VELAMMAL VIDHYASHRAM

MAMBAKKAM

ALL INDIA SENIOR SCHOOL

CERTIFICATE EXAMINATION
CHEMISTRY
INVESTIGATORY PROJECT REPORT
ON
REMOVAL OF NATURAL PIGMENTS BY THE
INTERACTION OF UV LIGHT AND OXYGEN
2023 - 2024

NAME: SHISHA LAEL. B. C

ROLL NO:
CLASS: XII SECTION - A
GROUP: BIOLOGY, MATHS
 BONAFIDE CERTIFICATE

This is to certify that this CHEMISTRY Investigatory Project on the

topic “Removal of Natural Pigments by the Interaction of UV
Light and Oxygen” has been successfully completed by Shisha
Lael. B. C of class XII-A, Roll No.: __________________ at Velammal
Vidhyashram, Mambakkam for the partial fulfilment of this project
as a part of All India Senior School Certificate Examination-CBSE,
New Delhi for the academic Year 2023– 2024.

Date: ……………………

Signature of Principal Signature of the Guide

Submitted for AISSCE 2023-2024, CHEMISTRY Practical examination

on ………………….

Signature of the Signature of the

Internal Examiner External Examiner
 ACKNOWLEDGEMENT

I have taken efforts to complete this project, however it would

have not been possible without the kind support and help of
many loving individuals and I would like to express my sincere
gratitude to all of them.
I hereby express my heartfelt thanks to our Senior Principal
for having given this opportunity to do the project and for her
constant encouragement and support.
I extend my sincere gratitude to my Chemistry teacher for her
valuable guidance offered to me. Her wholehearted
encouragement and constant stimulant inspiration, advice and
loving personality enabled me to complete the project
successfully.
I am also thankful to all our Teaching and Non-teaching staff
for their great help and support during my course of study.
I take pleasure in thanking my beloved family and friends,
especially my parents for being my pillars of strength and
always helping me and caring for me.
Above all, I thank the Lord Almighty for guiding me through
each and every step of my life.
Shisha Lael. B. C
XII-A
 INDEX

SL.NO. CONTENT PG. NO.

1 Introduction 1
2 Natural pigments 1
2.1 How are natural pigments obtained? 1
2.2 Examples of natural pigments 3
2.3 Classification of natural pigments 3
2.4 Applications of natural pigments 5
2.5 Extraction of natural pigments 5
3 Ultraviolet (UV) rays 6
4 Effects of UV radiation on natural pigments 8
5 Oxidation 10
5.1 Oxidation of pigments 10
6 Effect of oxygen on natural pigments 10
6.1 Do pigments oxidize? 12
7 Physical effects of pigment production in plants
(natural pigments) due to ultraviolent
radiation 12
8 Experiment [Removal of natural pigments by
the interaction of UV light and oxygen] 14
9 Pigment destruction due to UV rays and
oxidation in historical monuments 17
10 Conclusion 18
11 Bibliography 19
 Introduction
The aim of this investigatory project is to determine the role of UV
light and oxygen in the removal of natural pigments through an
experiment.

Natural pigments
Natural pigments are coloured substances derived from natural
sources, such as minerals, plants, and insects. The colorants may
be ground, washed, and sifted but otherwise are not chemically
modified.

Structure of natural pigment

How are natural pigments obtained?

Natural pigments are extracted from various biological sources
like plants, microorganisms—bacteria, fungi, and marine algae.

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Various colours made of natural pigments

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Examples of natural pigments:
Naturally occurring pigments such as Cochineal, Aeppo galls,
Annatto, Indigo, Ochres and Iron Oxides are known as Natural
Pigments.
Carmine is one of the most used pigments in the world.

Classification of natural pigments:

Before synthetic dyes and pigments were discovered, limited
number of natural colorant has been obtained from plants,
animals and minerals. The classification of colorants has become
mandatory due to huge increase in kind and number of colorants.
For this reason, colorants are classified based on their structure,

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source, colour, solubility and application methods. Dyes will be
investigated in two different groups as accordance with chemical
structures and application methods.

The basic classification groups were determined as azo,

anthraquinone, indigo, phthalocyanine, sulphur, nitro and nitroso
dyes by considering their chemical structures. According to
application method, they are grouped as reactive, disperse, acid,
basic, direct, and vat dyes. However, the classification of pigments
as organic and inorganic pigments is also regarded as an
appropriate way.

