• Title/Summary/Keyword: Reaction conditions optimization

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A Study on the Automation and Optimization of 9-(4-[$^{18}F$] Fluoro-3-hydroxymethylbutyl) Guanine Synthesis (9-(4-[$^{18}F$] Fluoro-3-hydroxymethylbutyl) guanine 합성의 자동화와 최적화에 관한 연구)

  • An, Jae-Seok;Hong, Sung-Tack;Kang, Se-Hun;Won, Woo-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.2
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    • pp.72-75
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    • 2011
  • Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage is that it is possible to monitor directly without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. This study was performed to automate 9-(4-[$^{18}F$] Fluoro-3-hydroxymethylbutyl) guanine ([$^{18}F$] FHBG) that are widely used as substrate for the HSV1-tk reporter gene in living organisms with positron emission tomography (PET) and find the optimized conditions of synthesis. Materials and Methods: Fully automated synthesis of [$^{18}F$] FHBG was performed using Explora-RN (CTI, USA) module. We have changed of reaction time (3, 5, 10 min) and temperature (110, 120, $130^{\circ}C$) for the optimized conditions of synthesis. Also we experimented to find the optimal concentration of precursor (5, 7, 10 mg). Results: [$^{18}F$] FHBG was purified by HPLC system and collected at around 10-12 min. Synthesis using Explora-RN module showed a $32.0{\pm}1.2%$ yield of radiochemical (decay corrected), the purity was greater than 98%. And the entire synthesis time was less than 48 min. Temperature of the highest synthesis yield was $130^{\circ}C$, reaction time was 5 minutes and concentration of precursor was 10 mg (recommended volume in manual) (n=36). In contrast to radiochemical yield of precursor 10 mg ($32{\pm}1.2%$), yield of 5 and 7 mg precursor was unstable. Conclusion: Automation of [$^{18}F$] FHBG synthesis at Explora-RN module has been completed. In addition, we were able to obtain optimized reaction time, temperature and concentration of precursor. Therefore this study would be provided more rapid synthesis and higher radiochemical yield.

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Development of Loop-mediated Isothermal Amplification (LAMP) for Detection of Vibrio alginolyticus (Loop-mediated isothermal amplification (LAMP)법을 이용한 Vibrio alginolyticus의 신속 진단법 개발)

  • Hong, Seung-hyun;Heo, Moon-Soo
    • Journal of Life Science
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    • v.25 no.8
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    • pp.903-909
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    • 2015
  • Loop-mediated isothermal amplification (LAMP), a PCR-based diagnostic method, is based on autocycling strand displacement DNA synthesis in the presence of exonuclease-negative Bst DNA polymerase under isothermal conditions. With the help of four specific primers that recognize six different sequences of a target DNA, LAMP has high specificity in pathogenic identification in a short time. Hence, in the present study, LAMP is used as a diagnostic tool in the identification of the most dreadful aquatic pathogenic species, Vibrio alginolyticus, and to develop species-specific LAMP primers and optimization of LAMP reaction conditions such as annealing temperature, elongation time, and other PCR chemical concentrations, including MgSO4, dNTPs, Betaine, and Bst polymerase. The optimized LAMP primers were also checked for specificity with other Vibrio species, which showed that the designed primers were very specific to V. alginolyticus After the first introduction of a species name like this one, the first part (“Vibrio” in this case) should be abbreviated to only the first letter.only. These are usually the most harmful pathogens of the Vibrio species that appear in shrimp and crabs. The results also revealed that the LAMP assay could be 10-fold more sensitive than conventional PCR in detecting V. alginolyticus. This could be the first report on using a rapid and highly sensitive technique, the LAMP assay, in the effective diagnosis of the pathogenic bacteria V. alginolyticus, which could help in the early detection of diseases, particularly in aquaculture.

