Evaluation of chemiluminescence reagents for selective detection of reactive oxygen species.
In order to evaluate the chemiluminescence (CL) reagents for selective detection of reactive oxygen species (ROS), we comprehensively measured the CL responses of 20 CL reagents (three luminol derivatives, two imidazopyrazinone derivatives, eight lophine derivatives, six acridinium ester derivatives and lucigenin) against six types of ROS (superoxide anion: O(2)(*-), hydroxyl radical: *OH, hydrogen peroxide: H(2)O(2), hypochlorite anion: ClO(-), singlet oxygen: (1)O(2), and nitric oxide: NO). As a result of the screening, it was found that nine CL reagents selectively detected O(2)(*-) while one CL reagent selectively detected *OH. However, no CL reagent had selectivity on the detection of H(2)O(2), ClO(-), (1)O(2) and NO. Our screening results could help to select the most suitable CL reagent for selective determination of different ROS. As an application study, 4-methoxyphenyl-10-methylacridinium-9-carboxylate (MMAC), one of the acridinium ester derivatives, showed high selectivity on the detection of O(2)(*-), and thus was applied to the assay of superoxide dismutase (SOD) activity. The dynamic range and detection limit of the developed CL assay were 0.1-10 and 0.06 U mL(-1), respectively. Significant correlation (r=0.997) was observed between the results by the CL assay using MMAC and the spectrophotometric assay using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt.
Yamaguchi
Shinya
S
Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Kishikawa
Naoya
N
Ohyama
Kaname
K
Ohba
Yoshihito
Y
Kohno
Maiko
M
Masuda
Toshinobu
T
Takadate
Akira
A
Nakashima
Kenichiro
K
Kuroda
Naotaka
N
D006861:Hydrogen Peroxide / Q000032:analysis; D017665:Hydroxyl Radical / Q000032:analysis; D006997:Hypochlorous Acid / Q000032:analysis; D049408:Luminescent Agents / Q000737:chemistry*; D009569:Nitric Oxide / Q000032:analysis; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000737:chemistry; D026082:Singlet Oxygen / Q000032:analysis; D013053:Spectrophotometry / Q000379:methods*; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000032:analysis
D049408:Luminescent Agents; D017382:Reactive Oxygen Species; D013481:Superoxides; D026082:Singlet Oxygen; D009569:Nitric Oxide; D017665:Hydroxyl Radical; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase