Laboratory Report

Full Laboratory Report

SIC2022 Basic Analytical Chemistry

Experiment No: 4
Title: Determination of Ca, Mg, Fe and Na in mineral waters by flame
atomic spectrophotometry
Name: Sharvani A/P Karthigesu
Matric number: U2103660/1
Name of Partners: 1. Low Ru Yi
2. Nur Eillia Shuhada Binti Ab Rahman
Date: 8/11/2022
Objective

1. To quantitatively determine concentration of Ca, Mg, Fe, and Na in mineral

water by Flame Atomic Spectrophotometry.
2. To perform elemental analysis of Ca, Mg, Fe and Na using Atomic Absorption
Spectrophotometry and Atomic Emission Spectrophotometry

Introduction

Residential water quality from different source varies due to the presence of various
trace elements that are present in the water. Though these trace elements often seem
to be very insignificant, they can lead to many interesting health problems. The trace
elements in our drinking water have to meet the Malaysian Drinking Water Standards.
If it crosses the limit, it may develop severe diseases for human, animals, and plants
and sometimes may cause death (Dkhar, E. N., Dkhar, P. S., & Anal, J. M. H., 2014).
The exposure of trace elements such as Fe, Cu, Zn, Mn, Ni, As and Pb to humans in
higher amount or the bio-accumulation of these elements in the human organ can
cause lethal health effects with various diseases, depending on the nature and
quantity of metal, through ingestion, dermal contact, and inhalation pathways
(Mirzabeygi et al., 2017). So, identification and qualification of these elements become
necessary.

In this experiment, the determination of Ca, Mg and Fe in mineral water sample is

done by atomic absorption spectroscopy (AAS) while atomic emission
spectroscopy (AES) is used to determine Na in mineral water samples. AAS measures
absorption of light at specific wavelength by elements in its free state, such as calcium
and magnesium. When monochromatic light irradiates a group of atoms, a specific
wavelength is absorbed by the trace element measured. The light wavelength is
absorbed by the electrons for excitation. The absorption is detected by recorder and
a graph is plotted on readout device. The instrumentation and working of the Atomic
Absorption Spectrometry is shown in figure 1.
 Figure 1: Instrumentation and working of the Atomic Absorption Spectrometry

A., J. (2022, September 16). Atomic absorption vs. atomic emission spectroscopy.
PSIBERG. Retrieved November 14, 2022, from https://psiberg.com/atomic-
absorption-vs-atomic-emission-spectroscopy/#google_vignette

Atomic emission spectroscopy measures the intensity of light emitted by the atoms in
excited state. When an excited atom returns to the ground level, it emits radiation
which is detected by detector. The instrumentation and working of the Atomic Emission
Spectrometry is shown in figure 2.

Figure 2: Instrumentation and working of the Atomic Emission Spectrometry

A., J. (2022, September 16). Atomic absorption vs. atomic emission spectroscopy.
PSIBERG. Retrieved November 14, 2022, from https://psiberg.com/atomic-
absorption-vs-atomic-emission-spectroscopy/#google_vignette
A linear calibration curve is plotted for absorbance vs concentration from measuring
the absorbance of a series of standard solutions of a specific range. The concentration
of the sample obtained will then be compared with the calibration plot to determine Fe,
Ca and Mg. For Na, a graph of emission intensity vs absorbance is plotted and the
similar steps are done.

Experimental Procedure

Preparation of standard solutions of Ca, Mg, Fe and Na

1. Concentration of standard solutions of Na, Fe, Mg, Ca and Na were recorded.

2. 5 standard solutions for each element were prepared using 50 mL volumetric

flask. The concentrations of the standard solutions prepared was within the
range as presented in the table below.

Element Concentration range (ppm)

Ca 0.10 – 0.50
Mg 0.02 – 0.15
Fe 0.25 – 3.0
Na 0.05 – 0.25
Table 1: The concentration range of standards
A sample of mineral water was diluted with dilution factor of 100/3 and 50/3.

Determination of calcium, magnesium and iron in water using AAS and

Determination of sodium in water using AES

1. The flame atomic spectrophotometer and atomic absorption photometer was

set up with the help of lab assistant.
2. Fe, Ca and Mg were measured in atomic absorption spectrophotometer while
Na was measured in Atomic Emission Spectrophotometer. The sample was
measured following each measurement of standards.
3. The data was recorded and graph was plotted. LOD and LOQ were
determined.

