Name: Nerissa Rajkumar

Student Number: 219003070

Date of Practical: 21 April 2021

ABSTRACT:

The aim of this experiment is to quantify the amount of manganese contained in a steel
sample by two methods viz., atomic absorption spectrophotometry and UV/VIS spectroscopy
(method of standard addition) and comparing the accuracy of the two methods. For the UV-
VIS method the correlation coefficient was found to be 0.9961 and a straight line was
obtained which implies that there is a directly proportional relationship between the
concentration and absorbance which is in accordance with the Beer-Lambert law. The
concentration of Mn in UV/Vis spectroscopy was found to be 76.47ppm in 10x dilution
which was higher than the true value and the % error was found to be 0.2476%. In AAS
technique, the calibration curve was used to determine the unknown concentration which was
found to be 22.96ppm in 10x dilution and the % error was found to be 0.0228% which is very
low. A correlation coefficient was found to be 0.9981; this tells us that the relationship is in
accordance with the Beer Lambert law as the absorbance and concentration are directly
proportional. From the correlation coefficients it can be seen UV/VIS provides more precise
data however from the %error we see that AAS is the better method.

AIMS:

The aim of this experiment is to quantify the amount of manganese contained in a steel
sample by two methods viz., atomic absorption spectrophotometry and UV/VIS spectroscopy
(method of standard addition) and comparing the accuracy of the two methods.
INTRODUCTION AND PRINCIPLES:

Steel is an alloy of iron which contains many different elements including manganese 1.
Manganese removes oxygen and sulphur when iron ore (an iron and oxygen compound) is
converted into iron 2. Manganese is added to steel in order to harden and toughen it enough
that it is corrosion-resistant 1. Without Manganese, it becomes difficult to forge or hot roll 1.
The determination of manganese in steel or any other metal alloys is a common
implementation in industry 1.

In this experiment, a steel sample was dissolved, and manganese was converted to a brown
coloured strongly absorbing form as it was oxidised to permanganate ion by potassium
periodate in order to absorb the intensity of light using spectrophotometer. Stainless steel
wasn’t used in this experiment because nitric acid does not have the ability to dissolve the
steel sample 1.

Spectroscopy is the study of the absorption and emission of light and other radiation by
matter, as related to the dependence of these processes on the wavelength of the radiation 3.
Spectrophotometers are defined as devices used to quantify sample solution. In this
experiment, a standard addition method is employed which requires addition of a constant
quantity of standard solution to all solutions was analysed using UV/Vis spectroscopy 1.
Atomic absorption spectrometry (AAS) is used to quantity atoms contained in the sample by
measuring the radiation absorbed by the analyte atom. The basis of these two techniques are
how chromophores within these atoms and molecules interact with light and the quantity of
light absorbed is measured as absorbance across a detector 4.

All this is encompassed by the Beer-Lambert law which states that there is a linear
relationship between the concentration and the absorbance of the solution, which enables the
concentration of a solution to be calculated by measuring its absorbance 5. The concentration
of the sample can be calculated using Beer-Lambert law by rearranging the following
equation:

A= ɛ c l

Where c = concentration, ɛ = molar absorptivity and l = is the path length through which the
radiation travels. Since absorbance and concentration are proportional the calibration plot
must be a straight line.
The Principle of UV-Visible Spectroscopy is based on the absorption of ultraviolet light or
visible light by chemical compounds, which results in the production of distinct spectra.

Spectroscopy is based on the interaction between light and matter. When the matter absorbs
the light, it undergoes excitation and de-excitation, resulting in the production of a spectrum
6
. When matter absorbs ultraviolet radiation, the electrons present in it undergo excitation.
This causes them to jump from a ground state (an energy state with a relatively small amount
of energy associated with it) to an excited state (an energy state with a relatively large amount
of energy associated with it). It is important to note that the difference in the energies of the
ground state and the excited state of the electron is always equal to the amount of ultraviolet
radiation or visible radiation absorbed by it 7.

