Atomic Absorption

Spectrophotometry (AAS)
Dr. Akepati S. Reddy
Associate Professor, Thapar University
Adjunct Scientist & Head, Dept. Analytical Services
TCIRD, Thapar Technology Campus
Patiala (PUNJAB) – 147 004
 Atomic Absorption Spectrophotometry
An Atomic Spectrophotometry technique to measure metal ion
concentration of samples using flame atomizer and light source
through the measurement of absorption of light by atoms
Atomic/flame emission spectrophtometry is a parallel technique
(measuring the light emitted at particular wavelength by excited
atoms by electrons falling from higher to lower energy level is basis)
Atomization: Process of conversion of metal ions of liquid solutions
into atoms – done in fuel/oxidant flame
Light source: Hollow cathode lamp/electrodeless discharge lamp -
light source of desired wavelength for absorption
Atomic absorption: Absorption or take up of light of a given
wavelength by atoms resulting in the excitation
Excitation: Promotion of electrons of an atom from lower energy
levels to higher energy levels
 Hollow
Cathode
Lamp

Gas Mixture Adjustment

Display Monochromator
Controls Flame
 Atomic Absorption Spectrophotometry
A quantitative technique introduced by Alan Walsh in 1955
Used to analyze samples for many metals and a few non-metals
– Water and wastewater, soils and even air samples
– Sludges and solid and hazardous wastes
– Foods
– Lubricating oils and greases
– Clinical samples
The method is insensitive to the molecular form of the metal and
measures the total metal concentration
The method is more specific – the atomic absorption bands are
very sharp (~0.003 nm) when compared with molecular
absorption bands (>20-25 nm)
 Flame Atomic Absorption
Solution containing metal cations is fed into flame (air/acetylene)
Flame atomizes the cations and the atoms are excited by the light
passing through the flame by absorption
– Wavelength of the light source should be matching to the energy
level of electronic transition
– Excitation involves atomic absorption and taking up of photons of
light and electronic transition
– Excited atoms quickly release the energy in the form of light
emission and this is basis for measurement in photometry (where
the flame, rather than light source, excites the atoms)
Amount of light absorbed is measured - Proportional to the
concentration of absorbing species of the sample
 Flame Atomic Absorption
A - absorbance due to the analyte in the flame
Io & I - light intensities striking the detector in the
 I0  absence and presence of analyte in the flame
A  log    E.b.c
 I  E - absorptivity coefficient
B - length of the flame
C - concentration of the atomic species

