OBJECTIVES

To optimize the burner system by flame atomic absorption spectroscopy using standard
solution.
To check the performance (sensitivity) of the atomic absorption spectroscopy using
standard solution.
To prepare a serial dilution and generate a standard calibration curve.
To determine amount of Copper (Cu) in sample.

INTRODUCTION

This experiment is all about the determination of Copper (Cu) by using Atomic
Absorption Spectroscopy (AAS). Atomic Absorption Spectroscopy uses flame to separate the
atoms of a liquid sample into gas atoms so that the light radiation can be used to excite the
atoms to a higher energy level. Thus, readings of the sample can be obtained. The purpose of
this experiment is to optimize the burner system by flame atomic absorption spectroscopy
using standard solution, to check the performance (sensitivity) of the atomic absorption
spectroscopy using standard solution, to prepare a serial dilution and generate a standard
calibration curve and to determine the amount of Copper (Cu) in a sample.

The experiment begins by preparing a serial dilution of 0.5 ppm, 1.0 ppm, 1.5 ppm, 2.0
ppm and 2.5 ppm copper standard solution mixed with 1mL concentrated acid and two
known samples of 2.0 ppm and 4.0 ppm copper standard solution. The prepared samples will
be put through the Atomic Absorption Spectroscopy (AAS) for reading. The readings will
show the amount of copper present in each of the sample prepared. Thus, a standard
calibration curve can be produced to show the relationship between the Absorbance (A) and
the Concentration (ppm).
 THEORY

Atomic absorption spectroscopy (AAS) is a spectroanalytical procedure for the

quantitative determination of chemical elements employing the absorption of optical radiation
(light) by free atoms in the gaseous state (Anonymous, 2013).

The Atomic absorption spectroscopy (AAS) technique is used for determining the
concentration of a particular element (the analyte) in a sample to be analysed. AAS can be
used to determine over 70 different elements in solution or directly in solid samples
employed in pharmacology, biophysics and toxicology research (Anonymous, 2013).

The technique makes use of absorption spectrometry to assess the concentration of an

analyte in a sample. It requires standards with known analyte content to establish the relation
between the measured absorbance and the analyte concentration and therefore relies on
the Beer–Lambert Law (Anonymous, 2013).

The Beer–Lambert law, also known as Beer's law or the Lambert–Beer law or the Beer–

Lambert–Bouguer law relates the absorption of light to the properties of the material through
which the light is travelling (Anonymous, 2013).

The law states that there is a logarithmic dependence between the transmission, T, of light
through a substance and the product of the absorption coefficient of the substance, a, and the
distance the light travels through the material the path length, b. The absorption coefficient
can, in turn, be written as a product of either a molar absorptivity of the absorber, ε, and
the molar concentration c of absorbing species in the material (Anonymous, 2013). The
formula in finding the Absorbance (A) is as shown below.

When working with A = abc

absorptivity, a

When working with

A = bc
molar absorptivity, 
 Experimental measurements are usually made in terms of Transmittance (T). The
Transmittance (T) of the solution is the fraction of incident radiation transmitted by the
solution, which is defined as:

T = I / Io

The Absorbance (A) of a solution is defined as the negative log of the Transmittance
(T) of the solution.

A = - log T = - log (I / Io) A = 2 - log %T

The formula used for the preparation of the serial dilution of copper standard solution
is as shown below:

M1V1 = M2V2

Three steps are involved in turning a liquid sample into an atomic gas:

Desolvation – the liquid solvent is evaporated, and the dry sample remains
Vaporizations – the solid sample vaporizes to a gas
Volatilization – the compounds making up the sample are broken into free atoms.

The flame is arranged such that it is laterally long (usually 10cm) and not deep. The
height of the flame must also be controlled by controlling the flow of the fuel mixture. A
beam of light is focused through this flame at its longest axis (the lateral axis) onto a detector
past the flame (Anonymous, 2013).

The light that is focused into the flame is produced by a hollow cathode lamp. Inside the
lamp is a cylindrical metal cathode containing the metal for excitation, and an anode. When a
high voltage is applied across the anode and cathode, the metal atoms in the cathode are
excited into producing light with a certain emission spectra. The type of hollow cathode tube
depends on the metal being analysed (Anonymous, 2013).
 For analysing the concentration of copper in an ore, a copper cathode tube would be used,
and likewise for any other metal being analysed. The electrons of the atoms in the flame can
be promoted to higher orbitals for an instant by absorbing a set quantity of energy (a
quantum). This amount of energy is specific to a particular electron transition in a particular
element. As the quantity of energy put into the flame is known, and the quantity remaining at
the other side (at the detector) can be measured, it is possible to calculate how many of these
transitions took place, and thus get a signal that is proportional to the concentration of the
element being measured (Anonymous, 2013). Below is a diagram showing the flow of the
operations in Atomic Absorption Spectroscopy (AAS).
 MATERIALS

