Instrumental analysis

PHC 321

Lecture (8)
Atomic spectroscopy

Course coordinators:
Assoc. Prof. Christine Maged
Dr.Heba Tarek
 Training Questions

A. Differentiate between the following:

• Atomic and molecular Absorption spectra.
• Laminar Flow Burner and Turbulent Flow burner.

B. K can be determined by flame photometry while Cr cannot. (rationalize)

C. Interpret the technique of fireworks.
 References:

• J. Mendham, R.C. Denney, J. D. Barnes, M.J.K. Thomas, Vogel's

Quantitative Chemical Analysis (6th edition, 2010), Prentice Hall, ISBN
0582226287.

• Donald L. Pavia, Gary M. Lampman, George S. Kriz, James A. Vyvyan,

Introduction to Spectroscopy (4th edition, 2012), Brooks Cole, ISBN
0495114782
Interactive teaching methods and activities

https://www.youtube.com/watch?v=rv9ge_lw6nk

https://www.youtube.com/watch?v=5fvWhCk7x6U

https://www.youtube.com/watch?v=x5Ga5SWIDNY

https://www.youtube.com/watch?v=tVULJAiEiaw

Interactive teaching
https://quizizz.com/admin/quiz/674c7659c26a3d995990367a
 Learning outcomes

At the end of the lecture the students will acquire knowledge about:

• Atomic Absorption Spectroscopy (AAS) Theory.

• Instrument of atomic absorption.
• Application of atomic absorption.
• Chromatography and its classification.
The main parts of atomic absorption spectrophotometer are:

1- Source of radiation 2- Chopper

3- Atomizer 4- Monochromator
5- Detector 6- Readout meter
1- Source of radiation: Hollow cathode lamp
A deuterium or hydrogen lamp can not be used WHY?
❑ Because the band width obtained from them is broad even when using very
efficient monochromator.
❑ In this case, the band width is much broader than the absorption line to be
absorbed by the sample, thus a hollow cathode lamp is used.

❑The hollow cathode lamp:

-Is a glass tube with a front quartz window.
-Contains an anode made of tungsten.
-A cylindrical cathode made of the same element of
the sample to be determined.
-The glass tube is filled with neon or argon (inert gas)
at a pressure of 1 to 5 torr.
Mechanism:
▪ When a low potential is applied between the two electrodes, ionization of the
inert gas occurs, ions move rapidly to the cathode and hit it causing dislodge of
metal atoms from the cathode and produce an atomic cloud in a process called
sputtering

▪ Some of the free atoms are in the excited states and emit their characteristic
radiation, then diffuse back to the cathode surface.

▪ The cylindrical configuration of the cathode is necessary WHY?

Because it tends to concentrate the radiation in a limited region of the tube with
the production of line beam of exactly the same wavelength to be absorbed by the
sample.
2- Chopper:
Its main function is to eliminate interferences due to flame emission by
modulation and fluctuation of the source output.

• The chopper is interposed between the hollow cathode lamp and the flame.
• It is a circular disc divided into four quarters two are mirrored and two are opened.
• The disc rotates at a constant high speed, when the mirrored quarter is in front of the lamp, the
radiations are reflected and only the light emitted by the flame is read by the detector.
• When the opened quarter is in front of the lamp, the lamp light is allowed to pass to the sample
and is absorbed. (both the flame emission and the lamp radiation are measured by the detector)
• So, the radiation reaches the detector in pulses. The detector converts this radiation to alternating
current (ac) signal and is amplified.
• In addition, the radiation coming from the flame itself and from atoms excited by the flame will
reach the detector continuously and converted to direct current signal.
• Both alternating and direct current signals pass through an (resistor-capacitor) RC filter
which allows the alternating signal only to pass and remove the dc signal which is due to
interfering flame emission.
The chopper of the AAS
3- Atomizer:
There are two types of atomizers:
1- Flame atomizer: the same a in flame emission but the burner must be rectangular.
2- Flameless or non flame atomizer:
It is a graphite furnace heated electrically up to 6000°C and contains a ribbon or boat in
which the sample is injected.
The sample is ashed then atomized by the action of heat.

Advantages of non flame atomizer:

1- The sample volume is small (few microliters).
2- unusual high senstivity 10-12.
3- Solid sample can be used directly.
4- Heat distribution is uniform and temperature is steady.
5- No need for fuel-oxidant mixture.
6- No flame noise.
4- Monochromator:
Grating to eliminate any resonance lines from the source or any radiation from the
flame or sample.

5- Detector:
Photomultiplier tube

6- Readout meter:
Absorbance or transmittance output.
• https://youtu.be/nGNGuWtt90A?si=LmsO9dIX8QVfbbdw
 Interference In AAS
Processes that affect the efficiency of producing gas-phase atom or the
interaction of these atoms with light which may be:
• Spectral Interferences: is related to spectral properties of components
other than atomized analyte
a) Spectral line Interference
b) Scattering
c) Broad Band Absorption