Optimization of Production Conditions of Biosurfactant from Bacillus sp. and its Purification (Bacillus sp.에 의한 생물계면활성제의 생산 및 그의 성질)

  • Kim, Jin-Sook;Song, Hee-Sang;Chung, Nam-Hyun;Bang, Won-Gi
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.109-114
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    • 2005
  • A bacterium capable of emulsifying hydrocarbon, n-hexadecane, and decreasing surface tension of the culture media using oil collapsing method was isolated. The bacterium was partially identified as Bacillus sp. and named BJS-51. n-Hexadecane was the most effective carbon source for production of biosurfactant. Surface tension was decreased from 76 dyne/cm to 31 dyne/cm and CMD (critical micelle dilution) had the highest value of 5.7 at 3% n-hexadecane. Ammonium phosphate was the most effective nitrogen source, when C/N ratio was 60, surface tension and CMD were 29 dyne/cm and 9.2, respectively. Optimum pH and temperature were 7.2 and $30^{\circ}C$, respectively. Produced biosurfactant was extracted and purified using organic solvent extraction method and preparative HPLC systems. After analysis by various color reaction, this biosurfactant was identified as lipopolysaccharide. Surface tension and CMC (critical micelle concentration) of purified biosurfactant were 27 dyne/cm and 0.08 g/l, repectively. CMD was 9.2, so the yield of biosurfactant was about 0.74 g/l at the optimal conditions. The biosurfactant was very stable at wide range of $pH\;2{\sim}12$ with surface tension $29{\sim}31\;dyne/cm$ and showed $29{\sim}30\;dyne/cm$ of surface tension after heat treatment at $100^{\circ}C$ for 60 min.

Optimization of Synthesis Condition for Nanoscale Zero Valent Iron Immobilization on Granular Activated Carbon (영가철이 고정된 입상활성탄 제조를 위한 최적 합성조건 도출)

  • Hwang, Yuhoon;Mines, Paul D.;Lee, Wontae;Andersen, Henrik R.
    • Journal of Korean Society of Environmental Engineers
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    • v.38 no.9
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    • pp.521-527
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    • 2016
  • Nanoscale zero valent iron (nZVI) has been intensively studied for the treatment of a plethora of pollutants through reductive reaction, however, the nano size should be of concern when nZVI is considered for water treatment, due to difficulties in recovery. The loss of nZVI causes not only economical loss, but also potential risk to human health and environment. Thus, the immobilization onto coarse or structured support is essential. In this study, two representative processes for nZVI immobilization on granular activated carbon (GAC) were evaluated, and optimized conditions for synthesizing Fe/GAC composite were suggested. Both total iron content and $Fe_0$ content can be significantly affected by preparation processes, therefore, it was important to avoid oxidation during preparation to achieve higher reduction capacity. Synthesis conditions such as reduction time and existence of intermediate drying step were investigated to improve $Fe_0$ content of Fe/GAC composites. The optimal condition was two hours of $NaBH_4$ reduction without intermediate drying process. The prepared Fe/GAC composite showed synergistic effect of the adsorption capability of the GAC and the degradation capability of the nZVI, which make this composite a very effective material for environmental remediation.

Effect of $CH_4$ addition to the $H_2O$ plasma excited by VHF ICP for production of $H_2$ (고주파유도결합에 의해 여기된 물플라즈마로부터 고효율 수소생산을 위한 메탄가스 첨가효과)

