Heparin-stabilized gold nanoparticles-based CUPRAC colorimetric sensor for antioxidant capacity measurement

Sensing of total antioxidant capacity (TAC) as an integrated parameter showing the collective action of various antioxidants is an important challenge in food, biochemical and drug analysis. A novel heparin-stabilized gold nanoparticles (AuNPs)-based 'cupric reducing antioxidant capacity' (CUPRAC) colorimetric sensor was designed for TAC measurement. Heparin, a sulfated polysaccharide, was both the reducing and stabilizing agent for distinct negatively-charged AuNPs synthesis. The stabilized AuNPs were added to the copper(I)-neocuproine (Cu(I)-Nc) solution formed from the reaction of Cu(II)-Nc with antioxidants, and the absorbance of the resulting Cu(I)-Nc-AuNPs (Cu(I)-Nc cationic chelate electrostatically adsorbed on gold nanoparticles) was measured at 455 nm. As opposed to other similar AuNPs-based sensors, the proposed nano-sensor exhibited excellent (1000-fold) tolerance toward inert electrolytes without aggregation. The linear range was wider than that of conventional CUPRAC, with lower LOD (0.2 µM for trolox) and higher molar absorptivity (8.36 × 10⁴ M^-1 cm^-1 for quercetin). The 'trolox equivalent antioxidant capacity' (TEAC) values and activity order for a number of antioxidants were in accordance with those of the reference CUPRAC assay. Antioxidant additions to black tea extract gave recoveries of 93-97% and RSD 2-6%. This green sensor significantly reduced reagent consumption, and operated in complex food samples with a simple, reliable and robust methodology.