Fluorescence response of 3'-(p-aminophenyl) fluorescein (APF), 3'-(p-hydroxyphenyl) fluorescein (HPF) and dichlorodihydrofluorescein diacetate (H2DCFDA) to various reactive oxygen species (ROS)—Table 18.4
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|Reactive Oxygen Species (ROS)||ROS Generation Method||APF *||HPF *||H2DCFDA *|
|Hydrogen peroxide (H2O2)||100 µM of H2O2||<1||2||190|
|Hydroxyl radical (HO•)||100 µM of ferrous perchlorate (II) and 1 mM of H2O2||1200||730||7400|
|Hypochlorite anion (–OCl)||3 µM (final) of –OCl||3600||6||86|
|Nitric oxide (NO)||100 µM of 1-hydroxy-2-oxo-3-(3-aminopropyl)-3-methyl-1-triazene (NOC-7) ||<1||6||150|
|Peroxyl radical (ROO•)||100 µM of 2,2'-azobis(2-amidinopropane), dihydrochloride (AAPH)||2||17||710|
|Peroxynitrite anion (ONOO–)||3 µM (final) of ONOO–||560||120||6600|
|Singlet oxygen (1O2)||100 µM of 3-(1,4-dihydro-1,4-epidioxy-1-naphthyl)propionic acid||9||5||26|
|Superoxide anion (•O2–)||100 µM KO2||6||8||67|
|Autooxidation||2.5 hours exposure to fluorescent light source||<1||<1||2000|
|* 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 ; dihydrofluorescein diacetates are colorless and nonfluorescent until both of the acetate groups are hydrolyzed and the products are subsequently oxidized to fluorescein derivatives.|
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