title: Determination of total plasma hydroperoxides using a diphenyl-1-pyrenylphosphine fluorescent probe.
abstract: Plasma hydroperoxides (HPs) are widely accepted to be good indicators of oxidative stress. By means of the method proposed here, which uses diphenyl-1-pyrenylphosphine (DPPP) as a fluorescent probe, all types of plasma HP were determined. The limits of detection and quantification of the method were 0.08 and 0.25 nmol of cumene hydroperoxide (CHP) equivalents in 40 μl of plasma, respectively. The method is satisfactory in terms of precision (5.3% for 14.5 μM CHP eq., n=8), and the recoveries were 91% and 92% after standard additions of 26 and 52 μM CHP, respectively. The selectivity of the proposed method is higher than 96%. Moreover, optimization of the reaction conditions and the addition of ethylenediaminetetraacetic acid (EDTA) disodium salt and 2,6-di-tert-butyl-4-methylphenol (BHT) prevented the formation of HP artifacts during the analysis. Therefore, the proposed method is useful for simple and quantitative determination of total plasma HPs.
authors.affiliation: Department of Nutrition and Food Science, XaRTA-INSA, University of Barcelona, 08028 Barcelona, Spain.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Santas
authors.forename: Francesc
authors.initials: F
authors.lastname: Guardiola
authors.forename: Magda
authors.initials: M
authors.lastname: Rafecas
authors.forename: Ricard
authors.initials: R
authors.lastname: Bou
mesh_terms: D000818:Animals; D002084:Butylated Hydroxytoluene; D004492:Edetic Acid / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006168:Guinea Pigs; D006861:Hydrogen Peroxide / Q000097:blood*; D007700:Kinetics; D009943:Organophosphorus Compounds / Q000737:chemistry*; D010636:Phenols / Q000737:chemistry; D011721:Pyrenes / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013050:Spectrometry, Fluorescence
chemical_list: D005456:Fluorescent Dyes; D009943:Organophosphorus Compounds; D010636:Phenols; D011721:Pyrenes; C066322:diphenyl-1-pyrenylphosphine; D002084:Butylated Hydroxytoluene; D004492:Edetic Acid; D006861:Hydrogen Peroxide