A Tandem Mass Spectrometric Method for Singlet Oxygen Measurement |
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| Authors: | Ping Huang Fikret Mamedov Stenbjörn Styring Esa Tyystjärvi |
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| Affiliation: | 1. Molecular Biomimetics, Department of Chemistry – ?ngstr?m Laboratory, Uppsala University, , Uppsala, Sweden;2. Molecular Plant Biology, Department of Biochemistry, University of Turku, , Turku, Finland |
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| 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. |
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