CM124-1L: ANALYTICAL CHEMISTRY LABORATORY

3rd Quarter SY 2018-2019

DETERMINATION OF METAL FROM VARIOUS SAMPLES USING ATOMIC

ABSORPTION SPECTROSCOPY
Santos, Nanette, D.1, Ortega, Mary Alyssa, T.2

1
Professor, School of Chemical, Biological, and Materials Engineering and Sciences, Mapúa University; 2Student, CM124-1L/A14, Mapúa University

ABSTRACT

The elemental analysis of Atomic Absorption Spectroscopy which is widely used to determine and analyze various
samples including the environment, metal, food, pharmaceutical and chemical samples. In this experiment, it
focuses on the determination of the metal content of the samples that were gathered from our surroundings and
from the essential things we need every day. Four types of samples were gathered by the researchers which are
the Water, Soil, Fruit and Vegetable sample. These were tested with different standard methods to be able to
determine the absorbance of the samples and to obtain the concentration of the metal content. The concentration
of the standard solution is directly proportional to the absorbances that were read by the machine. For the
determination of the concentration of metals in various samples, calibration curves were used to obtain it. The
results were 9 ppm in water sample, 0.6 ppm in soil sample, 3.272 ppm in fruit sample, and 0.16 ppm in vegetable
sample. These shows that the water sample has the greatest amount of metal content and vegetable sample has the
lowest concentration calculated.

Keywords: AAS, lead, copper, potassium, metal content

Introduction

Analysis of Heavy metals (Pb, Cu) in industrial
effluent using Atomic Absorption spectroscopy. A
heavy metal is a member of a loosely-defined subset of
elements that exhibit metallic properties. Heavy metals
occur naturally in the ecosystem with large variations
in concentration. Lead occurs naturally in soil, in levels
ranging from 10-50 ppm. Lead is also of concern in
soil, when it’s used for gardening. Plants take up lead
from the soil. Therefore, vegetables or herbs grown in
contaminated soil can lead to lead poisoning. All soils
naturally contain trace levels of metals. The presence
of metals in soil is not an indication that it is
contaminated. The metals detected by this method are
not all toxic. Fruits and vegetables are an important
component of diet. Different vitamins and minerals are
needed to intake for the body to have a normal growth
and maintenance. Fruit contains a high-water content
in general and addition to water, some fruits contains
different amount of Potassium.
Atomic Absorption Spectroscopy or AAS is one of the
most common instrumental methods for analyzing for
metals and some metalloids. This method could also
measure the concentration of metals in the sample.
AAS measure the amount of energy in the form of
photons of light that result to the change in wavelength
that is absorbed by the sample. Any atom has its own
distinct pattern of wavelengths at which it will absorb
energy. The sample is atomized in either a flame or a
graphite furnace that is lined up in a beam of light of
the appropriate wavelength. The atoms absorb
ultraviolet or visible light and make transitions from
the ground state to excited electronic states that is
caused by the flame (thermal energy). When the atoms
make their transition, they absorb some of the light
from the beam. The technique uses basically the
principle that free atoms generated in an atomizer can
absorb radiation at specific frequency. There are over
62 elements, mainly metals, are detectable by AAS.
Lead, Copper and Potassium are some of the metals
that were detected from the analyzed samples. Lead
and Copper are examples of heavy metals. These are
any metallic chemical element that has a relatively
high density and is toxic or poisonous at low
concentrations. The purpose of this experiment is to
determine the amount of metals from different samples