Results and Discussion

A) Determination of calcium, magnesium and iron in water using AAS

i) Tabulation and plot of absorbance vs concentration for the calcium,

magnesium, and iron measurement.
Table 2 : The concentration of calcium standards its absorbance.
 No. of Concentration of Ca Absorbanc
solution (ppm) e
1 0.10 0.0014
2 0.20 0.0125
3 0.30 0.0205
4 0.40 0.0285
5 0.50 0.0378

Table 3: The absorbance of Ca in sample

Sample Absorbance
Pure 0.864
Diluted(50/3) 0.143
Diluted(100/3) 0.064

Graph of Absorbance against Concentration of Ca

0.05

0.04

y = 0.0888x - 0.0065
0.03
R² = 0.9963
Absorbance

0.02

0.01

0
0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70
-0.01
Concentration of Ca (ppm)

Figure 3: The graph of absorbance against the concentration of Ca.

Table 4 : The concentration of magnesium standards its absorbance.

No. of Concentration of Mg Absorbanc
solution (ppm) e
1 0.04 0.0296
2 0.08 0.0521
3 0.12 0.0846
4 0.16 0.0969
5 0.20 0.1229
 Table 5: The absorbance of Mg in sample
Sample Absorbance
Pure 1.616
Diluted (50/3) 0.272
Diluted (100/3) 0.108

Graph of Absorbance against Concentration of Mg

0.1400

0.1200
Absorbance

0.1000
y = 0.5785x + 0.0078
0.0800 R² = 0.9866

0.0600

0.0400

0.0200

0.0000
-0.05 0.00 0.05 0.10 0.15 0.20 0.25
Concentration of Mg(ppm)

Figure 4: The graph of absorbance against the concentration of Mg.

Table 6 : The concentration of iron standards its absorbance.

No. of Concentration of Fe Absorbanc
solution (ppm) e
1 0.5 0.0399
2 1.0 0.0747
3 1.5 0.1075
4 2.0 0.1519
5 2.5 0.1632

Table 7: The absorbance of Fe in sample

Sample Absorbance
Pure 0.002
Diluted (50/3) 0.002
Diluted (100/3) 0.002
 Graph of Absorbance against Concentration of Fe
0.2500
y = 0.0648x + 0.0103
0.2000 R² = 0.9781
Absorbance

0.1500

0.1000

0.0500

0.0000
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5
Concentration of Fe(ppm)

Figure 5: The graph of absorbance against the concentration of Fe.

ii) Calculation of concentration of Ca, Mg and Fe in water

samples

By substituting the measured absorbance, y of sample into the equation y = mx + c,

we can obtain the concentration of sample, x.
x = concentration of element
y = absorbance of sample

The concentration of Ca in mineral water sample

A straight-line graph is plotted and y = 0.0888x - 0.0065 obtained from the

graph shown in Figure 1 is compared with y = mx + c.

Slope or m = 0.0888
The y-intercept = 0.0065

For Ca, the diluted sample of mineral water with dilution factor 100/3 has an
absorbance of 0.064 which falls outside of the calibration range (0.0014 – 0.038). The
concentration of Ca in mineral water could not be determined as the sample
absorbance is higher than the standards. Therefore the calibration equation y =
0.0888x - 0.0065 is not applicable to measure concentration of Ca.

To improve the result, we should reduce the dilution factor by 100/2 or 100/1 and
repeat the experiment till the value falls within the calibration range. It is said because,
as the volume of dilution increases, the number of molecules per unit volume
decreases.

The concentration of Mg in mineral water sample

A straight-line graph is plotted and y = 0.5785x + 0.0078 obtained from the

graph shown in Figure 1 is compared with y = mx + c.

Slope or m = 0.05785
The y-intercept = 0.0078

i) For the diluted mineral water,

Dilution Factor: 100/3
The measured absorbance, y is 0.108.
By substituting y = 0.108 into the equation y = 0.5785x + 0.0078
0.108 = 0.5785x + 0.0078
x = Mg concentration = 0.17 ppm
ii) For pure minetal water,
By multiplying x with dilution factor,
Concentration of Mg = 0.17 ppm x 100/3
=5.67 ppm

Table 8: The concentration and absorbance of Mg in sample

Sample Concentration,
ppm
Diluted(100/3) 0.17
Pure 5.67
For Mg, the diluted sample of mineral water with dilution factor 100/3 has an
absorbance of 0.108 which is within the calibration range (0.0296 – 0.1229).
Therefore, the concentration of Mg in mineral water could be determined.

The concentration of Fe in mineral water sample

A straight-line graph is plotted and y = 0.0648x + 0.0103 obtained from the

graph shown in Figure 1 is compared with y = mx + c.

Slope or m = 0.0648
The y-intercept = 0.0103
For the pure water sample,
The measured absorbance, y is 0.002.

