An improved amperometric creatinine biosensor based on nanoparticles of creatininase, creatinase and sarcosine oxidase

• Published in: Analytical biochemistry Vol. 537; pp. 41 - 49
• Main Authors: Kumar, Parveen, Jaiwal, Ranjana, Pundir, C.S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.11.2017
• Subjects: Amidohydrolases - chemistry; Amidohydrolases - metabolism; Ascorbic Acid - chemistry; Biosensing Techniques - instrumentation; Biosensor; biosensors; carbon; colorimetry; Creatinine; Creatinine - blood; detection limit; Dielectric Spectroscopy; Electrochemical Techniques - instrumentation; electrochemistry; Electrodes; Enzyme nanoparticles; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Fourier transform infrared spectroscopy; Glassy carbon electrode; Gold - chemistry; Humans; Limit of Detection; Metal Nanoparticles - chemistry; Microscopy, Electron, Scanning; nanoparticles; people; Sarcosine Oxidase - chemistry; Sarcosine Oxidase - metabolism; scanning electron microscopy; Serum; silver; silver chloride; sodium phosphate; transmission electron microscopy; Ureohydrolases - chemistry; Ureohydrolases - metabolism; Uric Acid - chemistry; Creatinine; Glassy carbon electrode; Serum; Enzyme nanoparticles; Biosensor
• ISSN: 0003-2697; 1096-0309; 1096-0309
• DOI: 10.1016/j.ab.2017.08.022

An improved amperometric biosensor for detection of creatinine was developed based on immobilization of nanoparticles (NPs) of creatininase (CA), creatinase (CI), and sarcosine oxidase (SOx) onto glassy carbon (GC) electrode. Transmission electron microscopy (TEM) and fourier transform infrared spectroscopy (FTIR) were employed for characterization of enzyme nanoparticles (ENPs). The GC electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectra (EIS) at different stages of its amendment. The biosensor showed optimum response within 2s at pH 6.0 in 0.1 M sodium phosphate buffer and 25 °C, when operated at 1.0 V against Ag/AgCl. Biosensor exhibited wider linear range from 0.01 μM to 12 μM with a limit of detection (LOD) of 0.01 μM. The analytical recoveries of added creatinine in sera were 97.97 ± 0.1% for 0.1 mM and 98.76 ± 0.2% for 0.15 mM, within and between batch coefficients of variation (CV) were 2.06% and 3.09% respectively. A good correlation (R2 = 0.99) was observed between sera creatinine values obtained by standard enzymic colorimetric method and the present biosensor. This biosensor measured creatinine level in sera of apparently healthy subjects and persons suffering from renal and muscular dysfunction. The ENPs electrode lost 10% of its initial activity within 240 days of its regular uses, when stored at 4 °C.