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Adsorptiona Partition Chromatography

The document discusses adsorption and partition chromatography. [1] Adsorption chromatography uses a stationary solid phase that adsorbs compounds from a liquid or gas mobile phase at different rates. [2] Partition chromatography separates compounds based on how they partition between two immiscible liquid phases. [3] The key difference is that adsorption chromatography relies on interaction with a solid adsorbent, while partition chromatography uses a liquid-liquid system.

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745 views

Adsorptiona Partition Chromatography

The document discusses adsorption and partition chromatography. [1] Adsorption chromatography uses a stationary solid phase that adsorbs compounds from a liquid or gas mobile phase at different rates. [2] Partition chromatography separates compounds based on how they partition between two immiscible liquid phases. [3] The key difference is that adsorption chromatography relies on interaction with a solid adsorbent, while partition chromatography uses a liquid-liquid system.

Uploaded by

SATYA SRIVASTAVA
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Amity Institute of Pharmacy

Adsorption & Partition


chromatography
Amity Institute of Pharmacy
36
Adsorption chromatography is the oldest types of chromatography technique. It

makes use of a mobile phase which is either in liquid or gaseous form. The mobile
phase is adsorbed onto the surface of a stationary solid phase.
Amity Institute of Pharmacy

Principle
Adsorption Chromatography involves the analytical separation of a chemical
mixture based on the interaction of the adsorbate with the adsorbent. The mixture
of gas or liquid gets separated when it passes over the adsorbent bed that adsorbs
different compounds at different rates.
Adsorbent – A substance which is generally porous in nature with a high surface
area to adsorb substances on its surface by intermolecular forces is called
adsorbent.
Some commonly used adsorbents are Silica gel H, silica gel G, silica gel N, silica gel
S, hydrated gel silica, cellulose microcrystalline, alumina, modified silica gel, etc.
Amity Institute of Pharmacy

Rf value?
The Rf value is defined as the ratio of the distance moved by the solvent
(i.e. the coloring or pigment being tested) and the distance moved by the
solvent (known as the Solvent front) along the paper, where both
distances are measured from the common origin or application baseline.
Amity Institute of Pharmacy

Stationary phase – Adsorbent is the stationary phase in adsorption


chromatography. The forces involved help to remove solutes from the adsorbent so
that they can move with the mobile phase.

Mobile phase – Either a liquid or a gas is used as a mobile phase in adsorption


chromatography. Forces involved help to remove solutes from the adsorbent so
that they can move with the mobile phase. When a liquid is used as a mobile phase
it is called LSC (Liquid-Solid Chromatography). When a gas is used as a mobile
phase it is called GSC (Gas-Solid Chromatography).
Amity Institute of Pharmacy

ADSORBENTS
An adsorbent is a substance, usually porous in nature and with a high surface
area that can adsorb substances onto its surface by intermolecular forces.

AN IDEAL ADSORBENT SHOULD HAVE FOLLOWING PROPERTIES-

The Ideal adsorbent must fulfil the following requirements:


1) Insoluble in mobile phase
2) Inert to solutes (adsorptive)
3) Colourless especially when work with coloured mixtures
4) Suitable particle size enough to give good separation and reasonable flow
rate
Amity Institute of Pharmacy

THE COMMON ADSORBENTS/ STATIONARY PHASE-

• Hydrated silica gel • Modified silica gel


• Silica gel G • Alumina
• Silica gel S • Kieselghur (Diatomaceous earth)
• Silica gel H • Cellulose MN 300
• Silica gel N • Cellulose microcrystalline
• Silica gel GF 254
• Silica gel HF 254
• Silica gel PF 254
Amity Institute of Pharmacy

Mobile phase:
Mobile phase may be a single liquid or a mixture of liquids and commonly used
mobile phases are-
1) Methanol
2) Ethanol
3) Ethyl acetate
4) Diethyl ether
5) Acetone
6) Chloroform
7) Hexane
Amity Institute of Pharmacy

