• Title/Summary/Keyword: Reaction monitoring

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Establishment and application of a qualitative real-time polymerase chain reaction method for detecting genetically modified papaya line 55-1 in papaya products (RT-PCR을 이용한 유전자변형파파야(55-1)검사법 확립 및 파파야가공식품의 적용 연구)

  • Kwon, Yu Jihn;Chung, So Young;Cho, Kyung Chul;Park, ji Eun;Koo, Eun Joo;Seo, Dong Hyuk;Kim, Eugene;Whang, Jehyun;Park, Seong Soo;Choi, Sun Ok;Lim, Chul Joo
    • Analytical Science and Technology
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    • v.28 no.2
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    • pp.117-124
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    • 2015
  • Genetically modified (GM) papaya line 55-1, which is resistant to PRSV infection, has been marketed globally. Prompt and sensitive protocols for specific detections are essential for the traceability of this line. Here, an event- and construct-specific real-time polymerase chain reaction (RT-PCR) method was established to detect 55-1. Qualitative detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% for the homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The method was applied in the qualitative detection of 55-1 in eight types of commercially processed papaya products. Additionally, papaya products were monitored to distinguish GM papaya using the P35S and T-nos RT-PCR detection methods. As expected, detection capacity was improved via modified sample preparation and the established RT-PCR detection method. Taking these results together, it can be suggested that a suitable method for the extraction and purification of DNA from processed papaya products was established for the detection of GM papaya.

A Rapid and Sensitive Detection of Aflatoxin-producing Fungus Using an Optimized Polymerase Chain Reaction (PCR)

  • Bintvihok, Anong;Treebonmuang, Supitchaya;Srisakwattana, Kitiya;Nuanchun, Wisut;Patthanachai, Koranis;Usawang, Sungworn
    • Toxicological Research
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    • v.32 no.1
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    • pp.81-87
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    • 2016
  • Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was $65^{\circ}C$. The optimized template and primer concentration were $1.5{\mu}L\;(50ng/{\mu}L)$ and $3{\mu}L\;(10{\mu}M/{\mu}L)$ respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at $88.0^{\circ}C$, $87.5^{\circ}C$, $83.5^{\circ}C$, and $89.5^{\circ}C$ respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.

Evaluation of Field Application and Estimation of Bedload Discharge in the Forest Watershed using the Hydrophone (하이드로폰을 이용한 산림유역 소류사 유출량 산정 및 현장 적용성 검토)

  • Seo, Jun-Pyo;Kim, Ki-Dae;Woo, Choong-Shik;Lee, Chang-Woo;Lee, Heon-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.12
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    • pp.807-818
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    • 2020
  • In this study, hydrophones using acoustic sensors were used to estimate the amount of bedload discharge in a forested watershed. The reaction characteristics were analyzed through hydrophone flume tests and field tests, and the quantitative bedload discharge was calculated and compared with that measured by a pit sampler. The hydrophone reaction changed the pulse according to the flow rate change, but did not react to standard sand. The pulse was different depending on the particle size and weight, and accordingly, there was a specific channel showing a suitable response. For a hydrophone installed in the field, by using an automatic impact device, the reaction characteristics of each channel were analyzed to confirm normal operation of the sensor and the suitability of the output value of each channel. In addition, a suitable channel was selected for the estimation of bedload discharge. The bedload discharge formula was developed using a hydrophone pulse and the average flow rate, and was compared with the measured data in the pit sampler in the study site. As a result of the study, if a hydrophone is used for monitoring the bedload in forested watersheds, it is considered effective in quantitatively estimating the weight of bedload discharge.

