Amperometric Biosensor for D-Lactate Assay

Miroslav Pohanka^1,2* and Petr Zbořil<sup>1

1</sup>Department of Biochemistry, Masaryk University, Kotláøská 2, CZ-611 37 Brno, Czech Republic
²Centre of Biological Defense, Central Military Institute of Health, CZ-561 66 Tìchonín, Czech Republic

Article history:
Received November 15, 2006
Accepted June 11, 2007

Key words:
D-lactate, D-LCR, carbon paste electrode, amperometric biosensor

Summary:
A carbon paste electrode biosensor for D-lactic acid analysis in biological samples was developed. D-lactate cytochrome c oxidoreductase from baker’s yeast (EC 1.1.2.4) was used as biorecognition element of the electrode in combination with phenazine methosulphate (PMS)–Reinecke’s salt precipitate as mediator. The enzyme was bound on paste due to Enzacryl using glutardialdehyde. The electrode response to D-lactate addition was linear in the concentration range of 0.015–100 mM. Limit of detection was equal to 56 μM when signal vs. noise criterium (S/N=3) is considered. Specificity for only D-lactate was proven. Relative standard deviation of 6.6 % for 12 mM D-lactate was found. The biosensor exerts long-term stability and also good stability during repeated measuring cycles. The electrode has a good potential for use in practical samples.

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