You are on page 1of 15

Chem126 Lab Instrumental Analysis

EXPERIMENT 5
Chemical Interferences in Atomic Absorption
Spectrophotometric Measurements
Vanessa Olga J. Dagondon and Ken M. Menez
Department of Chemistry, College of Arts and Sciences,
University of the Philippines Visayas, Miag-ao Iloilo

ABSTRACT
Calcium content in prepared samples containing other
species such as K, P, La, EDTA and Al was determined using
AAS or Atomic Absorption Spectroscopy. Four schemes were
adapted in the experiment: (1) absorbance of Ca standards
determined using air/acetylene flame; (2) absorbance of Ca +
K standards determined using air/acetylene flame; (3)
absorbance of Ca standards determined using N2O/acetylene
flame; and (4) absorbance of Ca + K standards determined
using N2O/acetylene flame. Addition of K and P contributes to
the chemical interference due to the incomplete dissociation
of compounds. This can be minimized by using a high
temperature
flame
N2O/acetylene
flame
instead
of
air/acetylene and by adding a releasing agent, La. Addition of
Al can cause another type of chemical interference using
interference due to effects of ionization. This can be
minimized by using a low temperature flame, air/ acetylene
and by adding a protective agent such as EDTA.

INTRODUCTION
Atomic spectroscopy is a series of different qualitative analyses
regarding the concentration of a specific substance in an analyte, with
each analysis applied based on the characteristics of the substance. 1 As
with any atomic spectroscopic method, the identity and concentration of a
substance in a sample solution could be determined by exciting the
molecules of the solution using a source, such as heat or strong light. This
excitation produces neutral atoms in the gas phase, which emit a specific
wavelength and intensity of light that gets captured and analyzed by
detectors.1 These processes are considered very successful in a wide array
Page 1 of 15

Chem126 Lab Instrumental Analysis


of applications in data analysis, with most processes already built into
special
automated

machines.10
This experiment uses the flame atomic
Figure 2. A schematic diagram for the process of
atomic absorption spectroscopy.1

Figure 1. Varian SpectrAA 55B atomic


absortion spectrometer. The user
interface is located top right, the light
source and detector at the lower
right, the nebulizer and aspirator
(clear tube) at the lower left, and the
covered burner at the top left which

spectroscopy, where an analyte is taken up


by an atomic absorption spectrometer to be
excited using different mixtures of acetylene
flame. The mixtures depend on the includes an exhaust chimney.1
substance being analyzed, as not all
substances can be atomized with the same type of flame. Each substance
has a certain temperature needed to atomize them in gas phase, as well
as a maximum speed for the flame to attain. In the case of this
experiment using metallic substances, too high a temperature can ionize
the metal while too low will have less metal atoms excited; both decrease
sensitivity.9 As only air acetylene and nitrous oxide acetylene torches
were used in this experiment, the maximum flame speed of air
acetylene is 160 cm s-1 and maximum temperature at 2300 C while
nitrous oxide acetylene attains a max flame speed of 180 cm s -1 and a
max temperature of 2955 C. 7 An example of an atomic absorption
spectrometer using the flame is shown in Figure 1, however a SHIMADZU
model was instead used for the experiment. This model had very similar
parts as the example, including an easy switch between two cathode
lamps.

The process of flame atomic spectroscopy needs the analyte to be


dissolved in a solvent; this analyte is then nebulized into the flame.
Nebulization is the application of an oxidant gas to force the dissolved
substance to spray evenly over the flame. 1 The flame will atomize, or turn
the sprayed analyte into neutral atoms in gas form by increasing the
temperature until the gas forms a plasma; in this plasma the ions stabilize
by bonding with free electrons and radicals to form neutral atoms. 1 A finer
Page 2 of 15

