INSTRUMENTAL ANALYSIS (I)

INTRODUCTION
Classification of Analytical Methods
Qualitative instrumental analysis is that measured property indicates presence of analyte
in matrix
Quantitative instrumental analysis is that magnitude of measured property is
proportional to concentration of analyte in matrix
Species of interest: All constituents including analyte and Matrix-analyte (concomitants)
Often need pretreatment - chemical extraction, distillation, separation, precipitation
(A) Classical:
Qualitative - identification by color, indicators, boiling points, odors
Quantitative - mass or volume (e.g. gravimetric, volumetric)
(B) Instrumental:
Qualitative - chromatography, electrophoresis and identification by measuring physical
property (e.g. spectroscopy, electrode potential)
Quantitative - measuring property and determining relationship to concentration (e.g.
spectrophotometry, mass spectrometry) Often, same instrumental method used for
qualitative and quantitative analysis

Types of Instrumental Methods

Property
Radiation emission

Example Method
Emission spectroscopy - fluorescence,
phosphorescence, luminescence
Radiation absorption
Absorption spectroscopy - spectrophotometry, photometry,
nuclear magnetic resonance, electron spin
resonance
Radiation scattering
Turbidity, Raman
Radiation refraction
Refractometry, interferometry
Radiation diffraction
X-ray, electron
Radiation rotation
Polarimetry, circular dichroism
Electrical potential
Potentiometry
Electrical charge
Coulometry
Electrical current
Voltammetry - amperometry, polarography
Electrical resistance
Conductometry
Mass
Gravimetry
Mass-to-charge ratio
Mass spectrometry
Thermal
Thermal gravimetry, calorimetry
Rate of reaction
Stopped flow, flow injection analysis
Radioactivity
Activation, isotope dilution
(Often combined with chromatographic or electrophoretic methods)

Example:
Spectrophotometry
Instrument:
Stimulus:
Analytical response:
Transducer:
Data:
Data processor:
Readout:
Data Domains:

spectrophotometer
monochromatic light energy
light absorption
photocell
electrical current
current meter
meter scale
way of encoding analytical response in electrical or
non-electrical signals.

Interdomain conversions transform information from one domain to another.

Detector (general): device that indicates change in environment

Transducer (specific): device that converts non-electrical to electrical data
Sensor (specific): device that converts chemical to electrical data
Non-Electrical Domains

Electrical Domains

Physical (light intensity, color)

Chemical (pH)
Scale Position (length)
Number (objects)

Current
Voltage
Charge
Frequency
Pulse width
Phase
Count
Serial
Parallel

Time - vary with time (frequency, phase, pulse width)

Analog - continuously variable magnitude (current, voltage, charge)
Digital - discrete values (count, serial, parallel, number*)

Digital Binary Data

Advantages (1) easy to store (2) not susceptible to noise
How to choose an analytical method? How good is measurement?

How reproducible? - Precision

How close to true value? - Accuracy/Bias
How small a difference can be measured? - Sensitivity
What range of amounts? - Dynamic Range
How much interference? - Selectivity
Precision - Indeterminate or random errors

Accuracy - Determinate errors (operator, method, instrumental)

Sensitivity

(larger slope of calibration curve m, more sensitive measurement)

Detection Limit
Signal must be bigger than random noise of blank

From statistics k=3 or more (at 95% confidence level)

Dynamic Range
At detection limit we can say confidently analyte is present but cannot perform reliable
quantitation
Level of quantitation (LOQ): k=10
Limit of linearity (LOL): when signal is no longer proportional to concentration

Selectivity:
No analytical method is completely free from interference by concomitants. Best method
is more sensitive to analyte than interfering species (interferent).

k's vary between 0 (no selectivity) and large number (very selective).
Calibration methods
Basis of quantitative analysis is magnitude of measured property is proportional to
concentration of analyte

Calibration curves (working or analytical curves)

Calibration expression is

Absorbance = slope [Analyte (ppm)] + intercept

SPECTROCHEMICAL TECHNIQUES
INTRODUCTION
Methods of analysis based on the interaction of LIGHT with matter. LIGHT
is an Electromagnetic (EM) radiation:

Speed of light, c = 2.99782 x 108 m / s

.Frequency is always fixed but velocity can vary!

-34

H = Planck's Constant = 6.626x10

3. Atoms, ions and molecules exist in certain energy states only
0
E = ground state
1
2
3
E , E , E = excited states
Excitation can be electronic, vibrational or rotational
Energy levels for atoms, ions or molecules different.

Js

2. When an atom, ion or molecule changes energy state, it absorbs or emits

energy equal to the energy difference
1

E = E E

3. The wavelength or frequency of radiation absorbed or emitted during a

transition is proportional to E

EMISSION SPECTRA
Have you noticed the bright yellow light emitted when crystals of NaCl fall in the
flame in your cooker at home? This is emission

Plot of emission intensity vs. or called emission spectrum

Atom:
line emission spectra Inner shell (core) electrons (1s2p) x-rays photons Outer
shell (valence) electrons (3d4p) UV/vis photons

Molecule:
vibrational and rotational transitions - band emission spectra

Continuum Spectra:
A piece of iron in the fire: first become red then white, Why?
Very broad band spectra in Emission from solids produced
radiation thermal excitation and relaxation of many vibrational
(and rotational) levels.

by blackbody

Blackbody Spectrum
Absorption Spectra
Plot of Absorbance vs. or called absorption spectrum just as in emission
spectra an atom, ion or molecule can only absorb radiation if energy matches
separation between two energy states

Atoms:
No vibrational or rotational energy levels - sharp line spectra with few features
For example:
Na 3s 3p 589.0, 589.6 nm (yellow)
Na 3s 5p 285.0, 285.1 nm (UV)
Visible enough energy for valence (bonding) excitations
UV and x-ray enough energy for core (inner) excitations
Molecules:
Electronic, vibrational and rotational energy levels - broad band spectra with
many features
E = Eelectronic + Evibrational + Erotational
For each electronic state - many vibrational states
For each vibrational state - many rotational states
many features
Absorption spectra affected by
(1) number of atoms in molecule
more features

(2) solvent molecules

blurred features

Effect of Chemical State

Emission
produces emission at
wavelength as absorption
(common for atoms)
Excitation methods:
EM radiation

same

SPECTROSCOPY AND INSTRUMENTATION

(IR, visible and UV)

Examples:
T=1.00 (100 %T), A=0.00
T=0.10 (10 %T), A=1.00
T=0.001 (0.1 %T), A=3.00
Determination of concentration from Absorbance measurements;
Absorbance is directly proportional to the concentration, c, and the path length, b,
of the absorbing species; BEER-LAMBERT'S LAW:

A c and A b
so A b c
A = a b c
proportionality constant, a = absorptivity - units (L/gcm)
If units of concentration are M (mol/L) then use molar absorptivity

Absorbance is additive

in a mixture of 2 components

So what is the BeerLambert law used to calculate?

