Chemically modified carbon nitride-chitin-acetic acid hybrid as a metal-free bifunctional nanozyme cascade of glucose oxidase-peroxidase for "click off" colorimetric detection of peroxide and glucose.

Hybrid nanomaterials-based artificial enzymes with numerous utilities are necessary to develop future bionic devices in mimicking physiological processes. This paper demonstrates bifunctional enzyme mimicking roles of a metal-free nanozyme hybrid of chemically modified graphitic carbon nitride (MGCN), chitin and acetic acid (AcOH). The MGCN exhibited glucose oxidase-mimicking activity and chitin-AcOH mirrored peroxidase. MGCN-chitin-AcOH when in contact with glucose, oxidised glucose to gluconic acid and hydrogen peroxide (HO) while the chitin-AcOH decomposed the generated HO, as proved separately, by concurrent oxidation of 3,3',5,5'-tetramethylbenzidine (TMB). The super-sensitive colorimetric process produced linear regression equation for HO as A = 0.00105C + 0.0630 (C:μM, R = 0.9961) with a detection limit of 0.052 μM, whereas for glucose, the linear relationship was A = 0.00084C + 0.0458 (C:μM, R = 0.9952) having a detection limit of 0.055 μM. The developed method was also successfully applied for assessment of HO and glucose in human serum and urine samples. Non-enzymatic glucose test strips from MGCN-chitin-AcOH based hydrogel were reported and verified for semi-quantitative analysis of glucose. These compared well with results from standard enzyme-based colorimetric procedure. The developed hybrid nanozyme provided feasible alternatives to the two natural enzymes (peroxidase and glucose oxidase) realized through real sample analysis. The developed hybrid nanozyme can be successfully used for colorimetric detection of peroxide and glucose in medical diagnostics.