Absorbance is proportional to the concentration of the absorbing

species in the sample
Calibration determines the proportionality between A and C
 Atomic Absorption Spectrophotometer
Has four major components
– Light source
– Burner head (sample is aspirated into the flame for atomizing)
– Monochromator (separates light into various wavelengths)
– Photomultiplier (detects the photons not absorbed)
 Atomic Absorption Spectrophotometer
Light source
• Hollow cathode lamp
– The element to be measured is contained within a sealed glass
lamp filled with low pressures of neon or argon (inert gas)
– Atoms are electrically excited to higher energy levels - emission
(photon beam) from excited atoms is used as light source
– Ensures matching of energies of the incident photons with those
needed for the excitation of atoms in the flame
– Multi-element hollow cathode lamps are now available
– Not all metals make good cathodes (volatile or not conducting)
 Atomic Absorption Spectrophotometer
Light source:
• Electrodeless discharge lamp
– A salt containing the metal of interest is sealed in quartz tube
along with inert gas
– Atoms are excited to higher energy states by radiofrequency coil
and emissions from the excited atoms is used as the light source
– Light intensity is higher but not stable
 Atomic Absorption Spectrophotometer
Burner Head and atomization of analyte:
• Acetylene is brunt with air or nitrous oxide as oxidant and the flame
generated is used to atomize the analyte
– Flame temp. is ~2300°C and depending on the metal to be analyzed it is adjusted
to be either a oxidizing (lean) or a reducing (rich) flame
– Light beam from the light source is passed through the flame in its way to the
monochromator – importance of alignment!
• Flame atomizing essentially involves
– Nebulization (transforming the sample into fine mist)
– Desolvation (drying the mist droplets into solid/gas aerosols)
– Volatilization (transforming the droplets into gaseous molecules)
– Dissociation (decomposition of molecules into atoms)
• Pressurized acetylene and air/nitrous oxide create vacuum in premix chamber
and this aspirates the sample into nebulizer
• Nebulizer transforms the sample into fine mist
• Part of this mist mixes with air/acetylene gases enters the flame for atomizing
 Atomic Absorption Spectrophotometer
Monochromator
• Light beam exiting the flame is passed through an entrance
slit into a monochromator for reflecting off a defraction
grating to separate the light into various wavelengths
Photomultiplier
• Light beam exiting the monochromator enters the
photochromator and strikes an electrically charged metallic
coil which results in emission of electrons
• The stream of electrons is amplified by successive
bombardments onto a series of dynodes
• Amplified electron beam is collected at an anode as a signal
and transmitted to the data processing system
 Performing Atomic Absorption
Performing atomic absorption can be considered to include
1. Sample preparation
2. Method review and PPE
3. Start equipment and perform system checks
4. Perform calibration
5. Run standard and sample
6. Flush out system and shutdown
 Performing Atomic Absorption
Sample preparation
• Standards and samples should be prepared as directed by the
procedures
• Samples may need dissolving and digestion in acid at high
temperature
• Samples are usually prepared in dilute nitric acid
– Acid used should be suitable for trace metal analysis
– Deionized water free from metals should be used
• For enhancing atomization reagents like lanthanum salts may be
added to the sample
• Both standards and sample should receive identical treatment
• Concentration in the sample should be within the range of the
standard concentration
 Performing Atomic Absorption
Method review and PPE
• Read and get familiar with the operating procedure
• Atomic absorption involves working with open flame and
handling samples – hence needs use of PPE (personal
protection equipment) like goggles and gloves
– Operating procedures regarding PPE use should be followed
 Performing Atomic Absorption
Start the equipment and perform system checks
• Turn on the ventilated fume hood
• Turn on instrument and set it up as per standard procedure
• Allow lamp to warmup
• Check the waste receptacle (should be at least 2/3rd full and not
overflowing)
• Ensure that the drain tube contains a trap and it is immersed in
the waste solution
• Check pressure of the acetylene tank (should be >100 psi)
• Open the valve on the air supply first before that of the
acetylene, and adjust pressure and flow rates as per
manufacturer’s instructions
• Check alignment of the lamp and the burner head
• Ignite the flame and allow it to thermally stabilize
 Performing Atomic Absorption
Perform calibration
• Aspire a blank solution and zero the instrument
• Aspire the standards to calibrate the instrument
• Generate a calibration curve
Run the standard and the sample
• Aspire the sample for concentration measurement
• On completion of sample run, aspire standard check solution to
ensure the system stability
Flush out the system and shut-down
• Aspire deionized water for >5 minutes to flush out and clean the
burner and chamber
• Extinguish the flame and shutdown the instrument
 Limitations of the Flame AAS and
addressing them
• Beer’s law plot must be linear over the concentration range
– Concentrated samples are diluted to bring the absorbance within
the linear portion of the Beer’s law plot
• Chemical interferences can result in incomplete atomization of
the metal ions of interest
– Stable refractory compounds may be formed
– Use of chelating agents (like EDTA) or release agents can be a solution
– Controlling the flame temperature or use of acetylene/nitrous oxide
flame can prove helpful
– Sulfate and phosphate ions can interfere with calcium
determination
– Addition of lanthanum salts can take care of these interferences
– Flame emission photometry can be used – can prove relatively
inexpensive
 Limitations of the Flame AAS and
addressing them
• Alkali metals have lower ionization energies and are easily
ionized in the flame and reduce the signal intensity
– Can be suppressed by adding an element that is more easily
ionized
– Analysis of Na or K can be improved by adding CsCl
• Certain samples exhibit interferences requiring background
correction due to the complex nature of the sample matrix
– Certain AAS are equipped with Duterium lamp (a continuous
source of UV radiation)
– Absorbance of light from Duterium lamp represents the
background correction needed
 Limitations of the Flame AAS and
addressing them
Variations in the output from a light source can result in noisy
output signal
• Double beam instruments allow overcoming this problem
– Light from the source is split into two beams one passing
through the flame and the other bypassing the flame
– Detector monitors both the beams and remove fluctuations due
to variations in the light source
• Solid state electronic components can now generate stable
light beams and modern AAS are single beam systems
 Limitations of the Flame AAS and their
addressing
Signal monitored by the photomultiplier includes
– Light not adsorbed by the atomic species
– Light emitted from the flame (flame emission) – error!
A light chopper is used between the light source and the flame
to alternately block out light source
The photomultiplier will alternately be measuring
– Only the flame emission
– Both the light not absorbed plus flame emission
Light due to flame emission is then subtracted from the total
light signal by the instrument’s electronics
Double-beam Design with Light Chopper
 Limitations of the Flame AAS and
addressing them
Interferences, poor reproducibility and poor detection limits for some
metals (metalloids like antimony, arsenic, selenium and tellurium)
necessitate use of Hydride Generation AAS (HGAAS)
• HGAAS, just as AAS, has a hollow cathode lamp, an air/acetylene flame
and an optical system
• Nebulizer is not used in HGAAS
• HGAAS uses
– A hydride generation system
– An optical cell
• Hydride generation system
– Aspirates the liquid sample at controlled rate and mixes with sodium
borohydride and HCl
– Creates a volatile hydride of the analyte metalloid that flows into the optical
cell
 Limitations of the Flame AAS and
addressing them
• Hydride generation system
– Production of a specific metalloid oxidation state prior to aspiring into the
hydride generation system is important
– Timing of the process (from reagent mixing to the volatile hydride
separation in the gas-liquid separator and sending to the optical cell) is
important
– High purity inert gas is used to purge the hydride together with the hydrogen
gas generated into the optical cell
– Most of the reagents introduced into the system flow out as highly acidic
waste into the waste container
• Optical cell and flame:
– It is fused silica glass tube through which the light beam passes (from Hollow
Cathode Lamp to Monochromator
– Placed usually above the air/acetylene flame
– The flame heat decompose/breakdown the volatile hydride and reproducibly
create analyte atoms -electrical heating alternative
 Optical cell