For Fe, the diluted sample of mineral water with dilution factor 100/3 has an
absorbance of 0.002 which falls outside of the calibration range (0.0399 – 0.1632).
The concentration of Fe in mineral water could not be determined as the sample
absorbance is very much lower than the standards. Therefore the calibration equation
y = 0.0888x - 0.0065 is not applicable to measure concentration of Ca.

To improve the results, we should heat the sample to concentrate it. It is because, at
100°C, only water molecules will evaporate while the metal ions don’t. As a
consequence, the number of mol per unit volume of metal ions increases and this
concentrated sample solution can be used to measure the absorbance of Fe.

B) Determination of sodium in water using AES

i) Tabulation and plot of emission intensity vs concentration for

sodium
Table 9 : The concentration of sodium standards its emission intensity.
No. of solution Concentration of Na Emission
(ppm) Intensity
1 0.05 0.1960
 2 0.10 0.3071
3 0.15 0.3985
4 0.20 0.5219
5 0.25 0.5492

Table 10: The emission intensity of Na in water sample

Sample Emission
Intensity
Pure 0.8456
Diluted (100/3) 0.3829

Graph of Emission Intensity against Concentration of Na

0.8000 y = 1.8424x + 0.1182
R² = 0.973
0.7000
Emission Intensity

0.6000
0.5000
0.4000
0.3000
0.2000
0.1000
0.0000
-0.05 0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.35
Concentration of Na(ppm)

Figure 6: The graph of emission intensity against the concentration of Na.

iii) Calculation for concentration of Na in water samples

By substituting the measured emission intensity,y of sample into the equation y = mx

+ c, we can obtain the concentration of sample, x.
x = concentration of element
y = emission intensity of sample

A straight-line graph is plotted and y = 1.8424x + 0.1182 obtained from the

graph shown in Figure 1 is compared with y = mx + c.

Slope or m = 1.8424
The y-intercept = 0.1182
 i) For the diluted mineral water,
Dilution Factor: 100/3
The measured absorbance, y is 0.3829.
By substituting y = 0.3829 into the equation y = 1.8424x + 0.1182,
0.3829 = 1.8424x + 0.1182
x = Na concentration = 0.14 ppm
ii) For pure mineral water,
By multiplying x with dilution factor,
Concentration of Mg = 0.14 ppm x 100/3
=4.67 ppm

Table 11: The concentration and emission intensity of Na in sample

Sample Emission Concentration,
Intensity ppm
Pure 0.8456 4.67
Diluted (100/3) 0.3829 0.14

For Na, the diluted sample of mineral water with dilution factor 100/3 has an emission
intensity of 0.3829 which falls within the calibration range (0.1960 – 0.5492). The
concentration of Na in mineral water can be determined.

i) Calculation of LOD and LOQ based on the calibration plot

The term LOD stands for limit of detection. It is the lowest amount of a
substance that can be distinguished from the absence of that substance (zero
point) with a stated confidence level. In our experiment, the confidence level
for LOD is 95%. We can estimate the detection limit from the standard
deviation of the blank and the slope of the calibration plot along with a defined
confidence factor.

The term loq stands for limit of quantitation. It gives the smallest concentration
of an analyte in a test sample that we can determine with acceptable
repeatability and accuracy. In other words, it is the concentration at which the
 entire analytical system must give a recognizable signal and acceptable
calibration point.
If the results lie between the range of LOD and LOQ, it means that a particular
analyte is present in a sample in a value that can be recorded. However, if the
value is lesser than LOD, it implies that the analyte is not detected which
means that the analyte is present in very small concentration. On the other
hand, if the value is more than LOQ, it indicates that the analyte is present in
a amount that can be detected (Madhu, 2020).

Sy
LOD = 3.3 ( )
S
Sy
LOQ = 10 ( )
S
Whereby,
Sy = SE × √N , where SE is the standard error of intercept and N is the number of tests
and
S = Slope of the graph y=mx+c

The LOD and LOQ for Ca, Fe and Na in water samples are calculated based on the
formulas above using the data analysis function in excel and the result obtained is
shown in table 10.

Noted that N=5,

Table 12: The LOD and LOQ values for Ca, Mg, Fe and Na in water sample
Ca Mg Fe Na
SE of intercept 0.00174 0.00516 0.00928 0.02941
SD of
intercept,Sy 0.00388 0.01153 0.02075 0.06576
LOD, ppm 0.144 0.066 1.057 0.118
LOQ, ppm 0.437 0.199 3.202 0.357
√N 2.23607 2.23607 2.23607 2.23607
Slope, S 0.0888 0.5785 0.0648
1.8424
➢ The concentration of Ca ions can be detected as the Ca ions are
present in a high concentration(absorbance higher than calibration
ramge)
➢ The Mg concentration of sample is 5.67 ppm which is higher than LOQ.
Therefore, it can be detected.
➢ The concentration of Fe ions can’t be detected as the Fe ions are
present in a very low concentration(absorbance lower than calibration
range) and the limitation of the instruments used causes it to be unable
to be detected.

