Rapid analysis of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) isotopologues in stable isotope-resolved metabolomics (SIRM) using direct infusion nanoelectrospray ultra-high-resolution Fourier transform mass spectrometry (DI-nESI-UHR-FTMS)

Joon Seon Yang, Teresa W.M. Fan, Jason A. Brandon, Andrew N. Lane, Richard M. Higashi

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

S-Adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are important metabolites in the one-carbon cycle that modulates cellular methylation required for proliferation and epigenetic regulation. Their concentrations, synthesis, and turnover are difficult to determine conveniently and reliably. We have developed such a method by coupling a simple and rapid purification scheme that efficiently captures both compounds, with high sensitivity, sample throughput direct infusion nanoelectrospray ultra-high-resolution Fourier transform mass spectrometry (DI-nESI-UHR-FTMS). This method is compatible with Stable Isotope-Resolved Metabolomic (SIRM) analysis of numerous other metabolites. The limits of detection for both SAM and SAH were <1 nM, and the linearity range was up to 1000 nM. The method was first illustrated for SAM/SAH analysis of mouse livers, and lung adenocarcinoma A549 cells. We then applied the method to track 13C1–CH3-Met incorporation into SAM and 13C6-glucose transformation into SAM and SAH via de novo synthesis. We further used the method to show the distinct effects on A549 and H1299 cells with treatment of anti-cancer methylseleninic acid (MSA), selenite, and selenomethionine, notably SAM depletion and increased SAM to SAH ratio by MSA, which implicates altered epigenetic regulation.

Original languageEnglish
Article number338873
JournalAnalytica Chimica Acta
Volume1181
DOIs
StatePublished - Oct 9 2021

Bibliographical note

Funding Information:
This work was supported in part by NIH grants 1U24DK097215-01A1 , P01CA163223-01A1 , 5P20GM121327 , 1P30ES026529-01A1 , and Shared Resource(s) of the University of Kentucky Markey Cancer Center P30CA177558 .

Publisher Copyright:
© 2021 Elsevier B.V.

Keywords

  • DI-nESI-UHR-FTMS
  • Methylseleninic acid
  • S-adenosylhomocysteine
  • S-adenosylmethionine
  • Selenite
  • Stable isotope-resolved metabolomics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

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