• Title/Summary/Keyword: Complex-mode reaction

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Determination of finasteride in human serum by LC-MS/MS (LC-MS/MS를 이용한 혈청 중 finasteride 분석)

  • Nam, Hye-Seon;Nam, Kyong-Hee;Jung, Su-Hee;Lee, Jang-Woo;Kang, Jin-Yeong;Hong, Soon-Keun;Kim, Tae-Sung;Kang, Tae-Seok;Yoon, Hae-Jung;Lee, Kwang-Ho;Rhee, Gyu-Seek
    • Analytical Science and Technology
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    • v.24 no.5
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    • pp.345-351
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    • 2011
  • A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) method was developed and validated for the determination of finasteride in human serum. Beclomethasone was used as internal standard (IS) and liquid-liquid extraction (LLE) using methyl tert-butyl ether (MTBE) was carried out to isolate analyte. The mass transitions monitored in multiple reaction monitoring (MRM) in positive ion mode were m/z 373.2${\rightarrow}$305.2 for finasteride and m/z 409.3${\rightarrow}$391.2 for IS. Retention times of finasteride and IS were 5.81 and 5.46 min, respectively. The limit of quantitation (LOQ) was 0.1 ng/mL and the calibration curve showed good linearity in the range of 0.1~20.0 ng/mL ($R^2$=0.9997). The intra-day assay precision and accuracy were in the range 6.3~10.6% and 97.3~103.6%, respectively, and the inter-day assay precision and accuracy were in the range 0.8~5.2% and 99.8~102.5%, respectively. The sample extract recovery of the method was 80~83%.

Computational Study on Unsteady Mechanism of Spinning Detonations

  • Matsuo, Akiko;Sugiyama, Yuta
    • 한국전산유체공학회:학술대회논문집
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    • 2008.03a
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    • pp.367-373
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    • 2008
  • Spinning detonations propagating in a circular tube were numerically investigated with a one-step irreversible reaction model governed by Arrhenius kinetics. Activation energy is used as parameter as 10, 20, 27 and 35, and the specific heat ratio and the heat release are fixed as 1.2 and 50. The time evolution of the simulation results was utilized to reveal the propagation mechanism of single-headed spinning detonation. The track angle of soot record on the tube wall was numerically reproduced with various levels of activation energy, and the simulated unique angle was the same as that of the previous reports. The maximum pressure histories of the shock front on the tube wall showed stable pitch at Ea=10, periodical unstable pitch at Ea=20 and 27 and unstable pitch consisting of stable, periodical unstable and weak modes at Ea=35, respectively. In the weak mode, there is no Mach leg on the shock front, where the pressure level is much lower than the other modes. The shock front shapes and the pressure profiles on the tube wall clarified the mechanisms of these stable and unstable modes. In the stable pitch at Ea=10, the maximum pressure history on the tube wall remained nearly constant, and the steady single Mach leg on the shock front rotated at a constant speed. The high and low frequency pressure oscillations appeared in the periodical unstable pitch at Ea=20 and 27 of the maximum pressure history. The high frequency was one cycle of a self-induced oscillation by generation and decay in complex Mach interaction due to the variation in intensity of the transverse wave behind the shock front. Eventually, sequential high frequency oscillations formed the low frequency behavior because the frequency behavior was not always the same for each cycle. In unstable pitch at Ea=35, there are stable, periodical unstable and weak modes in one cycle of the low frequency oscillation in the maximum pressure history, and the pressure amplitude of low frequency was much larger than the others. The pressure peak appeared after weak mode, and the stable, periodical unstable and weak modes were sequentially observed with pressure decay. A series of simulations of spinning detonations clarified that the unsteady mechanism behind the shock front depending on the activation energy.