Development of Mixed-bed Ion Exchange Resin Capsule for Water Quality Monitoring (수질 중 질소와 인 모니터링을 위한 혼합이온교환수지 캡슐의 개발)

  • Park, Chang-Jin;Kim, Dong-Kuk;Ok, Yong-Sik;Ryu, Kyung-Ryul;Lee, Ju-Young;Zhang, Yong-Seon;Yang, Jae-E
    • Applied Biological Chemistry
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    • v.47 no.3
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    • pp.344-350
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    • 2004
  • This study was conducted to develop and assess the applicability of mixed-bed ion exchange resin capsules for water quality monitoring in small agricultural watershed. Recoveries of resin capsules for inorganic N and P ranged from 96 to 102%. The net activation energies and pseudo-thermodynamic parameters, such as ${\Delta}G^{o\ddag},\;{\Delta}H^{o\ddag},\;and\;{\Delta}S^{o\ddag}$ for ion adsorption by resin capsules, exhibited relatively low values, indicating the process might be governed by chemical reactions such as diffusion. However, those values increased with temperature coinciding with the theory. The reaction reached pseudo-equilibrium within 24 hours for $NH_4-N\;and\;NO_3-N$, and only 8 hours for $PO_4-P$, respectively. The selectivity of resin capsules were in the order of $NO_3\;^-\;>\;NH_4\;^+\;>\;PO_4\;^{3-}$, coinciding with that of encapsulated Amberlite IRN-150 resin. At the initial state of equilibrium, the resin adsorption quantity was linearly proportional to the mass of ions in the streams, but the rate of movement leveled off, following Langmuir-type sorption isotherm. The overall results demonstrated that the resin capsule system was suitable for water quality monitoring in small agricultural watershed, judging from the reaction mechanism(s) of the resin capsule and the significance of model in field calibration.

Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선)

  • Cho, Yoon-Jae;Chae, Young-Sik;Kim, Jae-Young;Kim, Hyung-Soo;Kang, Ilhyun;Do, Jung-Ah;Oh, Jae-Ho;Kwon, Kisung;Chang, Moon-Ik
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.25-33
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    • 2013
  • Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Fenton Oxidation of Landfill Leachate by $Fe^0,\;Fe^{2+},\;Fe^{3+}\;/\;H_2O_2$ Systems ($Fe^0$, $Fe^{2+}$, $Fe^{3+}$ / $H_2O_2$ 시스템을 이용한 침출수의 Fenton 산화반응)

  • Park, Sung-Ho;Han, Ihn-Sup
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.4
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    • pp.402-408
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    • 2005
  • This investigation aimed at selecting the optimum catalyst and reaction conditions used in Fenton oxidation for landfill leachate treatment and was carried out at ambient temperature using a lab-scale experiment. The investigation led to the following results: 1) The optimum pH and dose for each iron catalyst were as follows: $Fe^{2+}\;=\;1,200\;mg/L$, $H_2O_2\;=\;1,200\;mg/L$, initial pH=3.0; $Fe^{3+}\;=\;1,200\;mg/L$, $H_2O_2\;=\;1,500\;mg/L$, initial pH=4.5; $Fe^0\;=\;1,200\;mg/L$, $H_2O_2\;=\;900\;mg/L$, initial pH=4.0, respectively. 2) The progress of Fenton oxidation could be instrumentally monitored by measuring redox potential evolution during leachate oxidation, thus, indicating the possibility of an on-line process monitoring. 3) A simple acid-base titration of Fenton-treated leachate proved that a relevant fraction of by- products formed during the treatment was made of acidic compounds in the optimum reaction condition for each catalyst used, thus demonstrating that the higher the extent of Fenton oxidation the greater was the amount of acids formed. 4) With the aim of selecting the optimum catalyst among $Fe^0$, $Fe^{2+}$ and $Fe^{3+}$, removal efficiency of each parameter in the optimum reaction conditions was considered. Although $Fe^{3+}$ was higher than other catalysts($Fe^0$, $Fe^{2+}$) in removal efficiency, $Fe^0$ was a optimum catalyst with a view of cost effectiveness.

