Hubble Asteroid Hunter III. Physical properties of newly found asteroids
Gel Filtration or Permeation Chromatography
1. GEL
CHROMATOGRAPHY
Mrs. Poonam Sunil Aher (M.Pharm, PhD)
Assistant Professor
Sanjivani College of Pharmaceutical Education and Research
(Autonomous),
Kopargaon, Ahmednagar-423603 (M.S.), INDIA
Email:poonamprakashpatil4174@gmail.com
2. Gel Chromatography, also called Gel Filtration,
in analytical chemistry, technique for separating chemical
substances by exploiting the differences in the rates at which they
pass through a bed of a porous, semisolid substance.
The method is especially useful for separating enzymes, proteins,
peptides, and amino acids from each other and from substances of
low molecular weight.
3. The separation of the components of a mixture by
gel chromatography is based on the differences in the molecular
sizes of the components. Small molecules tend to diffuse into the
interior of the porous particles so that their flow is restricted,
while large molecules are unable to enter the pores and tend to
flow unhindered. Thus, the components of highest molecular
weight leave the bed first, followed by successively smaller
molecules. The bed materials most extensively used are
polyacrylamide and a polymer prepared from dextran and
epichlorohydrin. The dry polymers are usually suspended in
suitable agents to form a homogeneous, semisolid mixture
4. • Principle:
• The gel filtration chromatography is based on the
molecular size and the hydrodynamic volume of the
components. The molecules are separated by the
differential exclusion or inclusion of solutes as they
pass through the stationary phase containing
heterosporous cross-linked polymeric gel or beads
5. APPLICATION
• Gel filtration chromatography can be used to
separate compounds such as small molecules,
proteins, protein complexes, polysaccharides, and
nucleic acids when in aqueous solution. When an
organic solvent is used as the mobile phase, the
process is instead referred to as gel permeation
chromatography.
6. TYPES
1.Gel Permeation Chromatography.
Gel permeation chromatography (GPC) or size-
exclusion chromatography (SEC) is a favored
technique for the analysis of polymers and
oligomeric compounds.
7. • Gel filtration is also known as size-exclusion
chromatography or molecular-sieve
chromatography. In this process, separation is
based on the differing ability (due to differing
molecular size) of molecules in the sample to enter
the pores of the gel-filtration medium.
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10. COLUMNS
• GPC columns are typically packed with porous
particles composed of polystyrene, poly(styrene-co-
divinylbenzene), silica, or cross-linked poly(methyl
methacrylate) of various diameter and pore sizes
• The pores of the packing material are sampled by
the analyte as it travels through the column
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13. • Gel Filtration Chromatography Applications
• Gel filtration chromatography, a type of size exclusion
chromatography, can be used to either fractionate
molecules and complexes in a sample into fractions
with a particular size range, to remove all molecules
larger than a particular size from the sample, or a
combination of both operations.
• Gel filtration chromatography can be used to separate
compounds such as small molecules, proteins, protein
complexes, polysaccharides, and nucleic acids when in
aqueous solution.
• When an organic solvent is used as the mobile phase,
the process is instead referred to as gel permeation
14. • Fractionation of molecules and complexes within a
predetermined size range
• Size analysis and determination
• Removal of large proteins and complexes
• Buffer exchange
• Desalting
• Removal of small molecules such as nucleotides,
primers, dyes, and contaminants
• Assessment of sample purity
• Separation of bound from unbound radioisotopes