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Applications of natural pigments in different fields:
1) Food colorant

2) Pharmaceuticals
3) Textile and Leather Industry

4) Cosmetics
5) Solar Cells

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Extraction of natural pigments:
Natural colorants from plant-based materials have gained
increasing popularity due to health consciousness of consumers.
Among the many steps involved in the production of natural
colorants, pigment extraction is one of the most important.
Soxhlet extraction, maceration, and hydrodistillation are
conventional methods that have been widely used in industry and
laboratory for such a purpose. Recently, various non-conventional
methods, such as supercritical fluid extraction, pressurized liquid
extraction, microwave-assisted extraction, ultrasound-assisted
extraction, pulsed-electric field extraction, and enzyme-assisted
extraction have emerged as alternatives to conventional methods
due to the advantages of the former in terms of smaller solvent
consumption, shorter extraction time, and more environmentally
friendly. Prior to the extraction step, pre-treatment of plant
materials to enhance the stability of natural pigments is another
important step that must be carefully taken care of. A
comprehensive review of appropriate pre-treatment and
extraction methods for chlorophylls, carotenoids, betalins, and
anthocyanins, which are major classes of plant pigments, is
provided by using pigment stability and extraction yield as
assessment criteria.

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 Ultraviolet (UV) Rays
UV-Ultraviolet is a form of electromagnetic radiation with
wavelength shorter than that of visible light, but longer than X-
rays. UV radiation is present in sunlight, and constitutes about
10% of the total electromagnetic radiation output from the Sun.

There are three types of UV rays:

1] Ultraviolet A rays (UVA)
 The atmosphere does little to shield these rays—most UVA
radiation reaches Earth’s surface.
 UVA rays cause skin aging and eye damage, and can lower
your body’s ability to fight off illness. UVA rays also
contribute to the risk of skin cancer.
2] Ultraviolet B rays (UVB):

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  The Earth’s atmosphere shields us from most UVB rays—the
amount of UVB rays that reach the Earth’s surface depends
on latitude, altitude, time of year and other factors.
 UVB rays cause sunburns, skin cancer, skin aging, and snow
blindness (a sunburn to your cornea that causes a
temporary loss of vision) and can lower your body’s ability
to fight illness.
3] Ultraviolet C rays (UVC):
 UVC rays do not reach the Earth’s surface because they are
completely absorbed by the atmosphere.
 Harmful effects from UVC rays are minimal

Effects of UV radiation on natural pigments:

Here photosynthetic plants are taken as an example as they
contain natural pigment chlorophyll.

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The objective of this study is to evaluate the effects of UV
radiation on photosynthetic pigments and UV absorbing
compounds of pepper plants in the greenhouse. Pepper plants
were grown in a uniform environment and after 35 days they
were exposed to UV-A and UV-C radiation for 15 and 8 days,
respectively. Changes in photosynthetic pigments, chlorophyll
and carotenoids and UV absorbing compounds, flavonoids and
anthocyanins were measured. It is found that the content of Chl-a
and Chl-b slightly decreased in UV-R exposed plants and although
this reduction was not significant, the total chlorophyll (Chl-T)
amount decreased significantly especially in UV-C exposed plants.
Carotenoids concentration was also reduced in UV-R exposed
plants and this reduction was significant in UV-C exposed plants.
UV absorbing compounds were increased. Concentration of
flavonoids was significantly increased in both UV-A and UV-C
treatments. Although anthocyanin concentration was also
increased in UV-R exposed plants, this rise was not significant.
UV-C radiation significantly reduced number of leaves per plant,
but no significant effect was found on leaf numbers of UV-A
treated plants. The fresh weight was significantly decreased in
shoot of UV-C treated plants but there was no significant change
in fresh weight in UV-A treated plants. In the root of UV-R treated
plants no significant changes were observed in fresh weight.

Also UV radiation induces immediate pigment darkening (IPD) by

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chemical modification of melanin, and possibly spatial
redistribution of melanosomes in keratinocytes and melanocytes.
UV exposure also leads to delayed tanning (DT) by new synthesis
of melanin {also a natural pigment present in human body} over
several days after UV exposure and persists for weeks leading to
skin cancer in the long run.

Oxidation
At its most basic level, oxidation is the loss of electrons. It
happens when an atom or compound loses one or more electrons.
Some elements lose electrons more easily than others. These
elements are said to be easily oxidized. Generally speaking, metals
including sodium, magnesium, and iron are easily oxidized.