Monitoring the Reoccurrence of Fire Blight and the Eradication Efficiency of Erwinia amylovora in Burial Sites of Infected Host Plants Using Sentinel Plants (미끼식물을 이용한 화상병 감염 기주 매몰지 내 화상병균 제거 효율 검증 및 병 재발 모니터링)

  • In Woong, Park;Yu-Rim, Song;Nguyen Trung, Vu;Eom-Ji, Oh;In Sun, Hwang;Hyeonheui, Ham;Seong Hwan, Kim;Duck Hwan, Park;Chang-Sik, Oh
    • Research in Plant Disease
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    • v.28 no.4
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    • pp.221-230
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    • 2022
  • The fire blight caused by Erwinia amylovora (Ea) was first reported in 2015 in Korea, and the disease has rapidly spread to 22 regions until 2021. In Korea, all host plants in the apple and pear orchards where fire blight occurred should be eliminated and buried by the Plant Protection Act. To prevent the spread of the disease, all burial sites were prohibited from planting the new host plants for the next three years. To confirm the eradication efficiency of Ea and the reoccurrence of fire blight, the surveillance facilities were established on three burial sites from 2019 to 2020 in Anseong-si, Gyeonggi-do, and Chungju-si, Chungcheongbuk-do. As host plants, five apple trees of fire blight-susceptible cultivar 'Fuji', were planted in each facility. All facilities were enclosed with fences and nets and equipped with two CCTVs, motion sensors, and several other sensors for recording weather conditions to monitor the environment of the sentinel plants in real-time. The sentinel plants were checked for the reoccurrence of fire blight routinely. Suspicious plant parts were collected and analyzed for Ea detection by loop-mediated isothermal amplification polymerase chain reaction and conventional polymerase chain reaction. Until November 2022, Ea has not been detected in all sentinel plants. These results might support that the burial control of infected plants in soil works efficiently to remove Ea and support the possibility to shorten the prohibition period of host plant establishment in the burial sites.

Rapid Detection of Radioactive Strontium in Water Samples Using Laser-Induced Breakdown Spectroscopy (LIBS) (Laser-Induced Breakdown Spectroscopy (LIBS)를 이용한 방사성 스트론튬 오염물질에 대한 신속한 모니터링 기술)

  • Park, Jin-young;Kim, Hyun-a;Park, Kihong;Kim, Kyoung-woong
    • Economic and Environmental Geology
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    • v.50 no.5
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    • pp.341-352
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    • 2017
  • Along with Cs-137 (half-life: 30.17 years), Sr-90 (half-life: 28.8 years) is one of the most important environmental monitoring radioactive elements. Rapid and easy monitoring method for Sr-90 using Laser-Induced Breakdown Spectroscopy (LIBS) has been studied. Strontium belongs to a bivalent alkaline earth metal such as calcium and has similar electron arrangement and size. Due to these similar chemical properties, it can easily enter into the human body through the food chain via water, soil, and crops when leaked into the environment. In addition, it is immersed into the bone at the case of human influx and causes the toxicity for a long time (biological half-life: about 50 years). It is a very reductive and related with the specific reaction that makes wet analysis difficult. In particular, radioactive strontium should be monitored by nuclear power plants but it is very difficult to be analysed from high-cost problems as well as low accuracy of analysis due to complicated analysis procedures, expensive analysis equipment, and a pretreatment process of using massive chemicals. Therefore, we introduce the Laser-Induced Breakdown Spectroscopy (LIBS) analysis method that analyzes the elements in the sample using the inherent spectrum by generating plasma on the sample using pulse energy, and it can be analyzed in a few seconds without preprocessing. A variety of analytical plates for samples were developed to improve the analytical sensitivity by optimizing the laser, wavelength, and time resolution. This can be effectively applied to real-time monitoring of radioactive wastewater discharged from a nuclear power plant, and furthermore, it can be applied as an emergency monitoring means such as possible future accidents at a nuclear power plants.

