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Chromatography Principle and its Types

What is Chromatography?

Chromatography is the technique for the separation, puriQcation,

and testing of compounds.

The term “chromatography” is derived from Greek, chroma meaning, “colour,” and graphein
meaning “to write.”

In this process, we apply the mixture to be separated on a stationary phase (solid or liquid)
and a pure solvent such as water or any gas is allowed to move slowly over the stationary
phase, carrying the components separately as per their solubility in the pure solvent.

Table of Contents
Important Chemistry Topics
Principles of Chromatography
Periodic Table Of Elements
Types of Chromatography
Aufbau Principle
Related Topics on Chromatography
What is Differential Extraction? Electron ConQguration

Applications of Chromatography Isomerism

Frequently Asked Questions Planck's Quantum Theory

Valence Bond Theory

Principles of Chromatography
Reactivity Series
Chromatography is a separation method where the analyte is combined within a liquid or
gaseous mobile phase., which is pumped through a stationary phase. Usually one phase is Named Reactions
hydrophilic and the other is lipophilic. The components of the analyte interact differently
Dipole Moment
with these two phases. Depending on their polarity they spend more or less time
interacting with the stationary phase and are thus retarded to a greater or lesser extent. Thermal Conductivity
This leads to the separation of the different components present in the sample. Each
Mole Concept
sample component elutes from the stationary phase at a speciQc time called as retention
time. As the components pass through the detector their signal is recorded and plotted in Tyndall Effect
the form of a chromatogram.
Balancing Chemical Equations

Types of Chromatography Electromeric Effect

The four main types of chromatography are Electrochemical Cell

Boyle's Law
1. Adsorption Chromatography
In the process of adsorption chromatography, different compounds are adsorbed on the
adsorbent to different degrees based on the absorptivity of the component. Here also, a
mobile phase is made to move over a stationary phase, thus carrying the components with
higher absorptivity to a lower distance than that with lower absorptivity. The main types of
CBSE Sample Papers
chromatographic techniques that are used in industries are given as under.
CBSE Sample Papers Class 8
Science
ADSORPTIONCHROMATOGRAPHY BBYJU'S
CBSE Sample Papers Class 9
Science

CBSE Sample Papers Class 10

Solvent
Science
Mixturecontaining
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components:XandY
Physics

Silica/Alumina CBSE Sample Papers Class 11

saturatedwith
solvent
Chemistry

CBSE Sample Papers Class 11

Biology

Ymovesdownthe CBSE Sample Papers Class 12

Solventflows
columnmorequickly
outofcolumn
thanXandsoleaves
Physics
thecolumnfirst
CBSE Sample Papers Class 12
Chemistry
2. Thin Layer Chromatography
CBSE Sample Papers Class 12
In the process of thin-layer chromatography (TLC), the mixture of substances is separated Biology
into its components with the help of a glass plate coated with a very thin layer of
adsorbent, such as silica gel and alumina, as shown in the Qgure below.

The plate used for this process is known as chrome plate. The solution of the mixture to be
separated is applied as a small spot at a distance of 2 cm above one end of the plate. The
plate is then placed in a closed jar containing a _uid termed as an eluant, which then rises CBSE Previous Year Question
up the plate carrying different components of the mixture to different heights. Papers
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Class 10 Science

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ICSE Sample Papers Class 8

3. Column Chromatography Chemistry
Column chromatography is the technique used to separate the components of a mixture
ICSE Sample Papers Class 8 Biology
using a column of suitable adsorbent packed in a glass tube, as shown in the Qgure below.
The mixture is placed on the top of the column, and an appropriate eluant is made to _ow ICSE Sample Papers Class 9 Physics
down the column slowly.
ICSE Sample Papers Class 9
Depending upon the degree of adsorption of the components on the wall adsorbent Chemistry
column, the separation of the components takes place. The component with the highest
ICSE Sample Papers Class 9 Biology
absorptivity is retained at the top, while the other _ow down to different heights
accordingly. ICSE Sample Papers Class 10
Physics

