EXERCISE NO.

4
Atomic Absorption Spectrometry

CHEM 137.1 2L
Group # 2

ROSALES, Abigail Jane C.

BRITANIA, Stephanie H.
PETERE, Mary Rose L.
SUERTE, Clark T.

Date performed: September 11 & 18, 2015

Date submitted: October 02, 2015

Submitted to: Sir Lloyd M. Lapoot

ABSTRACT

Atomic absorption/ emission spectroscopy is governed

by the absorption/emission of light to measure the
concentration of gas-phase atoms especially metals.
Using the working curve obtained from the standards
and
the
application
of
Beer-Lamberts
Law,
concentration of metals can be determined. The metals
Ca2+ and K+ in Bear Brand milk powder and Pb 2+ in a
water sample were studied. Results showed that the
obtained concentrations of the metals from the milk
sample were 555.23 ppm Ca2+ and 183.60 ppm K+
while 4.73 ppm Pb2+ using external calibration and4.34
ppm Pb2+ using standard addition method from the
water sample. Application of releasing agent, La, and
suppressant, Cs, which limits the activity of a certain
compound, was added to Ca2+ and K+ samples,
respectively.

I. INTRODUCTION

Atomic absorption spectrometry is used for the determination of the

concentration and presence of a metal in the sample. Metals absorb ultraviolet
light when they are excited and for each metal there is a characteristic
wavelength that can be absorbed. AAS measures the change in intensity.
Intensity is reduced when a metal is present and absorbs some light. The
change in intensity can be read in the computer as the absorbance.
Spectrophotometry, on the other hand, is the study of the absorption of light by
an analytical sample.

According to Skoog (2004), spectroscopic determination of atomic species

can only be performed on a gaseous medium in which the individual atoms (or
sometimes, elementary ions) are well separated from one another. Atomization
is the first and the most critical step in all atomic spectroscopic procedures
wherein the sample is volatilized and decomposed in such a way as to produce
an atomic gas.
The most widely used method to atomize the sample for atomic
spectroscopic studies is flame atomization. The sample produced atomic
absorption, emission and fluorescence spectra. Atomic absorption spectroscopy
(AAS) measures the radiation absorbed by the atom in ground state when it is
excited to a higher energy level while atomic emission spectroscopy (AES) is the
opposite which involves the quantification of discrete radiation that is emitted
by excited atoms upon returning to the ground state. Atomic fluorescence
spectroscopy (AFS), on the other hand, quantifies the amount of energy emitted
by atoms that have been excited via radiation from spectral source. AFS is
different from AES since the latter uses thermal, chemical or electrical
mechanism in order to excite electrons into a higher energy level. AAS or AES
differs from visible spectrophotometer since the former involves measurement
of the absorption/emission of light by vaporized ground state atoms while the
latter simply measures the % transmittance and absorbance of a solution as it
absorbs certain intensity of light at different wavelengths. The block diagram for
visible spectrophotometer, AAS and AES are shown below.

Figure 4.1 Block diagram of visible spectrophotometer.

Figure 4.2 Block diagram of AAS/AES.

The added steps in AAS and AES are very vital since these are the reason
why AAS and AES can detect concentrations of a specific element. The following
diagram shows the scheme in the atomization of a sample subjected to AAS and
AES.

Figure 4.3 Scheme for the atomization of sample in Atomic Absorption

Spectroscopy.
In this exercise, the different components of an AAS system: absorption
vs. emission modes was studied. Also, the effects of releasing agents and/or
ionization suppressants were determined and standard addition technique and
external calibration method for the trace metal analysis were used.

