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A Tandem Mass Spectrometric Method for Singlet Oxygen Measurement
Author(s) -
Karonen Maarit,
Mattila Heta,
Huang Ping,
Mamedov Fikret,
Styring Stenbjörn,
Tyystjärvi Esa
Publication year - 2014
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/php.12291
Subject(s) - singlet oxygen , chemistry , electron paramagnetic resonance , mass spectrometry , photochemistry , oxygen , thylakoid , molecule , tandem , analytical chemistry (journal) , singlet state , tandem mass spectrometry , nitroxyl , chromatography , nuclear magnetic resonance , organic chemistry , materials science , chloroplast , excited state , biochemistry , physics , nuclear physics , composite material , gene
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 .
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