Experiment 4:Flame Atomic Absorption Spectroscopy (AAS)

Theory/Background

Analytical chemistry is the study of the separation, identification and quantification of

the chemical components of natural and artificial materials. Qualitative analysis gives an
indication of the amount of one or more of these components. The separation of components
is often performed prior to analysis. Analytical methods can be separated into classical and
instrumental. Classical methods use separations such as precipitation, extraction, and
distillation and qualitative analysis by color, odor or melting point. Quantitative analysis is
achieved by measurement of weight or volume. Instrumental methods use an apparatus to
measure physical quantities of the analyte such as light absorption, fluorescence, or
conductivity. The separation of materials is accomplished using chromatography,
electrophoresis or Field Flow Fractionation methods.

Flame coloring was used for identification of sodium and potassium at 1758 by
MarGaff. Wollaston was the first who notes the presence of black line in spectrum of the sun
at 1802.The scientist called Fraunhoffer explained the presence of these line in 1815,where at
that time it was called fraunhoffer lines. In 1822 Hersel noticed that the spectrum of the flames
containing salts. This is the beginning of the spectroscopy. In 1859 Bunsen and Kirchoff
explained that when we transfer energy to the atoms or certain gases, they emit a characteristics
spectrum consisting of lines. These atoms can absorb light having the same wavelength as the
wavelength of the emitted ones. First flame spectrograph was built in 1928,which is for
quantitative determination of element. The advance spectroscopy was built in 1957 equipped
with electric part and proper light source based on the result of Walchs and coworkers.

In atomic absorption spectroscopy (AAS), light is absorbed by the atoms of an analyte.

If a solution containing a metal is aspirated in the form of aerosol into the hot flame, the
liquid/solvent is evaporated droplets and the metal is vaporized mainly as atoms. Alternatively
an atomic vapor can be produced by rapid electrochemical heating of graphite rod or tube on
which a drop of the sample has been placed. If the sample contains a particular element, its
atoms will selectively absorb some of the radiation thereby attenuating the beam and causing
the detector signal to fall. This absorbance is proportional to the concentration of that element
in the vapor state and hence in the original sample.
The atomization of a sample can be represented as follows:

Mo + h v  M* where o = ground state and * = excited state

In flame AAS, we aspirate a sample into a flame using nebulizer. The flame is lined up
in a beam of light of the appropriate wavelength. The flame (thermal energy) causes the atom
to undergo a transition from the ground state to the first excited state. When the atoms make
their transition, they absorb some of the light from the beam. The more concentrated the
solution, the more light energy is absorbed.

The light beam is generated by a lamp that Is specific for a target metal. The lamp must
be perfectly aligned so the beam crosses the hottest part of the flame and travels into the
detector. The detector measures the intensity of the beam light. When some of the light is
absorbed by a metal. The beam’s intensity is reduced. The detector records that reduction as an
absorption. That absorption is shown on a read out by the data system. The figure above shows
a schematic diagram of a flame AAS. As the diagram indicates, there are four primary parts to
the system – the light source, the flame apparatus, the detector, and the data system.

Fortunately, we do not need to separate solutions containing different metals. We

merely change lamps and adjust the detector wavelength.

We can find the concentrations of metals in sample running a series of calibration

standards through the instrument. The instrument will record the absorption generated by a
given concentration. By plotting the absorption versus the concentration of the standards, a
calibration curve can be plotted. We can then look at the absorption for a sample solution and
use the calibration curves to determine the analyte concentration in that sample.

Atomic absorption is a very sensitive technique compared with ordinary UV-Visible

spectrophotometry and can therefore be used for trace analysis. Flame atomic absorption is a
very common technique for detecting metals and metalloids in environmental samples. It is
very reliable and simple to use where the amount of light absorbed can be measured against a
standard curve to determine the concentration of the metal present

Objective:

1. To learn how to operate a flame AAS instrument

2. To plot a standard calibration curve for determination of Ca in a sample
3. To determine the amount of Ca in a sample using the standard calibration curve.