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using various AAS and to compare the different
methods of analysis.
Materials and Methods
Chemicals and Reagents
All the chemicals of analytical grade including HNO3,
Pb(NO3)2, Cu(NO3)2, KCl. Distilled water was also
used in this experiment.
Collection of Samples
Samples of soil were collected from a random
backyard soil in Manila and the group decided to use
pear as the fruit sample for this experiment. The leafy
vegetable used in this experiment is lettuce. Samples
were collected randomly in order to estimate the total
metal content (Pb, Cu, & K) and concentration in these
samples.
Sample Preparation and Treatment
Water Sample
200 mL of water sample was filtered using Whatman
#40 filter paper and transferred into a 400-mL beaker.
From this, 10 mL of conc. HNO3 was added to it and
was heated on a hot plate. The mixture was simmered
without boiling with a watch glass allowing the
mixture to undergo gentle reflux. The solution was
adjusted with the addition of HNO3 if it still does not
appear to a light color.
Fruit Sample
Samples were peeled off and washed with distilled
water. The peeled pear was sliced thinly and was put
on an aluminum foil. The sample was oven dried at
100-150ºC until it turns brown and to crisp that
indicates that it is dehydrated. After drying the sample,
2.0 g was weighed out from this and was put in a 250-
ml beaker. The dried sample was cut into smaller strips
and added 18 mL of concentrated nitric acid to it. From
this, the sample was heated on a hot plate for 30
minutes in medium heat. The sample was cooled after
heated and 10 mL of double distilled water was added.
The solution was filtered in a 100 mL volumetric flask
as the receiver with Whatman #40 filter paper and the
volume was made up to the mark with distilled water.
The pH of the filtered sample was measured.
Soil sample
20 g of soil sample was heated in an oven at 100-150
ºC for 45 minutes. After drying the sample, 2.0 g was
Experiment 06│ Group No.5-6│April 3, 2019
weighed out from this and was put in a 250-ml beaker.
Then, 18 mL of concentrated nitric acid was added to
it. From this, the sample was heated on a hot plate for
30 minutes in medium heat. The sample was cooled
after heated and 10 mL of double distilled water was
added. The solution was filtered in a 100 mL
volumetric flask as the receiver with Whatman #40
filter paper and the volume was made up to the mark
with distilled water. The pH of the filtered sample was
measured.
Vegetable Sample
500 g of leafy vegetable was heated in an oven at 100-
150ºC for 30 minutes. After drying the sample, 2.0 g
was weighed out from this and was put in a 250-ml
beaker. Then, 18 mL of concentrated nitric acid was
added to it. From this, the sample was heated on a hot
plate for 30 minutes in medium heat. The sample was
cooled after heated and 10 mL of double distilled water
was added. The solution was filtered in a 100 mL
volumetric flask as the receiver with Whatman #40
filter paper and the volume was made up to the mark
with distilled water. The pH of the filtered sample was
measured.
Preparation of Standards
0.2 g of Lead was used to make a stock solution.
Different volumes of stock solution were transferred to
50-mL volumetric flasks, which are 0.25 mL, 0.5 mL,
1.25 mL, 2.5 mL, 5 mL and blank. From this, 40 mL
of double distilled water was added to the volumetric
flasks and its initial pH was measured. 1-2 drops of
HNO3 was used to adjust the pH to be similar with the
sample. After adjusting the pH, the flasks were diluted
to mark.
For the Standard Addition Method, 2 ppm was the
concentration used as a standard solution for this
method. The standard solution was poured to five 50-
mL volumetric flasks with different volumes which
are: 2 mL, 4 mL, 6 mL, 8 mL and 10 mL and the sixth
flask served as a blank solution. From this, a uniform
volume of the sample, 5 mL, was poured to each flask
and was diluted up to the mark.
0.02 g of Cu was measured and transferred to 500-mL
volumetric flask and was dissolved in distilled water.
The solution was diluted up to mark. Several
concentrations were made from the analyte standard:
0.1 ppm, 0.2 ppm, 0.5 ppm, 1 ppm, and 2 ppm. Two
solutions were mixed together with same volumes but
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with different concentration of analyte standard and all Table 2. Absorbance Values of the Water sample
flask were diluted up to mark. Trial Absorbance
1 0.0193
2 0.0145
3 0.0136
Results and Discussion Mean 0.0158

There are four different samples that were used for this Table 2 shows the absorbance values of the water
experiment: Water, Soil, Fruit, and Vegetable sample. sample that were used to compare with pH values of
Every sample were tested to determine the the standard solutions. All trials resulted to a precise
concentration and amount of metals present. These reading and resulted to a calculated mean of 0.0158.
samples were tested in various methods in
determination of the concentration of the metal Table 3. Absorbance Values using Standard Addition Method
present: External Calibration method, Standard Volume Analyte Standard Absorbance
Addition Method, and Internal Standard Method. 0 0.0046
2 0.0048
4 0.0051
The table and figures below were the readings and data 6 0.0066
gathered for the water sample. 8 0.0061
10 0.0055
Table 1. Absorbance values using External Calibration Method
Concentration Absorbance Table 3 shows the readings gathered by the
0 -0.0008 spectrophotometer using a different method, Standard
0.1 0.001
0.2 0.0015
Addition Method. The more amount of the volume of
0.5 0.0027 standard added to the sample, the higher the
1 0.0048 absorbance reading is.
2 0.0072

0.007
Table 1 shows the relationship between the 0.006 y = 0.0002x + 0.0045
concentration of the external standard solution which
Absorbance

0.005
contains Lead and its resulted Absorbance. As shown 0.004
above, the blank solution or the 0-ppm external 0.003
standard resulted to a negative absorbance. The 0.002
absorbance being read by the machine increases as the 0.001
concentration of the solution increases. 0
0 2 4 6 8 10

