5/20/24, 11:29 PM Chromatography: Introduction, Principle, Classification and Applications

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CHROMATOGRAPHY: INTRODUCTION, PRINCIPLE, CLASSIFICATION AND

APPLICATIONS
November 23, 2020 by Adarsh Pandey

CHROMATOGRAPHY: INTRODUCTION, PRINCIPLE, CLASSIFICATION

AND APPLICATIONS

CONTENTS

Introduction
Principle
Classifications
Applications

INTRODUCTION

Chroma– “color” and graphein – “to write”.

Chromatography is a technique in which substance are separated, purified and identified from a
mixture for qualitative and quantitative analysis. On the basis of hydrophobic interactions, Polarity,
enzymes and net charges are separated by using chromatography.
Chromatography is a physical method of separation of compound.
TSWET, referred as the father of chromatography.
This technique was first used by Tswet, to separate various plants pigments such as chlorophylls and
xanthophylls by passing these pigments through glass column packed with finely divided calcium
carbonate.

PURPOSE OF CHROMATOGRAPHY

Analytical: To determine chemical composition of a sample.

Preparative: Used in purification of a substance.

PRINCIPLE OF CHROMATOGRAPHY

The principle of chromatography is very simple.

The sample to be examine or analyzing is termed as solute or analyte is allowed to interact or react
with two immiscible phases-
I. Mobile phase
II. Stationary phase

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5/20/24, 11:29 PM Chromatography: Introduction, Principle, Classification and Applications

Image source khan academy

The stationary phase which may be a solid is or a liquid supported on a solid, which is immobile it
means it does not move.
The mobile phase migrates the sample through stationary phase.
The mobile phase may be either liquid or gas.
All chromatographic methods involve separation of mobile phase through a stationary phase.

CLASSIFICATIONS OF CHROMATOGRAPHY

Chromatography technique can be explained into three fundamental ways:

I. Based on the shape of chromatographic bed.
II. Based on the physical nature of the stationary and mobile phases.
III. Based on the mechanism of the separation.

Based on the shape of chromatographic bed:

On the basis of the chromatographic bed, there are following two types of chromatography-
I. Planar chromatography.
II. Column chromatography.

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5/20/24, 11:29 PM Chromatography: Introduction, Principle, Classification and Applications

In planar chromatography, the stationary phase is spread on a flat, planar surfaces. The plane can
be paper acting as a stationary phase (paper chromatography), or stationary phase spread on glass,
metal or plastic plate (thin layer chromatography). Planar chromatography also known as open-bed
chromatography.
In column chromatography, the stationary phase is within a tube.

Image Source Khan Academy

Based on the physical nature of the mobile and stationary phase:

On the basis of physical nature of mobile phase, there are two types of chromatography:
I. Gas chromatography
II. Liquid chromatography
Further, on the basis of stationary phase, physical nature there are the following types:
I. Gas-solid chromatography
II. Gas-liquid chromatography
III. Liquid-solid chromatography
IV. Liquid-liquid chromatography

Based on the mechanism of separation:

The method of chromatography uses various types of mechanisms to separate analyte.

On the basis of mechanism of separation, it is of following types:
I. Partition chromatography
II. Adsorption chromatography
III. Size exclusion chromatography
IV. Affinity chromatography
V. Ion exchange chromatography

APPLICATION OF CHROMATOGRAPHY

In the pharmaceutical sector:

In detection of presence of trace elements or chemicals.

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5/20/24, 11:29 PM Chromatography: Introduction, Principle, Classification and Applications

This is also used for the separation of compound on the basis molecular weight and composition of
elements.
In the detection of unknown compound and in the purity of mixtures.
In the development of drug.

In the Chemical industry:

HPLC and GC are very much used for detecting various contaminants such as polychlorinated
biphenyl (PCBs) in pesticides and oils.

In the Food Industry:

In the detection of food spoilage and additives.

In the quantitative and qualitative analysis of nutrients in food.

In Forensic Science:

In forensic science, this technique is used in the analysis of crime scenes like hair and blood
samples.

In Molecular Biology Studies:

Various techniques in chromatography such as EC-LC-MS are involved in the study of metabolomics
and proteomics along with nucleic acid research.
HPLC is used in Protein Separation like Insulin Purification, Plasma Fractionation, and Enzyme
Purification and also used in the various industries like Fuel Industry, biochemical processes and
biotechnology.

Chromatography like HPLC is used in DNA fingerprinting and bioinformatics:

Assay & Content Uniformity.

HPLC in fingerprinting and Bioinformatics.
Petrochemicals and Catalysis.
Ebola Immunization.
Polymer Synthesis.
Clinical diagnosis of diseases and disorder.
LC-NMR (liquid chromatography (LC) and nuclear magnetic resonance).

Reference And Sources:

https://microbenotes.com/chromatography-principle-types-and-applications/
https://chromatography.conferenceseries.com/events-list/applications-of-chromatography
https://www.researchgate.net/publication/310994699_Fundamentals_and_Techniques_of_Biophysics
_and_Molecular_Biology

Also Read:

Classification of viruses on the basis of genome

Different Types of chromatography
Bacterial Growth Curve: Definition, Phases and Measurement
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5/20/24, 11:29 PM Chromatography: Introduction, Principle, Classification and Applications

Amino acids: Structure, Optical activity and Classifications

Gel electrophoresis: types, principles, instrumentation and applications

CHROMATOGRAPHY: INTRODUCTION, PRINCIPLE, CLASSIFICATION

AND APPLICATIONS

2 Thoughts On “Chromatography: Introduction, Principle, Classification And Applications”

tems
September 27, 2022 at 8:19 am

clearly explained, definitely easier than how school explains it. so helpful.

Reply

Saman
February 18, 2023 at 5:34 pm

It’s very much good web for study

Reply

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