  • Kim, Dae-Woon;Choo, Won-Il;Jang, Soo-Ouk;Jung, Yong-Ho;Lee, Bong-Ju;Kim, Young-Ho;Lee, Seung-Heun;Kwon, Sung-Ku
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.442-442
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    • 2008
  • Hydrogen was produced by water plasma excited in very high frequency inductively coupled tube reactor. Mass spectrometry was used to monitor gas phase species at various process conditions. Water dissociation rate depend on the process parameters such as ICP power, flow-rate and pressure. Water dissociation percent in ICP reactor decrease with increase of chamber pressure and $H_2O$ flow rate, while increase with increase of ICP power. In our experimental range, maximum water dissociation rate was 65.5% at the process conditions of 265 mTorr, 68 sccm, and 400 Watt. The effect of $CH_4$ addition to a water plasma on the hydrogen production has been studied in a VHF ICP reactor. With the addition of $CH_4$ gas, $H_2$ production increases to 12% until the $CH_4$ flow rate increases up to 15 sccm. But, with the flow rate of $CH_4$ more than 20 sccm, chamber wall was deposited with carbon film because of deficiency of oxygen in gas phase, hydrogen production rate decreased. The main roles of $CH_4$ gas are to reacts with O forming CO, CHO and $CO_2$ and releasing additional $H_2$ and furthermore to prevent reverse reaction for forming $H_2O$ from $H_2$ and $O_2$. But, $CH_4$ addition has negative effects such as cost increase and $CO_x$ emission, therefore process optimization is required.

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A optimization study on the preparation and coating conditions on honeycomb type of Pd/TiO2 catalysts to secure hydrogen utilization process safety (수소 활용공정 안전성 확보를 위한 Pd/TiO2 수소 상온산화 촉매의 제조 및 허니컴 구조의 코팅 조건 최적화 연구)

  • Jang, Young hee;Lee, Sang Moon;Kim, Sung Su
    • Journal of the Korea Organic Resources Recycling Association
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    • v.29 no.4
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    • pp.47-54
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    • 2021
  • In this study, the performance of a honeycomb-type hydrogen oxidation catalyst to remove hydrogen in a hydrogen economy society to secure leaking hydrogen. The Pd/TiO2 catalyst was prepared based on a liquid phase reduction method that is not exposed to a heat source, and it was showed through H2-chemisorption analysis that it existed as very small active particles of 2~4 nm. In addition, it was found that the metal dispersion decreased and the active particle size increased as the reduction reaction temperature increased. It was meant that the active metal particle size and the hydrogen oxidation performance were in a proportional correlation, so that it was consistent with the hydrogen oxidation performance reduction result. The prepared catalyst was coated on a support in the form of a honeycomb so that it could be applied to the hydrogen industrial process. When 20 wt% or more of the AS-40 binder was coated, oxidation performance of 90% or more was observed under low-concentration hydrogen conditions. It was showed through SEM analysis that long-term catalytic activity can be expected by enhancing the adhesion strength of the catalyst and preventing catalyst desorption. It is a basic research that can secure safety in a hydrogen society such as gasification, organic resource, and it can be utilized as a system that can respond to unexpected safety accidents in the future.

Exploration of alternative cell wall lysing enzymes and optimization of conditions for Ganoderma lucidum protoplast isolation (영지 원형질체 분리를 위한 대체 세포벽 분해 효소 탐색과 최적 조건 설정)

  • Minseek Kim;Min Ji Oh;Ji-Hoon Im;Eun-Ji Lee;Youn-Lee Oh
    • Journal of Mushroom
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    • v.22 no.3
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    • pp.87-91
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    • 2024
  • Recently, active research in Korea and worldwide has begun to focus on gene function and cultivar development using gene editing tools. This research, in addition to studies on edible mushroom, aims to enhance the physical and biochemical characteristics of mushrooms for applications in materials and substance production. For these studies, efficient isolation of protoplasts from the target mushroom is critical. However, several commercial cell wall-lysing enzyme cocktails, including Novozyme234, Glucanex, and Lysing enzymes, have recently been discontinued. In this study, we aimed to identify alternative enzyme systems to replace the discontinued cell wall-lysing enzymes for stable isolation of protoplasts from Ganoderma lucidum. To select an optimal osmotic buffer, enzyme function in 0.6 and 1.2 M Sorbitol, Sucrose, Mannitol, and KCl was assessed. The effect of reaction time was also evaluated. Protoplast isolation efficiency of each alternative enzyme was tested using lysing enzymes from Trichoderma harzianum, Chimax-N, and Yatalase, either individually or in combination. This matrix of studies identified enzymes and optimal conditions that could replace the discontinued lysing enzymes.

