KENDRIYA VIDYALAYA AFS

WADSAR
Chemistry investigatory project

NAME:- KIRTI ANAND CLASS =12th

YEAR:-2023-2024 ROLL NO. =14

CERTIFICATE
This is to certify that
KIRTI ANAND, a student of
class 12 A has successfully completed his chemistry
investigatory project entitled CHROMATOGRAPHY
under the guidance of MR. DEEPAK PANDYA (PGT
CHEMISTRY) during the academic year 2023-24

MR. DEEPAK PANDYA

(Internal Examiner) (External Examiner)

PRINCIPLE
 ACKNOWLADGEMENT

The successful completion of any task would be

incomplete without mentioning the names of
those people who helped to make it possible. I
take this opportunity to acknowledge my deep
senses of gratitude to my chemistry teacher MR.
DEEPAK PANDYA for his valuable support,
constant help and guidance at every stage
without which this project would not have come
forth.

I also register my sense of gratitude to my

parents for their immense encouragement and
support. I would also like to thank my friends for
encouraging me during the course of this project.

Index
1.Certificate
2.Acknowledgement
3.Introduction
4.Objective
5.Apparatus Required
6.Theory
7.Procedure
8.Observation
9.Result
10.Precautions
11Bibliography

INTRODUCTION
Chromatography is a technique used to affect the
separation of two or more dissolved solids contained
within a solution in very small quantities.In chemical
laboratories, where it is used for analysis, isolation, and
purification.

Terms used in Chromatography

a) Analyte: is the substance that is to be separated
from the mixture during chromatography.

b) Mobile phase: in Chromatography is the

component that moves with the sample. It is either
a gas or a liquid and is passed through the column
where the components of the mixture are absorbed.
c) Stationary Phase: Stationary Phase in
Chromatography is the one that doesn’t move
the sample. It is generally a porous solid that
 Principles of Paper Chromatography
absorbs components from the mobile phase.
The principle involved can be
partition chromatography or
adsorption chromatography. Partition
chromatography because the substances are partitioned or
distributed between liquid phases. The two phases are
water held in pores of the filter paper and the other phase
is a mobile phase which passes through the paper. When
the mobile phase moves, the separation of the
mixture takes place. The compounds in the mixture
separate themselves based on the differences in their
affinity towards stationary and mobile phase solvents
under the capillary action of pores in the paper.
Adsorption chromatography between solid and liquid
phases, wherein the solid surface of the paper is the
stationary phase and the liquid phase is the mobile phase.

 Paper Chromatography Applications

 To study the process of fermentation and ripening.

 To check the purity of pharmaceuticals.

 To detect the adulterants.

 To detect the contaminants in drinks and foods.

 To examine the reaction mixtures in biochemical

laboratories.
 To determine dopes and drugs in humans and
animals.
Aim: Separate the colored components present in the
mixture of red and blue inks by ascending paper
chromatography and find their Rf values.
 Theory
The compounds in the mixture separate themselves
based on the differences in their affinity towards
stationary and mobile phase solvents under the capillary
action of pores in the paper.

MATERIALS REQUIRED:
1. Chromatographic chamber with lid
2. Measuring jar
3. Whatman filter paper
4. Distilled water
5. Mixture of red and blue ink
6. Pencil
7. Scale
PROCEDURE
8. Glass rod
# Take a Whatman filter paper strip and using a pencil
draw a horizontal line 4cm from one end of the paper.
Then draw another line lengthwise (vertically) from the
centre of the paper. Name the point at which the two
lines intersect as P.

# using a fine capillary tube put a drop of the mixture of

red and blue inks at the point P. Let it dry in air.

# Put another drop on the same spot and dry again, so

that the spot is rich in the mixture.

# Pour equal amounts of isopropyl alcohol and distilled

water into a chromatographic chamber and mix it well
using a glass rod. This is used as the solvent.

# Suspend the filter paper vertically in the

chromatographic chamber containing the solvent in such a
way that the pencil line remains about 2cm above the
solvent level.

# Close the jar with its lid and keep it undisturbed.

# Notice the rising solvent along with the red and blue
inks. After the solvent has risen about 15 cm you will
notice two different spots of blue and red colors on the
filter paper.
# Take the filter paper out of the jar and using a pencil
mark the distance that the solvent has risen on the paper.
This is called the solvent front.

# Dry the filter paper and put pencil marks at the centre
of the red and blue ink spots.

# Measure the distance of the two spots from the original

line and the distance of the solvent from the original line.

# Calculate the Rf values of the red and blue inks using

the formula,

PRECAUTIONS

 The spot should be small and rich in mixture.

 Allow the spot to dry before putting the strip in the jar.

 Keep the strip erect. Do not let it curl.

 Do not allow the spot to dip in the solvent .

1. What does chromatography mean ?

Chromatography is a technique used to affect the separation of

two or more dissolved solids contained within a solution

2. **Introduction and Objective:**

QUES - What is the main objective of your chromatography project?

ANS: Objective: To separate and analyze components in a mixture using chromatography

QUES- Can you briefly explain the importance of chromatography in analytical chemistry?

ANS: Importance: Chromatography is crucial in identifying and quantifying substances in a

sample.

2. **Types of Chromatography:**

QUES- Describe the different types of chromatography.

ANS: Types: There are various types, such as gas chromatography, liquid chromatography,
and paper chromatography.

- Why did you choose a specific type of chromatography for your project?

Choice: Chose [specific type] for its suitability in separating the target components.

3. **Experimental Setup:**

- Can you explain the components of your chromatography setup?

Components: Setup includes a stationary phase, mobile phase, sample application point, and a
detection system

- What considerations did you take into account while setting up the experiment?

Considered factors like solvent choice, column material, and detection method.

4. **Principles of Chromatography:**

- Explain the basic principles behind chromatography.

Chromatography relies on differential partitioning of components between stationary and

mobile phases.
 - How does the separation of components occur in chromatography?

Separation: Components with different interactions for the phases separate as they travel
through the medium.

5. **Stationary and Mobile Phases:**

- Describe the role of the stationary phase in chromatography.

Role of Stationary Phase: Binds with sample components selectively.

- What is the significance of the mobile phase in the separation process?

Significance of Mobile Phase: Carries sample components through the stationary phase.

6. **Sample Preparation:**

- How did you prepare the samples for analysis?

7. **Rf Values:**

- What is the meaning of Rf (retention factor) in chromatography?

Meaning of Rf: Ratio of distance traveled by the solute to the distance traveled by the solvent.

- How do you calculate the Rf value, and what does it indicate about the separation?

Calculation: Rf = (Distance traveled by solute) / (Distance traveled by solvent).

8. **Identification of Components:**

- How do you identify the different components in the chromatogram?

Identification: Components identified based on their unique Rf values or using standards.

9. **Results and Analysis:**

- How did you interpret the chromatogram, and what conclusions can be drawn from it?
Interpretation: Chromatogram interpreted by analyzing peak shapes, sizes, and positions.

10. **Applications and Significance:**

- Can you explain the real-world applications of chromatography?

Applications: Widely used in pharmaceuticals, environmental analysis, and forensics.

 To check the purity of pharmaceuticals.

 To detect the adulterants.

 To detect the contaminants in drinks and foods.

 To examine the reaction mixtures in biochemical

laboratories.
 To determine dopes and drugs in humans and
animals.
- How can the results of your project be applied or contribute to scientific knowledge?

Significance: Provides accurate qualitative and quantitative information about a mixture.