Oxidation of pigments:
The oxidation reaction transforms the carotenoid pigments into
one or more degradation products that are colorless or of lower
coloring capacity than the initial reactant. The kinetic parameters
(order of reaction and kinetic constant κ) are determined
according to the integral method.

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 Effect of oxygen on natural pigments
Package oxygen transmission rate (OTR) can affect the stability of
natural color pigments such as anthocyanin, betalains and
chlorophylls in foods during storage. The oxygen sensitivity of
selected pigments in thermally pasteurized vegetable purees held
at a refrigeration temperature.

Photo-oxidation has been identified as a main cause of the light-

induced color and brightness changes to natural pigments in
historical objects in exhibitions may be subject. Therefore, a
possible approach to reduce the risk of light damage is the
reduction of the oxygen content in the atmosphere surrounding
these objects. In order to estimate the efficiency of this approach,
light induced changes in the reflectance spectra of a series of 30
historical colorants kept in normal ambient air with those
observed in a second series stored in nitrogen. Several of them
have not been studied in anoxia [absence of oxygen] yet.
Chemically inert binder and substrate were used for sample
preparation in order to minimize their influence on
photochemical processes. In contrast to previous studies on
radioactive damage in anoxia, the samples were irradiated by
various narrow band LED light sources covering the visible part
of the electromagnetic spectrum. It becomes obvious that for a
while, a few colorants exhibit no or only minor effects due to the

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reduced oxygen concentration the majority benefitted
significantly from this measure. Only four metal-containing
pigments suffered increased damage in the nitrogen atmosphere,
while three showed a complex behaviour and could not be
assigned to one of these groups.

Do pigments oxidize?
Pigments may oxidize, dissolve in acid or water, undergo phase
transitions, react with the binders in the paint, or degrade. For
example, eosin Y was a pigment historically favoured by many
artists, most notably Vincent van Gogh.

Physical effects of pigment production in plants

(natural pigments) due to ultraviolent radiation:
Plants exposed to prolonged UV radiation have a range of
different physical symptoms. Arabidopsis plants exposed to UV
light for five days showed a marked difference in dry weight,
Susceptible species of cucumbers developed chlorotic lesions or
spots [the yellowing or whitening of normally green plant tissue
because of a decreased amount of chlorophyll, often as a result of
disease or nutrient deficiency]. The chlorotic spots eventually
became necrotic [When plants or plant cell cultures are subjected
to abiotic stress they initiate rapid cell death with necrotic
morphology]. The leaves exposed to UV-B radiation where also

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thinner than the leaves grown with no UV-B radiation.

Peas and grape developed permanent stomatal damage after

exposure to UV-B radiation for several weeks reported
significantly more cuticular wax [a mixture of hydrophobic very-
long-chain fatty acids and their derivatives accumulated in the
plant cuticle] .However, plants grown at ambient UV light had the
highest amount of wax at both squaring and flowering. The wax
on these leaves was denser than the wax on the cotton grown at
high UV levels. This increase was on the adaxial surface of the leaf,
but not on the abaxial side showed similar results in pea leaf
cuticles. They also found that the UV-B radiation caused a shift in
cuticle composition, moving from alcohols to esters and
hydrocarbons.

Research says that smaller flowers on cotton plants exposed to

UV light. The cotton plants exposed to high and ambient levels of
UV-B light had 33% and 15% less, respectively, anthers compared
to the plants not exposed to UV-B light. In comparison to control
treatments (no UV radiation), research reported a stimulation in
cotton plant growth under exposure to ambient UV-B radiation
levels. However, cotton plants grown under elevated UV radiation
were much shorter than both the ambient UV-B light and no UV-B
treatment.

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The plants grown under full UV radiation blocking film had an
increased above ground dry weight 40% versus the plants grown
under film that did not block any UV radiation and 122%. The
plants also grown under the UV blocking film also had 28% and
66% more leaves in 2005 and 2006, respectively, than the lettuce
grown under the film that blocked no UV radiation. The
researchers also found that lettuce grown under standard
horticultural film had a dry weight increase over non-UV blocking
film of 10% and 34% in 2005 and 2006, respectively. However,
there were no significant changes found in the variable and
maximum photochemical efficiency of any UV treatment.
How plants become when they are exposed to UV rays constantly:

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 Experiment
Removal of Natural Pigments by the Interaction of
UV Light and Oxygen

Objectives:
This project's goal is to find out if laying fabric on grass increases
fading of a stain by the sun. I hypothesized that oxygen coming
from the grass could interact with UV light from the sun and form
ozone, a powerful oxidizing agent. Oxidation is an important
process of fading, especially for plant pigments.