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Computational Study on Unsteady Mechanism of Spinning Detonations

  • Matsuo, Akiko;Sugiyama, Yuta
    • 한국전산유체공학회:학술대회논문집
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    • 2008.10a
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    • pp.367-373
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    • 2008
  • Spinning detonations propagating in a circular tube were numerically investigated with a one-step irreversible reaction model governed by Arrhenius kinetics. Activation energy is used as parameter as 10, 20, 27 and 35, and the specific heat ratio and the heat release are fixed as 1.2 and 50. The time evolution of the simulation results was utilized to reveal the propagation mechanism of single-headed spinning detonation. The track angle of soot record on the tube wall was numerically reproduced with various levels of activation energy, and the simulated unique angle was the same as that of the previous reports. The maximum pressure histories of the shock front on the tube wall showed stable pitch at Ea=10, periodical unstable pitch at Ea=20 and 27 and unstable pitch consisting of stable, periodical unstable and weak modes at Ea=35, respectively. In the weak mode, there is no Mach leg on the shock front, where the pressure level is much lower than the other modes. The shock front shapes and the pressure profiles on the tube wall clarified the mechanisms of these stable and unstable modes. In the stable pitch at Ea=10, the maximum pressure history on the tube wall remained nearly constant, and the steady single Mach leg on the shock front rotated at a constant speed. The high and low frequency pressure oscillations appeared in the periodical unstable pitch at Ea=20 and 27 of the maximum pressure history. The high frequency was one cycle of a self-induced oscillation by generation and decay in complex Mach interaction due to the variation in intensity of the transverse wave behind the shock front. Eventually, sequential high frequency oscillations formed the low frequency behavior because the frequency behavior was not always the same for each cycle. In unstable pitch at Ea=35, there are stable, periodical unstable and weak modes in one cycle of the low frequency oscillation in the maximum pressure history, and the pressure amplitude of low frequency was much larger than the others. The pressure peak appeared after weak mode, and the stable, periodical unstable and weak modes were sequentially observed with pressure decay. A series of simulations of spinning detonations clarified that the unsteady mechanism behind the shock front depending on the activation energy.

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Mode of Action on EcoRI Restriction Endonuclease: EcoRI and EcoRI Variant N199H have Active Monomeric Forms

  • Kim, Jae-Jong;Koh, Suk-Hoon;Kim, Joong-Su;Lee, Dae-Sil
    • BMB Reports
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    • v.31 no.2
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    • pp.149-155
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    • 1998
  • The N199H variant of the EcoRI endonuclease has about twice the catalytic activity of the wild-type. A comparison of their biochemical characteristics, using synthetic oligonucleotides 5'-dAAAACTTAAGAAAAAAAAAAA-3' (KA) and 5'-dTTTTTGAATTCTTTTTTTTTT-3' (KT), helps to define the cleavage reaction pathway of these enzymes. Both EcoRI and EcoRI variant N199H were found to cleave singlestranded KA or KT about three times faster than the double-stranded forms, although the KT oligonucleotide was more susceptible. Using the ssDNA substrate in kinetic analyses, lower $K_m$ values were obtained for the N199H variant than for the wild-type at low (50 mM), as well as high (200 mM), sodium chloride concentrations. This difference between the endonucleases is attributed to a grealter accessibility for tbe substrate by the variant, and also a higher affinity for the DNA backbone. It also appears that the relative activities of the two enzymes, particularly at high ionic strength, are proportional to their populations in the monomeric enzyme form. That is, according to gel filtration data, half of the N199H molecules exist as monomers in 200 mM NaCl, whereas those of the wild-type are mainly dimeric. Consequently, the Asp199 residue of the EcoRI endonuclease may be implicated in the protein-protein interaction leading to dimerization, as well as in coupling to DNA substrates. In summary, it is proposed that active monomeric endonuclease molecules, derived from the dimeric enzyme, recognize and form a complex with a single stranded form of the DNA substrate, which then undergoes nucleophilic substitution and cleavage.

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Historical Consideration of Lignin Models for Native Lignin Structure (리그닌 화학구조 모델의 역사적 고찰)

