An amperometric biosensor based on poly(L-aspartic acid), nanodiamond particles, carbon nanofiber, and ascorbate oxidase-modified glassy carbon electrode for the determination of L-ascorbic acid.

An amperometric L-ascorbic acid biosensor utilizing ascorbate oxidase (AOx) immobilized onto poly(L-aspartic acid) (P(L-Asp)) film was fabricated on carbon nanofiber (CNF) and nanodiamond particle (ND)-modified glassy carbon electrode (GCE). Effects of AOx, ND, and CNF amounts were investigated by monitoring the response currents of the biosensor at different amounts of AOx, ND, and CNF. The electropolymerization step of L-aspartic acid on CNF-ND/GCE surface was also optimized. Scanning electron microscopy (SEM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) techniques were used to enlighten the modification steps of the biosensor. The effects of pH and applied potential were studied in detail to achieve the best analytical performance. Under optimized experimental conditions, the AOx/P(L-Asp)/ND-CNF/GCE biosensor showed a linear response to L-ascorbic acid in the range of 2.0 × 10-7-1.8 × 10-3 M with a detection limit of 1.0 × 10-7 M and sensitivity of 105.0 μAmM-1 cm-2. The novel biosensing platform showed good reproducibility and selectivity. The strong interaction between AOx and the P(L-Asp)/ND-CNF matrix was revealed by the high repeatability (3.4%) and good operational stability. The AOx/P(L-Asp)/ND-CNF/GCE biosensor was successfully applied to the determination of L-ascorbic acid in vitamin C effervescent tablet and pharmaceutical powder containing ascorbic acid with good results, which makes it a promising approach for quantification of L-ascorbic acid. Graphical abstract.