• Title/Summary/Keyword: Isotope analysis

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A Review on Metabolic Pathway Analysis with Emphasis on Isotope Labeling Approach

  • Azuyuki, Shimizu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.237-251
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    • 2002
  • The recent progress on metabolic systems engineering was reviewed based on our recent research results in terms of (1) metabolic signal flow diagram approach, (2) metabolic flux analysis (MFA) in particular with intracellular isotopomer distribution using NMR and/or GC-MS, (3) synthesis and optimization of metabolic flux distribution (MFD), (4) modification of MFD by gene manipulation and by controlling culture environment, (5) metabolic control analysis (MCA), (6) design of metabolic regulation structure, and (7) identification of unknown pathways with isotope tracing by NMR. The main characteristics of metabolic engineering is to treat metabolism as a network or entirety instead of individual reactions. The applications were made for poly-3-hydroxybutyrate (PHB) production using Ralstonia eutropha and recombinant Escherichia coli, lactate production by recombinant Saccharomyces cerevisiae, pyruvate production by vitamin auxotrophic yeast Toluropsis glabrata, lysine production using Corynebacterium glutamicum, and energetic analysis of photosynthesic microorganisms such as Cyanobateria. The characteristics of each approach were reviewed with their applications. The approach based on isotope labeling experiments gives reliable and quantitative results for metabolic flux analysis. It should be recognized that the next stage should be toward the investigation of metabolic flux analysis with gene and protein expressions to uncover the metabolic regulation in relation to genetic modification and/ or the change in the culture condition.

Analysis and Consideration of the Establishment of a Multiplexed Channel for Domestic RI Waste Nuclide Analysis (국내 방사성동위원소(RI) 폐기물 핵종분석 다중화채널 구축 성과 분석 및 고찰)

  • Han, Sang-Jun;Lee, Hong-Yeon;Kim, Bo-Gil;An, Eun-Mi
    • Journal of radiological science and technology
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    • v.44 no.4
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    • pp.351-358
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    • 2021
  • This research project is a program promoted to seek diversification of domestic radioactive waste analysis institutions, and seeks public development, win-win cooperation, and cooperation between the entrusted institution and the entrusted institution. Accordingly, the entrusted institution established a standard analysis procedure for establishing a quality control system for radioactivity analysis, establishing a radiation control zone, obtaining KOLAS accreditation, and performing proficiency tests, which are the performance ranges requested by the entrusted institution, and intersecting the radioactive isotope waste generated at the actual site. Verification was performed to confirm the analysis quality. In addition, facilities and equipment for radioactivity analysis were supplemented and expanded, and the basic technology foundation and technical skills were secured through securing professional technicians and education/training. It is judged that the entrusted institution will contribute to securing radiation safety through the smooth execution of treatment, disposal, and transportation through value creation and analysis of radioactive waste generated by radioactive isotope-using institutions (research institutes, hospitals, industries, etc.) by succeeding in this research project do.

First GC-IRMS in Korea and Its Application Fields (국내 최초로 도입된 GC-IRMS와 응용분야 소개)

  • Shin, Woo-Jin;Lee, Kwang-Sik;Ko, Kyung-Seok
    • Economic and Environmental Geology
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    • v.40 no.5
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    • pp.699-703
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    • 2007
  • Compound-specific isotope analysis (CSIA) by isotope ratio mass spectrometer (IRMS) interfaced with gas chromatography (GC) is a state of the art analytical technique for stable isotopes in earth sciences, environmental sciences and forensics. Since early 1990s, GC-IRMS has been widely used to investigate the authenticity of food in forensic science and to trace the sources of organic contaminants in environmental science. In Korea, a GC-IRMS was firstly installed at the Korea Basic Science Institute (KBSI) in early 2005. In this study, we introduce the GC-IRMS of the KBSI shortly to stimulate various isotope-related researches of Korea, and report preliminary CSIA results for BTEX of different manufacturers.

