Vanadium-Doped Porous Cobalt Oxide for Its Superior Peroxidase-like Activity in Simultaneous Total Cholesterol and Glucose Determination in Whole Blood Based on a Simple Two-Dimensional Paper-Based Analytical Device.

Vanadium-doped porous Co₃O₄ (V-porous Co₃O₄) was synthesized via a simple soft-templating method and used as a superior peroxidase mimic for the simultaneous colorimetric determination of glucose and total cholesterol (TC) in whole blood samples on a two-dimensional microfluidic paper-based analytical device (2D-μPAD). The large surface area and the presence of two metals in V-porous Co₃O₄ contributed to its excellent catalytic activity toward 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 3,3',5,5'- tetramethylbenzidine (TMB) with Michaelis-Menten constants (<i>K</i>_M) of 0.1301 and 0.0141 mM, respectively. The 2D-μPAD was fabricated using simple wax screen-printing and cutting techniques. The colorimetric reactions of both glucose and TC on 2D-μPAD were simultaneously performed by adding a single drop of a whole blood sample on the sample zone made of the LF1 membrane. After the enzymatic reactions, the generated hydrogen peroxide (H₂O₂) was oxidized by V-porous Co₃O₄ to produce hydroxy radicals (^•OH), inducing ABTS and TMB to generate colored products. The generated H₂O₂ was proportional to the intensities of the green and blue products of the glucose and TC systems, respectively. The developed 2D-μPAD required a short analysis time (∼5 min) with small volumes of samples (15 μL of whole blood) whereby no sample preparation was needed. Owing to several advantages including simplicity, low cost, long-term stability, and simultaneous readout, the novel V-porous Co₃O₄ coupled with 2D-μPAD proved to be promising for practical uses as a pioneering portable device for the determination of glucose, TC, and other important biomarkers without the need of technical supports.