Natural Baseline Groundwater Quality in Shingwang-myeon and Heunghae-eup, Pohang, Korea (포항시 신광면 및 흥해읍 일대 지하수의 배경수질 연구)

  • Lee, Hyun A;Lee, Hyunjoo;Kwon, Eunhye;Park, Jonghoon;Woo, Nam C.
    • The Journal of Engineering Geology
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    • v.30 no.4
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    • pp.469-483
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    • 2020
  • The results of long-term groundwater level and quality monitoring can be used not only as the basic data for evaluating the impact of various disasters including climate change and establishing responses, but also as key data for predicting and managing geological disasters such as earthquakes. Some countries use groundwater level and quality monitoring for researches to predict earthquakes and to assess the impacts of the earthquake disaster. However, a few cases in Korea report on individual groundwater quality factors (i.e., dissolved ions) observed before and after the earthquakes, being different from other countries. To establish the abnormality criteria for groundwater quality in Pohang, groundwater samples were collected and analyzed five times from 14 agricultural or private wells existing in Shingwang-myeon and Heunghae-eup. As a result of the analysis, it was found that Ca2+ was the dominant cation in Shingwang-myeon, while Na+ was the dominant cation in Heunghae-eup. The elevated NO3- concentration in Shingwang-myeon is contributed to the agricultural activity in the area. A high concentration of Fe was detected in a well on Heunghae-eup; the concentration exceeded the drinking water standard by nearly 100 times. Relatively higher dissolved ions were observed in the groundwater of Heunghae-eup, and it is considered as the result of the flow velocity difference and water-rock reaction accompanying the difference in bedrock and sediment characteristics. The groundwater of Shingwang-myeon appeared to be most affected by the weathering of granite and silicates, while that of Heunghae-eup was mainly affected by the weathering of silicates and carbonate. The background concentrations (baselines) of groundwater Shingwang-myeon and Heunghae-eup was identified through the survey; however, the continuous monitoring is required to monitor the possible changes and the repeatability of seasonal variation.

Characteristic Analysis of Tropospheric Ozone Sensitivity from the Satellite-Based HCHO/NO2 Ratio in South Korea (위성 기반 HCHO/NO2 비율을 통한 국내 대류권 오존 민감도 특성 분석)

  • Jinah Jang;Yun Gon Lee ;Jeong-Ah Yu;Kyoung-Hee Sung;Sang-Min Kim
    • Korean Journal of Remote Sensing
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    • v.39 no.5_1
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    • pp.563-576
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    • 2023
  • In this study nitrogen dioxide (NO2), formaldehyde (HCHO) from the Ozone Monitoring Instrument (OMI) and TROPOspheric Monitoring Instrument (TROPOMI), OMI/ Microwave Limb Sounder (MLS) tropospheric column ozone (TCO), and Airkorea ground-based O3 data were analyzed to examine the photochemical reaction relationship between tropospheric ozone and its precursors nitrogen oxides (NOx) and volatile organic compounds (VOCs). As a result of analyzing the trend of long-term changes from 2006 to 2020 using OMI satellite data, TCO showed an increasing trend, NO2 steadily decreased, and HCHO continued to increase in Northeast Asia. In addition, formaldehyde nitrogen dioxide ratio (FNR; HCHO/NO2 ratio), an indicator of ozone sensitivity, is gradually increasing, which means that the VOC-limited regime is decreasing. This study conducted a sensitivity analysis of ozone generation using TROPOMI FNR and ground-based ozone (O3) over the recent years (2019~2022) to identify the possible cause for the continuous increase of ozone in Korea. Similar to the previous studies, VOC-limited and transitional regimes appeared in megacities, and VOC-limited regimes also appeared in areas where major power plants were located. In VOC-limited regimes, in other words, areas where NOx is excessively saturated, the reduction in NOx emissions may have weakened the ozone titration and thus led to the increase of ozone. Therefore, VOC emissions should be reduced in the short term rather than NOx emissions to reduce ozone concentrations under the VOC-limited regime.