FLAME SPECTROSCOPY

OBJECTIVES:
⦿ 1.Identify the two widely used methods
in atomic spectroscopy and the nature
of the samples that can be analyzed by
each method
⦿ 2.Discuss the basic components of a
flame spectrophotometer
⦿ 3.Given the data, calculate the
concentration in parts per million of the
analyte from an AAS analysis
⦿ 4. Identify the principles behind
fluorometry, turbidimetry and
nephelometry and their respective
applications.
⦿ 5. Discuss the instruments required for
these spectrometric methods and
enumerate drug products assayed by
the above methods
⦿ 6. Appreciate the use of modern
instruments as improvement from
conventional methods
OUTLINE:
⦿ Atomic absorption (AAS) vs. atomic
emission (AES)
⦿ Basic components of an AAS and AES
⦿ Difference between a flame AA and
graphite furnace
⦿ Applications of AAS and AES in drug
assay
⦿ Principles and applications of
Fluorometry, Nephelometry, and
Turbidimetry
 Flame spectroscopic methods frequently
used for analysis:
1. Atomic emission
2. Atomic absorption
3. Inductively Coupled Plasma (ICP)
Atomic Emission Spectroscopy
Atomic emission
spectrophotometry (AES)
KEYPOINTS
Principles:
⦿ Atoms are thermally excited so that
they emit light and the radiation
emitted is measured
Applications:
⦿ Analysis of inorganic metals (Na, Pb, K,
Ca etc.)
⦿ Determination of metallic impurities in
some inorganic salts
 Other applications for atomic
absorption:

- Clinical analysis : Analyzing metals in

biological fluids such as blood and
urine.
- Environmental analysis : Monitoring our
environment – ex. finding out the levels
of various elements in rivers, seawater,
drinking water, air, and petrol.
- 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 – e g in concrete, where calcium is a
major
constituent, the lead level should be low because it
is
toxic.
- Mining: By using AAS the amount of
metals such as gold in rocks can be
determined to see whether it is worth
mining the rocks to extract the gold .

- Trace elements in food analysis

- Trace element analysis of cosmetics
- Trace element analysis of hair
Strengths:
⦿ Flame photometry provides robust,
cheap method; simple instrumentation

Limitations:
⦿ Only applicable to the determination of
alkali and some alkaline earth metals
 Atomic absorption
spectrophotometry (AAS)
KEYPOINTS
Principles:
⦿ Atoms of a metal are volatilized in a
flame and their absorption of a narrow
band of radiation produced by a hollow
cathode lamp, coated with the
particular metal being determined, is
measured
Applications:
⦿ Determination of metal residues

Strengths:
⦿ More sensitive than AES
⦿ Highly specific method of analysis

Limitations:
⦿ Only applicable to metallic elements
⦿ Each element requires a different hollow
cathode lamp for its determination
Atomic absorption spectroscopy (AAS)
⦿ is a spectroanalytical procedure for the
quantitative determination of chemical
elements employing the absorption of
optical radiation (light) by free atoms in
the gaseous state.
⦿ is an analytical technique that measures
the concentrations of elements. It makes
use of the absorption of light by these
elements in order to measure their
concentration .
- Concentration measurements are usually
determined from a working curve after
calibrating the instrument with standards
of known concentration.