Identification of location of separated components or spots-


1) Physical detection: we use UV light with certain wavelength.•
2) Chemical detection: we use a chemical reagent either by spraying or dipping in both
methods the chemical reagent will react with separated compound and give a
color.
3) Biological detection: by using certain microorganisms to detect the separated
compounds.
4) Radioactive detection: if the compound being separated have radioactivity, such
compound can be detected by using special instrument.
Amity Institute of Pharmacy

Applications involves separation and identification of;


1) Amino acids
2) Peptides and proteins
3) Alkaloids
4) Carbohydrates
5) Fats and fatty acids
6) Antibiotics
7) Narcotic analgesics
8) Glycosides
Amity Institute of Pharmacy

Partition chromatography
Amity Institute of Pharmacy

Principle-
The separation of the components from the sample mixture is carried out by the
process of partition of the components between 2 phases. Both phases are in
liquid form. In this process, the immiscible solid surface coated with the liquid
surface on the stationary phase is in the mobile phase.
The liquid surface is immobilized by a stationary phase which results in making its
a stationary phase. The mobile phase moves from the stationary phase and
components get separated. The separation depends on different partition
coefficient.
Amity Institute of Pharmacy

Types of Partition Chromatography


1.Liquid-liquid Chromatography – It is a chromatography technique where a sheet of
blotting paper, is used instead of adsorption column. The components are separated
based on their differential migratory velocities. On separating they are stained to make
the chromatogram visible.
2.Gas-liquid Chromatography – A chromatography technique in which the separation
of the mixture is done by an inert gas along a tube. The tube is filled with finely divided
inert solid. The solid is coated with a nonvolatile solvent. The migration of each
component occurs at a rate determined by its solubility in solvent as well as its vapor
pressure.
Amity Institute of Pharmacy

Applications of Partition Chromatography


1) To separate and detect color mixtures including pigments
2) To separate and determine proteins, lipids, alkaloids, glycosides, and other
biomolecules
3) To isolate and detect amino acids and carbohydrates
4) To examine the purity of pharmaceuticals
5) In DNA/RNA sequencing
6) To determine pollutants in cosmetics, food products and beverages
7) To isolate polar and non-polar compounds
Amity Institute of Pharmacy
Difference Between Adsorption and Partition
Chromatography Amity Institute of Pharmacy
Amity Institute of Pharmacy

Adsorption chromatography-
The analyte differently interacts with the stationary phase because every analyte has
a different affinity towards the adsorbent. Therefore, the separation of molecules
occurs at different retention times.
Advantages-
1) It has a wide range of mobile phases for the separation of compounds.
2) Adsorption chromatography is an important method to separate many components
that are not separated by other techniques.
3) The complex sample mixtures can be easily separated by this method.
4) Very few types of apparatus or types of equipment are required for isolation.
Amity Institute of Pharmacy

Disadvantages-
1) Obtained results from some methods of adsorption chromatography are
complex to reproduce.
2) This can cause catalytic variations.
3) Some solutes take retention time to separate this is a major disadvantage of
adsorption chromatography.
4) It is more complicated and costly when make automation.
Amity Institute of Pharmacy

The advantages of partition chromatography-


1) The advantages of partition chromatography consist of an easy-to-operate
and economical method for separation.
2) The partition chromatography technique can isolate both organic and
inorganic compounds.
3) It saves time because in a short duration it separates compounds.
4) It provides high efficiency.
5) This technique provides accurate results.
6) Selectivity is better in partition chromatography because a mobile phase can
be simply changed.
Amity Institute of Pharmacy

The disadvantages of partition chromatography-

1) The high volume of the mobile phase is sometimes required for separation
2) Data cannot be stored long in certain types of partition chromatography.
3) Automation has made it more difficult and costly.
4) In gas-liquid chromatography, only volatile compounds can be separated.
5) It is possible to separate only soluble analytes of sample mixtures.

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