Chem126 Lab Instrumental Analysis


aerosol will easily vaporize the analyte, and a hotter flame will easily
vaporize stable compounds; both reduce interferences at this step. 9 After
excitation, the released energy is converted as light, which travels
through a monochromator. The monochromator is a device that helps
select one certain wavelength to reach the detector. However, the light
coming from the neutral atomic gas is so narrow that it causes gaps in the
monochromator slit which greatly deviates the amount of light obtained
by the detector; this can be solved by activating a hollow cathode ray
tube that delivers radiation patterns similar to the substance being
analyzed.1 After the light reaches the detector, it is then analyzed. A
simplified process is shown as Figure 2.
Due to different substances having different properties, the metals
featured in this experiment have to be dissolved in certain concentrations
and have specific flames to atomize.
Aluminum was present in the form of aluminum chloride
hexahydrate. Being a more easily dissolvable form of aluminum, it was
added directly to the standard solution. The optimal wavelength for
aluminum is 396.1 nanometers while using a nitrous oxide acetylene
flame; due to it being partially ionized in the flame, a small amount of
potassium chloride was added to the solution as a suppressor. 2 Calcium
was used as a standard for all the other solutions in this experiment. Used
in the form of calcium carbonate, a small amount of nitric acid was added
before dilution of water to help it dissolve completely. The optimal
wavelength for aluminum is 422.7 nanometers, while using a nitrous oxide
acetylene flame; in air acetylene flames the interferences of calcium
can be reduced by adding lanthanum, while in nitrous oxide acetylene
flames potassium chloride is added to suppress ionization. 3 Potassium was
present as potassium chloride. The optimal wavelength is at 766.5
nanometers with an air acetylene flame; this same flame helps eliminate
interferences in the sample. 4 Lanthanum was present as lanthanum
chloride in the experiment. The optimal wavelength is at 441.7
nanometers, with a nitrous oxide acetylene flame; the interferences
caused by partial ionization were suppressed by the potassium present in
the solution.5 Phosphorus was present in the experiment as sodium
phosphate. The optimal wavelength is at 213.6 nanometers at a nitrous
oxide acetylene flame; due to being an uncommon substance
determined in AAS and having a lack of sensitivity in the process only a
few studies contain interference information about phosphorus.6
The standard solutions are prepared to obtain a common constant
between all other solutions. This standard was based around calcium. If
the concentration of the solution is plotted against the absorption of each
Page 3 of 15

Chem126 Lab Instrumental Analysis


standard, the slope of the resulting line equals the common constant or
molar absorptivity . This is used in the Beer Lamberts Law, which
relates absorbance (A) to the molar absorptivity, path length of light (b),
and concentration of the sample solution (c):9
A=bc
(1)
The obtained absorbance from the different solutions is then
converted to transmittance:9
A
T =10
(2)
In vice versa, absorbance could also equal the logarithmic function
of transmittance. To decrease interferences in the samples the same
suggestions that were mentioned earlier were used as well as adjusting
several aspects of the flame and samples to obtain the maximum
sensitivity in high precision.9
METHODOLOGY
Two series of standard solutions were prepared. The first series
contained only calcium carbonate. Five 100 milliliter volumetric flasks
were used to contain 0, 1, 2, 3, and 5 ppm of calcium carbonate,
respectively. These flasks were labeled from 1A to 5A in the manner of
increasing calcium concentration, and then diluted to mark with distilled
water. The second series was prepared similar to the first, however an
added approximate of 0.0381 grams of potassium chloride was added to
each flask. The second series flasks were labeled from 1B to 5B, same in
order as the first series.
A set of sample solutions were prepared in 100 mL volumetric
flasks, with different mixtures of substances for each. A stock solution of
250 mL calcium was prepared by dissolving 2.4976 grams calcium
chloride into 250 mL water in a 250 mL flask. This stock calcium solution
was used for each of the following sample solutions:
The first flask, labeled 1S, and all other flasks were each added with
3 mL of stock calcium solution. The second flask, labeled 2S, was added
0.0053 grams of sodium phosphate. The third flask, labeled 3S was added
0.0053 grams of sodium phosphate, and 0.3818 grams of potassium
chloride. The fourth flask, labeled 4S, was added with 0.0053 grams of
sodium phosphate, 0.3818 grams of potassium chloride, and 0.0017
grams of lanthanum chloride. The fifth flask, labeled 5S, was added with
0.0908 grams of aluminum chloride hexahydrate. The sixth flask, labeled
6S, was added with 0.0908 grams of aluminum chloride hexahydrate and
2.3146 grams of EDTA.