Figure 1: UV-VIS spectroscopy schematic

Source: https://www.microspectra.com/technical-support/248-spectrophotometer-design

The advantage of an Ultraviolet - Visible Light Spectrophotometer is its quick analysis ability
and easy to use. In astronomy research, a UV / Vis spectrophotometer helps the scientists to
analyze the galaxies, neutron stars, and other celestial objects. A UV spectrum can provide
rich information of the velocity and the elements of an astronomical object. In other
industries, UV / Vis spectrophotometer also brought the high-tech spectral analysis
possibilities. In the food industry, the quality and safety of foods are two most important
factor for consumers. In addition to the general sense of food quality factors, such as colour,
appearance, smell, taste, an UV / Vis spectrophotometer can further assist the food supplier
an instrumentation ways of analysis by chemistry, biology. And by the high standards of
quality control and production processes to improve the shelf life of the foods as well as
maintaining the safety. Besides, in the field of forensic analysis, medicine, or pharmacy, due
to the advantage of fast, easy analysis ability, a UV-Vis spectrophotometer is also widely
used in the researches.

For disadvantages, the stray light of UV-Vis spectrophotometer that caused by the faulty
equipment design and other factors could influence spectra measurement accuracy of the
absorption in substance, because the stray light will decrease linearity range and reduce the
absorbency of substance it measures. In addition, the electronic circuit design and the
detector circuit quality of spectrometer will affect the amount of noise that is coupled into the
measurement signal, thereby affecting the measurement accuracy and reduce the sensitivity
of the instrument 8.

UV-Visible spectroscopy is widely used in the field of analytical chemistry, especially during
the quantitative analysis of a specific analyte. For example, the quantitative analysis of
transition metal ions can be achieved with the help of UV-Visible spectroscopy. Furthermore,
the quantitative analysis of conjugated organic compounds can also be done with the help of
UV-Visible spectroscopy. It can also be noted that this type of spectroscopy can also be
carried out on solid and gaseous analytes in some conditions 6.

The method of standard addition is a type of quantitative analysis approach often used
in analytical chemistry whereby the standard is added directly to the aliquots of analyzed
sample. This method is used in situations where sample matrix also contributes to the
analytical signal, a situation known as the matrix effect, thus making it impossible to compare
the analytical signal between sample and standard using the traditional calibration
curve approach 9.

Advantages of Standard Addition Method 10:

 High quality measurement in unknown sample composition situations.

 Measurement without calibration.

 Quality of measurement can be stated as a function of the correlation coefficient.

Disadvantages of Standard Addition Method 11:

 Time consuming to perform a full calibration for one sample.

 It will not correct for additive interferences which cause a baseline shift.
 Inaccuracies in preparing the spiked samples can seriously affect the slope of the line,
causing errors to be made in the determining the original concentration.

Atomic absorption spectrometry (AAS) detects elements in either liquid or solid samples
through the application of characteristic wavelengths of electromagnetic radiation from a
light source. Individual elements will absorb wavelengths differently, and these absorbances
are measured against standards. In effect, AAS takes advantage of the different radiation
wavelengths that are absorbed by different atoms 14.

The absorption of light by atoms provides a powerful analytical tool for both quantitative and
qualitative analysis. Atomic absorption spectroscopy (AAS) is based upon the principle that
free atoms in the ground state can absorb light of a certain wavelength. Absorption for each
element is specific, no other elements absorb this wavelength.

AAS is a single-element method used for trace metal analysis of e.g., biological,
metallurgical, pharmaceutical and atmospheric samples. Spectroscopic determination of
atomic species can only be performed on a gasified sample in which the individual atoms
such as Ag, Al, Au, Fe and Mg are well-separated from each other 12.

