A tandem mass spectrometric method for singlet oxygen measurement.
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).
Laboratory of Organic Chemistry and Chemical Biology, Department of Chemistry, University of Turku, Turku, Finland.
Maarit
M
Karonen
Heta
H
Mattila
Ping
P
Huang
Fikret
F
Mamedov
Stenbjörn
S
Styring
Esa
E
Tyystjärvi
D002734:Chlorophyll / Q000819:agonists / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D028464:Cucurbita / Q000502:physiology / Q000528:radiation effects; D003497:Cyclic N-Oxides / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D008027:Light; D045332:Photosystem II Protein Complex / Q000378:metabolism*; D010880:Piperidines / Q000737:chemistry; D018515:Plant Leaves / Q000502:physiology / Q000528:radiation effects; D026082:Singlet Oxygen / Q000032:analysis* / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D020524:Thylakoids / Q000502:physiology* / Q000528:radiation effects
D003497:Cyclic N-Oxides; D045332:Photosystem II Protein Complex; D010880:Piperidines; D002734:Chlorophyll; D026082:Singlet Oxygen; C551336:2,2,6,6-tetramethylpiperidide; C003959:TEMPO