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Chemical Shift: Internal References and Solvent Effects01:17

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In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
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Stable isotope coded derivatizing reagents as internal standards in metabolite profiling.

Per Bruheim1, Hans Fredrik Nyvold Kvitvang, Silas G Villas-Boas

  • 1Department of Biotechnology, Norwegian University of Science and Technology, Trondheim, Norway. Per.Bruheim@ntnu.n

Journal of Chromatography. A
|May 1, 2013
PubMed
Summary

Isotope Coded Derivatization (ICD) offers a cost-effective solution for precise quantitative Metabolite Profiling. This method improves analytical performance for complex metabolite analysis, overcoming limitations of traditional methods.

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Area of Science:

  • Analytical Chemistry
  • Metabolomics
  • Biochemistry

Background:

  • Gas chromatography (GC) and liquid chromatography (LC) coupled with mass spectrometry (MS) are key techniques in Metabolomics.
  • Quantitative Metabolite Profiling aims to analyze more metabolites simultaneously, demanding high analytical precision.
  • Current methods face limitations in quantitative accuracy due to standard availability and ionization efficiency variability.

Purpose of the Study:

  • To review novel strategies for enhancing quantitative precision in Metabolite Profiling.
  • To highlight the development and application of isotopically labeled derivatizing reagents.
  • To introduce Isotope Coded Derivatization (ICD) as a viable alternative for comprehensive analysis.

Main Methods:

  • Utilizes derivatization with naturally labeled reagents for samples.
  • Employs separate derivatization of standards with isotopically labeled reagents.
  • Spikes derivatized standards into sample solutions before GC-MS or LC-MS analysis.

Main Results:

  • Isotope Coded Derivatization (ICD) provides a robust strategy to counteract variables affecting quantitative precision.
  • This approach is cost-effective and overcomes the lack of commercially available isotopically labeled standards for many metabolites.
  • ICD is particularly attractive for profiling over a hundred metabolites in a single analytical method.

Conclusions:

  • ICD significantly improves analytical performance and precision in quantitative Metabolite Profiling.
  • The reviewed isotopically labeled derivatizing reagents are crucial for advancing comprehensive metabolic analyses.
  • This strategy addresses the need for accurate quantification in complex biological samples.