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A simple 1H (12C/13C) filtered experiment to quantify and trace isotope enrichment in complex environmental and biological samples.
Steiner, Katrina; Bermel, Wolfgang; Soong, Ronald; Lysak, Daniel H; Jenne, Amy; Downey, Katelyn; Wolff, William W; Costa, Peter M; Ronda, Kiera; Moxley-Paquette, Vincent; Pellizzari, Jacob; Simpson, Andre J.
Affiliation
  • Steiner K; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Bermel W; Bruker Biospin GmbH, Rudolf-Plank-Str. 23, Ettlingen 76275, Germany.
  • Soong R; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Lysak DH; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Jenne A; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Downey K; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Wolff WW; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Costa PM; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Ronda K; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Moxley-Paquette V; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Pellizzari J; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada.
  • Simpson AJ; Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, ON M1C 1A4, Canada. Electronic address: andre.simpson@utoronto.ca.
J Magn Reson ; 361: 107653, 2024 Apr.
Article in En | MEDLINE | ID: mdl-38471414
ABSTRACT
Nuclear magnetic resonance (NMR) based 13C tracing has broad applications across medical and environmental research. As many biological and environmental samples are heterogeneous, they experience considerable spectral overlap and relatively low signal. Here a 1D 1H-12C/13C is introduced that uses "in-phase/opposite-phase" encoding to simultaneously detect and discriminate both protons attached to 12C and 13C at full 1H sensitivity in every scan. Unlike traditional approaches that focus on the 12C/13C satellite ratios in a 1H spectrum, this approach creates separate sub-spectra for the 12C and 13C bound protons. These spectra can be used for both quantitative and qualitative analysis of complex samples with significant spectral overlap. Due to the presence of the 13C dipole, faster relaxation of the 1H-13C pairs results in slight underestimation compared to the 1H-12C pairs. However, this is easily compensated for, by collecting an additional reference spectrum, from which the absolute percentage of 13C can be calculated by difference. When combined with the result, 12C and 13C percent enrichment in both 1H-12C and 1H-13C fractions are obtained. As the approach uses isotope filtered 1H NMR for detection, it retains nearly the same sensitivity as a standard 1H spectrum. Here, a proof-of-concept is performed using simple mixtures of 12C and 13C glucose, followed by suspended algal cells with varying 12C /13C ratios representing a complex mixture. The results consistently return 12C/13C ratios that deviate less than 1 % on average from the expected. Finally, the sequence was used to monitor and quantify 13C% enrichment in Daphnia magna neonates which were fed a 13C diet over 1 week. The approach helped reveal how the organisms utilized the 12C lipids they are born with vs. the 13C lipids they assimilate from their diet during growth. Given the experiments simplicity, versatility, and sensitivity, we anticipate it should find broad application in a wide range of tracer studies, such as fluxomics, with applications spanning various disciplines.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Protons / Isotopes Language: En Journal: J Magn Reson / J. magn. reson / Journal of magnetic resonance Journal subject: DIAGNOSTICO POR IMAGEM Year: 2024 Document type: Article Affiliation country: Canadá Country of publication: Estados Unidos

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Protons / Isotopes Language: En Journal: J Magn Reson / J. magn. reson / Journal of magnetic resonance Journal subject: DIAGNOSTICO POR IMAGEM Year: 2024 Document type: Article Affiliation country: Canadá Country of publication: Estados Unidos