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Atomic absorption spectroscopy (AAS) for elemental analysis of plant samples

Method · January 2017

DOI: 10.13140/RG.2.2.20976.15367

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 Manual of ICAR Sponsored Training Programme on “Physiological Techniques to Analyze the Impact of
Climate Change on Crop Plants” 16-25 January, 2017, Division of Plant Physiology, IARI, New Delhi

Atomic Absorption Spectroscopy (AAS) for Elemental Analysis of Plant Samples

Vijay Paul1*, Rakesh Pandey1, Ramesh K.V.2 and R.C. Meena1

1
Division of Plant Physiology, ICAR- Indian Agricultural Research Institute (IARI), New Delhi-110 012
2
Plant Physiology, ICAR-Indian Institute of Seed Science, Mau, U.P.
*E-mail: vijay_paul_iari@yahoo.com

Introduction This analytical method based on atomic absorption

Mineral elements particularly metals play important
role in biological system including plants. Magnesium
(Mg) is in chlorophylls, iron (Fe) is in hemoglobin
and cadmium (Cd) and aluminium (Al) are toxic to
plant metabolism. A technique referred as atomic
absorption spectroscopy (AAS) help in quantification
of these elements and thereby their importance. AAS
is therefore an important technique for agriculture
(soil, plant, water and fertilizers analysis) and other
areas including medical, feed stock, environment,
pollution, industrial effluents metallurgy, oil industry
and general inorganic analysis etc.
Principle and basic operation of AAS
The absorption spectrum of an element in its gaseous
atomic state consists of a series of well defined and
extremely narrow lines arising by the electronic
transitions of outer most electrons. In case of metals,
most of these transitions belongs to visible and UV
regions. Most obvious wavelengths at which
absorption or emission is observed are associated
with the transitions where minimal energy change
occurs e.g., 3s-3p transition in Na atom gives rise
the emission of yellow light and this is referred as
D-line transition. For the analysis, the sample
(element) is converted into atomic vapour and then
the absorption, either in visible and UV region (at a
selected wavelength), which is specific for a given
element is measured. For converting element into its
gaseous atomic state initial step in the whole
procedure of estimation involves spraying a solution
of the sample into the flame. This process is
accomplished by drawing the solution of the sample
as a fine mist into a suitable flame. In terms of
Functionality, the flame serves as analogous to cell
or cuvette containing the solution in the conventional
spectroscopy.
is highly sensitive because atomic absorption lines
are extremely narrow (0.02-0.5A) and transition
energies are unique for each of the element.
However, the limited line width creates a problem in
AAS measurement which is usually not the case in
the spectroscopy for a solution. In order to follow
the Lambert Beer’s law, it is necessary that the band
width of the source must be narrow to the band width
of the absorption peak. Even good quality
monocromators with effective band widths has
significantly greater bandwidth than the bandwidth
of the absorption peaks. So, when the atomic
absorption measurements are made with ordinary
monochromators equipped with continuous source
of radiations then also non-linear calibration curves
were obtained. The problem of limited width of the
atomic absorption peak is solved by the use of line
sources with band width narrower than the
absorption peaks. It is therefore, necessary to use
the exciting beam that contains high intensity of light
of the required wavelengths. This is achieved by the
use of hollow cathode lamp. For example, the
absorbance peak at 213.9 nm is to serve the basis of
determining zinc then the emission peak of the zinc
at the same wavelength is isolated and used. In this
instance, the line is produced by the zinc vapour lamp
exciting the zinc atoms by an electric discharge. The
other zinc lines emitted from the source are removed
with the help of filters or with a relatively cost-
effective monocromator (Figure 1).
Components of AAS
Fuels and oxidants
Fuels Natural gas- Hydrogen, propane, butane
and acetylene (acetylene is most
important and widely used)
Oxidants Air, oxygen rich air and nitrous oxide
(N 2O)

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 Manual of ICAR Sponsored Training Programme on “Physiological Techniques to Analyze the Impact of
Climate Change on Crop Plants” 16-25 January, 2017, Division of Plant Physiology, IARI, New Delhi

Flame

Light Source
Monochromator Photomultiplier
(Hollow cathode lamp) Detector

Fuel Oxidant

Sample
Figure 1: Schematic diagram showing the principle and working of atomic absorption spectroscopy (AAS)

Flame type Approximate Temp. (ºC) Element suitable for analysis

Air-coal gas 1800 ºC Cu, Zn, Pb, Cd, alkali metals
Air-propane 1900 ºC Same as above and volatile elements, noble metals
Air-acetylene (lean) 2300 ºC Alkaline earth methal
Air-acetylene (rich) 2300 ºC Sn, Ba, Cr etc.
N2O/acetylene 2955 ºC Al, V, Ti, Ta, Be, Se etc.

Low temperature flame (generated by normal LPG
gas) produces satisfactory results for elements which
are readily converted into atomic state (e.g., Cu, Zn,
Pb and Cd). High temperature flame (air-acetylene
mixture) is used for the elements which produce
refractory oxides and therefore require somewhat
high temperature for decomposition. Oxygen-
acetylene or N2O-acetylene may be employed for
elements which form stable oxides (e.g., Al, Be and
rare earth elements). Consumption of different fuels
during operation is quite variable. Air-acetylene flame
(1-4 lit/min), N2O-acetylene flame (4-9 lit/min) and
N2O flame (10-20 lit/min). N2O undergoes internal
freezing at the temperature lower than 20ºC and
therefore under these conditions infra red heat lamp
can be directed on the regulator for the normal and
required flow of the gas during the operation.
Burners
Two types of burners made up of corrosion resistant
material are used in AAS, stainless steel burner for
air-acetylene flame and titanium burner for N2O-
acetylene flame.
Analytical variables
There are different variables which are measured
by the instrument, they include; rate and flow of the
oxidant and fuel gases, position of the beam with
respect to the flame, nature of fuel and oxidant, nature
of anion present and rate of entry of the sample.
Among all these variables, the last one is of the
special value. If the rate is too low, the number of
atomic particles will be small, while the higher rate
of entry will cause an inordinate consumption of flame
energy in the evaporation process and very small
amount of energy is thus available for the formation

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 Manual of ICAR Sponsored Training Programme on “Physiological Techniques to Analyze the Impact of
Climate Change on Crop Plants” 16-25 January, 2017, Division of Plant Physiology, IARI, New Delhi
of the atoms. Viscosity of the solvent and the
presence of combustible organic solvents also
influence the atomic absorption peak. Two solutions
with same concentration of metal but with difference
in the other extraneous materials may give divergent
readings.
Advantages of AAS
 Minute amount of an element can be determined
even in the presence of high concentration of
other elements.
 The technique is more sensitive than emission
flame photometry.
 It is highly specific for a given element.
 It is rapid and requires only small amount of
material.
Limitations of AAS
 There is requirement of separate lamp for each
of the element to be determined.
 Elements such as Al, Ti, W, Mo, V, Si cannot be
detected. This is due to the fact that these
elements give rise oxides of the atoms in the
flame.
 The use of this technique is limited to metals
only. With non-metals difficulties arise from the
strong absorption of light by the oxygen in the
light path and from the flame gases themselves.
 If aqueous solutions are used the prominent anion
can affects the signal to a noticeable degree.
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