REVIEW ON BOOK ENTITLED

CHROMATOGRAPHY
AUTHORED BY RAJBIR SINGH

Submitted to
GUJARAT UNIVERSITY
For the degree of

MASTER OF SCIENCE
In

ORGANIC CHEMISTRY
By

GADHIYA AKSHAYKUMAR KANTIBHAI

University. No. ______, Roll No.: 07 HPP

Under the supervision of

Dr.Deepen S Gandhi

DEPARTMENT OF CHEMISTRY
GOVERNMENT SCIENCE COLLEGE GANDHINAGAR
GUJARAT UNIVERSITY
APRIL 2023

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PHYSICAL APPEARANCE OF THE BOOK

A book booth is very colorful and attractive. Its hard paper Cover and also
carrying the some graphics. It's typically composed of many pages bound
and protected by a cover.

BOOK SIZE : A5 size

BOOK DETAILS
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 First Edition 2002

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  Book of Cost :
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ABOUT THE OTHER

Rajbir Singh got his M.Sc. in physic from Meerut University, Meerut
and B.Ed. from Rajasthan University, Jaipur. He has long experience in
teaching science and research. He has to his credit number of book and
articles published in reputed journals. He is a present teaching Physics to
Post-graduate student at Central School for Tibetans in Mussoorie in
Uttaranchal. He is presently working on a gigantic project focusing on
modern organic/inorganic Chemistry series.

PREFACE

In this book a sincere efforts has been made to include the various types of
chromatographic techniques. The present book address itself-sufficient. Every
concept has been demonstrated by simple diagrams using simple mathematics
and elegant style. All explanation have well attended reasons and are
supplemented by appropriate data to bring home the point.

In many techniques more difficult concepts which may not of immediate need
to the student but would definitely add to his/her knowledge have also been
provided.

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SUBJECTIVE CONTENT OF THE BOOK

 CHAPTER-1. Chromatography

The first chapter covered by various types of Chromatography. Chromatography

is a technique used in analytical chemistry to separate and identify components of a
mixture. There are various types of chromatography such as adsorption Chromatography,
paper chromatography, thin-layer chromatography, counterfeiting chromatography, liquid-
liquid partition chromatography, column chromatography affinity chromatography, gel
chromatography, ion exchange chromatography, high performance liquid chromatography
and gas chromatography. Chromatography works based on the principles of adsorption,
partition, or distribution of the components of a mixture between a stationary phase and a
mobile phase. The choice of stationary and mobile phases, and the conditions of
separation, depend on the nature of the mixture and the components to be separated.
Chromatography is a powerful and versatile tool for separating, identifying and quantifying
the components of the mixture.

In chromatography, Rf  values are the most basic prerequisite of the experiment. Rf values
are used to determine polarity, relative masses, and relative solubilities, among other
things. The Rf (retardation factor) value is the ratio of the solute distance travelled to the
solvent is distance travelled.

One of the main benefits of chromatography is its versatility, as it can be used to separate
a wide range of compounds, including small molecules, proteins and nucleic acids.

 CHAPTER-2. Adsorption Chromatography

This chapter covered by Adsorption Chromatography. Adsorption chromatography is a

separation technique that is based on the interaction between a solid stationary phase and a
liquid or gaseous mobile phase. Adsorption chromatography is a type of separation
technique that uses adsorption, a process by which molecules adhere to the surface of a
solid or liquid, to separate mixtures of compounds. In adsorption chromatography, the

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mixture is passed through a column filled with an adsorbent material, such as activated
carbon or silica gel, that binds certain components of the mixture more strongly than
others. The adsorbed components then move through the column at different rates,
allowing them to be separated based on their adsorption characteristics.

Adsorption Chromatography follows that Langmuir adsorption isotherm and freundlich

adsorption isotherm.

 CHAPTER-3. Paper & Thin-layer Chromatography

Paper chromatography and Thin layer chromatography are two important techniques used
in the separation of mixtures of substances. Both techniques are used to separate the
components of a mixture based on their properties, such as solubility and polarity.

Paper chromatography involves the separation of components on a Whatman filter paper

that has been treated with a solvent. The mixture is applied to the Whatman filter paper,
and as the solvent moves up the paper, the components separate based on their solubility
and polarity. The components can then be visualized by staining or exposing the paper to a
light source.

Thin layer chromatography (TLC) is similar to paper chromatography, but it uses a solid
support, such as a glass or plastic plate coated with a thin layer of silica gel. The mixture is
applied to the plate, and as the solvent moves up the plate, the components separate based
on their solubility and polarity. The components can then be visualized by staining or
exposing the plate to a light source.

Paper chromatography and TLC (Thin-Layer Chromatography) are valuable tools in

analytical chemistry, and they continue to be widely used in various fields due to their
simplicity and cost-effectiveness.

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  CHAPTER-4. Countercurrent Chromatography

This chapter covered by Countercurrent Chromatography. Countercurrent chromatography

(CCC) is a separation technique that uses two immiscible liquid phases to separate and
purify a sample. This method differs from other chromatography techniques, such as
column chromatography, in that the sample and the mobile phase flow in opposite
directions, creating a countercurrent flow. This flow creates a centrifugal force that helps
to separate and purify the sample.