Apparatus Chemicals

Perkin Elmer Atomic Absorption 1000 ppm Copper (Cu) standard solution
Spectrometer model 100 AAnalyst Concentrated Nitric Acid (69 %)
Copper Hollow Cathode Lamp Deionized Water
7 volumetric flasks Distilled Water
Beakers
Pipettes (5 mL)
Stoppers
Droppers
Suction Pumps
Labelling Stickers

PROCEDURE

1. 10 mL of 1000ppm copper standard solution was poured into a small beaker and 100ppm
copper standard solution was prepared in 100ml volumetric flask by using the equation.
2. 1 mL of concentrated nitric acid was placed into a 100 mL volumetric flask and was filled
to the mark with deionized water.
3. The freshly prepared 100 ppm copper standard solution was used to prepare 2.0 ppm and
4.0 ppm of copper standard solution using 100 mL volumetric flask.
4. 0.5 ppm, 1.0 ppm, 1.5 ppm, 2.0 ppm and 2.5 ppm copper standard solution was prepared
from the 100 ppm copper standard solution.
5. 1 mL of concentrated nitric acid was added to each of the standard solution and was
marked up to the volume.
6. The serial dilution of 0.5 ppm, 1.0 ppm, 1.5 ppm, 2.0 ppm and 2.5 ppm copper standard
solution and the 2.0 and 4.0 ppm of copper standard solution was put through the Atomic
Absorption Spectroscopy (AAS).
7. The results were obtained and the amount of copper in each sample was determined.
 RESULTS

Instrument Parameters

Standard solution concentration (ppm) Absorbance (mean)

0.5 0.010
1.0 0.021
1.5 0.035
2.0 0.053
2.5 0.066
Unknown 1 (1.126 ppm) - (0.0275) 0.029
Unknown 2 (2.181 ppm) - (0.0565) 0.055

Comparison characteristic concentration value (2 ppm) = 0.077 mg/ L

Measured characteristic concentration value (2 ppm) = 0.147 mg/ L

Comparison characteristic concentration value (4 ppm) = 0.077 mg/ L

Measured characteristic concentration value (4 ppm) = 0.153 mg/ L

% RSD (mean) = 1.54

Correlation coefficient (final) = 0.9846

 DISCUSSION

This experiment is about the AAS which stands for Atomic Absorption Spectroscopy.
The AAS concerns more to the atomic information of a metallic sample, which is different to
that of the other two spectroscopies (UV-VIS and FTIR) which concern more to the
molecular information. This experiment explains on how to utilize the AAS in order to obtain
information about the metal scanned, in this case, the metal used is Copper (Cu).

The experiment first began by preparing a 100 ppm copper standard solution. Then,
appropriate volume was calculated using the dilution formula and the volume of stock was
used along with distilled water to fill the volumetric flask to the mark in order to prepare two
dilutions which are 2 ppm and 4 ppm. Later, a serial dilution of 0.5 ppm, 1.0 ppm, 1.5 ppm,
2.0 ppm, and 2.5 ppm were also prepared but with an addition of 1 % concentrated Nitric
Acid. All the dilutions were then brought to the AAS to undergo analysis. At the AAS, the 2
ppm and 4 ppm dilutions were analysed first, followed by the serial dilutions of 0.5 ppm, 1.0
ppm, 1.5 ppm, 2.0 ppm, and 2.5 ppm (with the addition of Nitric Acid). After the analysis
was completed, the results were printed out and interpreted.

Based on the results, the Absorbance readings obtained for the order 0.5 ppm, 1.0 ppm,
1.5 ppm, 2.0 ppm, and 2.5 ppm were 0.010, 0.021, 0.035, 0.053, 0.066, 0.029, and 0.055
respectively. For the 2 ppm dilution, a characteristic concentration value of 0.147 mg/ L was
obtained when compared to the standard of 0.077 mg/ L. As for the 4 ppm dilution, a
characteristic concentration value of 0.153 mg/ L was obtained when compared to 0.077 mg/
L. The RSD % (mean) obtained was 1.54, and the Correlation Coefficient (final) obtained
was 0.9846. The concentration value for the Unknown 1 and Unknown 2 obtained through
the calibration curve were, (1.126 ppm) - (A = 0.0275) and (2.181 ppm) - (A = 0.0565)
respectively, which were obtained through the calibration curve plotted based on the results
obtained. In conjunction to that, the value of the concentration obtained through the plotted
graph is similar to that of the printed results. Hence, it can be deduced that the objectives
have been achieved.
 During the experiment, some errors might occur. Among the errors is the volumetric
flasks used may have not been cleaned properly. This error may lead to misreading and may
also lead to obtaining a disrupted result. In conjunction to that, another error that may occur
while experimenting is the sample used. If the sample is contaminated, in which means an
impure substance was selected to scan through the AAS, then the results obtained will be less
precise and less accurate and the flow of the experiment will be less efficient. Not just that,
distilled water was supposed to be added to the sample until it reaches the mark, but less or
more water was added, thus leading to abnormal results.
 CONCLUSION