Introduction to a New Sample Preparation Apparatus (H/Device) for Measurement of Hydrogen Isotope Composition of Natural Water (신(新) H/Device를 이용한 자연수의 수소동위원소비 측정)

  • Park, Seong-Sook;Yun, Seong-Taek;So, Chil-Sup
    • Economic and Environmental Geology
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    • v.31 no.3
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    • pp.265-271
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    • 1998
  • In the hydrologic and hydrochemical studies of natural waters, oxygen and hydrogen isotope compositions of waters are very important to elucidate the origin and circulation pattern of water in the hydrologic system. The hydrogen isotope analysis of waters usually has been undertaken through the reduction of water to form hydrogen gas using pure metals (in general, zinc and uranium). In 1996, a new apparatus (H/Device) was developed to prepare the water samples (by the reduction with Cr metal) without some intrinsic problems that may yield incorrect and/or inaccurate data, and was installed at 1997 in the Center for Mineral Resources Research (CMR) in Korea University. However, the optimistic conditions of preparation and analysis of samples has not been established. In this paper, we introduce the efficiency of H/Device to obtain accurate hydrogen isotope values of water, and discuss both the optimum conditions including the effective reduction time and the probable mixing (memory) effect between successive samples. We obtained large amounts of a laboratory working standard (KUW; Korea University Water) with the average ${\delta}D_{SMOW}$ value of $-42.1{\pm}1.0$$(1{\sigma})$.

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Relative Quantification of Glycans by Metabolic Isotope Labeling with Isotope Glucose in Aspergillus niger

  • Choi, Soo-Hyun;Cho, Ye-Eun;Kim, Do-Hyun;Kim, Jin-il;Yun, Jihee;Jo, Jae-Yoon;Lim, Jae-Min
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.139-145
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    • 2022
  • Protein glycosylation is a common post-translational modification by non-template-based biosynthesis. In fungal biotechnology, which has great applications in pharmaceuticals and industries, the importance of research on fungal glycoproteins and glycans is accelerating. In particular, the importance of quantitative analysis of fungal glycans is emerging in research on the production of filamentous fungal proteins by genetic modification. Reliable mass spectrometry-based techniques for quantitative glycomics have evolved into chemical, enzymatic, and metabolic stable isotope labeling methods. In this study, we intend to expand quantitative glycomics by metabolic isotope labeling of glycans in Aspergillus niger, a filamentous fungus model, by the MILPIG method. We demonstrate that incubation of filamentous fungi in a culture medium with carbon-13 labeled glucose (1-13C1) efficiently incorporates carbon-13 into N-linked glycans. In addition, for quantitative validation of this method, light and heavy glycans are mixed 1:1 to show the performance of quantitative analysis of various N-linked glycans simultaneously. We have successfully quantified fungal glycans by MILPIG and expect it to be widely applicable to glycan expression levels under various biological conditions in fungi.

Ginseng authenticity testing by measuring carbon, nitrogen, and sulfur stable isotope compositions that differ based on cultivation land and organic fertilizer type

  • Chung, Ill-Min;Lee, Taek-Jun;Oh, Yong-Taek;Ghimire, Bimal Kumar;Jang, In-Bae;Kim, Seung-Hyun
    • Journal of Ginseng Research
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    • v.41 no.2
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    • pp.195-200
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    • 2017
  • Background: The natural ratios of carbon (C), nitrogen (N), and sulfur (S) stable isotopes can be varied in some specific living organisms owing to various isotopic fractionation processes in nature. Therefore, the analysis of C, N, and S stable isotope ratios in ginseng can provide a feasible method for determining ginseng authenticity depending on the cultivation land and type of fertilizer. Methods: C, N, and S stable isotope composition in 6-yr-old ginseng roots (Jagyeongjong variety) was measured by isotope ratio mass spectrometry. Results: The type of cultivation land and organic fertilizers affected the C, N, and S stable isotope ratio in ginseng (p < 0.05). The ${\delta}^{15}N_{AIR}$ and ${\delta}^{34}S_{VCDT}$ values in ginseng roots more significantly discriminated the cultivation land and type of organic fertilizers in ginseng cultivation than the ${\delta}^{13}C_{VPDB}$ value. The combination of ${\delta}^{13}C_{VPDB}$, ${\delta}^{15}N_{AIR}$, or ${\delta}^{34}S_{VCDT}$ in ginseng, except the combination ${\delta}^{13}C_{VPDB}-^{34}S_{VCDT}$, showed a better discrimination depending on soil type or fertilizer type. Conclusion: This case study provides preliminary results about the variation of C, N, and S isotope composition in ginseng according to the cultivation soil type and organic fertilizer type. Hence, our findings are potentially applicable to evaluate ginseng authenticity depending on cultivation conditions.

