Polymer Analysis

by UV - Visible
Spectroscopy
Experiment No: 07

PSC 313 1.0 Polymer

Practical Level 1B
G.A.N.K.Gunawardhana
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Date: 12/12/2023

Experiment No: 07

Experiment: Polymer analysis by UV-Vis spectroscopy

Objectives:
• To familiarize with the principles and instrumentation of UV visible
spectrophotometer.
• To gain knowledge on the use of that for polymer characterization.

Introduction:

UV-visible spectrometry is an analytical technique that measures the amount of

discrete wavelengths of UV or visible light that are absorbed by or transmitted through a
sample in comparison to a reference or blank sample. The composition of the sample affects
this property, which may reveal information about the substances and concentrations of the
sample. It is used for qualitative as well as quantitative analysis.

UV-Visible spectroscopy is based on the interaction between light and matter.

When light passes through or is absorbed by a molecule, it can cause the molecule to
vibrate. The wavelength of light that is most strongly absorbed by a molecule is called
the absorption maximum. By measuring the absorbance of light at different
wavelengths, it is possible to identify and characterize molecules. The principle of
operation for a spectrophotometer is that the wavelength of light is inversely proportional to
the size of its aperture. This means that shorter wavelengths will pass through a smaller
aperture than longer wavelengths. The monochromator in a UV-Vis spectrophotometer is a
disk with a series of slits that can be adjusted to select the desired wavelength.

There are two main methods for performing UV-Visible spectroscopy: absorption
spectroscopy and transmission spectroscopy. In absorption spectroscopy, a sample is placed
in the path of light and the absorbance of light at each wavelength is measured. In
transmission spectroscopy, the sample is placed in a cuvette and the transmitted light at each
wavelength is measured.

UV-Visible spectroscopy is used in a variety of applications, including analytical

chemistry, biochemistry, environmental science, and pharmaceuticals. It can be used to
identify and characterize molecules, measure the concentration of molecules in solution, and
determine the purity of a sample. UV-Visible spectroscopy is a powerful analytical tool that
can provide information about the structure, function, and dynamics of molecules. In
addition, UV-Vis spectroscopy can be used to monitor the progress of a reaction and to detect
impurities in a sample. By measuring the absorbance or transmission of light at specific
wavelengths, UV-Vis spectroscopy can be used to identify and characterize molecules. UV-
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visible spectroscopy can be used to measure the concentration of molecules in solution and
to determine the purity of a sample.

When considering the instrumentation of the UV–visible spectrometer, there are

several main components such as a light source, monochromator, wavelength filters, sample
holder, detector, and data recorder.

As a light-based technique, a steady source able to emit light across a wide range of
wavelengths is essential. The light source provides illumination at one or more specific
wavelengths. In the instruments tungsten or halogen lamp is commonly used for visible light,
whilst a deuterium lamp is the common source of UV light. As two different light sources are
needed to scan both the UV and visible wavelengths, the light source in the instrument must
switch during measurement.

UV-Vis spectroscopy requires a single wavelength for proper functioning

whereas the ideal output of a single wavelength is not possible. This is so because no
real wavelength selector is ideal. Although a single wavelength is not possible, a band
of radiation could be used. So an instrument with a narrow bandwidth would be
better.

There are two types of wavelength selectors named as filters and monochromators.
Filters are used to permit a certain band of wavelength. The simplest type of filter is the
absorption filter. Most commonly colored glass filters are used. They absorb a broad portion
of the spectrum (complementary colors) and transmit other portions (its own color). A
monochromator is an optical device that is used to select a narrow band of a wavelength of
light. It may be a quartz prism or grating. Monochromators are used for spectral scanning i.e.
varying wavelength of radiation over a range. They can be used for the UV-visible region.

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 Sample containers or cuvettes may be made up of Quartz, Borosilicate, or Plastic. Only
quartz is transparent in both UV & visible regions (200-700nm range). Glass & plastic are
suitable for the visible region only. Glass is not suitable for the UV region because it absorbs
UV radiation i.e. it is not transparent in the UV region. Plastic cells are not used for organic
solvents.