II. MATERIALS AND METHODS

The standards and samples were prepared prior to analysis. For the trace
analysis of lead, the water sample was prepared by accurately measuring 1.0 L
and around 400 mL of it was transferred to a 600-mL beaker. 5.00 mL
concentrated HNO3 was then added. The beaker was then placed in a hotplate
and the sample was evaporated to almost 100 mL. The remaining amount of
water was then added and concentrated by evaporating the sample until the
remaining amount was around 100 mL again. Another 5.00 mL concentrated
HNO3 was then added and the beaker was then covered with a watchglass. The
sample was concentrated to around 50 mL, cooled and filtered. The
concentration of lead in the water sample was determined using the external
calibration method and standard addition method. For the standard addition
method, 0.5 mL from the 1000 ppm Pb 2+ stock solution was obtained to prepare
50.0 mL of 10 ppm Pb2+ standard solution. Using this 10 ppm Pb 2+ standard
solution, 0, 1.0, 2.0, 4.0, 5.0 mL were pipetted into five different 50-mL
volumetric flasks. 5.0 mL of the water sample was then added into each of the
volumetric flask and was diluted to mark with deionized water.
On the other hand, for the analysis of calcium and potassium, 2.0 g of
Bear Band powdered milk was accurately weighed and transferred to an
evaporating dish. Three trials were made. The milk sample was placed in a hot
plate until almost black. The evaporating dish was then transferred to the
furnace at 550 for 3-4 hours or until a white/gray residue was observed.
The sample was then removed from the furnace and was cooled to room
temperature. 2 mL of concentrated HNO3 was then added followed by 10 mL of
distilled water. The solution was heated to boiling, cooled and transferred
quantitatively while filtering to a 50-mL volumetric flask and was diluted to mark
with distilled water. From the 1000 ppm Ca 2+ and K+ stock solution, 0.5 mL were
obtained from each to prepare 50.0 mL of 50 ppm Ca 2+ and 50 ppm K+ standard
solutions. Using this 50 ppm Ca 2+ and 50 ppm K+ standard solutions, 0, 1.0, 2.0,
3.0, 4.0, 5.0 mL were pipetted into six different 50-mL volumetric flasks. 5.00
mL of 2% La3+ solution was then added to volumetric flasks for calcium analysis
while 5.00 mL of 5% Cs solution was supplemented to volumetric flasks for
potassium analysis. Each volumetric flask was then diluted to mark using %
HNO3. Signals obtained from the working curve were then compared to the
signals obtained from the milk sample.

All prepared standards and samples were read in AAS. Absorption mode
was used for calcium and trace analysis of lead while flame emission mode was
used for potassium analysis. Figure 4.1 shows the instrument used in the
experiment. Blank readings of each metal analyzed were then subtracted to
correct the output absorbance.

Figure 4.4. Atomic Absorption/Emission Spectrometer.

III. RESULTS AND DISCUSSION

Atomic Spectroscopy is a technique for determining the concentration of a

particular metal element in a sample. This application took more than 200 years
to be well-developed. In 1752, Thomas Melville elaborated on the principles of
flame atomic emission spectroscopy. After 100 years, at around 1860, Kirchoff
and Bunsen were able to show an analytical relationship between the ground or
excited state atoms and the absorption or emission of discrete spectral
radiation. Analysis then was done using Bunsen burners and this became the
source of problem in terms of the reproducibility of the analytical signal. This
problem was solved in 1929 upon the introduction of new burners, nebulizers,
gas control devices, and detection systems by Lundegardh. This advancement in
spectroscopy was further improved by Sir Alan Walsh and his collaborators in
1955 through the development of hollow cathode lamp serving as the radiation
source. Lvov, Greenfield, Fassel, and West, then introduced the non-flame
atomizers that minimized the spectral and chemical interferences in the process
(Ahuja, 2006).
Flame atomic absorption spectroscopy is currently the most widely used
of all the atomic spectral methods because of its simplicity, effectiveness and
relatively low cost. As shown in the schematic diagram above, solutions are
aspirated into the oxidizer gas. Nebulizers, then, pass the liquid aerosol
generated into the flame. Upon passing through the flame, the aerosol particles
are desolvated, dissociated, and atomized.

