Electrochemical Determination of Creatinine Based on Multienzyme Cascade-Modified Nafion/Gold Nanoparticles/Screen-Printed Carbon Composite Biosensors

Abstract

Creatinine serves as a crucial diagnostic biomarker for assessing kidney disease. This work developed portable non-enzymatic and multienzyme-modified electrochemical biosensors for the detection of creatinine based on commercial screen-printed carbon electrodes (SPCEs). The non-enzymatic creatinine sensor was constructed by the electrochemical deposition of AuNPs onto the surface of a pre-activated SPCE by electrochemical activation, followed by the surface modification of a Nafion membrane. The developed AuNPs/SCPE exhibited excellent reproducibility, and the proposed Nafion/AuNPs/SPCE sensor showed excellent detection sensitivity and selectivity toward creatinine. In comparison, the enzymatic creatinine biosensor was gradually established by the electrodeposition of a Prussian blue (PB) membrane on the optimal AuNPs/SCPE surface, followed by multi-enzyme cascade modification (which consisted of creatinine amidohydrolase (CA), creatine oxidase (CI) and sarcosine oxidase (SOx)) and drop-casting the Nafion membrane to stabilize the interface. The introduction of a PB interlayer acted as the redox layer to monitor the generation of hydrogen peroxide (H2O2) produced by the enzymatic reaction, while the Nafion membrane enhanced the detection selectivity toward creatine, and the multi-enzyme cascade modification further increased the detection specificity. Both non-enzymatic and enzymatic creatinine sensors could detect the lowest concentrations of less than or equal to 10 μM. In addition, the efficiency and reproducibility of the proposed composite biosensor were also confirmed by repetitive electrochemical measurements in human serum, which showed a positive linear calibration relation of peak currents versus the logarithm of the concentration between 10 μM and 1000 μM, namely, ip (μA) = −7.06 lgC (μM) −5.30, R2 = 0.996. This work offers a simple and feasible approach to the development of enzymatic and non-enzymatic creatinine biosensors.