A novel electrochemical biosensor for antioxidant evaluation of phloretin based on cell-alginate/ʟ-cysteine/gold nanoparticle-modified glassy carbon electrode

Abstract Antioxidant evaluation of bioactive compounds is limited, since many methods lack a real physiological environment that can be used conveniently and intuitively. In this study, a simple, label-free and effective electrochemical biosensor method has been developed to evaluate the antioxidant effect of phloretin (Ph) by 3D cell modification on a glassy carbon electrode (GCE). In response to this, A549 cells were immobilized onto a self-assembled ʟ-cysteine/gold nanoparticle (AuNPs/ʟ-Cys)-modified GCE surface by a simple drop casting after encapsulated in alginate. The electrochemical impedance spectroscopy (EIS) results showed that the impedance value (R et ) increased with the concentration of H 2 O 2 in the range of 0–60 μmol/L with the correlation of 0.990 which acted as an oxidative stress model inducer. However, the EIS value decreased with the co-incubation of Ph ranging from 10 to 100 μmol/L, showing a dose-dependent manner and time effect, indicating that the variation of R et was responded to the antioxidant effect. The response impedance of the biosensor is linear to Ph concentrations from 20 μmol/L to 100 μmol/L with the detection limit (LOD) as 1.96 μmol/L. A significant correlation was observed between reactive oxygen species (ROS) values and R et values following the concentrations of Ph, thus demonstrating the good biological relevance of cell-based electrochemical method. The strategy has been used to evaluate Ph antioxidant capacity in real cells with satisfactory results, indicating the feasibility of biosensor analysis for antioxidant evaluation.