The aerosol particle size must be taken into consideration for aerosol
droplets that are too large can have insufficient exposure in the flame leading to
incomplete atomization aerosol droplets that are too small and can be easily lost
through wall collisions. The exposure of the sample in the flame is dependent on
the flow rate of the fuel and oxidizer, thats why optimization should be applied.
The flame also has an effect on the degree of atomization. The frequently used
flames used in analyses are air-acetylene and nitrous oxide-acetylene. The flame
temperature of the gases is 2300 oC and 2800OC, respectively. Nitrous oxideacetylene is recommended in analyses not solely due to higher flame
temperature but also to some excited cyanogen radicals that can reduce
refractory oxides to atoms. These refractory oxides has the ability to endure
flame without atomization and metals such as Al, Ba, Be, Ca, Sc, Si, Ta, Ti, U, V,
W, Zr can be in the form of refractory oxides. The air-acetylene, on the other
hand, is favored to those elements that have low ionization potentials like Li, Na,
K, Rb, and Cs since a flame with higher temperature can ionize the said
elements that can result to the reduction of the amount of the metal.
The ability of this technique to analyze up to 70 elements is due to the
atomizer. The electrons of the atoms in the atomizer can be promoted to higher
orbitals for a short amount of time by absorbing a set quantity of energy (i.e.
light of a given wavelength). This amount of energy (or wavelength) is specific
to a particular electron transition in a particular element, and in general, each
wavelength corresponds to only one element. This gives the technique its
elemental selectivity. And as the quantity of energy (the power) put into the
flame is known, and the quantity remaining at the other side (at the detector)
can be measured, it is possible, from Beer-Lambert law, 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.
The light source used in AAS is usually a discontinuous source since a
broad spectrum, produced by a continuous source, is not recommended in AAS
analyses. Discontinuous sources (line sources) used today are hollow cathode
lamps (HCL) and electrodeless discharge lamps. HCL is manufactured from the
element of interest and this lamp is filled with an inert under low pressure. The
mechanism of this source is through the production of inert gas discharge that is
responsible for the vaporization or sputtering of the element in the cathode.
These atoms are then excited via collision with the inert gas ions. Electrodeless
discharge lamp, on the other hand, has the same components inside like that of
HCL. The discharge is commenced by supplying electrons with a Tesla coil. This
type of lamp produces higher intensity atomic spectra than the HCL but the
spectral outputs are often unstable.

Like spectrophotometers, AA spectrometers use monochromators and

detectors for UV and visible light. The main purpose of the monochromator is to
isolate the absorption line from background light due to interferences. Simple
dedicated AA instruments often replace the monochromator with a bandpass
interference filter. Photomultiplier tubes are the most common detectors for
AAS.
Also, flame emission spectroscopy or flame photometry has found
widespread application in elemental analysis. The instruments are similar to that
of flame absorption except that the flame now acts as a radiation source; a
hollow- cathode lamp and chopper are therefore unnecessary.
The use of flame as atom reservoirs has some disadvantages and one of
them is the large consumption of the sample. Due to this, some non-flame
atomizers were developed. Electrothermal atomizers such as carbon rods,
carbon furnaces and tantalum ribbons are used for AAS and AFS. Atmospheric
pressure inductively coupled argon plasmas are used for AES analyses.
Quantitative atomic absorption/ emission analyses are based upon the
calibration curves involving a plot of absorbance/ emission versus concentration
of standards that have been prepared to mimic the solutions of the sample.
Standard addition methods are also used extensively in atomic spectroscopy in
order to try to compensate for differences between the composition of the
standards and the unknowns.
Trace metal analysis of lead of water sample via external calibration and
standard addition technique was also done in the experiment. Ideally, calibration
standards should approximate the composition of the samples to be analyzed
with respect to not only the analyte concentration but also the concentrations of
the other species in the sample matrix in order to minimize the effects of
various components of the sample on the measured absorbance. The standard
addition method can take several forms and the multiple addition method is
often chosen for photometric or spectrophotometric analyses. This technique
involves adding several increments of a standard solution to sample aliquots of
the same size. Each solution is then diluted to a fixed volume before measuring
its absorbance. When the amount of sample is limited, standard additions can
be carried out by successive addition of increments of the standard to a single
measured aliquot of the unknown. The measurements are made on the original
solution and after each addition of standard analyte. Results are shown below.
Table 4.1. Data for trace analysis of lead using external calibration.
[Pb2+], ppm

