Lesson 2: Atomic Absorption Spectroscopy (AAS)

Learning outcomes

Upon completion of this lesson, the learners will be able to

▪ Define atomic spectroscopy;

▪ Classify atomic spectroscopy;
▪ Describe the principles of AAS and AES spectroscopy;
▪ Describe the instrumentation and working procedure of AAS;
▪ Determine the concentration of unknown analyte by standard addition method.

2.1 Introduction
Atomic absorption spectroscopy (AAS) and atomic emission spectroscopy (AES) is a
spectroanalytical technique used to quantitative determination of chemical elements on the basis of
absorption/emission of radiation (light) by free atoms in the gaseous state. The more the number of
the atoms in a given sample, the higher is the intensity of absorption/emission and vice-versa. The
quantity of sample needed for analysis is low. Very low concentration of sample ranging from one
part per million (ppm) to one part per billion (ppb) can be detected. This is also known as metal
analysis spectroscopy as it is mainly used for the analysis of metals. Nearly all elements can be
detected in the sample by these methods (Figure 1).

Fig. 2.1 Elements detectable by AAS are highlighted in pink in this periodic table
2.2 Classification of atomic spectroscopy
Atomic spectroscopy can be classified in two major ways-
1. Atomic absorption spectroscopy
Atomic absorption spectroscopy is an analytical technique used for the determination of metal at
trace and ultra-trace levels by measuring how much light is absorbed by metal atoms in the ground
state.

2. Atomic emission spectroscopy

Atomic emission spectroscopy is an analytical technique used for the quantification of elements in
a given sample by measuring how much light is emitted by excited-state atoms.

2.3 Principle of AAS

It uses the principle that free atoms in gaseous state can absorb light at a specific, unique wavelength.
When this specific wavelength of light is passed through the atoms, the energy (light) is absorbed
by the ground state atoms. Electrons in the atom move from the ground state to an excited state. The
amount of light absorbed by the ground state atoms is measured and the intensity of absorption is
directly proportional to the concentration of the sample

2.4 Principle of AES

In AES, the electrons in the gaseous state atoms are excited when heated at high temperatures. The
excited atoms return to the ground states by emission of radiation in the form of discrete wavelength
packets. The concentration of atoms can be determined by measuring the intensity of the emitted
light (radiation). An atomic emission spectroscopy is identical to an AAS except
that no external light source is used to excite the atoms.

2.5 Working of AAS

1. Solid samples are converted into solution form.
2. Liquid sample is converted into fine mist by nebulization with the help of nebulizer.
3. Vaporized sample is mixed with fuel and oxidant.
 4. Sample is converted to atoms by heating.
5. Light characteristics to the atom are passed though it which leads to excitation of the atoms
by absorbing light.
6. Emission of light take place by excited atoms when it returns to ground state.
7. Concentration of analyte atom is determined by measuring the intensity of absorption or
emitted light.

2.6 Atomization
The process of converting an analyte to a free gaseous atom is called atomization. Converting an
aqueous analyte into a free atom requires to evaporate the solvent, volatilize the analytes, and, if
necessary, dissociate the analyte into free atoms.

Desolvating an aqueous solution of CuCl2, for example, leaves solid particulates of CuCl2.
Converting the particulate CuCl2 to gas phase atoms of Cu and Cl requires thermal energy.
CuCl2(aq)→CuCl2(s)→Cu(g)+2Cl(g)

2.7 Instrumentation of AAS

Any atomic absorption spectroscopy consists of the following types of components:

1. Atomizer
2. Light source
3. Monochromator
4. Detector
5. Recorder

Fig. 2.2: Schematic presentation of AAS

1. Atomizer
Atomizers are used for the conversion of solid or liquid samples to free gaseous atoms. The
temperature should be controlled very carefully for the conversion of atomic vapor. At too high
temperatures, atoms can be ionized.
Atomizers can be
a. Flame atomizer
b. Non-flame atomizer/Electrothermal atomizers
a. Flame atomizer
Fuel (acetylene) and oxidant (e.g., oxygen, air) gases are fed into a mixing chamber. A portion of
the sample solution is aspirated through a nebulizer and sprayed as a fine aerosol (~ 5 -10 nm) into
the mixing chamber and mixed with fuel and oxidant with the use of baffles prior to introduction
into the flame. The finest droplets of sample mist, or aerosol, are carried with the combustion gases
to the burner head, where atomization takes place. The excess sample is removed from the premix
chamber through a drain.
Nebulizer and burner heads are made of stainless. In case of high content of acid or other corrosive
reagents in the sample. nebulizers are constructed of a corrosion resistant material, such as an inert
plastic, platinum alloys or tantalum.

