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Vol. 73, Issue 11, November 2006, pp. 804-809

 

Bullet

 

Novel Fabrication of CA Membrane Bound Carbon Electrode for Bi-enzymatic Determination of Lactate

 

VIKAS1, HARISH2 and D. S. AHLAWAT3*

1Department of Biotechnology, 2Department of Chemistry, 3*Department of Physics,

Ch. Devi Lal University, Sirsa-125055, India

 3*Tel.: 00 91 09215570591; e-mail: technology_for@yahoo.co.in

 

Received: 17 September 2006   /Accepted: 30 November 2006   /Published: 4 December 2006

 

 

Abstract: Lactate oxidase from Pediococcus species has been immobilized onto cellulose acetate (CA) membrane to form an enzymatic membrane. HRP has been incorporated into carbon paste electrode. Enzymatic membrane was mounted over the HRP-carbon paste electrode with the help of dialyses membrane, which acts as working electrode. Lactate biosensor was constructed by connecting this fabricated working electrode to Ag/AgCl reference electrode along with platinum wire auxiliary electrode through potentiostate. The biosensor showed an excellent performance with a linear response range between 5M to 1mML-1 of lactate with a correlation coefficient (r) of .94 for n=30, when compared to standard colorimetric methods. The optimum pH of the biosensor is 6.5 and incubation temperature is 25C. This bi-enzyme electrode can be used for 150 determinants; over 45 days with out any considerable lose of activity, when stored at 4C in 0.5M sodium phosphate buffer (pH 6.5). The response time was 1 second and no major metabolic interference was observed.

 

Keywords: Lactate oxidase, Horseradish peroxidase, Carbon paste electrode, Cellulose acetate membrane

 

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