Atomic Absorption Spectrophotometer

Zool-711 (Practical)
Dr. M. Naeem
20 & 21 April 2020
• Atomic absorption is the determination of the presence and concentrations of
metals in liquid samples.

• Metals include Fe, Cu, Al, Pb, Ca, Zn, Cd and many more.

• Typical concentrations range in the low mg/L (ppm) range.

• In atomic absorption spectrometry, light of a specific wavelength is passed through

the atomic vapor of an element of interest, and measurement is made of the
decrease in the intensity of light as a result of absorption by atoms in the 'excited'
state
• Metals will absorb ultraviolet light in their elemental form when they are excited by heat, either by flame
or graphite furnace.

• Each metal has a characteristic wavelength that will be absorbed.

• The sample of interest is aspirated and atomized into the flame.

• If that metal is present in the sample, its atoms will absorb some of the light, thus reducing its intensity.

• This decrease in intensity of the light is the process of atomic absorption.

• The instrument measures the change in intensity.

• A computer data system converts this change into an absorbance.

Hollow cathode lamp
• The Hollow cathode Lamp is made with an element
(metal) as the cathode encased in a bulb with a low
pressure inert gas.

• It is imposed with a 10mA current such that when the

metal is excited electrically, characteristic spectral
line/lines are emitted and passed directionally through a
glass UV/VIS window of the lamp.

• This is the Source of the analytical light line for the

element which is to be measured. It gives a constant and
intense beam of the specific analytical line or
wavelength of the element.
 Nebulizer
• The nebulizer sucks up liquid sample at a controlled rate, create a fine aerosol that mixes
with fuel and oxidant for introduction into the flame.

• The nebulizer uses the combustion flames to atomize and introduce the sample into the
light path.

• The high velocity of the combustion gases cause a negative pressure inside the nebulizer
chamber and create a suction for an uptake tube by a process of aspiration.

• At the same time, the combustion gases rushing through the Nebulizer draws liquid
sample into the flow and is introduced into the flame as very fine droplets.

• Excess aspiration liquid is removed by gravity and the waste is collected through an exit
tube into a glass container. This waste is still highly acidic and care should be taken in its
handling and disposal.
 Flame

• An oxidant and a fuel gas are mixed together and lit to create the
flame.

• The Flame is the Atomizer in which the sample undergoes desolvation

and vaporization at high temperature.

• It destroys any analyte ions, break up complexes and creates the atoms
of the element of interest, eg. Feo, Cuo, Zno, etc.
 Monochromator

• The main purpose of the monochromator is to isolate a single atomic resonance line
(wavelength) from the lines emitted by the Hollow cathode lamp and transmit it to the
detector. It not only selects the specific analytical line, but excludes all other interfering
lines in that region.

• Polychromatic light from the light source enters the monochromater and a dispersive
device selects the wavelength to be measured and sends it through the exit slit to the
detector. The dispersive device is a defraction grating which is a glass block machine-
grooved in the order of 3000 grooves per millimeter and coated with highly reflected
aluminium. Light striking these grooves are reflected and dispersed according to
wavelength. By means of a selector knob on the monochromator a single analytical
wavelength of choice can be selected and focused through the exit slit via the second
mirror.
Samples
The samples and standards are often prepared in duplicate with acid to match the analyte's chemical matrix as closely as possible.
Acid contents of 1% to 10% are common. High acid concentrations help keep all dissolved ions in solution. The blank is a solution
representative of the matrix of samples and standards but without the analyte.

Autosampler
With the autosampling accessory, tedious and time-consuming operations can be avoided. The autosampler is a computer-controlled,
multi-purpose system. It automates standard and sample introduction for instrument calibration and analysis, and is a fully automated
analytical workstation.

Wastes
Liquid sample not flowing into the flame collects on the bottom of the nebulizer chamber and flows by gravity through a waste tube
to a glass waste container (highly acidic).

Shutdown
Before shutdown, aspirate deionized water through the system to flush out salts and acid. Then close off fuel first, then oxidant, and
bleed gas supply cylinders. Most modern instruments control the ignition and shutdown procedures automatically.

Maintenance
The Atomic Absorption Spectrophotometer is designed to give accurate and precise results. The instrument set up and operation
parameters should be observed as outlined in the Instrument Instruction Manual. However, occasional problems may arise that
frustrate the less experienced operator. Most of these problems arise in the area of sample introduction and improper maintenance.
Attention must be focused on lamps and burner, combustion gases, nebulizer tubing and drain assembly. A corrective diagnostic and
maintenance program should be etablished in order to solve the problems associated with the instrument and the analytical method.