The molar absorption coefficient, and max which is the wavelength at which
maximum absorption occurs.
PHOTOMETRIC ERROR
At low absorbance values,

I (incident) I (transmitted)
Thus, error is relatively large.
At high absorbance values,
I (transmitted) too small
not measured accurately.
Relative error in A vs %T of KMnO4
solutions at 523 nm where = 2400.
Lowest error needs %T = 38,6% = A value 0.43 A.
Working is possible within -/+ 15% which is absorbance 0.2 0.8 as best
measurement range.
INSTRUMENTS:

(1) Radiation Sources:

Tungsten Lamp
== Visible;
Deuterium lamp
== UV;
Globar Silicon Carbide
== IR.
(2) Sample Containers and Optics:
Cuvettes
Made of Glass 400-3000 nm (vis-near IR)
Silica/quartz 200-3000 nm (UV-near IR)
NaCl 200-15,000 nm (UV-far IR)
(3) wavelength selector
Filters:
Absorption filter - colored glass or dye between two glass plates;
Interference filter two thin sheets of metal sandwiched between glass plates,
separated by transparent material. Interference for transmitted wave through 1st
layer and reflected from 2nd layer
Monochromators:
consists of Entrance slit, Collimating lens or mirror; Dispersion element (prism or
grating), Focusing lens or mirror and Exit slit
Typical Prism Monochromator

(4) radiation detector

(A) Photovoltaic cells
(B) Phototube: radiation falls on a sensitive
Plate which produces electron.

(C) Photomultiplier tube (PMT)

(D) Thermal detectors

sensitive to IR (>750 nm)

(5) Signal processor and readout

Double Beam systems:
a. Light from the source is segmented by a rotating segmented mirror which
allows radiation through to the sample and reflects it through the reference
cell alternately.
b. The two beams are recombined by a rotating mirror in phase with the first
rotating mirror.
c. The combined beam is allowed to fall on the detector in which the
reference signal is electronically subtracted from the sample signal before
it is sent to the recording system.

Wavelengths of colors of visible light

Applications of UV-Vis Spectrometry:

UV/VIS IS USED FOR QUALITATIVE AND QUANTITATIVE DETERMINATIONS

Organic molecules
(bonding) molecular orbital as in

n (non-bonding) atomic orbital as in

Absorption of UV light energy cause transition of (bonding electrons) and n

(non-bonding) to * (Antibonding) molecular orbital

Inorganic Ions
Most transition metal ions are colored (absorb in UV-vis) due to d d electronic
transitions. Inorganic ions can be quantitatively determined by spectrophotometry
by reaction with some ligands to produce colored complexes which have clear
absorption in the visible region.

Remember:
Solution absorbs red appears blue-green
Solution absorbs blue-green appears red
Ligand Field Strengths:
Chromium can give different colors and hence absorb at different wavelengths
when reacts with different ligands.

Vis

UV
2-

I-<Br-<Cl-<F-<OH-<C2O4 ~H2O<SCN-<NH3<en<NO2-<CN
"Spectrochemical Series"
Chemical kinetics of a chemical reaction,
Reactant or product absorbs radiation at different wavelength. Measuring A with
time. Follow the formation of the product or the removal of the reactant from
solution.

Absorption of light by biological molecules: Chlorophyll a solutions absorb

blue and red light and are green in color. DNA solutions absorb light in the
ultraviolet and are colorless. Oxyhemoglobin solutions absorb blue light and are
red in color.
Solvent Effects:
Solvent effects mean UV-Vis not reliable for qualitative but excellent for
quantitative analysis.
For good analysis the sample must have these properties:
1. Stability in solution;
2. Adherence to Beer's law,
3. Large molar absorptivities,
4. Sufficient separation of the desired analyte absorbance wavelength from
interfering substances.
If Not, the substance is usually converted into a new species suitable for
quantitative spectrophotometry.
SAMPLE + CHROMOGENIC REAGENT UV-VIS ABSORBING PRODUCT

Example:
In the determination of iron we convert it into iron(III)-thiocyanate complex,
distinguished red color. We follow the procedure:
1. Pipet 5 mL of the stock Fe+3 solution (~0.001 M) and 3 mL of saturated
NH4SCN into a clean 100 mL volumetric flask and dilute to volume.
2. Place some of the solution in the cuvett of the UV/Vis;
3. Scan the solution with the UV-VIS (Change the wavelength at certain
speed).
4. Record the wavelength of maximum absorbance.
5. Measure the absorbance,
-3
6. Make 4 - 5 standard concentration solutions of Fe(III) (1.0 X 10 M to
-5
1.0 X 10 M and measure absorbance at the wavelength of maximum
absorbance.
7. Plot absorbance versus concentration.

8. Treat unknown sample solutions in the same way (Thiocyanate);

9. Measure the absorbance of unknown sample solutions.
10. Refer these absorbance values to their related concentrations in the
calibration graph.

ATOMIC ABSORPTION SPECTROMETRY AAS

Atomic absorption spectrometry (AAS) is an analytical technique that measures
the concentrations of metals. It is so sensitive that it can measure down to parts
per billion (g/L) in a sample. The technique makes use of the wavelengths of
light specifically absorbed by an element. The method is based on the absorption
of radiation by free atoms. Atomic absorption spectrometry has many uses:
Clinical analysis. Analysing metals in biological fluids such as blood and urine.
Environmental analysis. Monitoring our environment eg finding out the levels
of various elements in rivers, seawater, drinking water, air, petrol and drinks
such as wine, beer and fruit drinks.

