Amperometric biosensor for the determination of creatine.

An amperometric biosensor for the determination of creatine was developed. The carbon rod electrode surface was coated with sarcosine oxidase (SOX) and creatine amidinohydrolase by cross-linking under glutaraldehyde vapour. The SOX from Arthrobacter sp. 1-1 N was purified and previously used for creation of a creatine biosensor. The natural SOX electron acceptor, oxygen, was replaced by an [Fe(CN)(6)](3-) /[ Fe(CN)(6)](4-) redox mediating system, which allowed amperometric detection of an analytical signal at +400-mV potential. The response time of the biosensor was less than 1 min. The biosensor showed a linear dependence of the signal vs. creatine concentration at physiological creatine concentration levels. The optimal pH in 0.1 M tris(hydroxymethyl)aminomethane (Tris)-HCl buffer was found to be at pH 8.0. The half-life of the biosensor was 8 days in 0.1 M Tris-HCl buffer (pH 8.0) at 20 degrees C. Principal scheme of consecutively followed catalytic reactions used to design a biosensor for the determination of creatine.