Trial 1

Abs
Corrected
Trial 2

Corrected

0
2
4
6
8
10
AbsUnknown
Equation of the line
R2
Calculated Cx, ppm
Average Cx, ppm
Theoretical
unknown, ppm
Percent error (%)
Flow rate: 6.3mL/min

0.0002
0.0000
0.0361
0.0359
0.0655
0.0653
0.0992
0.0990
0.1325
0.1323
0.1597
0.1595
0.0764

0.0014
0.0000
0.0357
0.0343
0.0634
0.0620
0.0971
0.0957
0.1296
0.1282
0.1555
0.1541
0.0761

y = 0.016x + 0.002
y = 0.0155x + 0.0015
0.9989
0.9987
4.65
4.812903226
4.731451613
4.5
5.143369176 %

Trial 1
0.18
0.16

f(x) = 0.02x + 0
R = 1

0.14
0.12
0.1

Absorbance 0.08
0.06
0.04
0.02

10

12

[Pb2+], ppm

Fig. 4.5. Graph of Trial 1 for trace analysis of lead using external calibration.

Trial 2
0.18
0.16
f(x) = 0.02x + 0
R = 1

0.14
0.12
0.1

Absorbance 0.08
0.06
0.04
0.02

10

12

[Pb2+], ppm

Fig. 4.6. Graph of Trial 2 for trace analysis of lead using external calibration.

Table 4.2. Data for trace analysis of lead using standard addition
method.
Vol. of 10 ppm Pb2+,
mL
0
1
2
4
5

Trial 1
0.0095
0.0129
0.0164
0.0239
0.0261

Abs
Corrected
0.0000
0.0034
0.0069
0.0144
0.0166

Equation of the line

R2
Calculated Cx, ppm
Average Cx, ppm

y = 0.0034x + 7E-05
y = 0.0035x - 8E-05
0.9962
0.9968
4.117647059
4.571428571
4.344537815

Trial 2
0.0079
0.0112
0.0145
0.0223
0.0247

Corrected
0.0000
0.0033
0.0066
0.0144
0.0168

Trial 1
0.02
0.02

f(x) = 0x + 0
R = 1

0.01
0.01
0.01

Absorbance 0.01
0.01
0
0
0

Vol of Pb2+ added, mL

Fig. 4.7. Graph of Trial 1 for trace analysis of lead using standard addition
method.

Trial 2
0.02
0.02

f(x) = 0x - 0
R = 1

0.01
0.01
0.01

Absorbance 0.01
0.01
0
0
0

Vol of Pb2+ added, mL

Fig. 4.8. Graph of Trial 2 for trace analysis of lead using standard addition
method.

For calcium, the absorbance of the standards were obtained and plotted
against their respective concentrations generating the standard curve. From this
curve, interpolation was done to determine the concentration of Ca 2+ in the

sample. Data were summarized and presented in the tables and figure on the
next page.

Table 4.3. Data for Ca analysis.

Vol. of 50 ppm
Ca2+, mL
0
1
2
3
4
5
Equation of the
line
R2
Blank
Trial 1 (m = 2.002
g)
Trial 2 (m = 2.023
g)
Trial 3 (m = 2.013
g)
Average CCa, ppm
Wavelength Ca2+: 422.7 nm
50
50
DF trial for Ca2+: ( 1 ) ( 10 )

Corrected Abs [Ca2+]

Abs [Ca2+]
0.0019
0.0229
0.0474
0.0850
0.1174
0.1513

0.0000
0.0210
0.0455
0.0831
0.1155
0.1494
y = 0.0305x - 0.0072
0.9912

Absorbance
0
0.0684

Calculated CCa, ppm

0
619.4020535
525.1927098

0.0569

521.0966514

0.0564
555.2304716

Calcium
0.16
0.14

f(x) = 0.03x - 0.01

R = 0.99

0.12
0.1

Absorbance 0.08
0.06
0.04
0.02
0

Vol. of 50 ppm Ca2+, mL

Fig. 4.9. Linear regression plot for Ca analysis.