Apparatus

Beaker

Burette

Retort stand

Dropper

Volumetric flask, 50ml

Chemicals

Distilled water

1000ppm Ca/Cu/Cr/Zn stock solution (depending upon the lab instructor)

Unknown solution
Procedure

In this experiment, the liquid sample was introduced to the flame by aspirating it into the air
supplied to the flame. The place was taken at nebulizer chamber where sample solution was
broken up into very fine mist of droplets (aerosol) before entering the flame. In the flame, the
solvent evaporates and most of the element is converted in free atoms

The Ca was analyzed in a liquid sample.

A. Preparation of standard solutions

1. 10mL of 1000ppm stock solution was pipetted. It was transferred into 100mL
volumetric flask. And diluted with deionized water until calibration mark. The solution
was labelled ‘100ppm’
2. The 100ppm solution was used to prepare series of standard solution with concentration
of 1ppm,3ppm,5ppm,7ppm,9ppm by using 100ml volumetric flask.
3. The unknown solution was obtained
4. The absorbance of standard solution in (2) was measured, followed by the unknown

B. Operating of the AAS

Instrument: Perkin Elmer AAnalyst 700

Safety information

1. Do not leave flammable material near AA

2. Do not let acetylene tank level drop below 70psi
3. Do not stare into the lamp

Igniting and optimizing the Flame (Air / Acetylene operation)

1. The burner height was adjusted by turning the horizontal adjustment knob until the light
beam is centered over the burner slot.
2. The vertical positioning of the burner head was adjusted by rotating the knob until the
light is near but not hitting the top of the burner head.
3. The acetylene tank was opened and checked the flow, which to be from 85-100 kPa
4. The air valve was open and the flow was checked to be from 50-65 psi
5. The Oxidant switch was turned on to Air
6. The ignite button was pressed
7. The blank (water) was aspirated for 60-90 seconds. [A/Z] was pressed.
 8. A mid-concentration standard was aspirated. The horizontal and vertical was rotated
was adjusted to obtain maximum absorbance.
9. A blank was aspirated. The aspiration hose was leaved in the water.

Measurement of Atomic absorbance

1. The blank was aspirated by pressing [Cont],[A/Z]

2. The standards was aspirated and the absorbance was recorded for each one.
3. The samples was aspirated and the absorbance was recorded for each one

Shut down procedure

1. The water was aspirated for few minutes to clean the burner head.
2. The gas control switch on the pneumatics control panel was turned off
3. All the gas sources valves was closed
4. The gas control switch was turned to Air to allow the gasses to bleed. The gas control
was the turned off.
5. The instrument was turned off.

How to Run the AAS

1. The air compressor, stock cock air, acetylene regulator and acetylene tank was opened.
2. The compressed air must between 5-6kg/cm3 and the acetylene pressure is 100kPa.
3. Exhaust fan was switched on
4. Isolating switch was switched on
5. The aspiration hose should be in water or blank.
6. Instrument was turned on
7. The computer was turned on
8. WinLab 32 was double clicked until the calibrating equipment is ready.
9. The method was created by clicking on click file  new  method  starting
condition : Element Ca
10. Method editor : untitled (type in the date)
Method description: To determine the concentration of Ca
Calibration equation: linear through zero
Change unit (for standard and unknown)
11. Standard concentration
Blank: DI water
Standard: Std 1, Std 2, Std 3, Std 4 and Std 5
Concentration: 1ppm, 3ppm, 5ppm,7ppm, 9ppm
12. File  save as  method
Name:
Method:
13. File  new  sample info file  ok
Sample ID: unknown Ca
14. File  save as  sample info
File name:  save
15. Click workspace (Workspace) – highlight whatever is in the box  open  type in
your name
16. Click burner switch  analyze standard  analyze blank  analyze sample (unknown)
17. Switch off burner
18. Click tools  edit calibration
19. To print  click file  active window preview
20. To open the file (for future reference), click ‘file’  open workspace Filename 
ok  analyze  Recall calibration
Report

1. Using any available software, plot a standard calibration curve of absorbance versus
concentration of standard and determine the concentration of Ca in the unknown.