Pharmaceuticals. In some pharmaceutical manufacturing processes, minute

quantities of a catalyst used in the process (usually a metal) are sometimes
present in the final product. By using AAS the amount of catalyst present can be
determined.
Industry. Many raw materials are examined and AAS is widely used to check
that the major elements are present and that toxic impurities are lower than
specified eg in concrete, where calcium is a major constituent, the lead level
should be low because it is toxic.
Thus, the most important step is to produce free atoms of metals (M) from
sample solution (or Solid)

ATOMIZATION:
Flames
premix burner;
sample (12 mL) + oxidant + fuel :
air + acetylene; 24002700 K
acetylene + nitrous oxide;
29003100 K, refractory
elements

Nebulization; aerosol (narrow size distribution

and smallest sizes are desirable) rich flame;
excess C tends to reduce MO and MOH
into M lean flame; excess oxidant, hotter
need careful optimization
Flame aspiration
Typical burner and spray chamber. Aflame of
Acetylene/air (giving a temperature of 2200
2400 C) or Acetylene /dinitrogen oxide
(2600 2800 C) are often used. A flexible
capillary tube connects the solution to the
nebuliser. At the tip of the capillary, the
solution is nebulised ie broken into small
drops. The larger drops fall out and drain
off while smaller ones vaporise in the flame.
Only ca 1% of the sample is nebulised.

 simplest atomization
laminar flow burner
best for reproducibility (precision) (<1%)
low sensitivity, short time in beam

Electrothermal Atomizers:
For improved sensitivity atomization is
performed by a graphite furnace in
which a drop (1 100 L) of the sample
solution is placed. The tube is heated
electrically by passing a current
through it in a pre-programmed series
of steps:
30 40 sec at 150 C (Drying)
30 sec at 600 C (Ashing and charring of
organic material and convert sample
into oxides at fast heating rate
510 sec at 2000 2500 C (atomization)
To produce free atoms
Finally heating the tube to a still higher temperature ca 2700 C cleans it
ready for the next sample. During this heating cycle the graphite tube is flushed
with argon gas to prevent the tube burning away. Inelectrothermal atomisation
almost 100% of the sample is atomised. This makes the technique much more
sensitive than flame AAS.
Light source: Hollow Cathode Lamp, Each Element has specific Lamp.

300 V applied between anode (+) and metal (Na, Fe, Mn, ) cathode (-)
Ar ions bombard cathode and sputter cathode atoms
Fraction of sputtered atoms excited, then fluoresce

INSTRUMENTAL SET UP

Hydride generator
Some elements form volatile hydrides LIKE As, Se, Te, and Sn. They are useful
to improve sensitivity of AAS determination of these elements. The sample
solution reacted with NaBH4 in acid medium.
As3+ + NaBH4 AsH3 (Arsenic trihydride, Arsine)
The liberated hydride is transported to an atomizer by an inert carrier gas or by
other means. The advantage of this technique over flame AAS is the separation
and enrichment of the element to be determined and the significantly lower
detection limit resulting from greater efficiency of sample introduction. This
method may have more interference effects than the flame AAS system.

Mercury is converted first into Hg (II) and then allowed to react with a reducing
agent such as SnCl2 to produce elemental Hg. Air or nitrogen is allowed to pass
through the solution to carry the hg produced into the absorption cell (glass tube
with quartz windows placed in the radiation path and AA is measured.
TYPES OF INTERFERENCE
Spectral interference; overlapping signal from absorption or emission of
unwanted species (molecules or radicals)
Correction: choose another wavelength or use D2 lamp for flame signals
Ionization:
Higher atomization temperature cause ionization of analytes like Na or K;

Na Na+ + e
Correction: add ionization suppresser, easily ionizable atoms like CsCl (to
increase electrons in the flame to reverse the direction of ionization reaction of
the analyte ionization, Le Chteliers principle.
Atomic emission spectroscopy is very sensitive to temperature fluctuations.
Atomic absorption spectroscopy is less sensitive to temperature fluctuations.
Chemical interference;
Some metals cannot be determined in the presence of other species due to the
formation of thermally stable compounds and can not be atomized.
Example:
Ca can not be determined in the
presence of phosphate, because of
the formation of refractory calcium
phosphate and decreases the atomization
of Ca and the amount of atoms produced
are very low.
Correction:
A releasing agents is added like La3+,
salt solution LaCl3 to allow chemical exchange
Ca3(PO4)2 + 2LaCl3 3CaCl2 + 2LaPO4
CaCl2 is easily atomized

EDTA may also be used for other metals. Rich flame may also be used to give
high temperature able to atomize refractory materials.
Matrix modification
With electrothermal atomization, chemical modifiers can be added which react
with an interfering substance in the sample to make it more volatile than the
analyte compound. This volatile component vaporizes at a relatively low
temperature and is removed during the low and medium temperature stages of
electrothermal atomization.
Example:
For the determination of heavy metals in tobacco leaves, 0.5 1.0 g of the
tobacco leaves are digested with a mixture of 1 : 10 HClO4 : HNO3 until
complete dissolution. The resulted solutions are diluted into 100 mL with distilled
water, in volumetric flasks. The instrument is calibrated by analyzing standard
solutions of the metals: Na, K, Ca, Mg, Cu, Mn, Fe, Ni, Co and Zn. The standard
solutions are prepared by serial dilution from a 1000 ppm (mg/mL). A calibration
graph is prepared by aspiration of the standard solutions (1, 2, 4, 10 and 20 ppm)
and plotting the absorbance versus concentration. The sample solutions are
thensamples When the concentration is very low, preconcentration procedures
are applied like solvent extraction, evaporation. When the matrix is complex the
technique of standard addition is used.
Double beam spectrometers are successfully used in AAS.
SAMPLE PREPARATION
Sample preparation is often simple, and the chemical form of the element is
usually unimportant. This is because atomisation converts the sample into free
atoms irrespective of its initial state. The sample is weighed and made into a
solution by suitable dilution. Elements in biological fluids such as urine and blood
are often measured simply after a dilution of the original sample

CALIBRATION
Standard solutions are measured first
Aspirated and their AA signals are measured
The unknown sample solution is then
Analysed. The value of the unknown is used
To determine the concentration of the metal
in the unknown solution.

STANDARD ADDITION
In some cases, the composition of the
sample solution is very complicated and
the expected interferences are high. Here
a known amounts of the standard solution
are added to the sample solution. The AA is
measured for the untreated sample solution
and those treated with the standard added to
them. A calibration graph of the type shown
in the figure. The concentration of the sample
is represented by the negative intercept on the
concentration axis.

INFRARED SPECTROMETRY
~750-3000 nm
A spectrophotometric method for the determination of composition and structure
of compounds depending on the absorption of radiation in the INFRARED
-6
region of wavelengths, 2 50 m (m = 10 m). It is preferable to express the
region by frequency units:
-1
Wave number = Number of waves in one cm; 1 cm = 10000 / wavelength.