• Chemical Interferences: is related to the chemical processes occurring in

flames and electrothermal atomizers and their effects on signal.
a) Formation of compounds of low volatility

b) Ionization Effect
Spectral Interferences:
1- Spectral line Interference
Spectral line interference in flame emission refers to the phenomenon where different
spectral lines from various elements or compounds overlap or interact within the
emission spectrum of a flame, leading to broadening or distortion of the observed
emission lines.
• The first type of interference arises when two elements exhibit spectra, which
partially overlap,
and both emit radiation at some particular wavelength.
eg. - The Fe line at 3247.28Å overlaps with the Cu line at 3247.54Å
Solution: It can overcome either by taking measurements at an alternative
wavelength which has no overlap, if available, or by removing the interfering element
by extraction.
• The second type of spectral interference deals with spectral lines of two or more
elements which are close but their spectra do not overlap
Solution: It can be reduced by increasing the resolution of the spectral isolation
system.
Spectral Interferences:
2- Scattering
• Particulate matter from combustion products and sample materials scatter
radiation that will result in positive analytical error. The error from scattering
can be corrected for by making a blank measurement.
• Samples containing organic materials or organic solvents can form
carbonaceous (especially in cases of incomplete combustion) particles that
scatter radiation.
• Scattering phenomenon is most important when concentrated solutions
containing elements that form refractory oxides (like Ti, Zr, and W) are
present in sample matrix.
3- Broad Band Absorption
In cases where molecular species from combustion products or sample matrix
are formed in flames or electrothermal atomizers, a broad band spectrum will
result which will limit the sensitivity of the technique.
Eg.: Analysis of Ba in mixture of alkaline earth metals suffers interference from
Ca(OH)2 which has absorption band that overlap the resonance line of Ba.
 Application of Atomic absorption Spectroscopy (AAS)

AAS method is more applicable than flame emission, since all elements can be
determined by this technique. As about 99.8% of the atoms absorb the radiation,
but only 0.01% or less of them emit radiation in all directions.

Calibration curve method

The procedure is the same as under flame emission but measuring absorbance.
Chromatography
 What is chromatography?

• The term “ Chromatography” is composed of two

parts chroma means colour and graphien means
writing

• Chromatography is a separation technique firstly

used by a Russian botanist in 1906 to separate
coloured compounds like chlorophyll, carotene and
xanthene from plant extracts.

• Now chromatography is used to separate almost all

kinds of compounds in mixtures, either coloured or
not . But it kept the original name.
 Principle of chromatography

Depends on the separation of mixture of analytes

according to their different distribution between two
phases:

• Stationary phase (solid or liquid): fixed phase in which

the analyte mixture is placed on.
• Mobile phase (liquid or gas): moving on the fixed
phase to wash the analytes successively.

Separation depends on the equilibrium of the analyte

between the two phases (stationary and mobile)
 Mobile
phase

sample

Stationary
phase

Which analyte has stronger

Analyte 1 Analyte 2
affinity to the stationary phase?
Classification of Chromatographic Methods
I. According to the nature of mobile phase:
Liquid chromatography Gas chromatography

II. According to the stationary phase:

Normal phase chromatography Reversed phase chromatography

III. According to the technique used

Planner chromatography Columnar chromatography

IV. According to the mechanism of separation

Adsorption chromatography Partition chromatography
Ion exchange chromatography Size exclusion chromatography
Affinity chromatography
Classification of Chromatographic Methods
• Normal phase chromatography
Where polar stationary phase and non polar mobile phase are used. In this case,
polar components will be held more strongly (like dissolves like) and non polar
components are travelled faster.
• Reversed phase chromatography
Where non polar stationary phase and polar mobile phase are used. In this case, non
polar components will be held more strongly and polar components are travelled faster.

C18
III. According to the technique used
Planner chromatography Columnar chromatography
- Paper chromatography
- Thin-layer chromatography

Planner chromatography

1-Paper chromatography

• A method of partition chromatography using filter paper strips as

carrier or inert support.
• The factor governing separation of mixtures of solutes on filter
paper is the partition between two immiscible phases. One is
usually water adsorbed on cellulose fibers in the paper (stationary
phase). The second is the organic solvent flows past the sample on
the paper.
• Partition occurs between the mobile phase and the stationary phase
bound by the cellulose.
• The isolation depends on partition coefficient of the solute.
2-Thin- Layer chromatography

• Separation in TLC involves distributing a mixture of

two or more substances between a stationary phase
and a mobile phase.
• The stationary phase is a thin layer of adsorbent
(usually silica gel or alumina) coated on a plate.

• The mobile phase is a developing liquid which

travels up the stationary phase, carrying the
sample with it.

• Components of the sample will separate on the

stationary phase according to how much they adsorb
on the stationary phase versus how much they
dissolve in the mobile phase.
Identifying the spots (visualization)
Visualizing agents
Interpreting the data
High performance thin layer chromatography (HPTLC)
• HPTLC is an analytical technique based on TLC, but with
enhancements intended to increase the resolution of the
compounds to be separated and to allow quantitative
analysis of the compounds.

• Some of the enhancements such as the use of higher

quality TLC plates with finer particle sizes in the stationary
phase which allow better resolution.

• As a consequence, HPTLC offers better sensitivity and

resolution.
Column chromatography
• Stationary phase is held in a narrow tube through which
the mobile phase is forced under pressure or under the
effect of gravity

• When the mobile phase along with the mixture that

needs to be separated is introduced from the top of the
column, the movement of the individual components of
the mixture is at different rates.

• The components with lower adsorption and affinity to

stationary phase travel faster when compared to the
greater adsorption and affinity with the stationary phase.
• The components that move fast are removed first
whereas the components that move slowly are eluted
out last.
Factors affecting solutes separation in column chromatography