Flow injection spectrophotometric determination total antioxidant capacity in human serum samples based on response surface methodology to optimize synthesized peroxidase-like activity carbon dots

• Using RSM technology to guide the optimization of Co/N-CDs catalytic performance. • A flow injection method for TAC was developed using Co/N-CDs nanoparticles. • the surface charge of Co/N-CDs were characterized by gel electrophoresis. Total antioxidant capacity (TAC) is an important indicator for evaluating oxidative stress of the human body. Since TAC is related to the concentration of reducing substances, it can be detected by using peroxidase-like or oxidase-like activity of nanozyme materials. In this work, the cobalt and nitrogen co-doped carbon dots (Co/N-CDs) are fabricated for building stability and high peroxidase-like nanozyme through the Box-Behnken design of response surface methodology. The morphology and luminescence properties of obtained Co/N-CDs were characterized by TEM and fluorophotometer, respectively. Interestingly, the surface charge of Co/N-CDs are innovatively investigated by a simple and widespread gel electrophoresis, which holds the potential to be an alternative to Zeta potential analysis. In addition, a flow injection spectrophotometric assay to detect ascorbic acid is develop with a high sensitivity and automation based on a Co/N-CDs/guaiacol/H 2 O 2 catalytic reaction system. The proposed method is also responsive to other reducing substances such as cysteine and glutathione. Therefore, the presented sensor can realize the determination of TAC, and then, some actual human serum samples are detected accurately and quickly (the recovery rates are 93.46∼105.61 %).