- Atomic absorption is a very common

technique for detecting metals and
metalloids in environmental samples.
⦿ in analytical chemistry the technique is
used for determining the concentration
of a particular element (the analyte) in a
sample to be analyzed. AAS can be
used to determine over 70 different
elements in solution or directly in solid
samples employed in pharmacology,
biophysics and toxicology research.
⦿ Used to assay inorganic elements such
as K, Na, Ca, Pb, Cd and other metals
⦿ Assay of lithium carbonate
(antidepressant)
Theory:
- Knowledge in atomic theory
- Ex. color flame from a lit matchstick
- The predominant color, yellow, is due to
presence of Na in matchstick
- Na atoms (at. # = 11: e configuration) in
the unlit match are in ground state
- When atoms are excited by thermal
energy, the valence e is promoted to
higher level (excited state)
- Excited state of the atom is very unstable
and the e quickly drops back to the
ground state to resume condition
Sampling:
- Requires acetylene or other gas to
convert metal ions in soln. to atoms in
gaseous state
- Ex. NaCl soln
Na+ Cl- → NaCl → Na + Cl
soln gaseous atoms atoms
 The Atomic Absorption
Spectrometer
⦿ Atomic absorption spectrometers have 4
principal components
1 - A light source ( usually a hollow
cathode lamp )
2 – An atom cell ( atomizer )
3 - A monochromator
4 - A detector , and read out device .
 Schematic Diagram of an Atomic
Absorption Spectrometer

Detector and
Light source atomizer
monochromator readout device
(hollow cathode Lamp )
Parts of Atomic Absorption
Spectrophotometer:

- Just like flame emission but has a hollow

cathode lamp (contains He gas) elevate
the outer valence e to excited state
AAS advantage:
- ↑ sensitivity – only 0.01 % of the atoms in the
flame are in excited state and emit light which is
measured in Flame Emission method, the
remaining atom (99.99%) are in the ground state
and are unavailable in emission technique,
⦿ In atomic emission, a sample is subjected
to a high energy to produce excited
state, capable of emitting light
⦿ The energy source: electrical arc, flame,
plasma
⦿ In atomic absorption, light of just
wavelength impinges on free,
ground state atom, the atom may
absorb the light as it enters an
excited state
⦿ Capability of an atom to absorb very
specific wavelength of light
AES Basic Components:
AAS Basic Components:
⦿ Elements to be analyzed needs to be in
atomic state

⦿ Atomization is separation of particles

into individual molecules and breaking
molecules into atoms .This is done by
exposing the analyte to high
temperatures in a flame or graphite
furnace .
ATOMIZER:

Elements to be analyzed needs to be in

atomic state

Atomization is separation of particles into

individual molecules and breaking
molecules into atoms .This is done by
exposing the analyte to high
temperatures in a flame or graphite
furnace .
⦿ The role of the atom cell is to primarily
dissolvate a liquid sample and then the
solid particles are vaporized into their
free gaseous ground state form . In this
form atoms will be available to absorb
radiation emitted from the light source
and thus generate a measurable signal
proportional to concentration .

⦿ There are two types of atomization :

Flame and Graphite furnace
atomization .
 ATOMIC ABSORPTION ATOMIZER
1. Pre-mix Burner System
sample soln. aspirated → nebulize → fine
aerosol → mixing chamber (sample aerosol mix
w/ fuel + oxidant gases) → burner head (where
combustion and sample atomization occurs)
⦿ Only a portion of soln. is introduced to
burner chamber by nebulizer
⦿ Excess samples are drained
2. Impact Devices
⦿ Use to reduce droplet size
⦿ Types:
impact beads – improve nebulization
efficiency
- larger droplets convert to
smaller droplets

flow spoilers – to remove remaining large

droplets from aerosol
 LIGHT SOURCE:
1. Hollow cathode lamp
⦿ The light source is usually a hollow cathode
lamp of the element that is being measured .
Each element has its own unique lamp which
must be used for that analysis .
⦿ excellent, bright line source
⦿ Have finite lifetime
For example with lead, a lamp containing
lead emits light from excited lead atoms
that produce the right mix of wavelengths
to be absorbed by any lead atoms from the
sample .