The technique uses basically the principle that free atoms (gas) generated in an atomizer can
absorb radiation at specific frequency. Atomic-absorption spectroscopy quantifies the
absorption of ground state atoms in the gaseous state. The atoms absorb ultraviolet or visible
light and make transitions to higher electronic energy levels. The analyte concentration is
determined from the amount of absorption 13.
 Figure 2: atomic absorption spectrophotometer schematic

Source: http://delloyd.50megs.com/moreinfo/AA.html

The main advantages of AAS are given below:

 High sample throughput

 Easy to use

 High precision

 Inexpensive technique

The main disadvantages of AAS are as follows 15:

 only solutions can be analyzed

 less sensitivity compared to graphite furnace
 relatively large sample quantities are required (1-3 ml)
 problems with refractory elements

The are many applications for atomic absorption:

 Clinical analysis: Analyzing metals in biological fluids such as blood and urine.
 Environmental analysis : Monitoring our environment – e g finding out the levels of
various elements in rivers, seawater, drinking water, air, and petrol.
  Pharmaceuticals. In some pharmaceutical manufacturing processes, minute
quantities of a catalyst used in the process (usually a metal) are sometimes present
in the final product. By using AAS the amount of catalyst present can be determined.
 Industry: Many raw materials are examined and AAS is widely used to check that the
major elements are present and that toxic impurities are lower than specified – e g
in concrete, where calcium is a major constituent, the lead level should be low
because it is toxic.

EXPERIMENTAL:

Reagents:

 1000ppm Manganese solution

 4M Nitric acid
 Ammonium persulfate ((NH4)2S2O8)
 Sodium bisulphite (NaHSO3)
 Phosphoric acid, 85%
 Potassium periodate (KIO4)
 Steel Sample (E)

Instruments:

 Hot plates
 Spectrophotometer
Procedure:

Part i:

A standardised 1000ppm manganese solution was diluted in a 100mL volumetric flask to

produce a 100ppm working manganese standard. A 1.00g of sample steel was accurately
weighed into a 250mL beaker and dissolved with 50mL of 4M nitric acid boiled for about 10
minutes on hotplates in fume cupboard or until the sample steel dissolved. Ammonium
persulfate of 1.00g was added to the solution and this was boiled gently for a further 10
minutes- ~0.10g of sodium bisulphite was added to the solution if it had a pink colouration or
a brown deposit of MnO2 and boiled for a further 5 minutes. The solution was then cooled
and made up to 100mL in a volumetric flask. Thereafter, 5ml of the dissolved steel solution
was pipetted into 6 labelled beakers, and each solution was treated in the following manner:

Table 1: solution treatments

Solution: 85% H3PO4 / ml: Mn standard / ml: KIO4 / g:

Instrument zero 5 0 0
Blank 5 0 0.4050
Standard 1 5 1 0.4040
Standard 2 5 2 0.4050
Standard 3 5 5 0.4002
Standard 4 5 10 0.4026

Each solution was boiled for 5 minutes, cooled and transferred to a 100ml volumetric flask
and diluted to the mark with distilled water and mixing thoroughly. The solutions were then
analysed using spectrophotometer to measure the absorbance of each solution at the
wavelength of 525nm.
Part ii

The light absorption absorbed by Mn atoms was analysed using atomic absorption
spectroscopy (AAS). The dissolved steel sample prepared in part i was diluted to 100mL in a
volumetric flask using 10mL of the sample solution.

RESULTS:

---I used the W steel sample---

Table 2: The concentration and absorbance of Mn obtained using standard addition

method

Solution: Concentration / Absorbance:

ppm:
Run 1: Run 2: Run 3: Average:
Instrument zero 0 0.00 0.00 0.00 0.00
Blank 0 0.00 0.00 0.00 0.00
Standard 1 1 0.022 0.023 0.019 0.021
Standard 2 2 0.034 0.037 0.035 0.035
Standard 3 5 0.079 0.072 0.074 0.075
Standard 4 10 0.138 0.138 0.141 0.139
 Figure 3: Calibration curve for Manganese using UV/VIS Spectroscopy

Using Beer-Lambert law:

Atotal = (Ɛ .CS. VS + Ɛ. CU. VU) / VT

K = Ɛ. l / VT

Using the equation: y = 0.0136x + 0.0052

Slope = 0.0136 which is equal to k. CS

Intercept c = k. CU. VU = 0.0052

m/c = k. CS / k.CU.VU

CU = CS. b / m. VU

= (100ppm. 0.0052) / (0.0136. 5mL)

