Q:
|
What
is the difference between GPC, GFC, SEC and HPLC?
|
A:
|
Size
exclusion chromatography (SEC) is a chromatographic method in
which particles
are separated based on their size, or in more technical terms, their hydrodynamic volume.
It is usually
applied to large molecules or macromolecular
complexes such as
proteins and industrial polymers. Both molecular weight and three
dimensional shape
contribute to the degree of retention. (view the principle
as portrayed by General
Electric Corporation)
When an aqueous solution is used to
transport the sample through the column the technique is known as gel
filtration chromatography.
The name gel permeation
chromatography is used when an organic solvent is used as a
mobile
phase. GPC must operate free
of interactions to assure separation by size only. Only the entropy
effects should influence the separation. Normal HPLC methods
rely
on
interactions between sample and stationary phase;those interactions
result from ion exchange, bio-affinity, or chirality.
Gel
Permeation
Chromatography (GPC) is one of the most versatile and
powerful analytical techniques available for understanding and
predicting polymer performance. GPC is well established for
determining the molar mass of polymers. Compared to other
methods
of analysis such as osmometry or static light scattering, it has the
advantage that it determines complete distributions of molar masses as
opposed to merely an average molecular weight.
GPC separates the molecules in solution by their "effective size" or
Hydrodynamic volume. The sample is dissolved in an
appropriate
solvent and injected into a continually flowing stream of solvent
(mobile phase). The mobile phase flows through millions of
highly
porous, rigid particles (stationary phase) tightly packed together into
a column. As the mobile
phase flows through
the column the smaller molecules penetrate more pores than the larger
molecules. As a result, the smaller molecules take longer to emerge
from the column than the larger molecules
The width of the individual peaks reflects the distribution of the size
of molecules for a given polymer (macromolecule) and its components.
High
Performance Liquid
Chromatography (HPLC) is
an analytical technique for the separation and determination of organic
and inorganic solutes in any samples especially biological,
pharmaceutical, food, environmental, industrial, etc. In a liquid
chromatographic process a liquid permeates through a porous solid
stationary phase and elutes the solutes into a flow-through detector.
The stationary phase is usually in the form of small-diameter (5-10 mm)
uniform particles, packed into a cylindrical column. The typical column
is constructed from a rigid material (such as stainless steel or
plastic) and is generally 5-30 cm long and the internal diameter is in
the range of 1-9 mm.
|
|
|