An Amperometric Enzyme Electrode for Creatine Determination Prepared by the Immobilization of Creatinase and Sarcosine Oxidase in Poly(vinylferrocenium)

Abstract

A new enzyme electrode for the determination of creatine was developed by immobilizing creatinase (CI) and sarcosine oxidase (SO). The enzymes were co-immobilized in a poly(vinylferrocenium) matrix onto the surface of a platinum working electrode. Crosslinking with glutaraldehyte (GA) and bovine serum albumin (BSA) was selected as the best immobilization method for the enzymatic system. Determination of creatine was performed by the oxidation of enzymatically generated H₂O₂ at + 0.7 V vs. Ag/AgCl. The linear working range of the electrode was 2.0 × 10⁻⁵ − 3.2 × 10⁻⁴ M and the response time was about 50 s. The effects of pH, temperature, enzyme ratio and buffer concentration were investigated and optimum parameters were found to be 7.5, 37°C, 2.5:1 (CI:SO) and 0.05 M, respectively. The stability and reproducibility of the enzyme electrode have been also studied.

Keywords:

• Creatine
• Enzyme electrode
• Amperometry
• Poly(vinylferrocen)
• Creatinase
• Sarcosine oxidase