Table 2.

Comparison of different detection methods of mtROS

Methods	Drugs	Detected species	Principle	Limits
Fluorescent probes	MitoSOX	Superoxide	Attaching a hydrophobic cation to dihydroethidium.	Two fluorescent products, one of which is superoxide specific. The two products need to be separated by HPLC.
	MitoPY1	Hydrogen peroxide	Attaching a hydrophobic cation to a phenylboronate ester.	It also reacts with peroxynitrite.
	MitoAR MitoHR	Hydroxyl radical	Reaction with hydroxyl radical cleaves off the quenching motif.	They also react with peroxynitrite.
Mitochondrial inhibitors	Rotenone Antimycin	General ROS	Rotenone is complex I inhibitor and antimycin is complex III inhibitor.	Rotenone either decrease or increase ROS; Interruption of the ETC.
Mitochondrial antioxidant	MitoQ MitoTEMPO MitoVitE	General ROS	Attaching a hydrophobic cation to antioxidants.	-
Electron spin resonance	MitoDEPMPO	Free radicals	Free radicals have unpaired electrons, and thus are paramagnetic and detectable by ESR.	Lower sensitivity than fluorescent probes.