A beam of the electromagnetic radiation

emitted from excited lead atoms is passed
through the vaporized sample. Some of the
radiation is absorbed by the lead atoms in
the sample. The greater the number of
atoms there is in the vapor , the more
radiation is absorbed .
Hollow Cathode Lamp
Quartz window
cathode

Pyrex body

Anode
Anode

Cathode
2. Electrodeless Discharge Lamp
⦿ Brighter
⦿ More stable
⦿ More sensitive
 OPTICAL CONSIDERATIONS
Photometers – optical system w/c conveys
the light from the source to
monochromator:
1. Single beam
⦿ Single beam of light in a single optical
path
⦿ Fewer components
⦿ Less expensive
2, double beam
⦿ Additional optics to divide light from
lamp into a “sample beam” and
“reference beam”
⦿ Reference beam monitors lamp intensity,
prevent electronic anomalies,
compensate electronic drift
MONOCHROMATOR
⦿ Light from the source must be focused
on the sample cell and directed to the
monochromator, where the λ of light are
dispersed and the analytical line of interest is
focused on the detector.
⦿ A monochromator is used to select the
specific wavelength of light which is
absorbed by the sample, and to exclude
other wavelengths. The selection of the
specific light allows the determination of
the selected element in the presence of
others.
AUTOMATION OF ATOMIC ABSORPTION:

Automatic Sampler :
-freeing the analyst from task of manually
introducing each and every sample
Automated
Multi-Element AA:
-automatic set-up
-analyze a tray full
of samples for
multiple elements
without operator
intervention
Control of Analytical Interferences:

⦿ AA is known as very specific technique

with few interferences
 Flame Process:
1. Nebulization
2. Desolvation
3. Liquefaction
4. Vaporization
5. Atomization
6. Excitation
7. ionization
⦿ Since the thermal energy from the flame
is responsible for producing the
absorbing species, flame temperature is
an important parameter.
Temperature of Premix Flames:
OXIDANT - FUEL TEMP, OC

Air – methane 1850 – 1900

Air – natural gas 1700 – 1900

Air - hydrogen 2000 – 2050

Air – acetylene 2125 – 2400

N2O - acetylene 2600 - 2800

⦿ Cooler flames are subject to more
interference problem insufficient energy
for complete atomization
⦿ 2 premix flames now used almost
exclusively for AA are air-acetylene and
N2O-acetylene (hotter, for many
refractory forming elements)
INTERFERENCES
The concentration of the analyte element is
considered to be proportional to the
ground state atom population in the flame
,any factor that affects the ground state
atom population can be classified as an
interference .

Factors that may affect the ability of the

instrument to read this parameter can also
be classified as an interference .
1. MATRIX INTERFERENCE
⦿ Viscosity of sample, acid conc.
⦿ Any acid or other reagent added to the
sample should also be added to the
standards and blank
⦿ Use METHOD OF STANDARD ADDITIONS
to solve
⦿ Method of Standard Addition:
1st aliquot – sample only (no std added)
2nd aliquot – add 1mL of the std. (spike)
3rd aliquot – add 2 mL std.(spike)
2. CHEMICAL INTERFERENCE
Ex. effect of PO4 in Ca
Ca-PO4 – to eliminate:
add lanthanum to tie up
PO4 and allow Ca to be atomized or
use hotter flames
3. IONIZATION INTERFERENCE
⦿ Common w/ hotter N2O – acetylene
flame
⦿ If additional energy is applied, an
electron may be totally removed from
the atom, creating an ion
⦿ To eliminate: add ionization
suppressants: K, Rb, Cesium salts
CALIBRATION OF THE SPECTROPHOTOMETER:

⦿ Most modern AAS include microcomputer

based electronics (advanced calculation
capabilities – ability to calibrate and
compute conc. from absorbance data
even for non linear calibration curve)
⦿ In linear region, one std. and a blank
maybe sufficient to define relationship
between conc. and abs.
⦿ However, additional standards are used
to verify calibration accuracy
⦿ Accurate calibration can be achieved
w/ a minimum of 3 standards and a
blank
CALIBRATION CURVE
⦿ A calibration curve is used to determine
the unknown concentration of an element
in a solution. The instrument is calibrated
using several solutions of known
concentrations. The absorbance of each
known solution is measured and then a
calibration curve of concentration vs
absorbance is plotted.