Burner head

Peristaltic pump
waste

Mixing/reaction coil

Gas/liquid separator
 Good practices when working with AAS
• Never look directly at
– The light generated by the lamps
– The flame (it emits UV light)
• Handle the lamps carefully, and when not in use store them in
appropriate containers
• Do not use acetylene gas cylinder when pressure is <100 psi
(acetone present can enter into the gas lines and burner)
• There must be no gas leaks in tubing and connectors
• Polypropylene tube must be free from cracks, crimps and
blockages
– Periodically dismantle and clean the nebulizer
– Keep the solutions aspirated clear
 Good practices when working with AAS
• Periodically clean the burner head with the tools supplied by
the manufacturer
– Burner head can be removed and soaked overnight in deionized
water for the cleaning
– Take care while cleaning the burner head to prevent slit nicking
• Take measures to prevent flask-backs which occur when gases
are forced back into the drain lines
– Interlocks provided in the instrument prevent the flask backs
– Ensure trap on the drain tube
– Keep the drainage tube immersed in aqueous waste of the
waste receptacle
 Graphite Furnace AAS (GFAAS)
• Flameless atomizing replaces flame atomizing (otherwise similar in
all other aspects to Flame Atomizing AAS)
• An electrically heated graphite tube (graphite furnace) is used
• Sample is placed in the graphite tube for atomizing and the light
beam is passed through the tube for atomic absorption
• To ensure reproducibility, atomizing is done in 3 phases through
controlled heating (temperature-time programs)
– Dry to remove solvent by one fixed temperature-time program (50-
200°C)
– Pyrolysis and decomposition of the sample matrix by a second
temperature-time program (200-800°C)
– Atomize by rapidly increasing temperature in just few seconds to 2000-
3000°C by a third temperature program
 Graphite furnace AAS
Argon is used as
– Purge gas (to remove matrix components)
– Protective gas (to protect graphite tube from oxidative attack)
– New systems use two separate gas flows: an internal gas flow to
serve the purge gas function and an outer gas flow to serve the
protective gas function
Oxygen free environment and temporary argon gas flow
interruption make atomizing more efficient and increase the
atoms’ residence time in the atomic absorption volume
– Sensitivity is 2-3 orders more than that of the Flame AAS (metal
concentration at which light transmittance is reduced by 1%)
Physical properties of the sample (viscosity, surface tension,
density, etc.) do not influence the results
Slurries and solid samples can also be analyzed, and require
smaller quantity of sample
Lower precision, more time consuming, and demanding higher
levels of operator skills are the disadvantages
Thanks