INTRINSIC NMR ISOTOPE SHIFTS OF CYCLOOCTANONE AT LOW TEMPERATURE (저온에서의 싸이클로옥타논에 대한 고유동위원소 효과)

  • Jung, Miewon
    • Analytical Science and Technology
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    • v.7 no.2
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    • pp.213-224
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    • 1994
  • Several isotopomers of cyclooctanone were prepared by selective deuterium substitution. Intrinsic isotope effects on $^{13}C$ NMR chemical shifts of these isotopomers were investigated systematically at low temperature. These istope effects were discussed in relation to the preferred boat-chair conformation of cyclooctanone. Deuterium isotope effects on NMR chemical shifts have been known for a long time. Especially in a conformationally mobile molecule, isotope perturbation could affect NMR signals through a combination of isotope effects on equilibria and intrinsic effects. The distinction between intrinsic and nonintrinsic effects is quite difficult at ambient temperature due to involvement of both equilibrium and intrinsic isotope effects. However if equilibria between possible conformers of cyclooctanone are slowed down enough on the NMR time scale by lowering temperature, it should be possible to measure intrinsic isotope shifts from the separated signals at low temperature. $^{13}C$ NMR has been successfully utilized in the study on molecular conformation in solution when one deals with stable conformers or molecules were rapid interconversion occurs at ambient temperature. The study of dynamic processes in general requires analysis of spectra at several temperature. Anet et al. did $^1H$ NMR study of cyclooctanone at low temperature to freeze out a stable conformation, but were not able initially to deduce which conformation was stable because of the complexity of alkyl region in the $^1H$ NMR spectrum. They also reported the $^1H$ and $^{13}C$ NMR spectra of the $C_9-C_{16}$ cycloalkanones with changing temperature from $-80^{\circ}C$ to $-170^{\circ}C$, but they did not report a variable temperature $^{13}C$ NMR study of cyclooctanone. For the analysis of the intrinsic isotope effect with relation to cylooctanone conformation, $^{13}C$ NMR spectra are obtained in the present work at low temperatures (up to $-150^{\circ}C$) in order to find the chemical shifts at the temperature at which the dynamic process can be "frozen-out" on the NMR time scale and cyclooctanone can be observed as a stable conformation. Both the ring inversion and pseudorotational processes must be "frozen-out" in order to see separate resonances for all eight carbons in cyclooctanone. In contrast to $^1H$ spectra, slowing down just the ring inversion process has no apparent effects on the $^{13}C$ spectra because exchange of environments within the pairs of methylene carbons can still occur by the pseudorotational process. Several isotopomers of cyclooctanone were prepared by selective deuterium substitution (fig. 1) : complete deuterium labeling at C-2 and C-8 positions gave cyclooctanone-2, 2, 8, $8-D_4$ : complete labeling at C-2 and C-7 positions afforded the 2, 2, 7, $7-D_4$ isotopomer : di-deuteration at C-3 gave the 3, $3-D_2$ isotopomer : mono-deuteration provided cyclooctanone-2-D, 4-D and 5-D isotopomers : and partial deuteration on the C-2 and C-8 position, with a chiral and difunctional case catalyst, gave the trans-2, $8-D_2$ isotopomer. These isotopomer were investigated systematically in relation with cyclooctanone conformation and intrinsic isotope effects on $^{13}C$ NMR chemical shifts at low temperature. The determination of the intrinsic effects could help in the analysis of the more complex effects at higher temperature. For quantitative analysis of intrinsic isotope effects, the $^{13}C$ NMR spectrum has been obtained for a mixture of the labeled and unlabeled compounds because the signal separations are very small.

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Lead Isotope Analysis of Bronze Artefacts Excavated from Inyongsaji Site (납동위원소비 분석을 활용한 인용사지 출토 청동기의 원료 산지 연구)

  • Lee, Eun Woo;Kim, So Jin;Han, Woo Rim;Han, Min Su;Hwang, Jin Ju
    • Journal of Conservation Science
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    • v.30 no.4
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    • pp.345-351
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    • 2014
  • Chemical and Pb isotope analysis were performed in order to determine the origin of bronze artefacts excavated from the Inyongsaji site in South Korea. The result of ICP-MS shows that they are tin bronzes in which lead was not intentionally added during production. Pb isotope data analyzed by TIMS are plotted in the southern region of Korea and China of the distribution map drawn by Mabuchi(1985). On the other hand, the identical isotope data are plotted in the Taebaek basin and the Olcheon metamorphic belt correspondent to zone2 and zone3 respectively. It is believed that the isotope data on the tin bronzes which have very low lead content can be used to trace the origin of copper ore rather than those of either lead or tin ore. Pb istope analysis allows diverse interpretation as it can be applied to any object containing trace amounts of lead. In addition, accumulation of isotope data as well as further studies will improve reliability of the provenance studies.

Unveiling dietary lifestyle of a Joseon-era mummy by extracting isotopic information from hair and bone (조선시대 회곽묘 출토 미이라의 안정동위원소 분석으로 생애 주기에 따른 식생활 변화 추적)

  • Kang, Da Young;Shin, Ji Young
    • Analytical Science and Technology
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    • v.25 no.5
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    • pp.300-306
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    • 2012
  • This study reports the stable carbon and nitrogen isotope of mummy, the preserved human time capsule from Joseon Dynasty. The stable isotopic composition of biological tissue is expected to be a direct and constant function of diet because the stable isotope ratios do not decay or change in abundance through time. Over the past three decades, stable carbon and nitrogen isotope analysis has been extensively applied for archaeological topics. However, this is the first case in Korean mummy to extract palaeodietary information using stable isotope analysis. We extracted isotopic information from bone collagen and hair keratin, which reflects the lifetime average diet and short-term diet, respectively. The average values of ${\delta}^{13}C$ and ${\delta}^{15}N$ values are -19.0‰ and 11.4‰ (bone collagen), and -21.3‰ and 14.3‰ (hair keratin). Stable carbon isotope shows a mainly $C_3$-based diet from cradle to grave. On the other hand, there is significant difference in stable nitrogen isotope which implies a relatively enriched diet before death. We expect this study opens up a potential to understand the palaeodietary lifestyle in Joseon Dynasty.

Study on the Chemical Composition and Lead Isotope Ratios of Lead Glaze Used on Blue Tiles from Gyeoungbokgung Palace (경복궁 청기와에 사용된 납유의 화학조성과 납동위원소 특성연구)

  • So Jin Kim;Young Do Kim
    • Economic and Environmental Geology
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    • v.57 no.3
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    • pp.343-352
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    • 2024
  • Composition analysis and lead isotope ratio analysis were conducted to determine the coloring machanism on lead glaze used in Gyeongbokgung Palace and the provenance of the lead used as a flux. 31 blue tiles were classified into green, blue, and yellow. The chemical analysis of lead glazes on the blue tiles revealed that Pb, Si, and Cu were the main components, and trace amounts of Fe, Ca, Mg, and Al were detected. The Cu content was high in blue lead glaze, while Cu was not detected in yellow or brown lead glaze which instead had high Fe content. Therefore, it was found that lead was used as a flux and copper oxide as a coloring agent in the production of lead glaze. In addition, the lead isotope ratios of the lead glaze used in the blue tiles of Gyeongbokgung palace were plotted in zone 3 on the distribution map of lead isotope ratios on the Korean Peninsula, which includes Chungcheong-do and Jeolla-do. It is presumed that the flux for the lead glaze was sourced from galena found in these regions. The lead isotope ratios of the green glaze from the Three Kingdoms and Unified Silla period were mostly located outside the Korean Peninsula, showing that the provenance of lead had changed. In particular, the lead isotope ratios of the green glaze from the Three Kingdoms and Unified Silla period suggest exchange with neighboring countries. Also the lead isotope ratios of the green glazes from the same temple are different, so it is believed that they were made at different times or in different workshops.