Fluorophotometric determination of hydrogen peroxide with fluorescin in the presence of cobalt (II) and reaction against other reactive oxygen species.
A fluorophotometric method for the determination of hydrogen peroxide (H2O2) using fluorescin was developed. This method was based on the oxidative reaction of fluorescin, a colorless, non-fluorescent lactoid fluorescein, by H2O2 to give highly fluorescein fluorescence emission. In the determination of H2O2, the calibration curve exhibited linearity over the H2O2 concentration range of 1.5-310 ng mL(-1) at an emission wavelength of 525 nm with an excitation of 500 nm and with relative standard deviations (n = 6) of 2.51%, 2.48%, and 1.31% for 3.1 ng mL(-1), 30.8 ng mL(-1), and for 308 ng mL(-1) of H2O2, respectively. The detection limit for H2O2 was 1.9 ng mL(-1) six blank determinations was performed (rho = 6). This proposed method was applied to detection of other reactive oxygen species and nitrogen species (ROS/RNS) such as singlet oxygen (1O2), hydroxyl radical (*OH), peroxynitrite (ONOO-) etc., and it was possible to detect them with a high sensitivity. In addition, this proposed method was applied to the recovery tests of H2O2 in calf serum, human saliva, rain water, and wheat noodles; the results were satisfactory.
Nakahara
Ryosuke
R
Department of Clinical Chemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan. d06001@gly.oups.ac.jp
Kashitani
Satomi
S
Hayakawa
Kumi
K
Kitani
Yuuki
Y
Yamaguchi
Takako
T
Fujita
Yoshikazu
Y
D000818:Animals; D002417:Cattle; D003035:Cobalt / Q000737:chemistry*; D005433:Flour / Q000032:analysis; D005452:Fluoresceins / Q000032:analysis / Q000737:chemistry*; D015590:Fluorophotometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000737:chemistry; D026361:Reactive Nitrogen Species / Q000032:analysis; D017382:Reactive Oxygen Species / Q000032:analysis*; D012463:Saliva / Q000737:chemistry; D014867:Water / Q000737:chemistry
D005452:Fluoresceins; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D014867:Water; D003035:Cobalt; D006861:Hydrogen Peroxide