On the other hand, potassium analysis was employed using the Atomic
Emission technique. The emission of the standards were obtained and plotted
against their respective concentrations generating the standard curve. From the
curves, interpolation was done to determine the concentration of K + in the
sample. Data were summarized and presented in the table and figure below.
Table 4.4. Data for K analysis
Vol. of 50 ppm K+,
mL
0
1
2
3
4
5
Equation of the line
R2
Blank
Trial 1 (m = 2.002 g)

Emission [K+]
0.0000
0.0616
0.1060
0.1329
0.1592
0.1954
y = 0.037x + 0.0166
0.9728
Emission
Calculated CK, ppm
0
0
0.1908
235.4054054

Trial 2 (m = 2.023 g)
Trial 3 (m = 2.013 g)
Average CK, ppm
Wavelength K+: 404.0 nm
50
DFtrial for K+: ( 1 )

0.1367
0.1299

162.2972973
153.1081081
183.6036036

Potassium
0.25
0.2

f(x) = 0.04x + 0.02

R = 0.97

0.15

Emission

0.1
0.05
0

Vol. of 50 ppm K+, mL

Fig. 4.10. Linear regression plot for Ca analysis.

The constructed graphs show a direct proportionality between

concentration and absorption/emission, which is consistent with Beer-Lamberts
Law. However, the constructed graph is not perfectly linear due to instrumental
interferences. This deviation from linearity could be from disproportionate
decomposition of molecules at high concentration.

It was also experienced in the experiment that there was an over-reading

of the Ca absorption and K emission of samples based from the standard
solutions. This was troubleshot by diluting the sample. In the case of high
concentration of the analyte, other methods can be employed in order to permit
accurate determination of the concentration of the analyte. The use of an
alternative wavelength having a lower absorptivity as well as the reduction of
the path length by rotating the burner hand can be done (Sevostianova).

Another factor that caused deviation from the linearity is the presence of
interferences. Interferences can be classified into spectral, chemical, and matrix.
Spectral interferences occur due to other forms of radiation that overlaps the
light source. This is often observed when organic solvents are used and the
presence of magnesium in sodium analysis, copper in iron, and nickel in iron
analyses. Other spectral interferences are due to some effects that result to the
broadening of spectral line. Some of the broadening effects according to
Sevostianova are:
1 Doppler effect due to the different components of velocity in the line
of observation
2 Lorentz effect due to the presence of foreign atoms in the
environment of emitting or absorbing atoms. The magnitude of the
broadening varies with the pressure of the foreign gases and their
physical properties.
3 Quenching effect - In a low-pressure spectral source, quenching
collision can occur in flames as the result of the presence of foreign
gas molecules with vibration levels very close to the excited state of
the resonance line.
4. Self absorption or self-reversal effect - The atoms of the same kind as
that emitting radiation will absorb maximum radiation at the center of the
line than at the wings, resulting in the change of shape of the line as well
as its intensity.

Matrix interferences, on the other hand, occur due to the variation in the
surface tension and the viscosity of the analyte and the standard solution.
Chemical interferences, the most common of all, arise from the ionization of the
analyte and the formation of some compounds that do not dissociate in the
flame like calcium and strontium phospates, which both leads to a reduction in
the signal observed.
Treatment of the chemical interferences can be done by manipulating the
flame conditions and through the application of certain chemical agents. A low
temperature flame can inhibit ionization of the metals and a high temperature
flame can further atomize compounds. The other technique is the application of
releasing agents, suppressor, and protective agents. A releasing agent like La is
a competing ion that reacts with the interfering substance instead of the metal
of interest, hence releasing the analyte from interferences. Addition of
suppressor like Cs is useful to those elements with low ionization energy like Na.
These suppressors, usually with lower ionization energy than the metal of