Page 4 of 15

Chem126 Lab Instrumental Analysis


The spectral interference of the each solution was recorded using
atomic absorption spectroscopy by air/acetylene and N 2O/acetylene
flames. Each set of solutions were analyzed by air/acetylene and
N20/acetylene flames, with the standards being tested first before the
samples.
RESULTS

AND

DISCUSSION

Two sets of standards were prepared in the experiment: one is


composed of calcium in different concentrations and the other one is
composed of calcium and potassium in different concentrations. The
absorbances of each set of standards were analyzed in the AAS using two
flames: air/acetylene and N2O/ Acetylene flame. This resulted to four
schemes: Scheme 1 used air/acetylene flame to analyze the calcium
standards; scheme 2 used the same flame to analyze the calcium plus
potassium standards; scheme 3 used N2O/acetylene flame to analyze the
calcium standards; and, scheme 4 used the same flame to analyze the
calcium plus potassium standards. These four schemes will result to five
calibration curves used to obtain the concentration of calcium in the
prepared samples. This variation of conditions is done to identify the
effect of the added interference in the prepared samples and also the
effect of the type of flame used in the analyses.
Table 1 summarizes the result of absorbances obtained from each
schemes. Notice that in scheme 1 at the concentration of 1 mg/L, there
was no recorded absorbance. This is because the absorbance read was
negative and therefore would be erroneous. The data was discarded
instead. Figure 3 shows the calibration curves obtained from each
schemes. Linearity of the calibration curves was not satisfactory. The
calibration curve is somehow curved up or has an upward curvature.
While this may be due to the inaccurate preparation of the standards, it is
a fact that it is rare for atomic absorption calibration curves to show
ideality (i.e. linear plot). The upward curvature in the calibration curves
generated is usually observed on the standards of small concentration
range.11
Table 2 shows the absorbances recorded in each samples prepared.
As shown, there are six samples with the same concentration of calcium,
each of which contains different interferences. Table 3 shows the
concentration of calcium of the samples obtained from each scheme. It
can be observed the fluctuation of the values for the concentration of
calcium in each sample despite the fact that they are of the same

Page 5 of 15

Chem126 Lab Instrumental Analysis


concentration of calcium when prepared. This shows how much an
addition of interference affects the analysis done in AAS.
In the first sample which theoretically contains only 3mg/L calcium,
the obtained concentrations from the four calibration curves were much
lesser than the theoretical value (Table 3). This indicates that there are
indeed errors in the preparation of the calibration curves (i.e. preparation
of standards). All the other samples contain the same amount of calcium
as that of the first sample. However, the remaining samples contains
other components such as P, K, La, Al, and EDTA.
Interferences in atomic absorption fall into six categories: chemical
interferences, ionization interferences, matrix interferences, emission
interferences, spectral interferences, and background absorption. The
most common interferences are chemical interferences. A chemical
interference emerges when the sample being analyzed contains a
thermally stable compound with the analyte that is not totally
decomposed by the energy of the flame and thus, the number of atoms in
the flame capable of absorbing light is reduced. 12 There are to general
forms of chemical interferences: ionization and incomplete dissociation of
compounds.13
The effect of phosphorous and potassium in calcium, as in the
second and third sample, is an example of a chemical interference due to
incomplete dissociation of compounds. These interferents form
compounds which are not completely dissociated at the temperature of
the flame and hence prevent the formation of neutral ground state
atoms.13To overcome this interference, a higher temperature flame can be
used, as in in scheme 2 which uses N 2O/Acetylene flame, or La can be
added as a releasing agent, as in sample 4. A releasing agent, which can
be referred also as a competing cation, reacts with the interferent
releasing the analyte.12The presence of Al, as in samples 5, are another
example of a chemical interference. This time, the interference is due to
ionization. To overcome this inference, a lower temperature flame such as
air/ acetylene flame must be used, as in in scheme 1, because high
temperature flames such as nitrous N2O/acetylene may cause appreciable
ionization of the analyte element. The alkali and alkaline-earth metals
such as Al are more susceptible to ionization. To control this interference,
a suitable cation with an ionization potential lower than that of the analyte
is added. A protective agent such as EDTA can also be added to reduce
this effect, as in sample 6. A protective agent is a ligand reacts with the
analyte forming a relatively volatile complex.