Energy of IR cause vibrational or rotational excitation. Molecule must have

change in dipole moment due to vibration or rotation to absorb IR radiation.
Molecules with permanent dipole moments

() are IR active

HCl, H2O, NO
IR active

Atoms, O2, H2, Cl2

IR inactive

Types of Molecular Vibrations:

Stretch: change in bond length symmetric

asymmetric

Bend: change in bond angle: scissoring, wagging; rocking, twisting/torsion

Example in (C-O)

methanol 1034 cm-1

ethanol 1053 cm-1
butanol 1105 cm-1

INSTRUMENTATION:
Dispersive Grating IR Instruments:
Similar to UV-Vis spectrophotometer, BUT sample after source and before
monochromator in IR (sample after monochromator in UV-Vis - less incident
light)
Double beam is used and useful to eliminate atmospheric gas interference.
RADIATION SOURCES
Nernst Glower heated rare earth oxide rod
(~1500 K) 1-10 m

DETECTORS
Photoconducting PbS, HgCdTe light sensitive, resistance thermometer at
77 K
fast and sensitive

Now Fourier Transform (FTIR) Instruments

The main component in the FTIR spectrometer is an interferometer. This device
splits and recombines a beam of light such that the recombined beam produces a
wavelength-dependent interference pattern or an interferogram. The Michelson
interferometer is most commonly used.

Computer needed to turn complex signal into spectrum. They have two
advantages:
(1) Few optics, no slits == high intensity
(2) Simultaneously measure all spectrum at once saves time
APPLICATIONS
IR (especially FTIR) very widely used for qualitative analysis of gases, liquids
and solids
IR mostly used for rapid qualitative but not quantitative analysis
Sample preparation
Liquids are placed in liquid cell of short pathlength (0.015-1 mm). Solids are
dissolved in a solvent.
Solids (2 - 4 mg) is grinded and mixed with KBr to make semi-transparent pellet;
or: grind and mix with Nujol (hydrocarbon oil) to form mull. One drop is placed
between NaCl plates.
water (attacks windows)
Qualitative Analysis:
Step One Identify functional groups (group frequency region)
Step Two Compare with standard spectra with these groups. (fingerprint region)

Group Frequencies:
Approximately calculated from masses and spring constants
Variations due to coupling
Compared to correlation charts/databases

CHROMATOGRAPHIC SEPARATION
Introduction:
Chromatography is a separations method that relies on differences in partitioning
behavior between a flowing mobile phase and a stationary phase to separate the
the components in a mixture.
A column (or other support for TLC, see below) holds the stationary phase and the
mobile phase carries the sample through it. Sample components that partition
strongly into the stationary phase spend a greater amount of time in the column
and are separated from components that stay predominantly in the mobile phase
and pass through the column faster. The sample is separated into zones or bands
Elution Chromatography:
flushing of sample through column by continual mobile phase
(eluent) addition
migration rate fraction time spent in mobile phase
Planar chromatography - flat stationary phase, mobile phase
moves through capillary action or gravity
Column chromatography - tube of stationary phase, mobile phase
moves by pressure or gravity
Gas Chromatography
A method for the separation of components (mainly organic compounds) of a
homogeneous mixture and to determine the composition of the mixture.
It is based on:
- Injection of sample mixture into a column packed with certain materials
(stationary phase);
- Passing a fluid/gas (the mobile phase) through the column.
- Components of the mixture held strongly or weakly to the stationary phase.
- Mobile phase pass to other end of the column taking away the weakly bound
component first to the outlet of the column. Thus, the components will be
separated according to the boiling point, molecular weight, polarity, ability to for
hydrogen bonding with the stationary phase. The components will appear et the
end of the column at different times, tR , the retention time.
Two major types
Gas-solid chromatography -- (stationary phase: solid)
Gas-liquid chromatography -- (stationary phase liquid that is fixed or immobilized
on a certain support material)
Retention Volume, VR: The volume of the carrier gas which succeed to separate
the component.

F = average volumetric flow rate (mL/min)

F can be estimated by measuring flow rate exiting the column using soap bubble
meter (some gases dissolving in soap solution)
- often scales with vapor pressure (constant polarity analytes)
GC Instrumentation
Carrier gas:
He (common), N2, H2
F=25-150 mL/min packed column
F=1-25 mL/min open tubular column
Column:
2-50 m coiled stainless steel/glass/Teflon
Oven:

0-400 C ~ average boiling point of sample accurate to <1 C

Detectors:

FID, TCD, ECD, (MS)

Sample injection:
Sample in liquid form is injected using microsyringe. The injector must be kept at
some high temperature to change into vapour phase for easy analysis.
- direct injection into heated port (>Toven)
(i) 1-20 L packed column
(ii) 10-3 L capillary column

Carrier gas is allowed to enter the system after the injection port. This will lead the
vapours of the sample to move through the column. To make sure that the
components reached the end of the column (separated), a detector system must
be attached to the end of the column.
GC Detectors: for reasonable GC analysis the detector need to be:
Sensitive (10-8-10-15 g solute/s)
Operate at high T (0-400 C)
Stable and reproducible
Linear response
Desire
Wide dynamic range
Fast response
Simple (reliable)
Nondestructive
Uniform response to all analytes
Flame Ionization Detector (FID):
The separated components are allowed to enter a flame (Hydrogen/air) to be
burned and changed into radicals which will be ionized by the flame. The ions will
be detected by electrical conduction to give signal that a component is separated.
The FID is:
Sensitive (10-13 g/s)
Wide dynamic range (107)
Weakly sensitive to carbonyl, amine, alcohol, amine groups
Not sensitive to non-combustibles - H2O, CO2, SO2, NOx
Destructive
Thermal Conductivity Detector (TCD)
The conduction of heat from certain
filament by carrier gas (mobile phase)
He N2 or H2, will be lowered when the
organics reached the detector because
the thermal conductivity of them is lower
than that of the carrier gas. Thus, the
organics will cause Temperature rise in
filament. The TCD is characterized by:
Wide dynamic range (105)
Nondestructive
Insensitive (10-8 g/s) - nonuniform

Column Stationary Phases:

Packed
liquid coated silica particles
(<100-300 m diameter) in glass
tube
best for large scale but slow and
inefficient.
Immobilized Liquid Stationary
Phases:
low volatility and thermally stable
chemically inert (reversible
interactions with solvent)
chemically attached to support
(prevent "bleeding")
Many based on polysiloxanes or polyethylene glycol (PEG):

Some common Stationary phases for gas liquid chromatography

Stationary phases usually bonded by covalent linking to support and/or cross-linked by

polymerization reactions after bonding to join individual stationary phase molecules
non-polar analytes retained preferentially on Non-polar stationary phases
polar analytes retained preferentially on Polar stationary phases

Capillary/Open Tubular
wall-coated (WCOT) <1 m thick liquid coating on inside of silica tube
support-coated (SCOT) 30 m thick coating of liquid coated support on inside of silica
tube
best for speed and efficiency but only small samples
Film thickness (0.1-5 m) affects retention and resolution - thicker films for volatile
analytes, poorer resolutions
Chiral phases being developed for enantiomer separation
(pharmaceuticals)

Temperature Programming:
When a mixture of various boiling points (BP) components is to be analysed, at low
column temperature the High BP will elute after very long time because the vapor pressure
analyte increases as column temperature is raised. Elution will be faster. Meanwhile, low
BP components will elute rapidly at high column temperature without good resolution
(Separation is bad).