Detectors are devices that indicate the existence of some physical phenomenon. Some
examples of simple detectors are Transducers, Photodetectors, Photographic films.

Measuring a reference sample—also known as the "blank sample"—such as a cuvette

filled with the same solvent used to create the sample is essential for all studies. The
equipment then automatically uses the signal from the reference sample to assist in
determining the actual absorbance values of the analytes.

In UV-VIS spectroscopy, the transition of electrons at various levels by absorption of

radiation from ultraviolet to visible region is plotted in a graph. This line graph of various
absorptivities on specific levels of radiations is because of the absorption capacities of
compounds at certain levels. These levels are called regions of absorption and the compounds
are termed as chromophores.

The chromophores are present in almost every compound. This can be deduced by the
fact that almost all compounds and especially organic compounds can be identified and
quantified by the uv-vis spectroscopy.

The Beer-Lambert Law, also known as Beer's Law, is a fundamental principle used in
UV-VIS spectroscopy to relate the concentration of a solute in a solution to the absorbance
of light. Collectively Beer and Lambert’s laws state that the absorbance ‘A’ of an incident
monochromatic beam is directly proportional to concentration ‘c’ of the solution and path
length ‘ℓ’. The rate of decrease in intensity of monochromatic light is proportional to the
thickness of medium ‘ℓ’ and concentration ‘c’ of absorbing substance in dilution.

A∝c.ℓ

A=ε.c.ℓ
where,
ε is the molar absorptivity coefficient constant.
Or ε is the absorbance for a solution of concentration 1mole/dm -3 and a path length of
1cm.

The absorbance is the ratio of the intensity of incident electromagnetic radiation from
the source to that of refracted electromagnetic radiation detected by the detector.
Mathematical representation:
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 A = Log ℓo / ℓ
A=ε.c.ℓ

So,
Log ℓo / ℓ = ε . c . ℓ

To apply the Beer-Lambert Law, a calibration curve is often created. This involves
measuring the absorbance of known concentrations of the solute to establish a linear
relationship between absorbance and concentration. The slope of the line is related to the
molar absorptivity. Once a calibration curve is established, the Beer-Lambert Law can be
used to determine the concentration of an unknown sample. By measuring the absorbance of
the sample and applying the law, the concentration can be calculated.

UV-VIS spectrum is a graph that shows absorption at different wavelengths. The

relative maxima are known as lambda max (λmax). Spectra obtained by such techniques can
be useful for extracting information such as purity and composition. UV-Vis spectroscopic
spectra are frequently utilized to check the presence of different organic and conjugated
chemical species that have a wavelength in that range.

The study of effect of UV light on polymers has fascinated for polymer scientist for
many years. It is commonly used in both research and science as well as in industry
Polystyrene exhibits a new UV absorption band at around 290 nm, which is attributed to
associative interaction between pendant phenyl groups. Prepare a series of styrene solution
in different concentrations and observed their UV-Vis spectrums.

Figure 02: i) UV-Vis absorbance spectrum of styrene in methanol. (ii) Solid-state.

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 Figure 03: UV Visible Spectrophotometer

Apparatus:
• UV-VIS spectrophotometer
• beakers

Materials:
• Curcumin Extraction
• Distilled water

Methodology:

Firstly, the curcumin compound was dissolved in ethanol and a concentration series
was made by that. Then the solution series was analyzed one by one from the UV visible
spectrophotometer. Here same ethanol solution was used as the reference sample. The
absorbance values obtained for each concentration. The calibration curve was developed by
using that data. Finally, unknown samples were analyzed by the instrument supplying same
wavelength and the unknown concentration was calculated by the calibration curve.

