Table 18.4 Fluorescence response of 3′-(p-aminophenyl) fluorescein (APF), 3′-(p-hydroxyphenyl) fluorescein (HPF) and dichlorodihydrofluorescein diacetate (H2DCFDA) to various reactive oxygen species (ROS).

Reactive Oxygen Species (ROS) ROS Generation Method APF * HPF * H2DCFDA *
Hydrogen peroxide (H2O2)100 µM of H2O2<12190
Hydroxyl radical (HO•)100 µM of ferrous perchlorate (II) and 1 mM of H2O212007307400
Hypochlorite anion (OCl)3 µM (final) of OCl3600686
Nitric oxide (NO)100 µM of 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-methyl-1-triazene (NOC-7)<16150
Peroxyl radical (ROO•)100 µM of 2,2'-azobis(2-amidinopropane), dihydrochloride (AAPH)217710
Peroxynitrite anion (ONOO)3 µM (final) of ONOO5601206600
Singlet oxygen (1O2)100 µM of 3-(1,4-dihydro-1,4-epidioxy-1-naphthyl)propionic acid9526
Superoxide anion (•O2)100 µM KO26867
Autooxidation2.5 hours exposure to fluorescent light source<1<12000
* 10 µm of APF, HPF or DCF (2',7'-dichlorofluorescein) were added to sodium phosphate buffer (0.1 M, pH 7.4); ROS were generated as indicated; and fluorescence was measured using excitation/emission wavelengths of 490/515 nm (for APF and HPF) or 500/520 nm (for DCF). DCF was obtained by hydrolysis of H2DCFDA with base as described in ref; dihydrofluorescein diacetates are colorless and nonfluorescent until both of the acetate groups are hydrolyzed and the products are subsequently oxidized to fluorescein derivatives.
For Research Use Only. Not for use in diagnostic procedures.