Introduction
Enzyme Catalyzed redox reactions The function of the enzyme is to generate or
consume an electroactive species in a stoichiometric relationship with its substrate or target analyte
Amperometric transducer allows for the electrometrical reactions to proceed at the electrode surface, thus giving rise to a current
Introduction
S+C enzyme P + C’S = SubstrateC = Cofactor / CoreactantP = ProductC’ = Cofactor Product
Introduction
Three generations 1st generation: the normal product of the
reaction diffuses to the transducer and causes electrical response
2nd generation: involves specific mediators between reaction and transducer to generate improved response
Amperometric Glucose Biosensor
Developed by Updike and Hicks First reported use of enzyme electrode Glucose in blood-diagnosis of diabetes O2 detection H2O2 detection Enzyme mediation
Glucose Biosensor – O2
Central – Pt cathode, surrounding Ag/AgCl anode, KCl electrolyte solution
Teflon membrane Cathode (0.6V) O2 + 4H+ +4e- 2H2O Anode 4Ag + 4Cl- 4AgCl + 4e-
Glucose +O2 glucoseoxidase Gluconic acid + H2O
Glucose Biosensor – H2O2
Reverse polarity of electrodes for the O2 set up
Replace Teflon membrane with cellulose acetate membrane
Cathode: 2Ag + 2Cl-2AgCl + 2e-
Pt Anode (0.6V): H2O2 O2+2H+ + 2e-
Counter electrode: 4H++O2 2H2O – 4e-
Glucose Biosensor – redox mediators
Replace O2 with synthetic e- acceptor to shuttle e- from the flavin redox center of the enzyme to the electrode
Measurements become insensitive to O2 fluctuations, and can be carried out at lower potentials
Enzyme mediators – dimethyl ferrocene, medola blue
Glucose Biosensor – Redox mediators
Glucose + Gox(FAD) gluconic acid + Gox(FADH2)
Gox(FADH2)+2Mox Gox(FAD)+2Mred+2H+
2Mred 2Mox + 2e-
Common Enzyme electrodesMeasured Species
Enzyme Detected species
Lactate Lactate dehydrgenase
NADH
Lactate Lactate oxidase
H2O2
Phenol Tyrosinase Quinone
Cholesterol Cholesterol oxidase
O2
Some Commercial applications
YSI : Glucose/Lactate Prostate specific antigen (PSA) NOVA Bioprofile analyzer (PO2, Gluc,
Gln, Glu, Lactate)
Other Applications
Glucose: meat freshness, fermentation Rapid cell number monitor Monitor of herbicides in surface waters Amperometric and novel fluorescent
DNA probes Rapid electrochemical sensor for Iron