b. Electrothermal atomizers

Electrothermal atomizers, also known as a graphite furnace atomizer, that can produce temperatures
as high as 3,000°C.
The heart of the Graphite furnace is the atomization chamber which is made up of a graphite tube
open at both ends with a hole in the center for sample introduction. It is encased within graphite
electrical contacts at both ends and the sample is electrothermally vaporized into atoms. Water is
circulated outside of the tube to keep the furnace cool, and an external stream of inert gas flows
around the tube to prevent oxidation.
 To atomize sample, first, the sample is injected into graphite tube hole and dried at a low temperature
(120C). Then the sample is ashed at (~600oC) in order to remove matrix (anything other than
sample) in a graphite furnace, followed by a rapid temperature (2000-3000 oC) increase within the
furnace where the sample becomes a vapor containing atoms from the sample.
This atomizer used to detect trace and ultra-trace levels (down to low μg/L) while using only small
sample volumes (usually less than 100 μL).

2. Light source
a. Hollow cathode lamp (HCL)
The hollow cathode lamp has two electrodes: cathode (made of same element under test) and anode.
The anode and cathode are sealed in a glass cylinder normally filled with either neon or argon at
low pressure. At the end of the glass cylinder, there is a window transparent to the emitted radiation.
When an electrical potential is applied between the anode and cathode, some of the fill gas atoms
are ionized. The positively charged filled gas ions accelerate through the electrical field to collide
with the negatively charged cathode and dislodge some individual metal atoms in a process called
‘‘sputtering”. Sputtered metal atoms are then excited through kinetic energy transfer by filled gas
ions. When excited atoms come down to ground state, it emits wavelength of radiation characteristic
to the metal used to make the cathode. The emitted wavelengths are extremely narrow. Unlike
continuum radiation, the narrow-emitted radiation (wavelength) from the HCL can be absorbed
almost completely by unexcited atoms.

+
_-
b. Electrodeless discharge lamp (EDL)
The hollow cathode lamp is a good source of radiation for most of the elements to be analyzed. For
certain volatile elements (such as arsenic, germanium, or selenium) are very difficult to make stable
hollow cathode lamp due to low intensity and short lamp life. These problems can be solved by
using more stable light sources such as ‘‘electrodeless discharge lamp’’.
A small amount of the same metal or salt of the element to be determined is placed inside a sealed
quartz bulb with a low pressure of argon gas. This bulb is placed inside a small, radiofrequency (RF)
generator coil. When power is supplied to the coil, RF (microwave) (300-3000 MHZ) is generated.
The heat generated by RF vaporize and excite the atoms inside the bulb which emit characteristic
radiation to the element.

Advantages

• EDL produces higher emission intensity and sensitive than HCL.

• A lower contamination due to the absence of the electrodes, and a longer lifetime
3. Monochromator
A monochromator is used to select the specific wavelength of light which is absorbed by the sample,
and to exclude other wavelengths. It consists of a diffraction grating (dispersing element-
Prisms/Gratings), slits, and mirrors.

Fig. A typical monochromator design

Light from the source enters the monochromator at the entrance slit and is directed to the grating
where dispersion takes place. The diverging wavelengths of light are directed toward the exit slit.
By adjusting the angle of the grating and focusing mirror a selected emission light from the source
can be allowed to pass through the exit slit and fall onto the detector. All other lights are blocked
from exiting.
4. Detector
The wavelength of light which is isolated by the monochromator is directed onto the detector that
is typically a photomultiplier tube which converts light energy into electrical signal and amplified
in order to produce readable signals. A detector can be tuned to respond by a specific wavelength
or frequency.

5. Recorder
The recorder can receive electrical signals from the detector to convert them into a readable
response. Today, a computer system with suitable software is used for recoding signals coming from
the detector.
*

2.8 Atomic Absorption vs Atomic Emission Spectroscopy

Atomic Absorption Spectroscopy (AAS) Atomic Emission Spectroscopy (AES)

AAS is used to determine the concentration of
metal atoms on the basis of absorption of light.
Electromagnetic radiation is absorbed by atoms
present in ground state.
The obtained spectrum is dark lines with color
background.
It requires a light source.
It depends upon a number of ground-state
atoms
AES is used to determine the concentration of
metal atoms on the basis of emission of light.
Electromagnetic radiation is released by atoms
present in excited state.
The obtained spectrum is colored lines with
dark background.
It does not require a light source
It depends upon the number of excited-state
atoms

2.9 Types of interferences in AAS and their remedies

1. Spectral interferences
This type of interference normally takes place when the absorption lines of interfering species either
overlaps or lies very close to the analyte absorption. It may cause due to another element, radical,
molecule, combustion products (broadband absorption), sample matrix etc.

