Application of biosensor in urea

determination

Dr. Anushree Tripathi

 Overview
• Why urea determination?
• Urea biosensors
• Types of urea biosensors
 Why urea determination is required?
• Non-toxic
• Major nitrogenous end product of protein
metabolism
• Eliminated from body by kidney through urine
but is also secreted in body fluids such as blood
and saliva.
• Increase in urea level: Uremia
• Dysfunctioning of kidney: End-stage renal disease
(ESRD)
• To diagnose these diseases.
 Conventional methods
• Kidney function tests: Blood urea nitrogen,
glomerular filtration and serum creatinine.
• Complicated, time consuming and expensive
instrumental setup.
 Urea Biosensor

• Based on urease activity, catalyzes hydrolysis of

urea generating ammonium and bicarbonate
ions.
• Generated species influences the pH of the
surrounding environment which is directly
proportional to urea concentration.
• Ammonium ions in traces can be easily detected
by specific transducer.
Electrochemical Conductometric Thermal

Optical Manometric Piezoelectric

 Amperometric urea biosensor
• Urease catalysed reaction produces an
increase in pH which is directly proportional to
urea concentration.
• Biosensors fabricated by Polypyrrole and Poly
ortho-phenylenediamine( PoPD), affected by
pH changes and responded to ammonium
ions.
• Carbon nanotubes entrapment method was
used for construction of biosensors.
 Potentiometric urea biosensor
• Increase and decrease in potential based on
chemical reaction such as hydrolysis of urea
• Modes of detection of urea commonly used in
many biosensors, based on potentiometric
detection of ammonium ion, CO2 or pH changes
generated by enzymatic hydrolysis of urea in
accordance with urea concentration.
• Mostly based on pH detection
• Limitation: strongly depends on buffer capacity of
sample solution.
 Conductometric Urea biosensor
• Detect changes in solution resistance
(reciprocal of conductance)during enzymatic
reaction
• Advantages: Simple to construct, suitable for
miniaturization and mass production and
don’t require reference electrode
• Sol-gel immobilized urease on planar
interdigited electrode array for fabrication of
urea biosensor
 Thermal Urea biosensor
• Not commonly used
• Heat produced during a biochemical reaction
(Urease catalysed hydrolysis)
• Advantages: Principle of specificity
• Disadvantages: Non-specific heat effects from
changes, Sensitivity depends on molar
enthalpy, only more endo or exothermic
reactions are often considered for detection
 Optical Urea biosensors
• Involves immobilization of urease onto different
matrices such as Poly Vinyl Chloride (PVC)
• Limitation: None of the catalytic products
(Ammonia or CO2) or the analyte show optical
properties for adequate detection by optical
sensors. Due to this reason, an indicator dye is
often used to achieve adequate detection.
• Optical urea biosensors which doesn’t require the
addition of dye has been reported based on the
use of a conductive polymer (Polypyrrole).
 Manometric urea biosensors
• Measurement of the total quantity of a gas (NH3
or CO2) produced during the urease catalysed
hydrolysis of urea, have been made by simple
manometric methods.
• These are based on the ideal gas law which states
that volume occupied by a gas is directly
proportional to number of moles of the gas at
constant pressure and temperature.
• Disadvantage: Analysis of discrete samples in a
sealed system.
 Piezoelectric urea biosensors
• Detection of the change in mechanical
characteristics such as mass loading effect and
elastic characteristics due to immunity
reaction on a propagation plane or electrode
can be achieved with an acoustic device.
• Limitation: Acoustic sensor, which normally
detects change in mechanical properties
cannot detect an enzymatic reaction.