IP CHEM Shijin

PONGUMMOODU, TRIVANDRUM
SENIOR SCHOOL CERTIFICATE EXAMINATION – 2023-24
Report of
…………………………………………………………….Project
In……………………………………………………………………..
Titled…………………………………………………………………
Roll No:…………..
CERTIFICATE
This is to certify that Shijin S.S of Class XII has

completed the project titled "Study of
Chromatography" under my supervision.
She/he/they has/have shown great interest
and dedication in completing the project as
per the CBSE guidelines.
Internal Examiner External examiner
Principal
ACKNOWLEDGEMENT
I would like to take this opportunity to express my
gratitude to those who have been of great assistance in
finishing this project in the due course of the stipulated
deadline.
To begin with I would like to thank Principal Sr. Annie
M.I for her encouragement and for providing me with all
the required facilities for completing my project.
I extend my heartfelt thanks to Mrs. Sinu P, our
chemistry teacher who guided me in the successful
completion of this project.
I would also acknowledge my gratitude towards my
classmates for having been a source of inspiration and
for their patience in resolving what I couldn’t grasp
efficiently. I thank them all for expanding my interest in
the study that is programming.
CONTENT
1 INTRODUCTION
2 OBJECTIVE
3 EXPERIMENT 1:
SEPARATION OF PLANT
PIGMENTS
4 EXPERIMENT 2:
IDENTIFICATION OF INK
COMPONENTS
5 OBSERVATIONS
6 CONCLUSION
7 BIBLIOGRAPHY
INTRODUCTION
Chromatography is a technique used to separate

the different components of a mixture based on
their differential interactions with two phases: a
stationary phase and a mobile phase. The method
is widely used in chemistry to analyze mixtures of
substances and identify their components. Its
applications range from separating plant pigments
to analyzing ink and even complex forensic
investigations.
OBJECTIVE
The objective of this project is to explore and

demonstrate the process of chromatography, a
vital separation technique used to isolate
individual components from complex mixtures.
Chromatography is widely used in scientific
research, industries, and forensic analysis due to
its efficiency and precision. This project aims to:
● Understand the principles of how different

substances move at varying speeds through a
stationary medium, based on their molecular size,
solubility, and interaction with the mobile phase.
● Investigate how chromatography can be applied to
separate pigments from plants, as well as dyes
from common inks, using paper chromatography.
● Analyze the factors affecting the efficiency of
separation, such as solvent choice, particle size,
and their respective affinities for the stationary
and mobile phases.
● Provide insight into the practical applications of
chromatography in real-world scenarios, such as in
the pharmaceutical industry for drug purity testing,
environmental monitoring for pollutant detection,
and in the food industry for quality control.
EXPERIMENT 1
SEPARATION OF PLANT PIGMENTS
Aim:
To separate plant pigments using paper
chromatography.
Requirements:
Filter paper, Mortar and pestle, Fresh spinach
leaves, Ethanol, Beaker, Pencil, Capillary tube, Ruler,
Glass rod ,Solvent mixture (ethanol and water)
Procedure:
1. Crush fresh spinach leaves using a mortar and
pestle to extract the pigments.
2. Using a capillary tube, place a small drop of the
pigment extract on a filter paper about 2 cm from
the bottom edge.
3. Fill a beaker with a solvent mixture (ethanol and
water).
4. Suspend the filter paper in the beaker with the
pigment spot just above the solvent.
5. Allow the solvent to travel up the filter paper and
6. observe the separation of pigments.
7. Once the solvent front reaches near the top of the
paper, remove the paper and mark the solvent
front with a pencil.
8. Measure the distances traveled by each pigment
and calculate the Rf values.
Observation:
After the solvent (ethanol-water mixture) moved up
the filter paper, different bands of pigments were
observed.
The pigments separated based on their solubility in the
solvent and their interaction with the paper.
The following colors were visible:
● Chlorophyll: A green band observed at a distance
of approximately 5.2 cm.
● Carotenoids: A yellow-orange band observed at a
distance of approximately 3.8 cm.
The Rf values (Retention factor) were calculated as
follows:
Pigment Distance Moved Distance Rf Value

by Pigment (cm) Moved by
Solvent (cm)
5.2 8.0 0.65
Chlorophyll
Carotenoids 3.8 8.0 0.47

