title: Fluorophotometric determination of hydrogen peroxide and other reactive oxygen species with fluorescein hydrazide (FH) and its crystal structure.
abstract: Methods for the fluorophotometric determination of hydrogen peroxide (H(2)O(2)) and other reactive oxygen species (ROS) were proposed by using the fluorescence reaction between H(2)O(2) or other ROS and fluorescein hydrazide (FH). In the determination of H(2)O(2), the calibration curve exhibited linearity over the H(2)O(2) concentration range of 2.1-460 ng ml(-1) at an emission wavelength of 527 nm with an excitation of 460 nm and with the relative standard deviations (n=6) of 4.06%, 1.78%, and 2.21% for 3.1 ng ml(-1), 30.8 ng ml(-1), and for 308 ng ml(-1) of H(2)O(2), respectively. The detection limit for H(2)O(2) was 0.7 ng ml(-1) due to three blank determinations (rho=3). The calibration curves for ROS-related compounds were also constructed under the optimum conditions. This method was successfully applied in the assay of H(2)O(2) in human urine. In addition, we performed the characterization of FH, and interesting information was obtained with regard to the relationship between the chemical structure and fluorescence.
authors.affiliation: Osaka University of Pharmaceutical Sciences, Osaka, Japan. d06001@gly.oups.ac.jp
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Nakahara
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Fujimoto
authors.forename: Mitsunobu
authors.initials: M
authors.lastname: Doi
authors.forename: Kanako
authors.initials: K
authors.lastname: Morita
authors.forename: Takako
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Fujita
mesh_terms: D002138:Calibration; D005452:Fluoresceins / Q000737:chemistry*; D015590:Fluorophotometry / Q000379:methods*; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D005452:Fluoresceins; D017382:Reactive Oxygen Species; C052067:fluorescein hydrazide; D006861:Hydrogen Peroxide