⦿ The sample solution is fed into the

instrument, and the absorbance of the
element in this solution is measured .The
unknown concentration of the element is
then calculated from the calibration curve
 Calibration Curve
A 1.0 -
b 0.9 -
S 0.8 - .
o 0.7 - .
r 0.6 - .
b 0.5 - . .
a 0.4 - .
n 0.3 - .
c 0.2 -
e 0.1 -

10 20 30 40 50 60 70 80 90 100
Concentration ( g/ml )
 Determining concentration from
Calibration Curve

A 1.0 - absorbance measured

b 0.9 -
S 0.8 - .
o 0.7 - .
r 0.6 - .
b 0.5 - . .
a 0.4 - .
n 0.3 - . concentration calculated
c 0.2 -
e 0.1 -

10 20 30 40 50 60 70 80 90 100
Concentration ( mg/l )
Comparison Between Atomic
Absorption and Emission
Spectroscopy
Absorption Emission

- Measure trace metal - Measure trace metal

concentrations in concentrations in
complex matrices . complex matrices .

- Atomic absorption - Atomic emission

depends upon the depends upon the
number of ground number of excited
state atoms .
atoms .
- It measures the - It measures the
radiation absorbed by radiation emitted by
the ground state the excited atoms .
atoms.
- Presence of a light - Absence of the light
source ( HCL ) . source .

- The temperature in - The temperature in the

the atomizer is atomizer is big enough
adjusted to atomize to atomize the analyte
the analyte atoms in atoms and excite
the ground state them to a higher
only. energy level.
ACCESSORIES OF ATOMIC ABSORPTION:
COLD VAPOR
MERCURY
⦿ Since atoms for most AA elements
cannot exist in the free, ground state at
room temp, heat must be applied to
break the bonds
⦿ The only exception to this is MERCURY
⦿ Free Hg atoms can exist at room temp,
therefore Hg can be measured by AA
w/out heated sample cell
⦿ Procedure:
1. Hg + reducing agent (stannous chloride
or Na borohydride)
2. The volatile free Hg is driven by bubbling
Ar or air
3. Hg atoms are carried to absorption cell
4. Measure Absorbance
⦿ The detection limit – 0.02 µg/L
⦿ The most sensitive and reliable technique for
determining low conc. of Hg by AA
HYDRIDE GENERATION TECHNIQUE
⦿ Some resemblance to cold vapor Hg
system
⦿ The sample cell must be heated
(air-acetylene flame) or electrically
⦿ Elements analyzed:
As Sn Te
Pb Bi Se
GRAPHITE
FURNACE AA
⦿ Most advanced and widely used high
sensitivity sampling technique for AA
⦿ A tube of graphite is located in sample
compartment, w/ light path passing
through it
⦿ A small volume of sample soln. is placed in
the tube and then heated
⦿ Detection limit: ng/L – for trace metal
analysis
INDUCTIVELY
COUPLED
PLASMA (ICP)
EMISSION
⦿ An atomic emission technique using Ar
plasma as excitation source
⦿ Sample is introduced into premix spray
chamber then directed to central tube
of ICP “torch” – source of energy from
the system
⦿ No electrode/lamp to maintain and
replace
⦿ Many elements can be determined in
one automated analysis (speed
advantage over AA)
⦿ High cost
 General Information

☞Used for Qualitative Analysis

☞Used for Quantitative Analysis

☞Detection limit is in ppb range

☞Not possible to determine: H, N, O, C or Ar in trace

levels as they are used in solvents and plasma

☞Not possible to determine F, Cl and noble gases at

trace levels as they require high excitation energy

☞Not used for determining radioactive elements

ICP – MASS SPECTROMETER
(ICP – MS)
⦿ Exceptional sensitivity
⦿ High analysis speed
⦿ High cost
MASS SPECTROMETRY
MS is an analytical technique that measures:
⦿ the mass-to-charge ratio of charged
particles.
⦿ used for determining masses of particles,
for determining the elemental composition
of a sample or molecule, and
⦿ for elucidating the chemical structures of
molecules, such as peptides and other
chemical compounds.