title: Advantage of menadione-catalyzed chemiluminescent assay for the determination of viable mammalian cell number.
abstract: A chemiluminescent assay composed of TCPO [bis(2,4,6-trichlorophenyl)oxalate] and harmless rhodamine B is proposed to be superior in the determination of menadione-catalyzed hydrogen peroxide (H₂O₂) production by viable mammalian cells to that composed of TCPO and harmful pyrene [Anal. Biochem. 207 (1992) 255-260]. In tests, the proposed assay showed that the measurable concentration of H₂O₂ and the viable cell number ranged from 10⁻⁹ to 10⁻³ M and from 2 × 10² to 2 × 10⁶ cells/100 μl/well in the presence of 10% bovine serum, respectively. The measuring time was approximately 10 min. On the other hand, the measurable cell numbers by the colorimetric WST-1 and MTT assays requiring several hours ranged only from 10³ to 10⁴ cells/100 μl/well and from 10⁴ to 10⁵ cells/100 μl/well, respectively. The cytotoxicity of sodium dodecyl sulfate was also observed at intervals of 1 min by the proposed assay, but not by the above colorimetric assays.
authors.affiliation: Shizuoka Institute of Science and Technology, Fukuroi, Shizuoka 437-8555, Japan. yamasho@ms.sist.ac.jp
authors.forename: Shiro
authors.initials: S
authors.lastname: Yamashoji
authors.forename: Naoko
authors.initials: N
authors.lastname: Yoshikawa
authors.forename: Masayuki
authors.initials: M
authors.lastname: Kirihara
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Tsuneyoshi
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002384:Catalysis; D002448:Cell Adhesion; D006861:Hydrogen Peroxide / Q000378:metabolism; D049449:Luminescence; D051379:Mice; D024483:Vitamin K 3 / Q000378:metabolism*
chemical_list: D024483:Vitamin K 3; D006861:Hydrogen Peroxide