To solve this problem we must raise column temperature during separation temperature
programming - separates species with wide range of polarities or vapor pressures

Liquid Chromatography
A method for the separation of components (solids or high BP, high molecular
weight liquid organic compounds or other compounds) of a homogeneous mixture
and to determine the composition of the mixture.
It is based on the injection of the sample solution (i.e. solids must be dissolved in
certain solvent) mixture into a column packed with certain materials (stationary
phase) and passing a liquid (the mobile phase) through the column. The
components of the mixture will be held strongly or weakly to the stationary phase.
The liquid mobile phase will pass at high pressure through the column. It will take
away the weakly bound component first to the outlet of the column. Thus, the
components will be separated according to the boiling point, molecular weight,
polarity, ability to form hydrogen bonding with the stationary phase. The
components will appear et the end of the column at different times, tR , the
retention time.

Instrumentation for HPLC:

For reasonable analysis times, moderate flow rate required but small particles of
the stationary phase (1-10 m)
Solvent forced through column 1000-5000 psi (i.e. instrument is more
complicated than in GC.)
Solvents must be degasses to remove air bubbles "sparging"
High purity solvents
Single mobile phase composition - isocratic elution
Programmed mobile phase composition - gradient elution: Solvent polarity
(composition) continuously varied or stepped (See page 8a)
Up to 10,000 psi, small internal volumes
Sample injection

Similar to FIA, GC
Introduce small sample (0.1-100 L) without depressurization
Microsyringe/septum system (only <1500 psi)

HPLC COLUMNS:
Heavy-wall glass, stainless steel and plastic are among materials that can:
a. withstand high pressures.
b. resist the chemical action of the mobile phase.
c. 10-30 cm long;
d. 4-10 mm internal diameter;
e. 1-10 m particle size - 40,000-60,000 plates/m
f. Well-packed because channels would cause peak broadening and a
decrease in efficiency.
Usually a short guard column is placed in front of the analytical column. This
serves to increase the life of the analytical column by removing particulate matter
and contaminants from the solvents.
High
Speed
Separation

Isocratic

Detectors:
All properties previously discussed and
Bulk property detectors - measure property of mobile phase
(refractive index, dielectric constant, density)
Solute property detectors - measure property of solute not present in mobile phase
(UV absorbance, fluorescence, IR absorbance)
Normal phase HPLC nonpolar solvent/polar column
Interaction :
Adsorption
Packing materials :
Polar
ex. Silica gel
Silica-NH2
Silica-CN
Silica-OH
Mobile phase :
Non-polar
ex.n-Hex/CH2CL2
iso-Oct/IPA
iso-Oct/AcOEt
Sample :
Fat-soluble Different polarity

Reversed phase HPLC

Interaction :
Hydrophobic
Packing materials : Non-polar
ex. Silica-C18
Silica-C8
Polymer
Mobile phase :
Polar
ex.MeOH/H2O
CH3CN/H2O
MeOH/Buffer sol.
Sample :
Having different length of carbon chain
Comparison of Reversed Phase and Normal Phase

Stationary phase
Mobile phase
Interaction
Elution order

Normal phase

Reversed phase

High polarity
Low polarity
Adsorption
Low to High
(Polarity)

Low polarity
High polarity
Hydrophobic
Short to Long
(Length of Carbon chain)

Reversed-phase HPLC most common (high polarity solvent, high polarity solutes
elute first)
R is C8 or C18 hydrocarbon
faster elution
higher resolution

Column Optimization in HPLC:

More difficult than GC
- in GC mobile phase just transported solute
- in HPLC mobile phase interacts with solute
Analyte Polarity:
hydrocarbons< ethers< esters< ketones< aldehydes <amines<alcohols
Stationary Phase Choice:
Choose column with similar polarity to analyte for maximum interaction
Mobile Phase Choice:
Polar ("strong") solvent interacts most with polar analyte (solute) - elutes
faster but less resolution

Size Exclusion Chromatography (SEC)

GPC and GFC
Non-aqueous SEC :
Interaction :
Packing :

GPC (Gel Permeation Chromatography)

Gel permeation
Cross-Linked porous Polystyrene

Mobile phase:
Sample:

Organic solvent (THF, CHCl3, DMF)

Molecular weight distribution of polymer
SyntheticOligomerseparation

Aqueous SEC :
Interaction :
Packing :
Mobile phase:
Sample:

GFC (Gel Filtration Chromatography)

Gel permeation
Hydrophilic silica gel / Hydrophilic porous polymer
Buffer solution
Separation of Water-soluble polymers
(proteins, nucleic acid, sugar)
oligomers

SEC Separation mechanism

Amino Acid Analysis

Column:
Mobile phase:
Detection:
Sample:

AApakNaII-H
Sodium citrate buffer
Stepwise gradient
OPA post label
Ex 345nm Em 445nm
Sake

ELECTROCHEMICAL METHODS
Many electroanalytical methods are available. They are fast and inexpensive. They
are based knowledge about oxidation states, stoichiometry, rates, charge transfer
and equilibrium constants.
Electrochemical Cells:
The composition of electrochemical Cells is based on Oxidation and reduction
(redox) reactions. We need to separate species to prevent direct reaction (Fig 1).
Most electrochemical Cells contain external wires (electrons carry current), ion
solutions (ions carry current) and interfaces or junctions. All contain complete
electrical circuit and conducting electrodes (metal, carbon)

Fig. 1: Electrochemical Cells:

Electrons are transferred at electrode surface at liquid/solid interface. The potential

difference (voltage) is measure of tendency to move to Equilibrium.
Galvanic cell - cell develops spontaneous potential difference

Convention:

Reduction at Cathode
Oxidation at Anode

Galvanic cell - Zn anode (negative), Cu cathode (positive)

Electrolytic cells - require potential difference greater than galvanic potential
difference (to drive away from equilibrium)

Electrolytic cell
Electrolytic cell - Zn cathode (positive), Cu anode (negative)
Many chemically reversible cells are available. The short-Hand Cell notation places
the Anode on Left with liquid-liquid interface

Galvanic cell as written

Electrolytic cell if reversed
Not all cells have liquid-liquid junctions

Electrode Potentials:
Cell potential is difference between anode and cathode potential

Ecell =

Ecathode -

Eanode

when half-reactions written as reductions with electrons on left

Example:

Galvanic cell
Ecell = Ecathode -

Eanode

potential on each electrode Can't be measured independently - only

differences. The standard reference electrode is usually standard hydrogen
electrode (SHE), Fig 3. The SHE is assigned to 0.000 V.

