Analytical Technique

Biochemical application of potentiometric determination

  An electrochemical analysis { a technique that involves using an
electrochemical cell to measure the potential/or current of an analyte} based
on measurement of electrode potentials
 Used to determine the concentration of specific ions in a sample or conc of
a solute in solution by measuring the difference in electrode potentials
between two electrodes in an electrochemical cell
 It is made up of two electrodes (reference and test) inserted in two solutions

What is it? connected by a salt bridge

 A voltmeter is attached to the two electrodes and measures the potential
difference between them
 The test (indicator) electrode: usually a metal immersed in a solution of its
own ions whose concentration is to be determined
 It could also be a carbon rod electrode immersed in a solution of ions of
interest in two different oxidation states
 The reference electrode is that of known potential
  There are many biochemical uses of potentiometric analysis,
the following short list merely illustrating the range.
 Acid-base metabolism ; pH, PC0 2, bicarbonate etc. in blood;
titratable acidity in urine. Gastric analysis .. pH, chloride,
titratable acidity to any selected pH in gastric juice; pH fields
of gastric mucosa.

Uses  Fats ; fatty acids in serum or faecal extracts.

 Miscellaneous serum analyses .. chloride, sugar, copper, zinc,
cholinesterase, etc.
 pHstat ..maintenance of the pH of a solution at a predetermined
value for analytical or other purposes.
  For the estimation of urea, uric acid and glucose
 Involve measuring the potential of an electrode in a system
Potentiometric where the electrode and the solution is in

biosensors
  Differences between direct and indirect potentiometry
 Activities measured cannot be extrapolated readily to the
desired concentration
 A careful and accurate calibration
Factors  A suitable sample preparation
affecting the  Adjustment of the measuring conditions to the characteristics
results of the specimen and the matrix of the sample is necessary
before each measurement
 Hence the need for internal and external quality control
  Glass membrane
 Solid phase
Types of Ion  Fluid membrane
selective  Carrier
sensors  Gas-sensitive and
 Enzyme electrodes with immobilized enzymes