SCES2433
Electrochemistry
Potentiometry
Group Member :
Daniel Bong Chin Weng SEW120025
Lee Bee Heong
SEW120034
Lim Ee Kim
SEW120035
Lim Pei Hoong
SEW120036
Introduction
According to IUPAC recommendations, potentiometric biosensors have two
features(Koncki, 2007):
i. Biological component is an integral part of the sensor receptor, recognizing an analyte.
ii. The analytical signal generated by the sensor is a potential.
important
A potentiometric biosensor for analysis of an analyte from a fluid sample, comprising(Kost, Lindsay, &
Price, 1995):
iii. an indicating electrode
iv. a reference electrode
v.
a reagent of known amount overlaying a portion of the surface of the indicating electrode and
comprising an enzyme and a redox mediator. The redox mediator being in an amount to react with
all of the analyte at the surface of the indicating electrode, thereby causing a measurable change in
potential that correlates to the change in proportions of the oxidized and reduced forms of the
redox mediator at the surface of the indicating electrode
iv. a potentiometer in electrical connection with the indicating and reference electrodes.
This reaction may be monitored by monitoring the electrical potential at the indicating electrode
surface. Electrical potential at the indicating electrode surface changes as the concentration of oxidized
and reduced forms of the redox mediator change at the indicating electrode surface. The change in
potential that occurs may be correlated to the detection or measurement of analyte in the fluid sample .
Potentiometric Biosensor
Objective:
Develop a low cost screen printed and disposable type urea sensitive enzymatic sensor
system, based on potentiometry, for the detection and estimation of urea levels in milk
sample.
Potentiometric is selected for the purpose as it allows the determination of a wide spectrum
of ions and also as it applies portable and inexpensive equipment.
Sample :
Raw milk samples of caw collected from cattle farms situated around the contryside.
Experimental Set-up
1.
Indicating electrode fabrication
Potentiometry biosensor make use of ion selective electrodes in order to transduce the biological
reaction into electrical signal. Screen printed disposable NH4+ ion selective electrode developed
on the basis of double matrix membrane(DMM) technology as the transducer with a layer of
immobilized urease. For urea detection, the urease enzyme was immobilized by entrapping it in a
polymer matrix on the surface of the NH4+ ion selective membrane electrode.
2.
Reference electrode fabrication
Fabrication of a separate Ag/AgCl reference electrode for use with the working electrode by
adopting a simple thick film screen printing technique.
Figure 1. Schematic of a typical
fabricated working electrode for
potentiometric urea detection.
Principle of sensor operation
The urea sensor operation is based on the enzymatic decomposition of urea by urease:
CO(NH2)2 + H2OUrease CO2 + 2NH3
In the pH region where the enzyme is active (around pH 7), the products of the above
enzymatic reaction dissociate as:
CO2 + H2O HCO3 + H+
NH3 + H2O NH4 + + OH
Thus, it is possible to determine the urea concentration potentiometrically with the pNH4 +
electrode. Urease can be physically entrapped in a polymer gel and placed on the tip of an
electrode, which is sensitive to ammonium ions.
Results and Discussion
Adulteration of milk
The table shows clearly that the fabricated enzyme based biosensors are capable of detecting urea
presence in milk samples and more importantly, the biosensors can detect if there is any adulteration in
milk with urea.
Since urea is one of the ingredients in the manufacturing of Synthetic Milk , the present ammonium
ion sensitive based and urease immobilized screen printed biosensor system can detect the urea levels
and the adulteration of milk.
The present biosensor system adopted low cost and disposable screen printed ammonium ion sensitive
electrodes and the concentrations of the urea detected were found to be in good agreement with those
obtained by the sophisticated and expensive spectrophotometric technique.
References
Koncki, Robert. (2007). Recent developments in potentiometric biosensors for biomedical
analysis. Analytica Chimica Acta, 599(1), 7-15.
Kost, K.M., Lindsay, T.J., & Price, J.F. (1995). Potentiometric biosensor and the method of
its use: Google Patents.
Trivedi, UB, Lakshminarayana, D, Kothari, IL, Patel, NG, Kapse, HN, Makhija, KK, . . .
Panchal, CJ. (2009). Potentiometric biosensor for urea determination in milk. Sensors
and Actuators B: Chemical, 140(1), 260-266.