Conclusion:
Different plant pigments, such as chlorophyll and
carotenoids, are separated based on their solubility in
the solvent. Chlorophyll appears green, while
carotenoids show a yellow-orange band.
EXPERIMENT 2:
IDENTIFICATION OF INK COMPONENTS
Aim:
To identify the different components of black ink using
paper chromatography.
Requirements:
Black marker, Filter paper, Beaker, Ruler, Pencil, Water
or alcohol as a solvent
Procedure:
1. Draw a line on a filter paper using a black marker
about 2 cm from the bottom.
2. Fill a beaker with a small amount of water or
alcohol.
3. Place the filter paper in the beaker with the ink
spot above the solvent.
4. Let the solvent move up the filter paper and
separate the ink into its component dyes.
5. Remove the paper once the solvent has traveled
near the top, and mark the solvent front.
6. Identify the colors that have separated from the
black ink and calculate their Rf values.
Observation:
The ink separated into different colored components as
the solvent moved up the filter paper.
Different dyes were observed, showing that black ink is
a mixture of multiple dyes.
The following colors were separated:
● Blue dye: Moved to a distance of 6.5 cm.
● Red dye: Moved to a distance of 4.2 cm.
The Rf values were calculated as follows:
Pigment Distance Moved Distance Rf Value

by Pigment (cm) Moved by
Solvent (cm)
6.5 8.0 0.81

Chlorophyll
Carotenoids 4.2 8.0 0.53

Conclusion:
Black ink is composed of various colored dyes, which
are separated during chromatography based on their
solubility and affinity for the mobile and stationary
phases.
CONCLUSION
Chromatography is a highly effective and versatile
technique for separating components of complex mixtures
based on their differential solubility and interactions with
the stationary and mobile phases. The experiments
conducted demonstrated the fundamental principles of
chromatography and its practical applications in separating
both natural and synthetic substances.
In the first experiment, we successfully separated plant
pigments such as chlorophyll and carotenoids from
spinach leaves using paper chromatography. These
pigments were carried by the solvent at different rates due
to their varying affinities for the mobile and stationary
phases. This experiment highlighted the importance of
factors such as solubility and molecular structure in
determining how substances move through the stationary
medium. The distinct separation of green and yellow
pigments provided clear visual evidence of how
chromatography can isolate individual components from a
complex biological mixture.
In the second experiment, the components of black ink
were separated into different colored dyes. This
experiment emphasized how common commercial
products, such as ink, are made up of multiple
compounds, each with different chemical properties. The
separation of blue and red dyes showed how
chromatography can be used to break down seemingly
uniform substances into their individual parts, which is
crucial in various fields such as forensic analysis and
quality control in manufacturing.
Both experiments confirmed that chromatography can
efficiently separate components of a mixture based on
their movement through the stationary phase. The
experiments also demonstrated how factors such as the
choice of solvent, the type of stationary medium, and the
molecular properties of the substances being separated
play a critical role in the effectiveness of the separation.
In conclusion, chromatography is a vital tool in scientific
analysis, providing a reliable and accurate method for
separating and identifying substances. Through the
conducted experiments, it became clear that this method
not only allows for the separation of complex mixtures but
also enhances our understanding of the underlying
molecular interactions that govern these separations.
Chromatography remains a cornerstone of chemical
analysis due to its precision, versatility, and wide range of
applications, making it indispensable in research and
industry alike.

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PONGUMMOODU, TRIVANDRUM

SENIOR SCHOOL CERTIFICATE EXAMINATION – 2023-24

This is to certify that Shijin S.S of Class XII has

Internal Examiner External examiner

Chromatography is a technique used to separate

The objective of this project is to explore and

● Understand the principles of how different

SEPARATION OF PLANT PIGMENTS

Pigment Distance Moved Distance Rf Value

Carotenoids 3.8 8.0 0.47

IDENTIFICATION OF INK COMPONENTS

Pigment Distance Moved Distance Rf Value

6.5 8.0 0.81

Carotenoids 4.2 8.0 0.53

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