Page 6 of 15

Chem126 Lab Instrumental Analysis


It can be observed that the results of the experiment did not
coincide with the theory of the experiment. This can be accounted with
the inaccurate preparation of the standards and also of the samples.
Table 1. Absorbance Readings of the Standards
Air/Acetylene Flame
N2O/Acetylene Flame
Scheme 1
Scheme 2
Scheme 3
Scheme 4
Ca Standard
0 mg/L
0.0005
0.0002
1 mg/L
0.0020
2 mg/L
0.0778
0.0613
3 mg/L
0.0824
0.0661
5 mg/L
0.1963
0.1780
Ca Standard
+K
0 mg/L
1 mg/L
2 mg/L
3 mg/L
5 mg/L

0.0030
0.0068
0.0779
0.0824
0.1818

Table 2. Absorbance readings of the samples


Sample
Air/ Acetylene Flame
3 mg/L Ca
0.0824
3 mg/L Ca + P
0.0413
3 mg/L Ca +P + K
0.0519
3 mg/L Ca + P + K + La
0.0508
3 mg/L Ca + Al
0.0292
3 mg/L Ca + Al + EDTA
0.0002

Page 7 of 15

0.0029
0.0070
0.0897
0.0908
0.1367

N2O/ Acetylene Flame


0.0661
0.0140
0.0516
0.0693
0.0452
0.0200

Chem126 Lab Instrumental Analysis

Calibration Curve: Scheme 2


0.2000
0.1500
Absorbance

0.1000

f(x) = 0.04x - 0.01


R = 0.95

0.0500
0.0000
0 1 2 3 4 5 6
Concentration of Ca + K Standards (mg/L)

Calibration Curve: Scheme 1


0.2500
0.2000
f(x) = 0.04x - 0.02
R = 0.94

0.1500
Absorbance 0.1000
0.0500
0.0000

0 1 2 3 4 5 6
Concentration of Ca Standards (mg/L)

(a)

(b)

Page 8 of 15

Chem126 Lab Instrumental Analysis

Calibration Curve: Scheme 4


0.1500
f(x) = 0.03x + 0
R = 0.9

0.1000
Absorbance

0.0500
0.0000
0 1 2 3 4 5 6

Concentration of Ca + K Standards (mg/L)

Calibration Curve: Scheme 3


12.0000
10.0000
8.0000
Absorbance

6.0000
4.0000
2.0000
0.0000

0 f(x)
1 =2 3 4 5 6
R = 0
Concentration of Ca Standards (mg/L)

(c)

(d)
Page 9 of 15

Chem126 Lab Instrumental Analysis


Figure 3. Calibration curves generated from each scheme: (a) plot of the
absorbance obtained using air/acetylene flame against the concentration
of Ca standards, scheme 1; (b) plot of the absorbance obtained using
air/acetylene flame against the concentration of Ca + K standards,
scheme 2; (c) plot of the absorbance obtained using N 2O/acetylene flame
against the concentration of Ca standards, scheme 3; (4) plot of the
absorbance obtained using N2O/acetylene flame against the concentration
of Ca + K standards, scheme 4.

