A tandem mass spectrometric method for singlet oxygen measurement

Photochem Photobiol. 2014 Sep-Oct;90(5):965-71. doi: 10.1111/php.12291. Epub 2014 Jun 20.

Abstract

Singlet oxygen, a harmful reactive oxygen species, can be quantified with the substance 2,2,6,6-tetramethylpiperidine (TEMP) that reacts with singlet oxygen, forming a stable nitroxyl radical (TEMPO). TEMPO has earlier been quantified with electron paramagnetic resonance (EPR) spectroscopy. In this study, we designed an ultra-high-performance liquid chromatographic-tandem mass spectrometric (UHPLC-ESI-MS/MS) quantification method for TEMPO and showed that the method based on multiple reaction monitoring (MRM) can be used for the measurements of singlet oxygen from both nonbiological and biological samples. Results obtained with both UHPLC-ESI-MS/MS and EPR methods suggest that plant thylakoid membranes produce 3.7 × 10(-7) molecules of singlet oxygen per chlorophyll molecule in a second when illuminated with the photosynthetic photon flux density of 2000 μmol m(-2 ) s(-1).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chlorophyll / agonists
  • Chlorophyll / metabolism*
  • Chromatography, High Pressure Liquid
  • Cucurbita / physiology
  • Cucurbita / radiation effects
  • Cyclic N-Oxides / chemistry
  • Electron Spin Resonance Spectroscopy
  • Light
  • Photosystem II Protein Complex / metabolism*
  • Piperidines / chemistry
  • Plant Leaves / physiology
  • Plant Leaves / radiation effects
  • Singlet Oxygen / analysis*
  • Singlet Oxygen / metabolism
  • Tandem Mass Spectrometry / instrumentation
  • Tandem Mass Spectrometry / methods*
  • Thylakoids / physiology*
  • Thylakoids / radiation effects

Substances

  • Cyclic N-Oxides
  • Photosystem II Protein Complex
  • Piperidines
  • Chlorophyll
  • Singlet Oxygen
  • 2,2,6,6-tetramethylpiperidide
  • TEMPO