;정용진;이기동;권중호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.647-654
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    • 2000
  • Response surface methodology (RSM) was used for monitoring the changes in browning reaction and organoleptic quality of roasted Polygonatum roots under various of roasting conditions. Total free sugar decreased up to 13$0^{\circ}C$, but increased above 13 $0^{\circ}C$. The amounts of total free amino acids decreased in proportional to the roasting temperature and time. Theronine, glycine and serine decreased by about 91~94% under the roasting conditions. Browning color intensity of water extracts increased with the roasting time up to around 18 min, but decreased over 18 min. The optimum conditions based on overall palatability of the roasted Polygonatum roots were 13$0^{\circ}C$ and 15 to 25 min. Organoleptic qualities of the roasted samples showed higher correlations with the changes in free amino acids.

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Rapid Quantitative Analysis of Vancomycin in Human Plasma and Urine Using LC-MS/MS (LC - MS/MS를 이용한 혈장과 뇨중에서 Vancomycin의 빠른정량분석)

  • Kim, Hohyun;Roh, Hyeongjin;Han, Sang-Beom
    • Analytical Science and Technology
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    • v.15 no.5
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    • pp.410-416
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    • 2002
  • In this study, a new quantitative analytical method has been developed for the rapid determination of vancomycin in human plasma and urine using liquid chromatography/tandem mass spectrometry (LC - MS/MS). Chromatography was carried out on a $C_{18}$ XTerra MS column ($2.1{\times}30mm$) with a particle size of $3.5{\mu}m$. The mobile phase was 0.25% formic acid in 10% acetonitrile and the flow rate was $250{\mu}L/min$. Vancomycin and caffeine (internal standard) were detected by MS/MS using multiple reaction monitoring (MRM). Vancomycin gives a predominant doubly protonated precursor molecule ($[M+2H]^{2+}$) at m/z 725.0 and a corresponding product ion of m/z 100.0. Detection of vancomycin was good, accurate and precise, with a limit of detection of 1 nM in plasma. The calibration curves for vancomycin in human plasma was linear in a concentration range of $0.01{\mu}M$ - $100{\mu}M$ for plasma. This method has been successfully applied to determine the concentration of vancomycin in human plasma and urine from pharmacokinetic study and relative studies.

Simultaneous analysis of glucocorticoids in bovine muscle and liver by LC-MS/MS (LC-MS/MS를 이용한 소의 근육과 간 중에 잔류하는 glucocorticoids 동시 분석)

  • Shin, Sang-Eun;Cho, Hyun-Woo;Myung, Seung-Woon
    • Analytical Science and Technology
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    • v.23 no.4
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    • pp.405-413
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    • 2010
  • A new method for the simultaneous determination of six glucocorticoids (betamethasone, dexamethasone, prednisone, prednisolone, methylprednisolone, and flumethasone) in meats (bovine muscle, bovine liver) were established. Samples were effectively extracted using C18 cartridge with ethylacetate. Chromatographic separation was achieved using C18 column and negative electrospray ionization mass spectrometry was performed in the Multiple Reaction Monitoring mode for the effective quantitation and qualification of glucocorticoids. Acetonitrile and water (0.1% formic acid) were used as mobile phase and additive for effective electrospray ionization, and gave good chromatographic separation and mass spectrometric sensitivity. The limit of detection (LODs) and the limit of quantitation (LOQs) in spiked blank samples depending on types of matrix and pharmaceuticals were ranged from 0.2 to $1.0\;{\mu}g$/kg and 0.8 to $3.4\;{\mu}g$/kg, respectively. And the recoveries were between 89.5 to 119.6%. The established method showed good recoveries, accuracies, precisions and fast sample preparation and it will be applied to assay of glucocorticoids residues in meat.

Quantitative Analysis of Lysophosphatidic Acid in Human Plasma by Tandem Mass Spectrometry

  • Kim, Ho-Hyun;Yoon, Hye-Ran;Pyo, Dong-Jin
    • Bulletin of the Korean Chemical Society
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    • v.23 no.8
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    • pp.1139-1143
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    • 2002
  • Analysis of lysophosphatidic acids (LPAs) is of clinical importance as they can serve a potential marker for ovarian and other gynecological cancers and obesity. It is critically important to develop a highly sensitive and specific method for the early detection of gynecological cancers to improve the overall outcome of this disease. We have established a novel quantification method of LPAs in human plasma by negative ionization tandem mass spectrometry (MS-MS) using multiple reaction monitoring (MRM) mode without the conventional TLC step. Protein-bound lipids, LPAs in plasma were extracted with methanol : chloroform (2:1) containing LPA C14:0 as an internal standard under acidic condition. Following back extraction with chloroform and water, the centrifuged lower phase was evaporated and reconstituted in methanol. The reconstituted solution was directly injected into electrospray source of MS/MS. For MRM mode, Q1 ions selected were m/z 409, 433, 435, 437 and 457 which corresponds to molecular mass [M-H]- of C16:0, C18:2, C18:1, C18:0 and C20:4 LPA, respectively. Q2 ions selected for MRM were m/z 79, phosphoryl product. Using MS/MS with MRM mode, all the species of LPAs were completely separated from plasma matrix without severe interferences. This method allowed simultaneous detection and quantification of different species of LPAs in a plasma over a linear dynamic range of 0.01-25 ㎛olL-1 . The detection limit of the method was 0.3 pmol/mL, with a correlation coefficient of 0.9983 in most LPAs analyzed. When applied to the plasmas of normal and gynecological cancer patients, this new method differentiated two different groups by way of total LPA level.

Monitoring of Red Pepper Powder and Seasoned Red-Pepper Sauce using Species-Specific PCR in Conjunction with Whole Genome Amplification

  • Hong, Yewon;Kwon, Kisung;Kang, Tae Sun
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.146-150
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    • 2018
  • Red pepper is one of the most important spices popularly utilized in Korea. Because of the differences in tariff rates between red pepper powder and seasoned red-pepper sauce, seasoned red-pepper sauce is often therefore imported by consumers, then dried, ground, and added to red pepper powder for cost effective purposed to use the product the most effectively. In this study, we combined species-specific polymerase chain reaction (PCR) assays (for red pepper, garlic, onion, spring onion, and ginger) with whole-genome amplification (WGA). Thirty-nine red pepper powders were well in accordance with their labels. However, six red pepper powder and five seasoned red-pepper sauce products failed to meet their compliance requirements. As a consequence, our monitoring results revealed that the overall mislabeling rate detected in this study was identified at 22%. Thus, our findings showed that the species-specific PCR in conjunction with WGA was an ideal method to identify raw materials that are used in the manufacturing of red pepper powder and seasoned red-pepper sauce.