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Detection of free radicals generated from hydrogen peroxide, gallic acid and haemoprotein chemiluminescence system by electron spin resonance spectroscopy
Author(s) -
Kawane Masaaki,
Iida Tetsuo,
Yoshiki Yumiko,
Okubo Kazuyoshi,
Tsunakawa Mitsuaki
Publication year - 1999
Publication title -
luminescence
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.428
H-Index - 45
eISSN - 1522-7243
pISSN - 1522-7235
DOI - 10.1002/(sici)1522-7243(199911/12)14:6<321::aid-bio568>3.0.co;2-8
Subject(s) - chemistry , chemiluminescence , gallic acid , hemeprotein , hydrogen peroxide , radical , photochemistry , electron paramagnetic resonance , myoglobin , spin trapping , heme , organic chemistry , nuclear magnetic resonance , enzyme , antioxidant , physics
Low‐level chemiluminescence is produced in a hydrogen peroxide (H 2 O 2 )/gallic acid/haemoprotein system with single broad peaks around 520 nm, regardless of the biological role of the haemoprotein. The free haem iron systems (haemin and haematin systems) gave a higher photon intensity (1.5 × 10 4 and 2.0 × 10 4 cps) than that of the H 2 O 2 /gallic acid/haematoporphyrin system. These results indicated that haem iron plays a significant role in the photon emission of haemoprotein systems. A free radical with a g value of 2.0058 was detected through a direct electron spin resonance (ESR) method. The photon intensity of the H 2 O 2 /gallic acid/haemoprotein system decreased in the order: HRP > cytochrome c > myoglobin > haemoglobin, and this corresponded to the decrease in radical intensity. These results indicated that the formation of the free radical with a g value of 2.0058 may be the key step for chemiluminescence in the H 2 O 2 /gallic acid/haemoprotein system. A quartet line similar to DMPO‐OH adducts and uncomplexed free radicals (g = 2.0058) was detected using the ESR spin‐trapping method in the H 2 O 2 /gallic acid/cytochrome c system. Copyright © 1999 John Wiley & Sons, Ltd.