A study on the Degradation and By-products Formation of NDMA by the Photolysis with UV: Setup of Reaction Models and Assessment of Decomposition Characteristics by the Statistical Design of Experiment (DOE) based on the Box-Behnken Technique (UV 공정을 이용한 N-Nitrosodimethylamine (NDMA) 광분해 및 부산물 생성에 관한 연구: 박스-벤켄법 실험계획법을 이용한 통계학적 분해특성평가 및 반응모델 수립)

  • Chang, Soon-Woong;Lee, Si-Jin;Cho, Il-Hyoung
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.1
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    • pp.33-46
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    • 2010
  • We investigated and estimated at the characteristics of decomposition and by-products of N-Nitrosodimethylamine (NDMA) using a design of experiment (DOE) based on the Box-Behken design in an UV process, and also the main factors (variables) with UV intensity($X_2$) (range: $1.5{\sim}4.5\;mW/cm^2$), NDMA concentration ($X_2$) (range: 100~300 uM) and pH ($X_2$) (rang: 3~9) which consisted of 3 levels in each factor and 4 responses ($Y_1$ (% of NDMA removal), $Y_2$ (dimethylamine (DMA) reformation (uM)), $Y_3$ (dimethylformamide (DMF) reformation (uM), $Y_4$ ($NO_2$-N reformation (uM)) were set up to estimate the prediction model and the optimization conditions. The results of prediction model and optimization point using the canonical analysis in order to obtain the optimal operation conditions were $Y_1$ [% of NDMA removal] = $117+21X_1-0.3X_2-17.2X_3+{2.43X_1}^2+{0.001X_2}^2+{3.2X_3}^2-0.08X_1X_2-1.6X_1X_3-0.05X_2X_3$ ($R^2$= 96%, Adjusted $R^2$ = 88%) and 99.3% ($X_1:\;4.5\;mW/cm^2$, $X_2:\;190\;uM$, $X_3:\;3.2$), $Y_2$ [DMA conc] = $-101+18.5X_1+0.4X_2+21X_3-{3.3X_1}^2-{0.01X_2}^2-{1.5X_3}^2-0.01X_1X_2+0.07X_1X_3-0.01X_2X_3$ ($R^2$= 99.4%, 수정 $R^2$ = 95.7%) and 35.2 uM ($X_1$: 3 $mW/cm^2$, $X_2$: 220 uM, $X_3$: 6.3), $Y_3$ [DMF conc] = $-6.2+0.2X_1+0.02X_2+2X_3-0.26X_1^2-0.01X_2^2-0.2X_3^2-0.004X_1X_2+0.1X_1X_3-0.02X_2X_3$ ($R^2$= 98%, Adjusted $R^2$ = 94.4%) and 3.7 uM ($X_1:\;4.5\;$mW/cm^2$, $X_2:\;290\;uM$, $X_3:\;6.2$) and $Y_4$ [$NO_2$-N conc] = $-25+12.2X_1+0.15X_2+7.8X_3+{1.1X_1}^2+{0.001X_2}^2-{0.34X_3}^2+0.01X_1X_2+0.08X_1X_3-3.4X_2X_3$ ($R^2$= 98.5%, Adjusted $R^2$ = 95.7%) and 74.5 uM ($X_1:\;4.5\;mW/cm^2$, $X_2:\;220\;uM$, $X_3:\;3.1$). This study has demonstrated that the response surface methodology and the Box-Behnken statistical experiment design can provide statistically reliable results for decomposition and by-products of NDMA by the UV photolysis and also for determination of optimum conditions. Predictions obtained from the response functions were in good agreement with the experimental results indicating the reliability of the methodology used.