Standard concentration vs Volume of added standard

30

23.93
25
20.57
Concentration(ppm)

20
14.69
15
11.65

10
5.72
5

0
0 5 10 15 20 25 30
Volume standard added(mL)

Absorbance vs Concentration
0.7 0.66

0.57
0.6

0.5
0.41
Absorbance

0.4
0.32
0.3

0.2 0.16

0.1

0
0 5 10 15 20 25 30
Concentration(ppm)

2. Attach all printouts obtained from your experiment in the lab report
3. Discuss the principle of operation of the AAS you used and include the function of each
component.(In discussion)
Questions
In a standard addition method, you will prepare a series of solutions in which you add different
increments of standard solutions to fixed aliquots of an unknown sample X i.e. 10.00mL of X.
All solution were prepared using 50mL volumetric flasks. Suppose that the analysis of standard
solutions of X gave the following results:

Solution Concentration of added standard,ppm Absorbance

Standard 1 0.00 0.201
Standard 2 2.44 0.292
Standard 3 4.88 0.378
Standard 4 7.32 0.467
Standard 5 9.76 0.554

1. Plot the standard addition graph

Absorbance vs Concentration
0.6 0.554

0.5 0.467

0.378
0.4
Absorbance

0.292
0.3
0.201
0.2

0.1

0
0 2 4 6 8 10 12
Concentration(ppm)

2. Determine the concentration of X(in ppm) in the unknown sample.

 DATASHEET EXPERIMENT 4:

FLAME ATOMIC ABSORPTION SPECTROSCOPY (AAS)

Name: Marvellis anak Machillies Date: 17 October 2018

Student ID: 2016248128 Group: AS1205B

Name and model of instrument:

Table 4.1: Table of added volume and concentration of diluted standard

Solution Volume Of added standard,mL Concentration of added

standard, ppm
1 5 5.72
2 10 11.65
3 15 14.69
4 20 20.57
5 25 23.93

Unknown Number:

Name of element: Iron

Concentration of unknown: 11.43ppm

Lecturer’s signature:

_______________________
Discussion:

The purpose of the experiment is to:

Atomic Absorption Spectroscopy is an analytical technique used for the qualitative and
quantitative determination of the elements present in different samples like food,
nanomaterials, biomaterials, forensics, and industrial wastes. The technique is based on the
principle that ground state metals absorb light at specific wavelength. Metal ions in a solution
are converted to atomic state by means of a flame. When light of the correct wavelength is
supplied, the amount of light absorbed is measured and a reading can be obtained. Flame atomic
absorption is a very accurate quantitative technique and also a good qualitative technique. This
is one of the main reasons it is the most widely used of the atomic absorption methods. The set
up for most Flame atomic absorption spectrometers are relatively simple in design. The most
complicated part of the instrument is the nebulizer. The nebulizer system is highly important
in FAAS. The nebulizer converts the sample solution into a mist or aerosol. The nebulized
sample is then carried into the flame. Radiation from the specific light source is focused on the
atomic vapor in the flame. The radiation then enters the monochromator, which isolates the
line in the vapor which then the light is measured by a photomultiplier tube (detector). The
signal is then process and the computer system prints the output on screen. Flame atomic
absorption spectroscopy is a sensitive technique for the quantitative determination of more than
sixty metals as it is used for determining the concentration of metals it can be applied in
Environmental Analysis which can also detect if there is trace of metal present in food.
Conclusion:

In the nutshell, during this experiment and after this experiment was conducted we were
able to learn on how to handle and operate Flame Atomic Absorption Spectroscopy instrument
well and at the same time was able to plot a standard calibration curve for determination of Ca
in a sample and determine the amount of Ca in a sample using the standard calibration curve.

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