ICSE Sample Papers Class 10

Chemistry

ICSE Sample Papers Class 10

Biology

ISC Sample Papers Class 11 Physics

ISC Sample Papers Class 11

Chemistry

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ISC Sample Papers Class 12

Chemistry

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Column Chromatography

4. Partition chromatography ICSE Previous Year Question

In this process, a continuous differential partitioning of components of a mixture into a Papers
stationary phase and mobile phase takes place. The example of partition chromatography ICSE Previous Year Question Papers
can be seen in paper chromatography. In this process, chromatography paper is used as a Class 10 Physics
stationary phase which is suspended in a mixture of solvents that act as a mobile phase.
ICSE Previous Year Question Papers
Here, we put a spot at the base of the chromatographic paper with the mixture to be Class 10 Chemistry
separated and as the solvent rises up this paper, the components are carried to different
degrees depending upon their retention on the paper. The components are thus separated ICSE Previous Year Question Papers
at different heights. Class 10 Maths

ISC Previous Year Question

Papers class 12
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Class 12 Physics

ISC Previous Year Question Papers

Class 12 Chemistry

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Class 12 Biology

Related Topics on Chromatography

Adsorption Chromatography
Thin Layer Chromatography
Column Chromatography
Partition Chromatography
Applications of Chromatography

What is Differential Extraction?

Differential extraction is the method of separation of any organic component present in an
aqueous solution. In this process, we use an organic solvent for which the solubility of the
desired compound is more than compared to that in water. Also, the organic solvent is
chosen such that it is immiscible with the aqueous solution so that it can form layers and
can be separated easily using a separating funnel.

The organic compound is later recovered by the process of distillation or evaporation. The
process of continuous extraction is used in cases when the solubility of the compound is
less in the organic solvent.

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Applications of Chromatography Submit

In bio analytical chemistry, chromatography is mainly used for the separation, isolation and
puriQcation of proteins from complex sample matrices. In cells for example, proteins occur
alongside numerous other compounds such as lipids and nucleic acids. In order to be
analysed, these proteins must be separated from all the other cell components. Then the
proteins of interest might have to be isolated from other proteins and puriQed further.

Chromatography is an essential part of almost any protein puriQcation strategy. A number

of different chromatographic techniques are used for the puriQcation and analysis of
proteins. They can be classiQed according to the physical principle involved in the
separation process. Typical examples include reversed phase chromatography, ion
exchange chromatography, abnity chromatography and size exclusion chromatography.

Frequently Asked Questions – FAQs

What is the basic principle of chromatography?
Chromatography is based on the concept of separating molecules in a mixture added to
the ground or solid and liquid stationary state (stable phase) when travelling with the aid of
a mobile phase.

What is the Rf value in chromatography?

RF stands for retention factor in paper chromatography, or the distance a _uid compound
moves up a plate of chromatography. For each particular solvent, all compounds have a
common RF value, and RF values are used to equate unknown samples with known
compounds.

How is RF value useful?

The distance travelled by sample divided by the distance travelled by the solvent. It is a
component characteristic and can be used to classify components for a given system at a
known temperature.

Where is chromatography used?

Chromatography is used in industrial processes to purify materials, test trace amounts of
contaminants, isolate chiral compounds and quality control test products.
Chromatography is the physical process of separating or analysing complex mixtures.

To learn more about differential extraction and chromatography and other separation
techniques, download BYJU’S The Learning App.

Test your Knowledge on Differential

extraction chromatography!

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3 Comments

Shravan FEBRUARY 2, 2020 AT 10:43 AM

It was wonderful time I spent

Excellent message

REPLY

Levi Armstrong JUNE 22, 2020 AT 7:49 AM

It’s interesting to know that chromatography is used in an industrial process to purify

materials and test trace amounts of contaminants to control the quality of the
products. My father is looking for a portable HLPC system for his work, and I was
curious about what it was for, which is why I’ve been researching about it. That’s why
I really appreciate this article because I learned a lot about chromatography. Thanks!

REPLY

E.LEELAVATHY MAY 2, 2022 AT 11:47 AM

VERY USEFUL TOPIC IN PHARMA AND OTHER INDUSTRY

REPLY

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