Total antioxidant capacity test (ABTS rapid method)

The total antioxidant capacity (T-AOC) of clusterzyme and contrast (trolox and anthocyanin) was determined by the rapid ABTS method using the T-AOC Assay Kit (S0121, Beyotime). Please refer to the specification for specific sampling methods. The antioxidant capacity was evaluated by measuring the absorption value at 414 nm. In the process of reaction kinetic analysis, we adjusted the concentration of different concentration of ABTS^•+ (the molar extinction coefficient of ABTS^•+: ɛ_414 nm = 3.6 × 10⁴ mol⁻¹ cm⁻¹) by changing the concentration of H₂O₂. The reaction kinetic analysis process was reflected by the change of absorbance at 414 nm monitored by the UV-vis spectrophotometer under kinetic mode. The steady-state kinetic parameters were determined by varying the concentration of ABTS^•+ in the presence of clusterzymes (5 ng/µL). The maximum reaction velocity (V_max) and Michaelis–Menten constant (K_m) were calculated using the Lineweaver–Burk equation.