It can be anode or cathode. Pt does not take part in reaction. The Pt electrode is
coated with fine particles (Pt black) to provide large surface area. However, the SHE
is cumbersome to operate.
Alternative reference electrodes are:
Ag/AgCl electrode

Calomel electrode

Electrode and Standard Electrode Potentials (E and E0):

How do we know which way reaction will go spontaneously?
Use electrode potentials, E (potential of electrode versus SHE) to find Eanode and
Ecathode. Then find Ecell.
But electrode potential varies with activity of ion
aX = X . [X]
aX = activity X =activity coefficient and [X] = concentration
X varies with presence of other ions (ionic strength)

Note: activity of pure liquid or solid in excess=1.00

Note: use pressure (atm) for gases
If a=1.00 M, the electrode potential, E, becomes standard electrode potential, E0

Cell containing Cu/Cu2+ and Cd/Cd2+ called couple

(1) Cu2+ + 2e- Cu spontaneously forward
Cd2+ + 2e- Cd spontaneously backward (Cd Cd2++2e-)
(2) e- flow towards Cu electrode (cathode/positive electrode)
e- flow away from Cd electrode (anode/negative electrode)
(3) Cu2+ good electron acceptor (oxidizing agent)
Cd good electron donor (reducing agent)
The most positive E or E0 spontaneously forward forming cathode
Calculation of Cell Potentials, Ecell:
Eanode
Ecell = Ecathode when written as reductions
Example:

Zn reaction spontaneously backward - forms negative electrode - place of oxidation

anode

If a = 1.00 M, E = E0:

Ecell =

Ecathode -

=+
=+

0.337 1.100 V

Eanode
( 0.763)

Spontaneous reaction is galvanic

Ecell indicates if reaction is spontaneous as written

Ecell positive - reaction forward
Ecell negative - reaction backwards
Electrode potential is related to position of equilibrium

If reaction is long way from thermodynamic equilibrium, K will change with time
Eventually, concentrations reach equilibrium values and K stops changing (true
equilibrium constant Keq)
In general:

In principle, can calculate E and Ecell from E0 for any activity from Nernst equation:

E= E0 when log quotient in Nernst equation is unity

E0 is relative to SHE
E0 is measure of driving force for half-cell reduction
Limitations of Standard Electrode Potentials:
(1) E0 is temperature dependent
(2) Substitution of concentration for activity always introduces error. Error is
worse at high ionic strength
(3) Formation of complexes, association, dissociation alter E0
Formal potentials (E0') apply for specific reactions when specifying ALL
concentrations

POTENTIOMETRY
REFERENCE ELECTRODES:
reversible
little hysteresis
follows Nernst equation
stable potential with time

Saturated Calomel
Electrode (SCE):

Half-cell for Calomel Electrode:

Position of equilibrium affected by aCl- from KCl so E0 depends on aClMost common saturated calomel electrode SCE ([Cl-]~4.5 M)
Silver/Silver Chloride Electrode: Similar construction to calomel
Ag wire coated with AgCl
solution of KCl sat'd with AgCl

Again depends on aCl-, but commonly sat'd (~3.5 M)

Potential vs. SHE

Which one?
Ag/AgCl better for uncontrolled temperature (lower T coefficient)
Ag reacts with more ions
Precautions in Use:
Level of liquid inside reference electrode above analyte level to minimize
contamination
Plugging problematic if ion reacts with solution to make solid
(e.g. AgCl in Cl- determination)

Measuring Concentration using Electrodes:

Indicator Electrodes for Ions:
Electrode used with reference electrode to measure potential of unknown
solution
potential proportional to ion activity
specific (one ion) or selective (several ions)

Ecell = Eindicator - Ereference

Two general types - metallic and membrane electrodes
Metallic Indicator Electrodes:
I. Electrodes of the first kind
- respond directly to changing activity of electrode ion
Example: Copper indicator electrode

BUT other ions can be reduced at Cu surface

- those with higher +ve E0 (better oxidizing agents than Cu) Ag, Hg, Pd...

In general, electrodes of first kind:

simple
not very selective
some metals easily oxidized (deaerated solutions)
some metals (Zn, Cd) dissolve in acidic solutions

Electrodes of the second kind - respond to changes in ion activity through

formation of complex
Example: Silver works as halide indicator electrode if coated with silver halide
Silver wire in KCl (sat'd) forms AgCl layer on surface

Electrodes of the third kind - respond to changes of different ion than metal
electrode
Membrane (or Ion Selective) Electrodes:
Membrane:
Low solubility - solids, semi-solids and polymers
Some electrical conductivity - often by doping
Selectivity - part of membrane binds/reacts with analyte
Two general types - crystalline and non-crystalline membranes
Non-crystalline membranes:
Glass - silicate glasses for H+, Na+
Liquid - liquid ion exchanger for Ca2+
2+
3Immobilized liquid - liquid/PVC matrix for Ca and NO
Crystalline membranes:
Single crystal - LaF3 for FPolycrystalline or mixed crystal - AgS for S2- and Ag+

GLASS MEMBRANE ELECTRODES:

Combination pIon electrode (ref + ind)

Contains two (reference) electrodes - glass membrane is pH sensitive
Glass Membrane Structure:
SiO44- framework with charge balancing cations
- SiO2 72 %, Na2O 22 %, CaO 6 %
In aqueous solution, ion exchange reaction at surface

H+ carries current near surface

Na+ carries current in interior
Ca2+ carries no current (immobile)

Surface where more dissociation occurs becomes negatively charge with respect to
other surface
Alkaline Error:
At high pH, glass electrode indicates pH less than true value
Low [H+] means membrane exchanges with alkali metal ions in solution too

Most accurate 0-10 (0.01-0.03 pH units)

Interference in Glass Membrane Electrodes:
Sensitive to
H+
alkali metal ions
Selectivity coefficients (kX/Y) measure sensitivity to other ions
Range between 0 (no interference) to 1 (as sensitive to alkali and hydrogen ions)
to >1 (large interference)

Glass Electrodes for Other Ions:

Maximize kH/Na for other ions by modifying glass surface (usually adding Al2O3
or B2O3)
Possible to make glass membrane electrodes for
Na+, K+, NH4+, Cs+, Rb+, Li+, Ag+ ...