0.01 Volume of Standard Added

0.008 y = 0.0037x + 0.0004 Figure 2. Linear graph of Absorbance values and volume of
standard solution added
Absorbance

0.006

0.004 Table 4. Absorbance Values using Internal Standard Method

Concentration Absorbance
0.002 0.1 0.0026
0.2 0.0058
0 0.5 0.0043
0 0.5 1 1.5 2 2.5 1 0.007
-0.002
Concentration 2 0.0108

Figure 1. Linear graph of absorbance values and external Table 4 shows the readings gathered by the
standard concentration spectrophotometer using a different method, Internal
Standard Method. The results in this method is
different with the two methods but still it shows a

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relationship between the two values. The concentrated Table 6. Absorbance values of the Soil sample
the solution of the analyte standard is, the higher the Trial Absorbance
absorbance being read. 1 0.0018
2 0.0027
3 0.0029
7 Mean 0.00247
6
5 Table 6 shows the absorbance values of the soil sample
Volume

4 that were used to compare with pH values of the

3 standard solutions. All trials resulted to a precise
2 reading and resulted to a calculated mean of 0.00247.
1 y = -2.3923x + 5.641
0 Table 7. Absorbance Values using Standard Addition Method
0 0.5 1 1.5 2 2.5 Volume Analyte Standard Absorbance
Concentration of analyte stadard, ppm 0 0
2 0.0029
Figure 3. Linear graph of Absorbance values and different 4 0.0007
concentrations of analyte standard 6 -0.0004
8 0.0021
10 0.0074
The table and figures below were the readings and data
gathered for the soil sample.
Table 7 shows the readings gathered by the
spectrophotometer using a different method, Standard
Table 5. Absorbance values using External Calibration Method
Addition Method. The more amount of the volume of
Concentration Absorbance
0 0 standard added to the sample is not directly
0.1 0.0005 proportional to the readings of absorbance.
0.2 0.0008
0.5 0.0045
1 0.0088
2 0.1678
0.008
Table 5 shows the relationship between the
0.006
concentration of the external standard solution which
Absorbance

contains Lead and its resulted Absorbance. As shown 0.004

above, the blank solution or the 0-ppm external
0.002 y = 0.0006x - 0.0009
standard resulted to a 0 absorbance. The absorbance
being read by the spectrophotometer increases as the 0
concentration of the solution increases. 0 5 10 15
-0.002
Volume of Standard Added
0.02
Figure 5. Linear Graph of Absorbance values and the volume
0.015 of the analyte standard added
Absorbance

0.01 Table 8. Absorbance Values using Internal Standard Method

y = 0.0093x - 0.0004 Concentration Absorbance
0.005
0.1 0.0076
0.2 0.0083
0 0.5 0.0103
0 0.5 1 1.5 2 2.5 1 0.0135
-0.005 2 0.0222
Concentration

Figure 4. Linear graph of Absorbance Values and Table 8 shows the readings gathered by the
Concentration of the External Standard spectrophotometer using a different method, Internal
Standard Method. The results in this method is

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different with the two methods but still it shows a Table 10. Absorbance values of Fruit sample
relationship between the two values. The concentrated Trial Absorbance
the solution of the analyte standard is, the higher the 1 0.7518
2 0.7867
absorbance being read. 3 0.7712
Mean 0.7699
0.4
Table 10 shows the absorbance values of the fruit
Absorbance

0.3 sample that were used to compare with pH values of

0.2 the standard solutions. All trials resulted to a precise
reading that ranges from 0.75-0.77 and resulted to a
0.1 calculated mean of 0.7699.
0 y = -0.1083x + 0.3133
0 0.5 1 1.5 2 2.5 Table 11. Absorbance values using Standard Addition Method
Volume Analyte Standard Absorbance
Concentration of analyte standards, ppm 0 0.1227
2 0.1288
Figure 6. Linear Graph of the absorbance and the 4 0.1639
concentration of Analyte Standard concentration 6 0.1846
8 0.2215
10 0.253
The table and figures below were the readings and data
gathered for the fruit sample.
Table 11 shows the readings gathered by the
Table 9. Absorbance values using External Calibration Method spectrophotometer using a different method, Standard
Concentration Absorbance Addition Method. The results above show the
0 0 relationship between the amount of the standard
0.1 0.246 solution and its readings. The more amount of the
0.2 0.056
0.5 0.193
volume of standard added to the sample, the higher its
1 0.714 absorbance is.
2 0.764
0.3
Table 9 shows the relationship between the 0.25
Absorbance

concentration of the external standard solution which 0.2

contains Lead and its resulted Absorbance. As shown 0.15
above, the blank solution or the 0-ppm external 0.1
standard resulted to a negative absorbance. The
0.05 y = 0.0136x + 0.1112
absorbance being read by the machine increases as the
0
concentration of the solution increases. 0 5 10 15
Volume of Standard Added, mL
1
Figure 8. Linear graph of Absorbance values and the volume
of the standard solution added
0.8
Absorbance

