Chromatography:

Chromatography is a laboratory technique used for the separation, identification, and

quantification of components in a mixture. It involves passing the mixture dissolved in a "mobile
phase" through a "stationary phase," leading to the separation of the mixture's components
based on their differing interactions with these two phases.

Principle:
The principle of chromatography relies on the differential partitioning between the mobile phase
and the stationary phase. Here's how it works:

Stationary Phase: This is the phase that does not move. It can be a solid or a liquid supported
on a solid surface.
Mobile Phase: This is the phase that moves through the stationary phase, carrying the
components of the mixture with it. It can be a liquid (in liquid chromatography) or a gas (in gas
chromatography).

Paper chromatography:
Paper chromatography is a simple and cost-effective analytical technique used to separate and
identify mixtures of substances. It is a type of partition chromatography where the stationary
phase is a sheet of paper, typically filter paper, and the mobile phase is a liquid solvent.

Principle of Paper Chromatography

The principle of paper chromatography is based on the differential partitioning of compounds
between the stationary phase (paper) and the mobile phase (solvent). Here's how it works:

Stationary Phase: The paper acts as the stationary phase. It contains water molecules that are
adsorbed onto the cellulose fibers, effectively making it a solid-liquid chromatography system.
Mobile Phase: A solvent or mixture of solvents that moves through the paper by capillary
action, carrying the components of the mixture with it.

paper chromatography

Column chromatography:
Column chromatography is a widely used technique in laboratories for purifying individual
chemical compounds from mixtures

Principle of Column Chromatography

The principle of column chromatography relies on the differential partitioning between the
stationary phase and the mobile phase. Components of the mixture travel through the column at
different rates due to their varying interactions with the stationary phase, leading to separation.

Key Components
Stationary Phase: Usually a solid adsorbent material such as silica gel or alumina packed
inside a vertical glass or plastic column.
Mobile Phase: A liquid solvent or a mixture of solvents that moves through the column by
gravity or pressure carrying the sample in it.

column chromatography

TLC:
Thin-Layer Chromatography (TLC) is a simple, rapid, and cost-effective analytical technique
used to separate, identify, and quantify components in a mixture. It involves a stationary phase,
typically a thin layer of silica gel or alumina coated onto a glass, plastic, or aluminum plate, and
a mobile phase, which is a solvent or mixture of solvents.

Principle of TLC
The principle of thin-layer chromatography is based on the differential adsorption of compounds
onto the stationary phase and their solubility in the mobile phase. As the mobile phase moves
up the plate by capillary action, different components of the mixture travel at different rates and
thus separate from one another.

Stationary Phase: A thin layer of adsorbent material (such as silica gel or alumina) coated onto
a flat, inert support (such as glass, plastic, or aluminum).

Mobile Phase: A solvent or solvent mixture that moves by capillary action through the
adsorbent layer, carrying the sample with it.

TLC

GLC:
Gas-Liquid Chromatography (GLC), often simply referred to as Gas Chromatography (GC), is
a powerful analytical technique used to separate, identify, and quantify volatile compounds in a
mixture. It involves the use of a gas as the mobile phase and a liquid stationary phase coated
onto a solid support.

Principle of GLC
The principle of gas-liquid chromatography is based on the differential partitioning of compounds
between the mobile phase (carrier gas) and the stationary phase (liquid coated on a solid
support). As the mixture is carried through the column by the gas, different components interact
with the stationary phase to varying degrees, causing them to separate as they travel through
the column.

Stationary Phase: A liquid phase coated onto the surface of an inert solid support (e.g.,
diatomaceous earth or silica gel) contained within a column.
Mobile Phase: An inert gas (typically helium or nitrogen) that carries the sample through the
column.

GLC

HPLC:
High-Performance Liquid Chromatography (HPLC) is an advanced form of liquid
chromatography used to separate, identify, and quantify components in complex mixtures. It
operates under high pressure to push the mobile phase through a column packed with the
stationary phase, enabling faster and more efficient separations compared to traditional liquid
chromatography.

Principle of HPLC
The principle of HPLC is based on the differential partitioning of compounds between the mobile
phase (a liquid solvent) and the stationary phase (a solid or liquid supported on a solid).
Components of the mixture interact differently with these phases, leading to their separation as
they travel through the column.

Stationary Phase: Typically consists of small porous particles packed into a column. The
particles are usually coated with a stationary phase material that interacts with analytes based
on various properties such as polarity, size, or charge.

Mobile Phase: A solvent or solvent mixture that carries the sample through the column. The
composition of the mobile phase can be adjusted to optimize separation based on the analytes'
properties.

HPLC

Ionexchange chromatograph:

Ion Exchange Chromatography is a powerful technique used to separate ions and polar
molecules based on their affinity to ion exchangers. It is widely used in biochemistry and
analytical chemistry for purifying proteins, peptides, nucleic acids, and other charged
biomolecules.

Principle of Ion Exchange Chromatography

The principle of ion exchange chromatography is based on the reversible interaction between
charged molecules in the sample and oppositely charged groups on the stationary phase (ion
exchange resin). Molecules are separated based on their charge and their affinity for the ion
exchange resin.

Stationary Phase: Consists of a solid support with charged groups (either positively or
negatively charged), known as ion exchangers.
Mobile Phase: Includes a buffer solution and the sample containing the mixture of molecules.

Ion exchange chromatography .

Molecular sieve chromatography:

Molecular sieve chromatography is a technique used for separating molecules based on their
size and shape using a porous stationary phase. It's commonly used in biochemistry and
analytical chemistry to separate proteins and other large biomolecules.

The principle of molecular sieve chromatography relies on the differential exclusion of

molecules based on their size and shape. Larger molecules are excluded from entering the
pores of the stationary phase beads and therefore travel more quickly through the column, while
smaller molecules enter the pores and take longer to elute. This separation is based solely on
size, making it a valuable tool for fractionating biomolecules without altering their chemical
structure.

Stationary Phase: Consists of porous beads with uniform pore sizes.

Mobile Phase: Typically a buffer solution that maintains optimal conditions for separation.

molecular sieve chromatography

Affinity chromatography:
Affinity chromatography is a powerful technique used for the selective separation and
purification of biomolecules based on their specific interactions with ligands immobilized on a
stationary phase.

Principle:
Principle: The principle of affinity chromatography is based on the specific interactions between
a target molecule (analyte) and a ligand immobilized on a solid support (stationary phase). The
ligand is chosen based on its ability to bind specifically to the target molecule. When the sample
containing the target molecule is applied to the column, it selectively binds to the ligand while
other molecules pass through or are washed away. After the non-specifically bound molecules
are removed, the target molecule is eluted under conditions that disrupt the binding interaction.

Stationary Phase: Consists of a solid support with an immobilized ligand that selectively binds
the target molecule.

Mobile Phase: Includes a buffer solution and the sample containing the mixture of molecules.

Affinity chromatography