Fluorophotometric determination of hydrogen peroxide and other reactive oxygen species with fluorescein hydrazide (FH) and its crystal structure.
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.
Nakahara
Ryosuke
R
Osaka University of Pharmaceutical Sciences, Osaka, Japan. d06001@gly.oups.ac.jp
Fujimoto
Tsuyoshi
T
Doi
Mitsunobu
M
Morita
Kanako
K
Yamaguchi
Takako
T
Fujita
Yoshikazu
Y
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
D005452:Fluoresceins; D017382:Reactive Oxygen Species; C052067:fluorescein hydrazide; D006861:Hydrogen Peroxide