Crystalline Membrane Electrodes:

Usually ionic compound
Single crystal
Crushed powder, melted and formed
Sometimes doped (Li+) to increase conductivity
Operation similar to glass membrane

Presence of F- analyte pushes equilibrium right, reduces +ve charge on

electrode surface

Liquid Membrane Electrodes:

Based on potential that develops across two immiscible liquids with
different affinities for analyte
Porous membrane used to separate liquids
Example: Calcium dialkyl phosphate insoluble in water, but binds Ca2+ strongly
Molecule Selective Electrodes:
Gas Sensing Probes
Biocatalytic Membranes
Gas Sensing Probes:
Simple electrochemical cell with two reference electrodes and gas
permeable PTFE membrane
allows small gas molecules to pass and
dissolve into internal solution
Analyte not in direct contact with either electrode - dissolved

POTENTIOMETRIC TITRATIONS
Titrations can be followed potentiometrically;
Reaction involves removal or addition of some ion for which an electrode is
available.

Changing Potential of the indicating electrode during titration (Nernst Eq.).

Titration curve: Potential vs. mL titrant.
At Eq. point, slope is steepest.
Concentration cell is used: Two sides, identical electrodes, in the reference side
a solution similar to that expected at the Eq. point;
with addition of titrant the potential difference decreases until reach zero.

INSTRUMENTATION
Nernst eq. is valid when no current passes through the cell. Precision
potentiometer Now the PH meters or p-ion meters are used for this purpose.

VOLTAMMETRY
Electrochemistry techniques based on current (i) measurement as function of voltage
(Eappl)

THREE ELECTRODES
Working electrode
Reference electrode
Counter electrode
Supporting electrolyte

(microelectrode) place where redox occurs

surface area few mm2 to limit current flow
constant potential reference (SCE)
inert material (Hg, Pt) plays no part in
redox but completes circuit
alkali metal salt (e.g. KNO3) does not react with
electrodes but has conductivity

Why not use 2 electrodes?

OK in potentiometry - very small currents.
Now, want to measure current (larger=better) but
potential drops when current is taken from electrode (IR drop)
must minimize current withdrawn from reference electrode surface
Potentiostat (voltage source) drives cell
supplies whatever voltage needed between working and counter
electrodes to maintain specific voltage between working and reference
electrode
NOTE:
Almost all current carried between working and counter electrodes
Voltage measured between working and reference electrodes
Analyte dissolved in cell not at electrode surface!

Microelectrodes
C, Au, Pt, Hg each useful in certain solutions/voltage ranges

At -ve limit, oxidation of water

At +ve limit, reduction of water

Varies with material/solution due to different overpotentials
Overpotential always reduces theoretical cell potential when current is flowing

= Ecurrent - Eequilibrium
Overpotential (overvoltage) develops as a result of electrode polarization:
concentration polarization - mass transport limited
adsorption/desorption polarization - rate of surface attach/detachment
charge-transfer polarization - rate of redox reaction
reaction polarization - rate of redox reaction of intermediate in redox
reaction
Overpotential means must apply greater potential before redox chemistry occurs

Hg particularly useful
high overpotential at -ve limit
easy to prepare clean surface

Hg Microelectrodes:
Dropping Mercury Electrode
(DME)

Voltammograms
The potential applied to the cell in continuously increased, usually in the negative
direction (DME becoming more negative with respect to SCE) and the current is
recorded. The voltammetric waves are graphs of current (i) vs. applied
voltage (Eappl). The position of the wave (half wave potential, E1/2) is
characteristic of the analyte (reduced) and the height (Diffusion Current) is
directly proportional to Concentration (See the Fig.).

Hg microelectrode is cathode -ve terminal in above

Increase in current at potential at which A can be reduced (reaction demands

electrons, supplied by potentiostat)
Two important points
0
Half wave potential (E1/2) is close to E for reduction reaction

Limiting current (il) proportional to analyte concentration (really, activity)

il = k cA
Current is just measure of rate at which species can be brought to electrode
surface
Two methods:
Stirred - hydrodynamic voltammetry
Unstirred - polarography (dropping Hg electrode)
Single voltammogram can quantitatively record many species provided enough
separation between waves

Problems with dissolved O2 - must purge (sparge) solutions

POLAROGRAPHY
First voltammetric technique
Differs from hydrodynamic
unstirred (diffusion dominates)
DME is used as working electrode current varies as drop grows then falls
off

Linear Scan Polarography:

Unstirred - only diffusion - currents smaller

Diffusion Current (Ilkovic Equation)

n = number of electrons; cA = analyte conc (mM); t = drop time (s)

2 -1
D = diffusion coefficient (cm s ); m = flow rate of Hg (mg/s)
Residual Current
redox reactions of impurities in solution
charging of Hg drop
(non-faradaic current/non-redox current)

Advantages of DME (compared to planar electrodes):

clean surface generated
clean surface generated
rapid achievement of constant current during drop growth
remixing of solution when drop falls
0
high Hg overvoltage means even metals with high -ve E can be
measured without H2 formation
Disadvantages of DME:
Hg easily oxidized, limited use as anode (E< +0.4 V)

nonfaradaic residual currents limit detection to >10-5 M

cumbersome to use (toxic mercury)
sometimes produce current maxima for unclear reasons (use maxima
suppressor)

Pulse Polarography: Differential Pulse Polarography (DPP):

Conventional polarography somewhat limited by nonfaradaic currents
DPP relies on two measurements when difference between faradaic and
-7
-8
nonfaradaic currents are largest. Detection limits 10 -10 M.