0.6

0.4 y = 0.42x + 0.0346

0.2

0
0 0.5 Concentratinn
1 1.5 2 2.5
Figure 7. Linear graph of the absorbance values and the
concentration of external standard

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The table and figures below were the readings and data Table 14. Absorbance Values using Standard Addition Method
gathered for the vegetable sample. Volume Analyte Standard Absorbance
0 0.0006
2 0.0403
Table 12. Absorbance values using External Calibration 4 0.0667
Method 6 0.0922
Concentration Absorbance 8 0.1225
0 -0.0066 10 0.1503
0.1 0.071
0.2 0.1456
0.5 0.3531 Table 14 shows the readings gathered by the
1 0.5159 spectrophotometer using a different method, Standard
2 0.8753 Addition Method. The results above show the
relationship between the amount of the standard
Table 12 shows the relationship between the solution and its readings. The more amount of the
concentration of the external standard solution which volume of standard added to the sample, the higher its
contains Lead and its resulted Absorbance. As shown absorbance is.
above, the blank solution or the 0-ppm external
standard resulted to a negative absorbance which is -
0.2
0.0066. The absorbance being read by the machine
increases as the concentration of the solution increases. 0.15

Absorbance
0.1
1
0.8 0.05
Absorbance

0.6 0
0 5 10 15
0.4 y = 0.4291x + 0.0539
Volume of standard added, mL
0.2
0 Figure 10. Linear graph of Absorbance values and the volume
0 0.5 1 1.5 2 2.5 of the standard solution added
-0.2
Concentration
Table 15. Absorbance values using Internal Standard Method
Concentration Absorbance
Figure 9. Linear graph of Absorbance values and the
0.1 0.0276
concentration of the external standard solution
0.2 0.025
0.5 0.0271
Table 13. Absorbance Values of Vegetable Samples 1 0.0273
Trial Absorbance 2 0.0283
1 0.0005
2 0.0008
3 0.0012 Table 15 shows the readings gathered by the
Mean 0.00083 spectrophotometer using a different method, Internal
Standard Method. The results in this method is
Table 13 shows the absorbance values of the vegetable different with the two methods but still it shows a
sample that were used to compare with pH values of relationship between the two values. The concentrated
the standard solutions. All trials resulted to a precise the solution of the analyte standard is, the higher the
reading ranging from 0.0005-0.001 and resulted to a absorbance being read.
calculated mean of 0.00083.

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0.031 y = -0.0005x + 0.0307 References

0.0305
A(a)/A(IS)

Ahmed, Moustafa. (2012). Atomic Absorption

0.03 Spectroscopy (AAS). 10.13140/RG.2.2.29580.51844.
0.0295
0.029
Atomic Absorption Spectroscopy by James W.
0 0.5 1 1.5 2 2.5 Robinson
Concentration of analyte standards, ppm
Schrenk, W.G., 1975. Analytical Atomic
Figure 11. Linear graph of Absorbance values and the Spectroscopy. Plenum Press, New York
concentration of analyte standards
“Spectrochemical Analysis by Atomic Absorption and
After gathering the readings of the absorbance from the Emission” by L.H.J.Lajunen, Royal Society of
spectrophotometer, the calculated concentration of the Chemistry, 1992
metals determined each sample are shown in the table
below: Varma, A., 1985. Handbook of Atomic Absorption
Analysis. Vol. I. CRC Press, Boca Raton.
Table 16. Concentration of metals from each sample
Sample Concentration of metal present
Water 9 ppm
Soil 0.6 ppm
Fruit 3.272 ppm
Vegetable 0.16 ppm

The results above show that the water sample has the
highest amount of metal present and the leafy
vegetable sample has the lowest amount of metal
present.

Conclusion

The absorbance read is directly proportional to the

concentration of atomic vapor in the flame released by
the spectrophotometer. As the results and data were
observed, it shows that all the four samples contain
heavy metals that could harm the people or the
environment. Based from the data gathered, the water
sample contains the highest amount of metal content
that can be due to the several waste that are being
thrown in the water. This could be the main reason why
it has the greatest amount of concentration. On the
other hand, the vegetable sample have the least amount
of metal content. It is important to know the
components of the food or water we intake because
even though these concentrations might appear small,
it could still be a hazard to our health.

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