Impedimetric Paper-Based Enzymatic Biosensor Using Electrospun Cellulose Acetate Nanofiber and Reduced Graphene Oxide for Detection of Glucose From Whole Blood

The excellent characteristics of nanocellulose fiber-based papers have made them one of the most attractive and innovative materials for the development of electrochemical paper-based analytical devices (ePADs). Here, the authors describe a new ePAD based on cellulose nanofibers (CNs) for the determination of glucose concentration from whole blood samples. Cellulose acetate (CA) nanofibers were prepared by the electrospinning method. Then, the paper layer regenerated to cellulose by deacetylation in alkaline solution. To obtain a smooth and continuous CNs layer, it was treated with trimethyl chitosan (TMC). The formation of CA layer, its alkaline regeneration and TMC modification was confirmed by atomic force microscopy, scanning electron microscopy, Fourier-transform infrared spectroscopy, and energy dispersiveX-ray. Screen-printed three electrodes (SPEs) were fabricated by sputtering a thick layer of gold (Au) on the TMC/CNs substrate. Subsequently, reduced graphene oxide (rGO) was used to modify the surface of the working electrode. The fabrication steps of rGO-Au-SPE were characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscop. The results showed that design, construction and modification process was well achieved. Next, glucose oxidase is successfully immobilized onto the regenerated CNs layer which has been previously located in the sensing area of the rGO-Au-SPEs. The ePAD glucose assay exhibited high sensitivity of 9.9 × 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">-4</sup> KΩ <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">-1</sup> · mM <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">-1</sup> to glucose in the range 3.3 - 27.7 mM (R2 = 0.99) with a detection limit of 0.1 mM, high reproducibility (RSD = 0.57-1.59%) and outstanding selectivity as well as stability. This biosensing motif represents a general platform for the analysis of other biomarkers.