POLAROGRAPHY
Qualitative Analysis:
Since the half wave potential is characteristic
of the species undergoing reduction or
oxidation at the DME, it can be used for
its identification (fig). Portion AB and DF are
extended, a tangent is drawn to the curve
at point C. Line GH is bisected, and a line JK
is drawn parallel to AB and DF. The abscissa
of the point of intersection of JK with the curve
gives the value E1/2. The selection of the
supporting electrolyte is very important
because of the different tendency of ions to
complex formation:

Quantitative Analysis
The magnitude of diffusion current is related to the concentration of the reducible
species through the Elkovic Equation. It is impractical to measure the factors in
the Equation and a comparison method is preferable. The equation may be

id = K C
K is evaluated for a standard solution, and then the value of id for the unknown
solution is measured to evaluate the concentration.
For example, Suppose Cd is to be determined
in a sample of Zn. A 0.1 g sample is dissolved
in HCl, a few drops of Triton X-100 added,
and the solution was diluted to a known
volume with 1.0 M KCl. A portion is placed in
the polarographic cell and sparged with N2 to
remove dissolved oxygen. Polarogram is
recorded. The potential range from about -0.4
to 0.8 V versus SCE is suitableA wave at
about -0.64 V in this experiment (Qualitative:
Cd is present or not). The reduction
potential of Zn is far negative (No interferences). Another polarogram is recorded for
a standard Cd solution. The values of id for
both standard and unknown solutions from the
graph. The conc. Of the unknown can be
calculated.

TYPES OF ELECTRODES
1. Metal in equilibrium with its ions: The oxidized form Aox is the cation, and
the reduced form Ared is the free metal. The E0 values are given relative to
saturated Calomel electrode, SCE, at 25oC. The electrolyte is a solution of a
salt of the metal with anion of a strong mineral acid, SO4, NO3, and ClO4 are
suitable.
Ea = Ea0 + RT/nF Ln(Aox)
Because the activity of a pure solid element is taken = 1.0.
The more active metals (Na, K, .) cannot be used because they are not
stable in water. Less active metals, (Ag) may not give good response in dil.
Solutions.
2. A metal in eq with a saturated solution of a slightly soluble salt
Ag/ AgCl (s)
Mostly as reference electrodes
3. Ametal in Eq. with two slightly soluble salts with a common anion
Ag2S
CdS

2 Ag + S2 Cd + S2-

This half cell can be used to measure the activity of Cd ions. The second
salt must be slightly more soluble than the first. On e widely used
electrode has eq. between EDTA, Hg2+ ions and the ions of a di, tri, or
tetravalent metal, the slightly dissociated complexes play the same roles
as the slightly soluble sulfide salts in the above example.

CONCENTRATION CELL:
Cell with two identical electrodes dipping into solutions similar in every thing
except the concentration of ion to which the electrodes aare sensitive.
Ag/AgCl electrode in an unknown solution of chloride
Other Ag/AgCl electrode in standard soln of chloride, = 0.1 M
Es = + 0.222 0.0591 Log 0.1

= + 0.281 V

For the Unknown if the conc. 0.15 M

Ex = 0.222 0.0591 Log 0.15

= +0.271 V

Ecell = Es Ex = 0.01 V by subtracting the more negative from the more

positive potential.

POTENTIOMETRY
Suppose the meter indicates a 0.450 V difference in potential between the
electrode:
ESCE = + 0.246 V
By subtraction
Log [Ag] = (Ecell E0Ag + 0.246)/2,303 (RT/nF)
= (0.450 0.799 + 0.246)/0.059 = - 1.74
Thus, [Ag] = 0.018 M
Consider a cell made of Pt electrode dipping into a solution 0.1 M in Fe(II)SO4.
The KCl salt bridge from an SCE is inserted directly into the same solution, since
no harmful interaction can occur. A potential difference of 0.395 V is observed ar
25oC. The goal is to fine the percentage of Fe(II) that has been converted by air
oxidation to Fe(III). Starting with the two half cell potentials, and for n = 1:
Ept = E0Fe3+/Fe2+ + RT/nF Ln[Fe3+]/[Fe2+]
ESCE = + 0.246 V
Ecell = Ept - ESCE
= E0Fe3+/Fe2+ - ESCE + 0,0591 Log [Fe3+]/[Fe2+]
From which Log [Fe3+]/[Fe2+] was calculated = 0.0063 = 0.63%.
Therefore, 0.63 percent of Fe(II) has been oxidized by air.

Potentiometric Titration:
Titration of Ce(IV) with Fe(II):
Fe3+ + e
Ce4+ + e

Fe2+
Ce3+

Reference half cell is SCE

Therefore
2 Fe3+ + 2Hg + 2Cl- 2 Fe2+ + Hg2Cl2
And
2 Ce4+ + 2Hg + 2Cl- 2 Ce3+ + Hg2Cl2
Fe2+ in solution and Ce4+ is the titrant

CONDUCTOMETRY
A non-Faradaic quantity that can be used successfully for analysis, the Electric
Conductance.
A cell consisting of two Pt electrodes in an ionic solution. If a constant DC
potential is applied on this network, nothing will happen if the voltage is small
enough that no electrochemical process can occur. If the voltage is higher then
current will flow through the resistance of the solution. The conductance of a
solution in Siemense is the reciprocal of the resistance. It is related to the ratio of
the area (a) of the electrodes and to the distance between them, (d), and to the
total ionic concentration:
-3

L = 10 (a/d) ziCi i
a/d is a geometric property of the cell, and its reciprocal is cell constant, , with
-1
units of cm . Ci is the molar conc. Of ion i with charge zi and i I is the equivalent
ionic conductivity. The summation covers all ions of both signs.
is a property of ions that gives quantitatively information about their relative
contribution to the conductance of a solution. 0 is the limiting value of at zero
concentration, infinite dilution. It differs from ionic mobility by the factor F
(Faraday constant). For some applications, it is necessary to know the cell
constant . It is generally determined from measurements on solutions of known
specific conductance L = K.
For calibration purposes:
KCl Conc., g KCl/Kg of solution
71.1352
7.41913
0.74526

K, S.cm
0.11134
0.01285
0.0014088

Instrumentation:
To measure electrolytic conductance, we use the wheatstone bridge modified for
ac operation. The circuit is supplied by different resistances to be selected by a
switch to give precise ratios of 0.1, 1 or 10. The unknown resistance is that of the
conductance cell.
Applications:
Most applications are concerned with aqueous solutions. Water is very poor
conductor. Water stills and deionizers are usually provided with conductance
monitors.
Solutions of strong electrolytes show a nearly linear increase of conductance with
concentration up to about 10 to 20% by weight. At higher concentrations, the

conductance decrease again, as interionic attraction hinders free movement of

ions through the solution.
Conductometric Titrations
1. Significant difference in conductance between original solution and the
reagent;
2. The distance between electrodes does not change during titration;
3. Necessary to correct for the dilution due to addition of reagents besides
the consumption of the reactants.
4. Hydrolysis or partial solubility of precipitates may cause departure from
linearity.
5. No need to determine the cell constant, since relative values are enough
to determine end point.
Example:
Titration of HCl by NaOH

H+

OH-

L
L
Na+
Cl-

mL NaOH
Overall shape

mL NaOH
Details of the conductance