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Results:
Table 01: Absorbance values for each concentration of curcumin solution from UV spectrophotometer

Curcumin Absorbance
concentration(mg/L)
57.1428 0.749
52.1739 0.6772
47.9999 0.62
44.4444 0.5777
41.3793 0.5417
38.7096 0.5137
36.3636 0.477
34.2857 0.4444
32.4324 0.4227
30.7692 0.405
29.2682 0.384
27.9069 0.3661
25.5318 0.3443
24.4897 0.3237
23.5293 0.3106
21.8181 0.287
21.0525 0.2736
19.672 0.2579
18.4614 0.246
The calibration curve

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Calculations:

Absorbance Concentration (mg/L)

0.33 24.816
0.461 35.406
0.554 42.404

Discussion:

In this experiment, curcumin sample was analyzed by using UV visible

spectrophotometer. But from our instrument, the spectrum could not be obtained. Therefore,
only the absorbance values for each concentration of curcumin solution series were obtained.
By using that absorbance values calibration curve was plotted. The concentrations of
unknown samples can be identified by obtaining their absorbance from UV visible
spectrophotometer. Here, the same ƛmax value should be supplied for both samples with
known concentrations (using to develop the calibration curve) and samples with unknown
concentrations (need to analyze). Here, ethanol was used as the solvent. Therefore, the same
ethanol solution should be taken as the reference sample.

Figure 04: Chemical structure of curcumin

This is the chemical structure of curcumin. According to the functional groups in this
compound, specific wavelengths should be absorbed. Therefore, to obtain correct data, the
ƛmax value should be identified by using spectrum and that wavelength should be supplied to
obtained absorbance values for samples with known concentrations.

According to the literature, following spectrum is obtained for different concentration

of curcumin solutions.

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 Figure 05: Absorption spectra of curcumin—ethanol solutions with different curcumin
concentration from 1, 2.5, 5, 10 μg and 20 μg/mL

The curcumin solution shows absorbance at λ = 425 nm and linear increasing

absorption at peak 425 nm when the curcumin concentration increased. The inset is linear
relation of the absorption intensity and curcumin concentration in this concentration range

The UV–Vis spectra of curcumin represent transitions between electronic energy

levels. These transitions are generally between a bonding or lone-pair orbital and an unfilled
non-bonding or anti-bonding orbital. The absorption spectra of curcumin dissolved in ethanol
is presented in Fig 5. with curcumin concentration varying from 1, 2.5, 5, 10 and 20 μg. The
absorption spectrum of curcumin is a broad band with maximum absorbance peak at a
wavelength ~425 nm, which could be assigned to low energy π–π* excitation of the curcumin

This peak is typical for curcumin dissolved in organic solvent. We see that the more the
curcumin solution is diluted, the more the UV–VIS absorption intensity decreases. The more
diluted the curcumin solution is, the more the UV–visible absorption intensity decreases. The
main reason the absorbance decreases in aqueous solution when concentration decreases is
the decrease in the number of absorption centers, while on the other hand, it is also the
degradation of curcumin in water medium by a reaction at the keto-enol group.

Conclusions:

By using UV visible spectrophotometer, an unknown concentration of the curcumin

sample can be calculated. The absorbency increases with increasing concentration.
The concentration values of A, B, C were obtained as 24.816 mg/L, 35.406 mg/L, 42.404
mg/L respectively.

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References:

1) Tom, J. (2023, December 15). UV-Vis Spectroscopy: Principle, Strengths and

Limitations and Applications. Analysis & Separations From Technology Networks.
https://www.technologynetworks.com/analysis/articles/uv-vis-spectroscopy-
principle-strengths-and-limitations-and-applications-349865 (16/12/2023)

2) Team, P. (2022, July 15). UV-Vis Spectroscopy: Principle, Instrumentation, and

Applications - Instrumental Analysis - PSIBERG. PSIBERG.
https://psiberg.com/uv-vis-spectroscopy/ (17/12/2023)

3) Van Nong, H., Hung, L. X., Thang, P. N., Chinh, V. D., Vu, L. V., Dung, P. T., Van
Trung, T., & Nga, P. T. (2016, July 22). Fabrication and vibration characterization
of curcumin extracted from turmeric (Curcuma longa) rhizomes of the northern
Vietnam. SpringerPlus, 5(1). https://doi.org/10.1186/s40064-016-2812-2

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