Difference between revisions of "Hydrogen peroxide"
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=== Measurement of hydrogen peroxide === | === Measurement of hydrogen peroxide === | ||
Two techniques to measure H<sub>2</sub>O<sub>2</sub> concentration and H<sub>2</sub>O<sub>2</sub> production can be combined with the O2k: | Two techniques to measure H<sub>2</sub>O<sub>2</sub> concentration and H<sub>2</sub>O<sub>2</sub> production can be combined with the O2k: | ||
# [[O2k- | # [[O2k-Fluo LED2-Module|O2k-Fluorometry]] | ||
# Electrochemical measurement with an amperometric H<sub>2</sub>O<sub>2</sub> sensor. | # Electrochemical measurement with an amperometric H<sub>2</sub>O<sub>2</sub> sensor. | ||
==== Fluorescence ==== | ==== Fluorescence ==== | ||
The fluorometric measurement of H<sub>2</sub>O<sub>2</sub> in the O2k is based on the [[O2k- | The fluorometric measurement of H<sub>2</sub>O<sub>2</sub> in the O2k is based on the [[O2k-Fluo LED2-Module]], using [[Amplex red]] (or one of its brand names).<ref>Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. - [[Makrecka-Kuka 2015 Biomolecules |»Bioblast link«]].</ref> | ||
::» [[Amplex red]] -» [[MiPNet18.06_Amplex-Yeast]] -» [[MiPNet18.05_Amplex-Mouse-heart]] -» [[MiPNet17.17_Amplex-Mouse-brain]]. | |||
==== Electrochemical measurement ==== | ==== Electrochemical measurement ==== | ||
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It is sometimes stated that the available H<sub>2</sub>O<sub>2</sub> electrodes may not have the required sensitivity for most ROS studies. WPI states a lower detection limit of "<100 nM" for their 2 mm "macro" electrode and "<10 nM" for their 0.1 mm "mini sensor". The latter value is quite similar to the stated detection limit for fluorescence based methods, however it is not clear if the "mini sensor", probable developed for direct tissue insertion, works well in macroscopic chambers. A direct comparison would be helpful. Please add your experiences! | It is sometimes stated that the available H<sub>2</sub>O<sub>2</sub> electrodes may not have the required sensitivity for most ROS studies. WPI states a lower detection limit of "<100 nM" for their 2 mm "macro" electrode and "<10 nM" for their 0.1 mm "mini sensor". The latter value is quite similar to the stated detection limit for fluorescence based methods, however it is not clear if the "mini sensor", probable developed for direct tissue insertion, works well in macroscopic chambers. A direct comparison would be helpful. Please add your experiences! | ||
Picomolar detection ranges have been reported for a method combining Amplex Red (R) with square wave voltammetry.<ref> | Picomolar detection ranges have been reported for a method combining Amplex Red (R) with square wave voltammetry.<ref>Lyon JL, Stevenson KJ (2006) Picomolar peroxide detection using a chemically activated redox mediator and square wave voltammetry. Anal Chem 78:8518-25. - [[Lyon_2006_Anal_Chem |»Bioblast link«]]</ref> | ||
<references/> | <references/> | ||
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=== Reoxygenation of the respiration medium with H<sub>2</sub>O<sub>2</sub> === | === Reoxygenation of the respiration medium with H<sub>2</sub>O<sub>2</sub> === | ||
:» [[Re-oxygenation|More details]] |
Revision as of 18:53, 10 January 2016
Hydrogen peroxide |
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
Description
Hydrogen peroxide, H2O2 or dihydrogen dioxide, is one of several reactive oxygen intermediates generally referred to as reactive oxygen species (ROS). It is formed in various enzyme-catalyzed reactions (e.g. superoxide dismutase) with the potential to damage cellular molecules and structures. H2O2 is dismutated by catalase to water and oxygen. H2O2 is produced as a signaling molecule in aerobic metabolism and passes membranes more easily compared to other ROS.
Abbreviation: H2O2
MitoPedia methods:
Respirometry,
Fluorometry
MitoPedia topics:
Substrate and metabolite
Applications in HRR
Measurement of hydrogen peroxide
Two techniques to measure H2O2 concentration and H2O2 production can be combined with the O2k:
- O2k-Fluorometry
- Electrochemical measurement with an amperometric H2O2 sensor.
Fluorescence
The fluorometric measurement of H2O2 in the O2k is based on the O2k-Fluo LED2-Module, using Amplex red (or one of its brand names).[1]
Electrochemical measurement
For electrochemically (H2O2 electrode) measuring H2O2 (and all other amperometric methods) please see our O2k-NO Amp-Module and its manual MiPNet15.05 NO-Manual. All stated there about measuring NO is technically also correct for using a H2O2 electrode together with the oxygraph. In this manual you will also find which accessories you need to use a H2O2 electrode together with the O2k-MultiSensor System (there are different requirements for different generations of O2ks). The electrode itself has to be bought from a third party.
It is sometimes stated that the available H2O2 electrodes may not have the required sensitivity for most ROS studies. WPI states a lower detection limit of "<100 nM" for their 2 mm "macro" electrode and "<10 nM" for their 0.1 mm "mini sensor". The latter value is quite similar to the stated detection limit for fluorescence based methods, however it is not clear if the "mini sensor", probable developed for direct tissue insertion, works well in macroscopic chambers. A direct comparison would be helpful. Please add your experiences!
Picomolar detection ranges have been reported for a method combining Amplex Red (R) with square wave voltammetry.[2]
- ↑ Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. - »Bioblast link«.
- ↑ Lyon JL, Stevenson KJ (2006) Picomolar peroxide detection using a chemically activated redox mediator and square wave voltammetry. Anal Chem 78:8518-25. - »Bioblast link«