  • 황병호
    • Journal of Korea Foresty Energy
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    • v.23 no.1
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    • pp.45-68
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    • 2004
  • The word of lignin is derived from the Latin word 'ligum' meaning wood. Lignin is complex polymer consisting of coniferyl alcohol, sinapyl alcohol and p-coumaryl alcohol unit and has an amorphous, three dimensional network structure which is hard to be hydrolyzed by acid. Lignin is found in the cell wall of plants lignified. The mode of polymerization of these alcohols in the cell wall lead to a heterogeneous branched and cross-linked polymer in which phenyl propane units are linked by carbon-carbon and carbon-oxygen bonds. This polymerization of precursors, p-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol to lignin is formed by enzymic dehydrolyzation. The reaction is initiated by an electron transfer which results in the formation of resonance-stabilized phenoxy radical. The combination of these radicals produces a variety of dimers, trimers and oligomers and so on. Lignin research has been divided into basic and practical application field. The basic studies contains biosynthesis, chemical structure, distribution in the cell wall and reactivity by reductants, oxidants and organic solvents. The application research will be approached the reaction of lignin in various pulp making involving pulp bleaching and its effect on pulp qualities. Lignin also will be studied for the production of fine chemicals, polymer products and the conservation into an energy source like petroleum oil because the amount of lignin produced in pulp making process is more than 51,000,000 tons per year in the world. Both basic and application research must lay emphasis on the development for the utilization of lignin and the pulping process. But these researches can not be completed without understanding lignin structure containing functional groups. Therefore, this paper was focused on the review of lignin formulation which has been studied since 1948 in chronological order. This review was based on monomers, dimers, trimers and tetramers of phenyl propane unit structures which were isolated and identified by different methods from various wood.ious wood.

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Crystal Structure of (S)-3-Hydroxybutyryl-CoA Dehydrogenase from Clostridium butyricum and Its Mutations that Enhance Reaction Kinetics

  • Kim, Eun-Jung;Kim, Jieun;Ahn, Jae-Woo;Kim, Yeo-Jin;Chang, Jeong Ho;Kim, Kyung-Jin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1636-1643
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    • 2014
  • 3-Hydroxybutyryl-CoA dehydrogenase is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA, in which acetoacetyl-CoA is reduced to 3-hydroxybutyryl-CoA. To understand the molecular mechanisms of n-butanol biosynthesis, we determined the crystal structure of 3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD). The monomer structure of CbHBD exhibits a two-domain topology, with N- and C-terminal domains, and the dimerization of the enzyme was mostly constituted at the C-terminal domain. The mode of cofactor binding to CbHBD was elucidated by determining the crystal structure of the enzyme in complex with $NAD^+$. We also determined the enzyme's structure in complex with its acetoacetyl-CoA substrate, revealing that the adenosine diphosphate moiety was not highly stabilized compared with the remainder of the acetoacetyl-CoA molecule. Using this structural information, we performed a series of site-directed mutagenesis experiments on the enzyme, such as changing residues located near the substrate-binding site, and finally developed a highly efficient CbHBD K50A/K54A/L232Y triple mutant enzyme that exhibited approximately 5-fold higher enzyme activity than did the wild type. The increased enzyme activity of the mutant was confirmed by enzyme kinetic measurements. The highly efficient mutant enzyme should be useful for increasing the production rate of n-butanol.

Development of an 1-Dimensional Dynamic Numerical Model for BTX Removal Process Analysis by Gaseous-Biofilm Filtration (기체상-생물막 여과 공법의 BTX 제거 공정 해석을 위한 1차원 동적 수치모델 개발)

  • Kim, Yeong-Kwan;Choi, Sung-Chan;Kim, Seog-Ku;Lee, Yong-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.37 no.12
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    • pp.689-695
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    • 2015
  • A biofilm filtration for the removal of gaseous pollutants has been recognized as a process with a complex interaction between the gas flow characteristics and the process operating variables. This study aims to develop an one dimensional dynamic numerical model which can be utilized as a tool for the analysis of biofilm filtration process operated in plug flow mode. Since, in a plug flow system, minor environmental changes in a gaseous unit process cause a drastic change in reaction and the interaction between the pollutants is an influencing factor, plug flow system was generalized in developing the model. For facilitation of the model development, dispersion was simplified based on the principles of material balance. Several reactions such as competition, escalation, and control between the pollutants were included in the model. The applicability of the developed model was evaluated by taking the calibration and verification steps on the experimental data performed for the removal of BTX at both low and high flow concentration. The model demonstrated a correlation coefficient ($R^2$) greater than 0.79 under all the experimental conditions except for the case of toluene at high flow condition, which suggested that this model could be used for the generalized gaseous biofilm plug flow filtration system. In addition, this model could be a useful tool in analyzing the design parameters and evaluating process efficiency of the experiments with substantial amount of complexity and diversity.