This experiment revolves around the usage of Atomic Absorption Spectroscopy (AAS).
The results were obtained in a printed page. Calculations were made using the dilution
equation and serial dilutions were made to act as a standard as well as to undergo the analysis
in order to obtain a calibration curve. Through this experiment, I have learned that the atomic
information of a compound (metal), was able to be obtained and in a rather short period when
using the AAS. I have also learned that the AAS was able to determine the concentration of
the unknown samples through calibration of the serial dilutions. The calibration done by the
AAS with the serial dilutions will lead to the production of a calibration curve which
provides information of the scanned sample. Based on the results obtained, a graph of
absorbance against concentration (ppm) was plotted and a comparison has been made with
the theoretical graph from the results. For unknown 1, the absorbance reading obtained from
the theoretical graph was 0.029, and the reading from the plotted graph was 0.0275. On the
other hand for unknown 2, an absorbance reading of 0.055 was obtained from the theoretical
graph, and a reading of 0.0565 was obtained from the plotted graph. It can be observed that
the values obtained from the graph plotted and the theoretical graph was similar. Therefore,
based on the results, the objectives of this experiment were achieved. Other than that, the
experience gained from this experiment will be very useful and will definitely aid us in the
future in terms of efficiency and competency.
 RECOMMENDATIONS

To avoid errors during the experiment, early precautions must be done. Reading the lab
manual beforehand will give you a brief idea on how the experiment works and definitely
reduce the chances of making errors. The volumetric flasks that have been used in the
experiment must be clean properly so that the sample will not be mixed with other samples.
Furthermore, the sample prepared for the use of experimenters in carrying out the experiment
must not be contaminated. For instance, the Copper (Cu) prepared has to be pure and must
not mixed with other substances as the results acquired from the Atomic Absorption
Spectroscopy (AAS) will be far off from the expected results. Asking the technician to check
on the samples before conducting the experiment is highly recommended. Moreover, while
preparing the serial dilution, the addition of distilled water has to reach the mark and it should
not be over the limit as the dilution process is a permanent process. Thus, using a dropper in
conducting the serial dilution will prevent this error from appearing and aid in obtaining
maximum precision and accuracy in results. Other than that, parallax error might occur
during the experiment. Hence, managing the serial dilution on a flat and stable surface is one
of the best ways to prevent errors from occurring.
 REFERENCES

1. [Mohd Zulkhairi Abdul Rahim, 2013],

Analytical Chemistry Laboratory Manual
[20th March 2013]

2. [Anonymous, 2013],
http://en.wikipedia.org/wiki/Atomic_absorption_spectroscopy
[20th March 2013]

3. [Anonymous, 2013],
http://en.wikipedia.org/wiki/Beer%E2%80%93Lambert_law
[20th March 2013]

4. [Anonymous, 2013],
http://www.galbraith.com/spectroscopy.htm
[20th March 2013]

5. [mike_pisay2005, 2008]
http://malaysia.answers.yahoo.com/question/index?qid=20080828143645AA0fiXm
[20th March 2013]
 APPENDICES

Pre Laboratory Questions

[1] What is the function of Monochromator?

Monochromator acts as a wavelength selector. It isolates analytical lines’ photons that
pass through the flame as well as to remove scattered light of other wavelengths from
the flame. In doing so, only a narrow spectral line impinges on the PMT.

[2] You are given a 100 ppm mercury stock solution. What is the volume needed in mL to
prepare a 15 ppm standard stock solution in 50 mL volumetric flask?

M1V1 = M2V2

(100 ppm)V1 = (15 ppm)(50 mL)

V1 = 750/ 100

V1 = 7.5 mL

[3] How would you determine the performance of the Atomic Absorption Spectrometer?

Based on my experience in the laboratory, the three key performances of the AAS are:

 Simultaneous analysis of different elemental species can be done.

 It can be used for trace analysis (where the levels of the analyte is in ppm).
 Sample preparation and analysis is extremely fast.
[4] Calculate the Characteristic Concentration Value for 2 ppm copper standard that has the
absorbance value of 0.194 absorbance.

Characteristic Concentration Value = 0.0044  known concentration used

Absorbance for known concentration used

Characteristic Concentration Value = (0.0044)  (2 ppm)

0.914

= 9.628  10-3 ppm

Post Laboratory Questions

1. Ignator 4. Horizontal adjustment knob

2. Burner 5. Vertical adjustment knot

3 No bulizer adjustment nut