interest, are ionized instead of the metal of interest. On the other hand,
protective agents prevent interference by preferentially forming stable but
volatile species with the analyte. Application of these techniques leads to a
greater amount of sample detection. The application of either a releasing agent
or suppressor led to an increase in the concentration of the metal which is
consistent with the effect of such agents. These agents act as shields in order to
protect the metal of interest from various reaction conditions such as ionization
and formation of compounds that are not decomposed in flame.

IV. SAMPLE CALCULATION

Trace metal analysis of Lead

Plot absorbance versus volume of aliquot
Working equation:
C b
Cx = s
Vxm

Where:
Cx = concentration of unknown
b = y-intercept (from the graph of absorbance versus volume
of aliquot)
Cs = concentration of standard solution
= 10 ppm
m = slope (from the graph of absorbance versus volume of
aliquot)
Vx = final volume of unknown
= 50 mL or 5x10-3 L

Cx =

Considering Trial 1:
From the standard solutions:
0.0034
slope
interce
7 x 105
pt
Cs b

Vxm

10 ppm x 7 x 105
Cx=
=4.117647059 ppm
0.0034 x 5 x 103 L

Average Cx =

4.117647059+ 4.571428571
=4.344537815 ppm
2

Determination of the concentration in the unknown

K analysis

Using the equation of the line obtained from Beers Law plot: A = abc
Plot absorbance (A) vs. concentration (c)

y = mx + b
A = ac + b
Where

c =

A = absorbance
a = absorptivity/slope
b = y-int
c = concentration

A-b
a

Considering Trial 1:
0.1908- 0.0166
c =
=4.708108108 ppm
0.037
Calculation of the concentration of K+ in milk sample
50
= 4.708108108 ppm x DF ( 1 = 235.4054054 ppm
V. CONCLUSIONS

For this experiment, the different components of an AAS system:

absorption vs. emission modes was studied. Also, the effects of releasing agents
and/or ionization suppressants were determined and standard addition
technique and external calibration method for the trace metal analysis were
used.

Certain considerations must be done in order to have an accurate

measurement of the concentration. The aerosol particle size must be taken into
consideration for aerosol droplets that are too large can have insufficient
exposure in the flame leading to incomplete atomization aerosol droplets that
are too small can be easily lost through wall collisions. The flame must be
matched with respect to the analyte. A low flame temperature is used for
elements with low ionization energy and a high flame temperature is used in
elements capable of forming compounds that cannot be decomposed in flame.
The atomic absorption spectroscopy still relies on Beer-Lamberts Law but
there are some deviations from the linearity. These deviations from linearity
could be from disproportionate decomposition of molecules at high
concentration as well as the presence of certain interferences, especially
chemical interferences.
Chemical interferences can be eliminated through the use of suppressors
and releasing agents. Releasing agents are cations that react preferentially with
the interferent. The releasing agent that was used in the experiment was
Lanthanum. Suppresants are those that limit the activity of a certain compound.
The suppressant used in the experiment was Cesium. These agents act as
shields in order to protect the metal of interest from various reaction conditions
such as ionization and formation of compounds that are not decomposed in
flame. Flame manipulation can also be used to treat interferences.
Standard addition technique was used in trace metal analysis. This
technique involves adding several increments of a standard solution to sample
aliquots of the same size. Each solution is then diluted to a fixed volume before
measuring its absorbance.

VI. REFERENCES

AHUJA S., N. Jespersen. 2006. Comprehensive Analytical Chemistry 47.

Elsevier B.V.
SEVOSTIANOVA, E. Atomic Absorption Spectroscopy. Retrieved October 1,
2015.
SKOOG D.A., D.M. West, F.J. Holler, S.R. Crouch. 2004. Fundamentals of
Analytical Chemistry. 8th ed.Brooks/Cole. USA.