NEW MIDDLE EAST INTERNATIONAL
SCHOOL
CHEMISTRY PROJECT
2024-25
Name: Mubarakah Oakasa
Class: XII-A
Admission Number: 14320
Project report submitted in fulfilment of class
XII syllabus by AISSCE.
CERTIFICATE
This is to certify that this record of bonafide
project work is carried out by Mubarakah Oakasa
Admission No: 14320 of class XII in the
laboratory of this school during 2024-25.
External examiner Teacher in Charge
Principal
DECLARATION
I hereby declare that the project work entitled
“Chromatography” is submitted in partial
fulfillment of the study of class XII. Under the
scheme in Chemistry is a record of bonafide
work carried out by Mubarakah Oakasa under
the guidance of Mrs. Linu, and no part of it has
been submitted for any other study.
ACKNOWLEDGEMENT
I would like to extend my heartfelt gratitude for the
completion of this project which was motivated by
the encouragement and guidelines of our respected
teachers and respected principal ma’am. On that
note, I am grateful to our chemistry teacher, Mrs.
Linu, for aiding with all possible resources and
knowledge required for this project. My sincere
gratitude is owed to Mrs. Tabassum Farooqui, the
principal and Mrs. Grace Thomas, the principal in
charge of New Middle East International School, for
her part in counseling and providing required
resources for the completion of our academic
assignments. Finally, I regard a final
acknowledgement to CBSE, for granting us with
assignments that expand our curiosity to hover and
immerse ourselves into the main purposes and
awareness of the topics we consider for these
assignments.
INDEX
Introduction 6
Principle 7
Types of chromatography 8
1. Column chromatography 9-10
2. Thin layer chromatography 11
3. Gas chromatography 12
4. Paper chromatography 13-16
Conclusion 17
Bibliography 18
INTRODUCTION
Chromatography is an important biophysical
technique that enables the separation, identification,
and purification of the components of a mixture for
qualitative and quantitative analysis. Proteins can be
purified based on characteristics such as size and
shape, total charge, hydrophobic groups present on
the surface, and binding capacity with the stationary
phase. Four separation techniques based on
molecular characteristics and interaction type use
mechanisms of ion exchange, surface adsorption,
partition, and size exclusion. Other chromatography
techniques are based on the stationary bed, including
column, thin layer, and paper chromatography.
Column chromatography is one of the most common
methods of protein purification.
PRINCIPLE
Chromatography is based on the principle where
molecules in mixture applied onto the surface or into
the solid, and fluid stationary phase (stable phase) is
separating from each other while moving with the
aid of a mobile phase. The factors effective on this
separation process include molecular characteristics
related to adsorption (liquid-solid), partition (liquid-
solid), and affinity or differences among their
molecular weights. Because of these differences,
some components of the mixture stay longer in the
stationary phase, and they move slowly in the
chromatography system, while others pass rapidly
into mobile phase, and leave the system faster.
TYPES OF CHROMATOGRAPHY
Types of chromatography on the basis of difference
in the moving phase and the stationery phase:
1. Column chromatography
2. Thin-layer chromatography
3. Gas chromatography
4. Paper chromatography
1. COLUMN CHROMATOGRAPHY
Column chromatography is a technique which is used to
separate a single chemical compound from a mixture
dissolved in a fluid.
Types:
i) Adsorption chromatography
Adsorption chromatography is the oldest type of
chromatography technique. It makes use of a mobile
phase which is either in liquid or gaseous form. The
mobile phase is adsorbed onto the surface of a stationary
solid phase.
ii) Partition chromatography
Partition chromatography is based on differential
partitioning of components of a sample mixture between
two phases – stationary phase and mobile phase.
2. THIN LAYER CHROMATOGRAPHY
Thin Layer Chromatography is a technique used to isolate
non-volatile mixtures. The experiment is conducted on a
sheet of aluminium foil, plastic, or glass which is coated
with a thin layer of adsorbent material. The material
usually used is aluminium oxide, cellulose, or silica gel.
3. GAS CHROMATOGRAPHY
Gas chromatography (GC) is a common type of
chromatography used for separating and analyzing
compounds that can be vaporized without decomposition.
Typical uses of GC include testing the purity of a
particular substance, or separating the different
components of a mixture.
4. PAPER CHROMATOGRAPHY
Paper chromatography is an analytical method used to
separate colored chemicals or substances. It is now
primarily used as a teaching tool, having been replaced in
the laboratory by other chromatography methods such as
thin-layer chromatography (TLC).
The setup has three components. The mobile phase is a
solution that travels up the stationary phase by capillary
action. Here paper is used to support the stationary phase,
water.
Rƒ value:
The retention factor (Rƒ) may be defined as the ratio of
the distance travelled by the solute to the distance
travelled by the solvent. It is used in chromatography to
quantify the amount of retardation of a sample in a
stationary phase relative to a mobile phase. Rƒ values are
usually expressed as a fraction of two decimal places.
If Rƒ value of a solution is zero, the solute remains in the
stationary phase and thus it is immobile.
If Rƒ value = 1 then the solute has no affinity for the
stationary phase and travels with the solvent front.
A solvent in chromatography is the liquid the paper is
placed in, and the solute is the ink which is being
separated.
Rƒ value depends on:
• Absorbent
• Solvent
• The chromatography plate itself
• Application technique
• Temperature of the solvent and plate
So, several solvents offer several Rƒ values for the same
mixture of compound.
Types of paper chromatography:
I. Descending
Here, the mobile phase is placed in a solvent holder at the
top. The spot is kept at the top and solvent flows down the
paper from above.
II. Ascending
Here the solvent travels up the chromatographic paper.
Both descending and ascending paper chromatography are
used for the separation of organic and inorganic
substances. The sample and solvent move upward.
III. Ascending-descending
This is the hybrid of both of the above techniques. The
upper part of ascending chromatography can be folded
over a rod in order to allow the paper to become
descending after crossing the rod.
IV. Two-dimensional
In this technique a square or rectangular paper is used.
Here the sample is applied to one of the corners and
development is performed at a right angle to the direction
of the first run.
V. Circular chromatography
A circular filter paper is taken and the sample is
deposited at the center of the paper. After drying the
spot, the filter paper is tied horizontally on a Petri
dish containing solvent, so that the wick of the paper
is dipped in the solvent. The solvent rises through the
wick and the components are separated into
concentric rings.
CONCLUSION
Initially, chromatographic techniques were used to
separate substances based on their color as was the
case with herbal pigments. With time, its application
area was extended considerably. Nowadays,
chromatography is accepted as an extremely
sensitive, and effective separation method. Column
chromatography is one of the useful separation, and
determination methods. Column chromatography is
a protein purification method realized especially
based on one of the characteristic features of
proteins. These methods are also used to control
purity of a protein.
BIBLIOGRAPHY
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5206
469/
• https://byjus.com/chemistry/adsorption-
chromatography/
• https://www.vedantu.com/chemistry/partition-
chromatography
• https://testbook.com/chemistry/partition-
chromatography
• https://en.wikipedia.org/wiki/Gas_chromatography
• https://en.wikipedia.org/wiki/Paper_chromatography