Fig. 2: Absorption spectra of a mixture of arsenic (As) as analyte (A) and cadmium (Cd) as the
interferent.
This problem can be solved by choosing alternative wavelength using smaller slit width or tune the
monochromator to a different spectral line for the element of interest so that there is no overlap.

2. Scattering
This is caused by scattering of the incident radiation from the source to matrix impurities.
Incomplete combustion of organic materials which give rise to molecular species that exhibit broad
band spectra. For example, refractory oxides formed by Ti, Zr, and W due to atomization of high
concentration solutions. This problem can be solved by using blank.

3. Chemical interferences
Chemical interference occurs when an analyte is not totally decomposed in flame. There is less
atoms present, and therefore a reduced absorbance of the analyte. Calcium signal is depressed due
to formation of CaSO4 or Ca3(PO4)2. This problem can be solved by the addition of EDTA which
form complexes with the calcium and eliminates these interferences or adding excess releasing agent
such as strontium or lanthanum ions which complex with the phosphate and sulphate ions and
improve the sensitivity of the calcium measurement.

4. Ionization interference
At elevated flame and furnace temperatures, atoms with low ionization potentials become ionized.
Any ionization reduces the population of both the ground state and the excited state of neutral free
atoms, thus lowering the sensitivity of the determination.
This problem is readily overcome by adding an excess (ca. 100-fold) of a more easily ionized
element such as K, Cs, or Sr to suppress ionization of analyte. Ionization of the suppressor forms
more electrons and greater charges of positive ions that suppress the ionization of the analyte
species. The addition of suppressants is even more important in analyses that require the hotter
acetylene/nitrous oxide flames.

5. Background absorption of source radiation: This is caused by the presence of a particle from
incomplete atomization. This problem is overcome by increasing the flame temperature.

2.10 Determination of metal analyte concentration by standard addition method

The standard addition method analyzes an unknown sample and the same unknown sample spiked
with a known amount of target component, then uses absorbances of unknown and spiked samples
to determine quantity of unknown.
The matrix is everything in the sample, other than analyte. This method is used when analyte
contains matrix (substances other than analyte) that interferes with the signal (increase or decrease)
of analyte. Analyte present in biological fluids, food, soil samples contains complex matrix. The
idea is to add standard to the sample ("spike" the sample) to monitor the change in instrument
response.
Standard addition method can be classified in two ways-
1. Single point standard addition
2. Multiple point standard addition

1. Single point standard addition

a. Add a fix volume of sample (e.g., 10 ml) in two volumetric flasks (e.g. 100 ml).
b. Now add a fix volume of standard (same to
analyte) (e.g., 10 ml of 100 ppm) in any one of the
above flasks and mix well. Calculate
concentration of standard in the solution
(S1V1=S2V2 or 10100= S2100 2 or S2=10 ppm)
c. Now adjust the volume up to the mark (100 ml) by
distilled water and mix well.
d. Take absorbance of the both samples by AAS.
Calculation of concentration of analyte
Let’s the concentration of analyte and standard in
diluted samples are Cx and Cs and absorbances are Ax
and AT, respectively. According to Beer’s law-
Ax Cx or Ax=k Cx ………………………..(a)
AT  (Cx+Cs) or AT=k (Cx+Cs)……………(b)

As long as Vs is small relative to Vx, the effect of the standard’s matrix on the sample’s matrix is
insignificant. Under these conditions the value of the k is same in both equations.

Dividing equation (a) by (b), we get

Ax kCx Ax AT Ax
= or = or Cx = Cs  … … … … … … … … … . (c)
Cx k(Cx + Cs ) Cx (Cx + Cs ) (AT −Ax )
Now multiply the value of Cx by dilution factor to get concentration of the analyte in the sample.

