title: High-throughput total cupric ion reducing antioxidant capacity of biological samples determined using flow injection analysis and microplate-based methods.
abstract: High-throughput cupric ion reducing antioxidant capacity (CUPRAC) methods were developed for assessment of total antioxidant capacity (TAC) in urine and serum, based on reduction of Cu(II)-neocuproine complex to highly colored Cu(I)-neocuproine complex, measured spectrophotometrically at 450 nm. The reaction time was significantly reduced from 30 to 4 min by application of a calibration compound (uric acid) with kinetic behavior similar to that shown by urine samples. The method was implemented in a microformat (96 well plates) and also in an automatic fashion (flow injection analysis, FIA). A determination throughput value of 288 h(-1) (microplate method) or of 15 h(-1) (automatic FIA) was attained. Application of both methods to human serum (SRM 909b, level I) and urines (n = 9) provided TAC values in agreement with those of the end-point batch method.
authors.affiliation: REQUIMTE, Departamento de Química, Faculdade de Farmácia, Universidade do Porto, Portugal.
authors.forename: Joana P N
authors.initials: JP
authors.lastname: Ribeiro
authors.forename: Luís M
authors.initials: LM
authors.lastname: Magalhães
authors.forename: Salette
authors.initials: S
authors.lastname: Reis
authors.forename: José L F C
authors.initials: JL
authors.lastname: Lima
authors.forename: Marcela A
authors.initials: MA
authors.lastname: Segundo
mesh_terms: D000975:Antioxidants / Q000032:analysis*; D003300:Copper / Q000737:chemistry*; D017022:Flow Injection Analysis; D057166:High-Throughput Screening Assays; D006801:Humans; D007477:Ions / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D010084:Oxidation-Reduction; D012016:Reference Values
chemical_list: D000975:Antioxidants; D007477:Ions; D003300:Copper