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Singlet molecular oxygen ((1)O2) generation from biomolecule oxidation was confirmed using (18)O-labeled compounds and HPLC-MS/MS. This study demonstrates (1)O2 involvement in physiological and pathophysiological processes.

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Spectroscopy

Background:

  • Ultraweak chemiluminescence from biomolecule oxidation is linked to singlet molecular oxygen ((1)O2) and excited carbonyls.
  • Dioxetane intermediates are implicated in the generation of (1)O2 and excited triplet carbonyls.

Purpose of the Study:

  • To unequivocally demonstrate the generation of (1)O2 during biomolecule oxidation.
  • To investigate the role of (1)O2 in physiological and pathophysiological mechanisms.

Main Methods:

  • Utilized (18)O-labeled hydroperoxide and triplet dioxygen ((18)[(3)O2]) for isotopic labeling.
  • Employed High-Performance Liquid Chromatography coupled to tandem Mass Spectrometry (HPLC-MS/MS) for compound detection.
  • Performed direct spectroscopic detection and characterization of (1)O2 light emission at 1,270nm.

Main Results:

  • Confirmed (18)O-labeled (1)O2 ((18)[(1)O2]) formation via chemical trapping with anthracene-9,10-diyldiethane-2,1-diyl disulfate disodium salt (EAS).
  • Detected (18)O-labeled EAS endoperoxide using HPLC-MS/MS.
  • Observed characteristic light emission at 1,270nm, confirming singlet delta state monomolecular decay.

Conclusions:

  • Photoemission and chemical trapping provide clear evidence that hydroperoxide and excited carbonyl production generates (18)[(1)O2].
  • The findings strongly suggest the involvement of (1)O2 in physiological and pathophysiological processes.