Advantages:
✓ This method is routinely used when the expected range for the analyte’s concentrations is small.
✓ Simple, less expensive and faster than the traditional multi-level standard additions:
2. Multiple point standard addition
In this method, several solutions containing the same amount of sample and different amounts of
standard solutions are prepared in order to determine the concentration of sample. At least three
standards with increasing concentration, although more are preferable should use in this method. It
is needed to have some idea of the concentration in the sample prior to analysis. The optimal size of
each standard addition is that which gives a signal 1.5 to 3 times that of the sample.
Procedure:
1. Take four volumetric flasks of 100 ml capacity (for example) and mark 1 to 4.
2. Then add a constant volume (Vunk=20 ml) of the analyte (unknown solution) to each of the
four volumetric flasks.
3. In first flask, do not add any extra standard and adjust volume up to 100 ml by solvent and
mix well. So, the concentration of standard in first flask is zero and spike= 0 ppm.
4. In 2nd flask, add 5 ml of 400 ppm standard solution (same as analyte) and adjust volume up to
the mark and mix well. Spike=20 ppm (S1V1=S2V2 or 5400= S2100 2 or S2=20 ppm).
5. In 3rd flask, add 10 ml of 100 ppm standard solution and adjust volume up to the mark and
mix well. Spike=40 ppm (S1V1=S2V2 or 10400= S2100 2 or S2=40 ppm).
6. Similarly, in 4th flask, add 15 ml of 400 ppm standard solution and adjust volume up to the
mark and mix well. Spike=60 ppm (S1V1=S2V2 or 15400= S2100 2 or S2=60 ppm).
7. Similarly, in 5th flask, add 20 ml of 400 ppm standard solution and adjust volume up to the
mark and mix well. Spike=80 ppm (S1V1=S2V2 or 20400= S2100 2 or S2=80 ppm).
8. Now, aspirate each sample and measure the absorbance by AAS.
Ideally, the highest spike concentration should be approximately equal to the concentration of
analyte in the unknown.
Preparation of calibration curve and determination of concentration of unknown
a. Preparation of calibration curve
1. Prepare a standard addition calibration curve by plotting the concentrations of added standard
(spiked) on the x-axis and their corresponding absorbance along the y-axis. A plot of the signal
intensities of the solutions vs. the added concentrations yields a straight line (y = mx + b,
where y is the absorbance, x = concentration of the spiked, b = y-intercept, m=slope).
2. To calculate the concentration of the unknown analyte, extrapolate backward until it intercepts
with the x-axis. This extrapolated value (|xE |) is the concentration of unknown sample
(ignoring negative sign).
3. xE can be calculated from equation, y=mx+b. When y=0, x= xE, therefore, 0=mxE + b or xE
=|b/m|) (ignoring negative sign).
Advantage: The standard addition method tends to compensate for variations caused by physical
and chemical interferences in the sample solution.

Numerical problems:
In order to determine the concentration Fe in a food sample, 10 ml of sample is added separately in
4 (1 to 4) 100 ml volumetric flask. 10, 20 and 30 ml of 10 mM standard Fe solution is added 2, 3
and 4 flasks, respectively and the volume of all 4 flasks was adjusted to 100 ml by distilled water
and mixed. Well. Now, absorbance of all four flasks were measured by AAS and were 1.36, 1.88,
2.35 and 2.89, respectively. Calculate the concentration of Fe in the sample solution.

Solution:
For 1st standard: M1V1=M2V2 or 1010= M2  100 or M2=1 mM, Spiked=1 mM
For 2nd standard: M1V1=M2V2 or 1020= M2  100 or M2=2 mM, Spiked=2 mM
For 3rd standard: M1V1=M2V2 or 1030= M2  100 or M2=3 mM, Spiked=3 mM

Spiked (mM) Absorbance

(x) (y)
0.00 1.36
1.00 1.88,
2.00 2.35
3.00 2.89

We know, y=mx + b, where concentration of Fe, xE= b/m, when, y=0. The value of m and b can be
calculated from the above data in the following ways-

xi yi xi yi xi2
0.00 1.36 0.00 0.00
1.00 1.88 1.88 1.00
2.00 2.35 4.70 4.00
3.00 2.89 8.67 9.00
xi=6.00 yi=8.48 xiyi=15.25 xi2=14

Concentration of Fe = b/m=1.361/0.506=2.69 mM (diluted sample)

Concentration of Fe = 102.69=26.9 mM (original sample)

2.11 Applications of Atomic Absorption Spectroscopy

1. Water analysis (e.g. Ca, Mg, Fe, Si, Al, Ba content)
2. Food analysis (e.g. Ca, Mg, Fe, Zn, Cu etc)
3. Analysis of animal feedstuffs (e.g. M n , F e , C u , C r , S e , Zn)
4. Analysis of additives in lubricating oils and greases (Ba, C a , N a , L i , Z n ,
Mg)
5. Analysis of heavy metals in waste (Pb, Hg, As, Cr, Ni etc)
6. Analysis of blood electrolytes (C a , M g , N a , K , F e )
 Evaluation at the end of the lesson

1. Define atomic spectroscopy;

2. Classify atomic spectroscopy;
3. Describe the principles of AAS and AES spectroscopy;
4. Describe the instrumentation and working procedure of AAS;
5. Determine the concentration of unknown analyte by standard addition method.
6. Write down the applications of AAS.