Fluorescence and HPLC Detection of Hydroxyl Radical by a Rhodamine-Nitroxide Probe and its Application in Cell Imaging
Authors:
- 28 November 2013
- journal article
- Published by Springer Nature in Journal of Fluorescence
- Vol. 24 (2) , 313-318
Abstract
A rhodamine nitroxide probe was designed to detect the hydroxyl radical (·OH), which presented high selectivity for ·OH over other reactive oxygen species (ROS) and linear fluorescence response to ·OH produced by Fenton reaction. The product was detected by HPLC-MS, indicating that the main product of the reaction was O-methylhydroxylamine and the product peak areas measured by HPLC-UV/vis and HPLC-FLD both enhanced proportionally with the increase of ·OH concentration. The application of the probe in biological system was explored to trace the production of ·OH in cells under oxidative stress condition induced by rotenone which can inhibit the mitochondria respiratory chain complex I and we found that appropriate rotenone may induce the normal human liver cells (L02) and human hepatoma cells (HepG2) to produce ·OH at different degrees.Keywords
This publication has 27 references indexed in Scilit:
- Two-Photon Fluorescence Microscopy Imaging of Cellular Oxidative Stress Using Profluorescent NitroxidesJournal of the American Chemical Society, 2012
- New Rhodamine Nitroxide Based Fluorescent Probes for Intracellular Hydroxyl Radical Identification in Living CellsOrganic Letters, 2011
- Development of a DNA-binding TEMPO derivative for evaluation of nuclear oxidative stress and its application in living cellsFree Radical Biology and Medicine, 2010
- A New Highly Selective and Sensitive Assay for Fluorescence Imaging of .OH in Living Cells: Effectively Avoiding the Interference of PeroxynitriteChemistry – A European Journal, 2010
- 2′,7′-Dichlorodihydrofluorescein as a fluorescent probe for reactive oxygen species measurement: Forty years of application and controversyFree Radical Research, 2010
- Perylenebisimide-linked nitroxide for the detection of hydroxyl radicalsAnalytica Chimica Acta, 2009
- Novel mitochondria-localizing TEMPO derivative for measurement of cellular oxidative stress in mitochondriaBioorganic & Medicinal Chemistry Letters, 2007
- Evidence of ROS generation by mitochondria in cells with impaired electron transport chain and mitochondrial DNA damageMitochondrion, 2007
- Mitochondrial Complex I Inhibitor Rotenone Induces Apoptosis through Enhancing Mitochondrial Reactive Oxygen Species ProductionJournal of Biological Chemistry, 2003
- Chemically mediated fluorescence yield switching in nitroxide-fluorophore adducts: optical sensors of radical/redox reactionsJournal of the American Chemical Society, 1988