Table 3. Concentration of Ca in the samples in mg/L


Air/ Acetylene Flame
N2O/ Acetylene Flame
Sample
Scheme 1
Scheme 2
Scheme 3
Scheme 4
3 mg/L Ca
2.4630
2.5266
2.2004
2.2448
3 mg/L Ca + P
1.4433
1.4098
0.6848
0.4231
3 mg/L Ca +P + K
1.7063
1.6978
1.7786
1.7378
3 mg/L Ca + P + K + La
1.6790
1.6679
2.2935
2.3566
3 mg/L Ca + Al
1.1430
1.0809
1.5924
1.5140
3 mg/L Ca + Al + EDTA
0.4235
0.2929
0.8594
0.6329

CONCLUSION
Atomic Absorption Spectroscopy (AAS) is a technique for measuring
quantities of chemical elements present in a sample by measuring the
absorbed radiation by the chemical element of interest. The sample is
excited by radiation making its atoms absorb ultraviolet or visible light
and make transitions to higher energy levels. In this experiment, flame
atomic spectroscopy is used to excite the analyte using different mixtures
of acetylene flame. The analyte in this experiment is calcium. Calcium
content of 6 samples containing interferences and other components were
determined. To determine the calcium content in the samples, two sets of
calcium standards (Ca standards and Ca+K standards) were prepared to
create calibration curves. The absorbances of the standards were
determined using air/acetylene and N2O/acetylene flames as indicated in
the four schemes followed in the experiment. Four calibration curves were
obtained in the experiment. Effects of interference in the determination of
calcium in the sample were examined. Potassium and phosphorus caused
a chemical interference due to incomplete dissociation of compounds. This
interference can be aided by using a higher temperature flame
(N2O/acetylene flame instead of air/acetylene) and also by adding
Lanthanum which is a releasing agent. Aluminum can cause another type
Page 10 of 15

Chem126 Lab Instrumental Analysis


of chemical interference due to effects of ionization. To aid this, a lower
temperature flame such as air/ acetylene flame must be used because
high temperature flames such as nitrous N 2O/acetylene may cause
appreciable ionization of the analyte element and also by adding a
protective agent such as EDTA. These theories were not reflected in the
results of the analysis because of the inaccurate preparation of standards
and samples.

1.

2.

3.

4.

5.

6.

7.

8.

LITERATURE CITED
Introduction. Determination of Calcium by Atomic Spectroscopy.
Chem 334: Quantitative Analysis Laboratory, Colorado State
University. March 24, 2016. p. 1 2.
Standard Conditions: Al (Aluminum). Flame Atomic Absorption
Spectrometry: Analytical Methods. Agilent Technologies, Australia.
13th ed. November 2015. p. 16.
Standard Conditions: Ca (Calcium). Flame Atomic Absorption
Spectrometry: Analytical Methods. Agilent Technologies, Australia.
13th ed. November 2015. p. 24.
Standard Conditions: K (Potassium). Flame Atomic Absorption
Spectrometry: Analytical Methods. Agilent Technologies, Australia.
13th ed. November 2015. p. 42.
Standard Conditions: La (Lanthanum). Flame Atomic Absorption
Spectrometry: Analytical Methods. Agilent Technologies, Australia.
13th ed. November 2015. p. 43.
Standard Conditions: P (Phosphorus). Flame Atomic Absorption
Spectrometry: Analytical Methods. Agilent Technologies, Australia.
13th ed. November 2015. p. 54.
Amos, M. D. & Willis, J. B. Choice of Flame. Spectrochimica Acta:
Use of high temperature pre mixed flames in atomic absorption
spectroscopy. Pergamon Press Ltd., Northern Ireland. vol. 22. 1966.
p. 1327.
Martizano, J. Materials & Procedures. Chemical Interferences in
Atomic Absorption Spectrophotometric Measurements. University of
the Philippines Visayas Miagao Campus, Philippines. March 2016.
p. 1.
Page 11 of 15

