Reduced Fluoresceinamine for Peroxynitrite Quantification in the Presence of Nitric Oxide

• Type: Article in International Scientific Journal
• Date: 2012

Authors

Eliana F C Simoes (Author) Other; Joao M M Leitao (Author) Other; Joaquim C G E Esteves da Silva (Author) FCUP

Journal

Title: Journal of Fluorescence

Vol. 22

Pages: 1127-1140

ISSN: 1053-0509

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 5 Citations

Scopus - 5 Citations

Scientific classification

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-002-8BQ

DOI: 10.1007/s10895-012-1051-3

Abstract (EN): A new fluorescent analytical methodology for the quantification of peroxynitrite (ONOO-) in the presence of nitric oxide (NO) was developed. The quantification of ONOO- is based in the oxidation of the non-fluorescent reduced fluoresceinamine to a high fluorescent oxidized fluoresceinamine in reaction conditions where the interference of NO is minimized. Screening factorial experimental designs and optimization Box-Behnken experimental design methodologies were used in order to optimize the detection of ONOO- in the presence of NO. The factors analysed were: reduced fluoresceinamine concentration (C (Fl) ); cobalt chloride concentration (C (CoCl2) ); presence of oxygen (O (2) ); and, the pH (pH). The concentration of sodium hydroxide (C (NaOH) ) needed to diluted the initially solution of ONOO- was also evaluated. An optimum region for ONOO- quantification where the influence of NO is minimal was identified - C (Fl) from 0.50 to 1.56 mM, C (CoCl2) from 0 to 1.252 x 10(-2) M, pH from 6 to 8 and C (NaOH) 0.10 M. Better results were found in the presence of NO at pH 7.4, C (Fl) 0.5 mM, without oxygen, without cobalt chloride and with a previous dilution of peroxynitrite solution with C (NaOH) 0.1 M. This methodology shows a linear range from 0.25 to 40 mu M with a limit of detection of 0.08 mu M. The bioanalytical methodology was successfully applied in the ONOO- quantification of fortified serum and macrophage samples.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 14