title: Evaluation of chemiluminescence reagents for selective detection of reactive oxygen species.
abstract: 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.
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Naoya
authors.initials: N
authors.lastname: Kishikawa
authors.forename: Kaname
authors.initials: K
authors.lastname: Ohyama
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Ohba
authors.forename: Maiko
authors.initials: M
authors.lastname: Kohno
authors.forename: Toshinobu
authors.initials: T
authors.lastname: Masuda
authors.forename: Akira
authors.initials: A
authors.lastname: Takadate
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Nakashima
authors.forename: Naotaka
authors.initials: N
authors.lastname: Kuroda
mesh_terms: 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
chemical_list: 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