Optimization of Cellulase Production from Paenibacillus jamilae BRC 15-1 (Paenibacillus jamilae BRC15-1의 Cellulase 생산 최적화)

  • Cha, Young-Lok;Yoon, Young-Mi;Yoon, Ha-Yan;Kim, Jung Kon;Yang, Ji-Young;Na, Han-Beur;Ahn, Jong-Woong;Moon, Youn-Ho;Choi, In-Hu;Yu, Gyeong-Dan;Lee, Ji-Eun;An, Gi Hong;Lee, Kyeong-Bo
    • KSBB Journal
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    • v.30 no.6
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    • pp.283-290
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    • 2015
  • In this study was selected the cellulolytic microorganism and investigated optimum condition of cellulase production for the cellulosic bioethanol production. A bacterial strain Paenibacillus jamilae BRC15-1, was isolated from soil of domestic reclaimed land. For optimizing cellulase production from the selected strain, various culture parameters were investigated such as culture medium, pH (pH 4~10), temperature ($25{\sim}50^{\circ}C$) and culture time (2~72 h). As a result, P. jamilae BRC15-1 efficiently produced cellulase from cellulosic biomass under following conditions: 24 h of culture time (pH 7, $40^{\circ}C$) in manufactured media of CMC (carboxymethyl cellulose) with peptone. Optimum saccharifying condition of crude enzyme produced from P. jamilae BRC15-1 was identified on pH 6 and $40^{\circ}C$ of reaction temperature, respectively. This crude enzyme from P. jamilae BRC15-1 was used for saccharification of pretreated sweet sorghum (Sorghum bicolor var. dulciusculum Ohwi) bagasse under the optimal condition. Finally, pretreated sweet sorghum bagasse including 0.1 g of glucan was saccharified by crude enzyme of P. jamilae BRC15-1 into 2.75 mg glucose, 0.79 mg xylose and 1.12 mg arabinose.

The development and validation of a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) procedure for the determination of fluoroquinolones residues in chicken muscle using modified QuEChERS (quick, easy, cheap, effective, rugged and safe) method

  • Park, Sunjin;Kim, Hyobi;Choi, Byungkook;Hong, Chung-Oui;Lee, Seon-Young;Jeon, Inhae;Lee, Su-Young;Kwak, Pilhee;Park, Sung-Won;Kim, Yong-Sang;Lee, Kwang-jick
    • Korean Journal of Veterinary Service
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    • v.42 no.4
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    • pp.289-296
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    • 2019
  • A novel rapid procedure with liquid chromatography tandem mass spectrometry (LC-MS/MS) detection has been developed by changing various conditions including sample preparation such as QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) methodology. This work has been involved the optimization and validation of detection method for fluoroquinolones which are widespread used in livestock especially in the chicken. Five grams of homogenized chicken muscle were extracted with QuEChERS EN and acetonitrile containing 5% formic acid and cleaned with anhydrous magnesium sulfate and C18 sorbent. The separation was performed on Acquity UPLC HSS T3 (2.1 mm×100 mm, 1.8 ㎛) column. The mobile phase A and B were composed of water containing 0.1% formic acid and acetonitrile containing 0.1% formic acid, respectively. Flow rate was 0.25 mL/min and column temperate was 40℃. LC-MS/MS with multiple reaction monitoring has been optimized for ten fluoroquinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, marbofloxacin, norfloxacin, ofloxacin, orbifloxacin, pefloxacin and sarafloxacin). The method developed in this study has been presented good linearity with correlation coefficient (R2) of 0.9971~0.9998. LOD and LOQ values ranged from 0.09 to 0.76 ppb and from 0.26 to 2.29 ppb, respectively. The average recoveries were from 77.46 to 111.83% at spiked levels of 10.0 and 20.0 ㎍/kg. Relative standard deviation (%) ranged 1.28~11.90% on intra-day and 3.10~8.38 % on inter-day, respectively. This analysis method was applicable to the livestock residue laboratories and was expected to be satisfactory for the residue surveillance system.