Chem126 Lab Instrumental Analysis


9. Melville, J. Theory. Atomic Absorption Spectroscopy of Metal
Alloys. Chemistry 105: Instrumental Methods in Analytical
Chemistry, Berkeley College of Chemistry, University of California,
California. March 3, 2014. p. 2.
10.
Walsh, A. Introduction. Spectrochimica Acta: The application
of atomic absorption spectra to chemical analysis. Chemical Physics
Section, Division of Industrial Chemistry, Commonwealth Scientific
and Industrial Research Organization, Melbourne, Australia.
Pergamon Press Ltd., London. vol. 7. 1955. pp. 108 177.
11.
Harvey, D. Modern Analytical Chemistry. United State of
America: The McGraw-Hill Companies, Inc.; 2000 [cited 2016
February]. Available from: http://elibrary.bsu.az/
12.
Skoog D. A., West D. M., Holler F. J., Crouch S. R. 2014.
Fundamentals of Analytical Chemistry Ninth Edition. Canada: Nelson
Education, Ltd. 1026p.

APPENDICES
I. Tables
Table 4. Statistical Data for the Schemes 1, 2,3 and 4
Statistical
Parameter
M
B
sr
sm
sb
sc
r

Scheme 1

Scheme 2

Scheme 3

Scheme 4

0.040304054
-0.016868919
2.28x10-2
5.93x10-3
1.66x10-2
6.22x10-1
0.939

0.036798649
-0.01057703
1.96 x10-02
5.10 x10-03
1.43 x10-02
6.42 x10-01

0.036439189
-0.01916622
2.28 x10-02
5.92 x10-03
1.65 x10-02
6.91 x10-01

0.0286
0.0019
2.15 x10-02
5.60 x10-03
1.56 x10-02
8.34 x10-01

0.9454

0.9267

0.8969

II. Sample Calculations


Least Square Method
Page 12 of 15

Chem126 Lab Instrumental Analysis


*For Scheme 1
Let x be the concentration of the Ca standards and y be the absorbance
x

x2

y2

xy

0.0005

0.0000002
5

0.0020

0.000004

0.002

0.0778

0.0824

0.1963

25

sum

11

0.3590

39

ave

2.2

0.0718

7.8

0.0060528
4
0.0067897
6
0.0385336
9

0.051380
54
0.010276
10

( x )2
( 11 )2
S xx = x
=39
=14.8
n
5
2

( y )2
( 0.3590 )2
S yy = y
=0.05138054
=0.02560434
n
5
2

( xy )
[ ( 11 ) ( 0.3590 ) ] =0.5965
S xy =xy
=1.3863
n
5

m=

b=

S xy 0.5965
=
=0.040304054
S xx
14.8

y
x 0.3590
11
m
=
( 0.02560434 )
=0.01686892
n
n
5
5

( )

( )

Determination of calcium in sample where y = 0.0824


equation of line : y=0.040304054 x0.01686892

Page 13 of 15

0.1556
0.2472
0.9815

1.3863
0.27726

(y1mx1+b)2
0.0003016
79
0.0004594
65
0.0001977
06
0.0004684
3
0.0001356
91

0.001562
97
0.000312
59

Chem126 Lab Instrumental Analysis


x=

0.0824+0.01686892
=2.4630 mg/ L
0.040304054

Uncertainty of Measurement
0.040304054
( 2)(14.8)
0.02560434
=2.28 102
52
S yy m2 S xx
=
N2
s r=

sr
(2.28 102 )2
=
=5.93 103
S xx
14.8
s m=
xi

N
1

s b=s r

Page 14 of 15

Chem126 Lab Instrumental Analysis

y
n

0.040304054
0.3590
0.0824
5
1 1
+ +
1 5
( 2)(14.8)
2.28 102

0.02560434

2
( m S xx |=

1 1
+ +
M N
sr

m
sc =

uncertainty=(2.28 102 )2+(5.93 103 )2+(1.66 102 )2+(6.22 101 )2 =6.22 101
*All calculations for the remaining schemes are done is the same way
shown.

Page 15 of 15