Amperometric creatinine biosensor based on covalently coimmobilized enzymes onto carboxylated multiwalled carbon nanotubes/polyaniline composite film.

A mixture of commercial creatinine amidohydrolase (CA), creatine amidinohydrolase (CI), and sarcosine oxidase (SO) was coimmobilized covalently via N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxy succinimide (NHS) chemistry onto carboxylated multiwalled carbon nanotube (c-MWCNT)/polyaniline (PANI) nanocomposite film electrodeposited over the surface of a platinum (Pt) electrode. A creatinine biosensor was fabricated using enzyme/c-MWCNT/PANI/Pt as working electrode, Ag/AgCl as reference electrode, and Pt wire as auxiliary electrode connected through potentiostat. The enzyme electrode was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and electrochemical impedance spectroscopy (EIS). The biosensor detected creatinine levels as low as 0.1 μM, estimated at a signal-to-noise ratio of 3, within 5s at pH 7.5 and 35°C. The optimized biosensor showed a linear response range of 10 to 750 μM creatinine with sensitivity of 40 μA/mM/cm(2). The fabricated biosensor was successfully employed for determination of creatinine in human serum. The biosensor showed only 15% loss in its initial response after 180 days when stored at 4°C.

[1]  S. Iijima.  Helical microtubules of graphitic carbon , 1991, Nature.

[2]  Kazuaki Sawada,et al.  Development of amperometric glucose biosensor based on glucose oxidase co-immobilized with multi-walled carbon nanotubes at low potential , 2009 .

[3]  Eun Ju Kim,et al.  Disposable creatinine sensor based on thick-film hydrogen peroxide electrode system , 1999 .

[4]  B. J. Venton,et al.  Review: Carbon nanotube based electrochemical sensors for biomolecules. , 2010, Analytica chimica acta.

[5]  T. Smith-Palmer,et al.  Separation methods applicable to urinary creatine and creatinine. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[6]  B. D. Malhotra,et al.  Polyaniline Langmuir-Blodgett film based cholesterol biosensor. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[7]  D. Walsh,et al.  Comparison of electrochemical, electrophoretic and spectrophotometric methods for creatinine determination in biological fluids , 2002 .

[8]  R. Renneberg,et al.  Hydrogel matrix for three enzyme entrapment in creatine/creatinine amperometric biosensing , 1996 .

[9]  S. Khokhar,et al.  Electrochemical creatinine biosensors. , 2008, Analytical chemistry.

[10]  A. V. van Zanten,et al.  Interferences in current methods for measurements of creatinine. , 1991, Clinical chemistry.

[11]  Sandeep Yadav,et al.  An amperometric oxalate biosensor based on sorghum oxalate oxidase bound carboxylated multiwalled carbon nanotubes-polyaniline composite film. , 2011, Journal of biotechnology.

[12]  S. C. Lo,et al.  Glucose interference in Jaffé creatinine method: effect of calcium from peritoneal dialysate. , 1994, Clinical chemistry.

[13]  Jing-Juan Xu,et al.  Direct electrochemistry and electrocatalysis of heme proteins immobilized on gold nanoparticles stabilized by chitosan. , 2005, Analytical biochemistry.

[14]  Kyeongjae Cho,et al.  Thermal Expansion and Diffusion Coefficients of Carbon Nanotube-Polymer Composites , 2002, cond-mat/0203349.

[15]  F. Matzkies,et al.  Amperometric creatinine biosensor for hemodialysis patients. , 2001, Clinica chimica acta; international journal of clinical chemistry.

[16]  C. Kochansky,et al.  Determination of uremic toxins in biofluids: creatinine, creatine, uric acid and xanthines. , 2000, Journal of chromatography. B, Biomedical sciences and applications.

[17]  Richard P. Buck,et al.  Microfabricated amperometric creatine and creatinine biosensors , 1996 .

[18]  Julian W. Gardner,et al.  Sensors and Sensory Systems for an Electronic Nose , 1992 .

[19]  R. L. Elsenbaumer,et al.  Handbook of conducting polymers , 1986 .

[20]  H. Nam,et al.  A planar amperometric creatinine biosensor employing an insoluble oxidizing agent for removing redox-active interferences. , 2001, Analytical chemistry.

[21]  Hsuan‐Jung Huang,et al.  A creatinine deiminase modified polyaniline electrode for creatinine analysis , 1999 .

[22]  W. Wernet,et al.  A highly sensitive amperometric creatinine sensor , 1997 .

[23]  A. Tiwari,et al.  Chitosan-g-polyaniline: a creatine amidinohydrolase immobilization matrix for creatine biosensor , 2009 .

[24]  F. van Lente,et al.  Diagnostic and prognostic utility of phospholipase A activity in patients with acute pancreatitis: comparison with amylase and lipase. , 1991, Clinical chemistry.

[25]  K. Yoda,et al.  Multi-enzyme membrane electrodes for determination of creatinine and creatine in serum. , 1983, Clinical chemistry.