➢ The concentration of Na is 4.67 ppm. It falls within the LOD and LOQ
value which shows that Na can be detected in the sample mineral
water.

Figure 7 : The data analysis (regression) for Ca.

 Figure 8: The data analysis (regression) for Mg.

Figure 9 : The data analysis (regression) for Fe.

Figure 10 : The data analysis (regression) for Na.

Discussion

Table 12: Maximum acceptable value and concentration of element in mineral water
for Ca, Mg, Fe and Na.

Elements Maximum Acceptable Concentration of element

Value, ppm in mineral water, ppm
Calcium 500 N/A
Magnesium 125 5.67
Iron 0.03 N/A
Sodium 50 4.67
From table 12, we can see that the concentration of magnesium and sodium fall
below the maximum acceptable value which indicates that the mineral water sample
measured is safe to be consumed.

There are a few other precautions that should be taken while conducting this
experiment. First, we must use a new micropipette tip to draw out different solutions
to prevent contamination. We must also ensure that there are no air bubbles at the
tip of micropipette. It is also important to ensure that there are no solution adhering
to the wall of the micropipette to obtain a very precise reading. Next, the micropipette
should always be in a upright position to prevent its dysfunctions. Then, all the
volumetric flasks must be calibrated with distilled water prior to use. When
measuring the absorbance of samples in AAS and AES, we must take the
measurement of blank twice.

Conclusions

Question and Answer

1. Why is flame emission a more sensitive technique for some cations, mainly
the alkaline and earth alkali cations, while atomic absorption has greater
sensitivity for other cations, such as the transition metal ions?

Flame emission spectroscopy uses intensity of light emitted from flame at

particular wavelength to determine the quantity of element in a sample. The
temperature of the flame produced is relatively low which is only suitable to
dissociate very reactive metals, specifically alkali metals and alkali earth
metals.
However, in atomic absorption spectroscopy, stable elements like
transition metals absorb light in the beam to excite to a higher energy level.
The substance's absorption is measured to determine the wavelength and
amount of radiation. Alkali metals are not measured in AAS because they
will drop to the ground state after getting excited due to instability.

2. Explain why AAS is so selective, i.e. why do other elements not usually
interfere in the analysis?
 The atoms will absorb electromagnetic radiation at a specific wavelength. The
wavelength of light source absorbed by atoms is the same as element that is
being evaluated. Since other elements in the sample won't absorb the
wavelength of interest, this technique becomes highly selective towards the
element of interest only.

3. Why LOD and LOQ are important in chemical analysis.

The limits of detection (LOD) and quantification (LOQ) are parameters used
to determine the lowest analyte concentration that can be detected with a
certain analytical method with acceptable precision. For this reason, it is
crucial to do both calculations in chemical analysis to obtain a very precise
measurement of element present in a sample.

References

Madhu, M. (2020, August 9). Difference between LoD and LoQ. Compare the
Difference Between Similar Terms. Retrieved November 13, 2022, from
https://www.differencebetween.com/difference-between-lod-and-loq/

Armbruster, D. A., & Pry, T. (2008). Limit of blank, limit of detection and limit of
quantitation. The Clinical biochemist. Reviews, 29 Suppl 1(Suppl 1), S49–S52.

Department of Chemistry, Faculty of Science, University of Malaya. (n.d.). Basic

Analytical Chemistry Second-Year Laboratory Manual. Retrieved from
https://kimia.um.edu.my/

What is atomic absorption spectroscopy (AAS). SciMed. (n.d.). Retrieved November

13, 2022, from https://www.scimed.co.uk/education/what-is-atomic-absorption-
spectroscopy-aas/

Mirzabeygi, M., Abbasnia, A., Yunesian, M., Nodehi, R. N., Yousefi, N., Hadi, M., &
Mahvi, A. H. (2017). Heavy metal contamination and health risk assessment
in drinking water of Sistan and Baluchistan, Southeastern Iran. Human and
Ecological Risk Assessment: An International Journal, 23(8), 1893-1905.
https://doi.org/10.1080/10807039.2017.1322895

Dkhar, E. N., Dkhar, P. S., & Anal, J. M. H. (2014). Trace Elements Analysis in
Drinking Water of Meghalaya by Using Graphite Furnace-Atomic Absorption
Spectroscopy and in relation to Environmental and Health Issues. Journal of
Chemistry, 2014, 975810. https://doi.org/10.1155/2014/975810