= 7.647 ppm

Dilution Factor = 100mL /10mL

= 10
Concentration of Mn in sample (ppm) = 7.647 mg L-1 x 10

= 76.47 mg L-1

Mass of Mn (g) = 76.47 mg L-1 x 0.100L

= 7.647 mg

= 0.007647 g

Mass % Mn = (0.007647g / 1.0094g) x 100%

= 0.7576%

Absolute error = ǀx-measured – x-true|

= ǀ0.7576– 0.51ǀ

= 0.2476%

Table 3: The concentration and absorbance of Mn obtained using the AAS method

Standard Concentration Absorbance: Standard %RSD:

solution: / ppm: deviation:
Run Run Run Average:
1: 2: 3:
1 2 0.044 0.046 0.047 0.046 0.001528 3.32
2 4 0.097 0.096 0.096 0.096 0.000577 0.60
3 6 0.131 0.134 0.132 0.132 0.001528 1.16
4 8 0.174 0.175 0.176 0.175 0.001000 0.57
5 10 0.218 0.220 0.220 0.219 0.001155 0.53
Sample 2.30 0.057 0.055 0.052 0.055 0.002517 4.58
 Figure 4: Calibration of Manganese using AAS

y = 0.0213x + 0.0061

Absorbance of sample= 0.055

0.055 = 0.0213x + 0.0061

Concentration of sample= x = 2.30 ppm

Dilution Factor = 100mL /10mL

=10mL

Concentration of Mn in 100mL = 2.30mg L-1 x 10

= 22.96 mg L-1

Mass of Mn (g) = 22.96 mg L-1 x 0.100 L

= 2.30 mg
= 2.30 x10-3 g

Mass % of Mn = (2.30 x10-3 g / 1.0094g) x 100%

= 0.5328%

Absolute error = ǀx-measured – x-true|

= ǀ0.5328 – 0.51ǀ

= 0.0282%

DISCUSSION:

The aim of this experiment was to quantify the amount of manganese contained in a steel
sample by two methods viz., atomic absorption spectrophotometry and UV/VIS spectroscopy
(method of standard addition) and comparing the accuracy of the two methods. Standard
addition method in UV/Vis spectrophotometry was used so as to decrease the matrix effect; if
this technique was not used the final concentration of manganese in a sample steel would
have been affected from the calibration curve equation.

From the calibration curve in Figure 3, the correlation coefficient was found to be 0.9961
which implies that there is a directly proportional relationship between the concentration and
absorbance which is in accordance with the Beer-Lambert law. The concentration of Mn in
UV/Vis spectroscopy was found to be 76.47 ppm in 10x dilution which was higher than the
true value and the % error was found to be 0.2476%.

In AAS technique, the calibration curve was used to determine the unknown concentration
which was found to be 22.96 ppm in 10x dilution and the % error was found to be 0.0228%
which is very low. The instrument was calibrated using solutions of known concentrations. A
correlation coefficient was found to be 0.9981; this tells us that the relationship is in
accordance with the Beer Lambert law as the absorbance and concentration are directly
proportional. From the correlation coefficients it can be seen UV/VIS provides more precise
data however from the %error we see that AAS is the better method.
To prepare a 1000ppm of Mn standard solution, we can roughly use about 1000mg of sample
comprising of manganese and dissolve it in 1 litre volumetric flask and dilute to the mark
with deionised water. The desired mass to make 250Ml of 1000ppm standard solution would
be 250mg which is obtained in the following manner: 1000ppm x 0.250L. When analysing a
solution or sample using UV/Vis spectrophotometry or AAS turbid solutions are not
necessary because the light may not be able to pass through a sample solution and it will
affect the analysis. Potassium periodate was added in manganese sample in this experiment to
oxidise it to permanganate in. which would give the purple colour that could be measured.

CONCLUSION:

The aim of this experiment was to quantify the amount of manganese contained in a steel
sample by two methods viz., atomic absorption spectrophotometry and UV/VIS spectroscopy
(method of standard addition) and comparing the accuracy of the two methods and this was
successfully achieved. Using the UV-VIS method the absolute error was found to be
0.2476% and using the AAS method the absolute error was found to be 0.00228%. We can
conclude that the AAS method is a better method to use for more accurate results. The
correlation coefficients for both methods shows very high accuracy. This indicates a linear
relationship between concentration and absorption.

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