Amperometry: Principles and Applications

Amperometry is an electrochemical technique in which the current is measured as

a function of time under a constant potential. It is widely used to study redox
reactions, analyze concentrations of analytes, and monitor reaction kinetics.

Amperometry is used in electrophysiology to study vesicle release events using

a carbon fiber electrode. Unlike patch clamp techniques, the electrode used for
amperometry is not inserted into or attached to the cell but brought nearby of the
cell. The measurements from the electrode originate from an oxidizing reaction of
a vesicle cargo released into the medium. Another technique used to measure
vesicle release is capacitive measurements.

It is the electric current measured between a pair of electrodes. The measured

current is directly proportional to the concentration of the analyte. Example: Blood
glucose monitor

Carbon is used as a working electrode which is coated with mediator and glucose
oxidase. Ag/AgCl is used as the reference electrode. The enzyme oxidase catalyzes
the reaction of glucose with oxygen. Hydrogen peroxide concentration is measured
by oxidation which occurs at +0.6V.

H2O2——>O2+2H+ + 2e-

Current is directly proportional to H 2O2 concentration this in turn is directly

proportional to glucose concentration. If O2 is low, complete conversion is
inhibited. Hence mediator ferrocene is introduced. Now, current is directly
proportional to the concentration of ferrocene which in turn is directly proportional
to glucose concentration.
Detection methods

Single-potential amperometry

Any analyte that can be oxidized or reduced is a candidate for amperometric

detection. The simplest form of amperometric detection is single-potential,
or direct current (DC), amperometry. A voltage (potential) is applied between
two electrodes positioned in the column effluent. The measured current changes as
an electroactive analyte is oxidized at the anode or reduced at the cathode. Single-
potential amperometry has been used to detect weak acid anions, such
as cyanide and sulfide, which are problematic by conductometric methods.
Another, possibly more important advantage of amperometry over other detection
methods for these and other ions, such as iodide, sulfite, and hydrazine, is
specificity. The applied potential can be adjusted to maximize the response for the
analyte of interest while minimizing the response for interfering analytes .

Pulsed amperometry (pulsed amperometric detection, PAD)

An extension of single-potential amperometry is pulsed amperometry, most

commonly used for analytes that tend to foul electrodes. Analytes that foul
electrodes reduce the signal with each analysis and necessitate cleaning of the
electrode. In pulsed amperometric detection (PAD), a working potential is applied
for a short time (usually a few hundred milliseconds), followed by higher or lower
potentials that are used for cleaning the electrode. The current is measured only
while the working potential is applied, then sequential current measurements are
processed by the detector to produce a smooth output. PAD is most often used for
detection of carbohydrates after an anion exchange separation, but further
development of related techniques show promise for amines, reduced sulfur
species, and other electroactive compounds.

Principles of Amperometry

1. Faradaic Current: The current observed is primarily due to the transfer of

electrons during oxidation or reduction at the electrode.
2. Electrode Potential Control: A fixed potential is applied to the working
electrode relative to a reference electrode.
3. Mass Transport: The current is influenced by the rate of mass transport of
the analyte to the electrode, involving:
o Diffusion (movement due to concentration gradient)
o Migration (movement due to electric fields)
o Convection (movement caused by stirring or flow).
4. Steady-State vs. Transient Behavior:
o At steady state, the current becomes constant as the system reaches
equilibrium.
o In transient amperometry, the current changes with time due to
evolving reaction conditions.

Electrode Setups in Amperometry

1. Three-Electrode System:
o Working Electrode (WE): Where the reaction of interest occurs.
Common materials: platinum, gold, glassy carbon.
 o Reference Electrode (RE): Maintains a stable potential (e.g.,
Ag/AgCl, saturated calomel electrode).
o Counter Electrode (CE): Completes the circuit and allows current
flow (usually inert materials like platinum).
2. Electrode Configurations:
o Stationary Electrodes: Often used in batch experiments.
o Rotating Disk Electrode (RDE): Enhances mass transport and
ensures uniform analyte diffusion.
o Microelectrodes: Allow for high sensitivity, fast response times, and
reduced solution resistance.

Amperometric Techniques

1. Chronoamperometry

 Definition: The measurement of current as a function of time after a step

change in potential.
 Applications:
o Determining diffusion coefficients.
o Studying reaction kinetics.
o Measuring concentrations of electroactive species.
 Experimental Steps:

1. The potential of the working electrode is stepped from an initial value

to a target value.
2. Current is recorded over time as the electroactive species reacts.

2. Cyclic Voltammetry (CV)

  Definition: A technique in which the potential of the working electrode is
cycled linearly back and forth while recording the resulting current.
 Applications:
o Studying redox processes and reaction mechanisms.
o Determining formal potentials.
o Analyzing electron transfer kinetics.
 Key Features:
o The current response forms characteristic peaks for reduction
(cathodic peak) and oxidation (anodic peak).
o The peak separation (ΔEp\Delta E_pΔEp) gives information on the
reversibility of the reaction.
 Experimental Steps:

1. Potential is ramped from an initial value to a vertex potential and

back.
2. Current is recorded as the potential changes.

Comparison Between Chronoamperometry and Cyclic Voltammetry

Feature Chronoamperometry Cyclic Voltammetry

Potential Linear sweep and reverse
Single-step change
Control potential cycling
Current vs. Current decays over time Current forms peaks based on
Time (Cottrell decay) redox events
Diffusion studies, concentration Reaction mechanism, redox
Applications
analysis studies
Advanced Techniques

1. Hydrodynamic Amperometry:
o Use of a flow system (e.g., flow injection analysis) or rotating
electrodes to enhance mass transport.
2. Differential Pulse Amperometry:
o Involves applying potential pulses for greater sensitivity and
resolution.
3. Square Wave Amperometry:
o Combines potential square waves with amplitude modulation to detect
low concentrations.

Applications of Amperometry

1. Biochemical Analysis:
o Glucose sensors (e.g., glucose oxidase-based amperometric
biosensors).
2. Environmental Monitoring:
o Detection of heavy metals and pollutants.
3. Pharmaceuticals:
o Quantification of active ingredients and drug metabolites.
4. Food Industry:
o Monitoring antioxidants and additives.