Chemiluminescence assay for tetrahydrobiopterin based on the generation of hydrogen peroxide using isoluminol-microperoxidase in the presence of 1-methoxy PMS.
We developed a novel highly sensitive chemiluminescence (CL) method for BH(4). The principle of the proposed method is based on active oxygen formation induced by 1-methoxy-5-methyl phenazinium methyl sulphate (1-methoxy PMS) in the presence of dissolved oxygen. Furthermore, active oxygen is determined by a CL assay involving the luminol reaction with microperoxidase. In this report, we examined the mechanism of formation and identified the reactive oxygen species derived from BH(4) employing 1-methoxy PMS. Additionally, optimum conditions for the CL assay of BH(4) were established.
Arakawa
Hidetoshi
H
School of Pharmaceutical Sciences, Showa University, Tokyo 142-8555, Japan. arakawa@pharm.showa-u.ac.jp
Masuda
Kouichi
K
Tajima
Noriko
N
Maeda
Masako
M
D001708:Biopterins / Q000031:analogs & derivatives* / Q000032:analysis; D004578:Electron Spin Resonance Spectroscopy; D006861:Hydrogen Peroxide / Q000138:chemical synthesis*; D049449:Luminescence; D008165:Luminol / Q000031:analogs & derivatives* / Q000737:chemistry; D008773:Methylphenazonium Methosulfate / Q000031:analogs & derivatives* / Q000737:chemistry; D010544:Peroxidases / Q000737:chemistry*
D001708:Biopterins; D008773:Methylphenazonium Methosulfate; C032487:isoluminol; D008165:Luminol; C023530:1-methoxy-5-methylphenazinium methyl sulfate; D006861:Hydrogen Peroxide; D010544:Peroxidases; C021799:microperoxidase; C003402:sapropterin