Countercurrent Chromatography has become a popular option for the purification of

natural products and medicinal compounds. It can also be combined with other separation
techniques, such as liquid chromatography, to enhance the separation efficiency

 CHAPTER-5. Liquid-liquid Partition Chromatography

This chapter covered by liquid-liquid Partition Chromatography. The basic principle of this
technique is the distribution of solutes between two immiscible liquid phases, where the
stationary phase is a liquid coating on a solid support and the mobile phase is another
liquid.

In this technique, the sample mixture is dissolved in a suitable solvent and then passed
through a column packed with an immiscible liquid phase. The immiscible liquid phase is
usually an organic solvent. As the sample mixture is passed through the column, the
different compounds in the mixture partition between the two liquid phases, with some
compounds preferentially partitioning into the stationary phase and others into the mobile
phase.

The most common stationary phases used in LLPC are water, while the most commonly
used mobile phases organic solvents. It is a simple and efficient method that can be used
for the separation of a wide range of biological compound.

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  CHAPTER-6. Column & Affinity Chromatography

This chapter covered by Column Chromatography & Affinity Chromatography.

Column chromatography is a separation technique that involves passing a mixture of

components through a stationary phase, such as a column packed with a solid material, and
then collecting the components as they emerge from the column. The separation is based
on differences in the interactions between the components and the stationary phase.
Column chromatography is widely used for purification and fractionation of proteins,
enzymes, antibodies, DNA, RNA, and other biomolecules.

Affinity chromatography is a type of column chromatography that separates molecules

based on their specific biochemical interactions. It makes use of the specific binding
between a ligand, which is covalently attached to the solid support, and its target molecule.
The target molecules bind to the ligand, while other components of the mixture pass
through the column, resulting in the selective purification of the target molecule. Affinity
is widely used for purification and isolation of specific proteins, biomolecules, protein-
DNA interactions, and other biological interactions.

 CHAPTER-7. Gel Chromatography

This chapter covered by Gel Chromatography. Gel chromatography, also known as

size-exclusion chromatography, is a type of chromatography used to separate
molecules based on their size. In this method, a sample is passed through a column
filled with a gel matrix of specific pore sizes, which allows smaller molecules to enter
the pores and travel through the column more slowly than larger molecules that
cannot enter the pores. As a result, the larger molecules are eluted first, followed by
progressively smaller ones. Gel chromatography is a widely used technique in
biochemistry and molecular biology for purifying proteins, nucleic acids, and other
biomolecules, and can be used for both analytical and preparative purposes.

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  CHAPTER-8. Ion Exchange Chromatography &
Electrophoretic Techniques

This chapter covered by Ion Exchange chromatography and Electrophoretic Techniques.

Ion exchange chromatography is a technique used to separate and purify molecules based
on their net charge. It works by using a column filled with beads containing charged
functional groups, such as positively charged or negatively charged functional groups. It is
also known as cation-anion exchange chromatography. The sample mixture is loaded onto
the column, and the charged molecules in the mixture interact with the functional groups
on the beads, causing them to be temporarily trapped in the column. The non-charged
molecules pass through the column and are collected separately. There are two types of ion
exchanger. 1. Cationic exchangers, 2. Anionic exchangers.

Electrophoretic techniques are based on the principle that charged particles, such as
proteins or nucleic acids, can be separated based on their size, charge, and shape under the
influence of an electric field. There are various type of Electrophoresis such as Low
Voltage Thin Sheet Electrophoresis(TLE), High voltage Electrophoresis(HVE).
Electrophoretic technique that is used to separate biological molecules such as amino
acids, peptides, nucleotides, proteins and nucleic acid.

 CHAPTER-9. High Performance Liquid

Chromatography & Gas Chromatography

This chapter covered by High Performance Liquid Chromatography and Gas

Chromatography. High performance liquid chromatography (HPLC) and gas
chromatography (GC) are both analytical techniques used to separate, identify, and
quantify the components in a mixture.

HPLC is a liquid chromatography technique that uses a high-pressure pump to force the
sample through a stationary phase, where the components are separated based on their
chemical and physical properties. The stationary phase is typically a column packed with a

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material that interacts with the analytes of interest, while the mobile phase is a liquid that
carries the sample through the column. HPLC is particularly useful for the separation of
polar compound such as drugs and their metabolites, peptides, vitamins, polyphenols and
steroid.

Gas chromatography (GC) is a technique used to separate and analyze the components of a
mixture based on their different distribution between a stationary phase (typically a high
boiling point liquid) and a mobile phase (a carrier gas). The mixture is injected into the
instrument and carried by the carrier gas through a long, thin column that contains the
stationary phase. As the mixture passes through the column, different components interact
differently with the stationary phase, causing them to travel at different rates and separate
from each other. The separated components are then detected and quantified by a detector
at the end of the column. GC is a two types of chromatography. 1. Gas Solid
Chromatography, 2.Gas Liquid Chromatography. GC is a powerful and versatile analytical
technique that has many applications in fields ranging from environmental science to
forensic analysis.

Merits of book

This book Is very useful for the graduate and post graduate students .This book covered all
Chromatography’s techniques and his basic information. This book is a comprehensive
guide to chromatography and provides a clear explanation of the principles,
instrumentation, and applications of various types of chromatography. Every concept has
been demonstrated by simple diagrams using simple mathematics and elegant style. The
order of the techniques included in this book is conventional.

Demerits of book

This book provides short information about the some chromatography. The heading size
of particular topic is not properly set it must be in bold and large in size so reader can
easily recognize.

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