Materials:
 UV light
 Plastic trays
 Green, blue, red, black and white fabric (100% cotton)
 Red wine or beetroot
 Cellophane wrap
 Oxygen and nitrogen gas.

Methods:
(1) Prepare fabric samples and dye white cloth with red wine
or beetroot.

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(2) Make 4 trays, 2 with holes for gas inflow/outflow [trays
A/B].
(3) Mount 2 samples [cards with one square of every fabric
color and stained fabric] onto trays A/B and one onto
trays C/D.
(4) Attach nitrogen tank to tray A and oxygen tank to tray B.
(5) Fill trays C/D with oxygen and seal with cellophane and
rubber bands.
(6) 15 hours later take one sample out of trays A/B and
remove sample from tray C.
(7) After 24 hours remove remaining samples from tray A, B,
and D.
(8) Scan samples into computer using a colour scanner.

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Results:
Fading was increased for the stained samples that were in the
sealed oxygen trays compared to samples exposed to continuous
oxygen or nitrogen flow. The differences were observed after 15
and 24 hours of UV light exposure.

The black fabric showed a small amount of fading which did not
seem to be affected by the conditions. The stained fabric
subjected to sealed oxygen and UV light showed the most fading. I
had hypothesized that this is because ozone was able to build up
in this tray and oxidize the wine stain along with the effects of the
UV light. Less fading was seen with the continuous flow of oxygen
because ozone did not have a chance to build up. Nitrogen is inert
so any fading was most likely due to the UV light itself. For the
black fabric, the condition did not appear to matter suggesting
that the fading was also due to the UV light. Stain trick on the
grass may work because UV light from the sun causes the
formation of ozone near the grass, which increases the fading of
the stain. This project is about how pigments in a stain are
removed through the interaction of ultraviolet light and oxygen.

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 Pigment destruction due to UV rays and
oxidation in historical monuments
The wonders of the world and the most loved and saved art
pieces created by our ancestors which remain as the only way to
remind us of all the historic moments that happened before we
existed and are saved today as artifacts for the future to have as
symbols of past. Knowing that these pieces are slowly
deteriorating and slowly losing their originality because the
pigments in them are naturally fading away due to their
interaction with UV rays and oxygen and currently there is no
way to safeguard them is heart wrecking. Let’s take one of the
seven wonders of our world ‘Taj Mahal’ which is slowly but
steadily degrading mainly because of pollution but also due to the
loss of saturation in pigments in the marble due to constant
interaction with the UV rays and oxygen. Even masterpieces like
“The Mona Lisa” are estimated to be fully destroyed within the
next century due to oxidation that is oxygen reacting with the
pigments in the painting and in monuments the constant
exposure to UV rays the natural pigments present in them are
slowly getting removed. Thus these artifacts our symbols of past
are slowly losing their color and form due to this phenomenon.
These are real life examples for the removal of natural pigments
due to UV rays and oxidation.

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 Conclusion

In conclusion, natural pigments all around the world in any shape

or form will eventually get destroyed or removed or degraded
due to its long time exposure to UV rays and oxygen. Keeping this
in mind, ways to protect natural pigments need to be found.

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 Bibliography
1. https://www.cdc.gov/nceh/features/uv-radiation-
safety/index.html#:~:text=Ultraviolet%20(UV)%20radiatio
n%20is%20a,The%20sun
2. https://www.naturalpigments.com/natural-non-toxic-
pigments
3. https://link.springer.com/chapter/10.1007/978-981-15-
4638-
9_9#:~:text=Natural%20pigments%20are%20extracted%2
0from,and%20marine%20algae%20%5B3%5D.
4. https://www.epa.gov/radtown/ultraviolet-uv-radiation-
and-sun-exposure
5. https://scialert.net/fulltext/?doi=ijb.2008.486.490
6. https://trace.tennessee.edu/cgi/viewcontent.cgi?referer=&
httpsredir=1&article=1540&context=utk_gradthes
7. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3400707/
#:~:text=UV%20radiation%20induces%20immediate%20pi
gment,persists%20for%20weeks%20(8).
8. http://csef.usc.edu/History/2005/Projects/J0530.pdf
9. https://www.tandfonline.com/doi/abs/10.1080/00393630.
2020.1762401?journalCode=ysic20
10. https://pubmed.ncbi.nlm.nih.gov/31206676 /

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