Gel Permetion Chromatography 1

GEL PERMETION CHROMATOGRAPHY:
Gel permeation chromatography (GPC) is a type of size exclusion

chromatography (SEC), which separates analytes on the basis of size. The
technique is often used for the analysis of polymers. As a technique, SEC
was first developed in 1955 by Lathe and Ruthven.
Size-exclusion chromatography (SEC) a chromatographic method in which
molecules in a solution is separated by their size and in some
cases molecular weight. It is usually applied to large molecules or
macromolecular complexes such as proteins and industrial polymers.
Typically, when an aqueous solution is used to transport the sample
through the column, the technique is known as gel-filtration
chromatography, versus the name gel permeation chromatography,
which is used when an organic solvent is used as a mobile phase.
The gel permeation chromatography was developed by J.C. Moore of
the Dow Chemical Company in 1964.
HOW GPC WORKS?

GPC separates based on the size or hydrodynamic volume (radius of
gyration) of the analytes. Radius of gyration or gyradius refers to the
distribution of the components of an object around an axis. This differs from
other separation techniques which depend upon chemical or physical
interactions to separate analytes. Separation occurs via the use of porous
beads packed in a column.
The smaller analytes can enter the pores more easily and therefore spend
more time in these pores, increasing their retention time. Conversely, larger
analytes spend little if any time in the pores and are eluted quickly. All
columns have a range of molecular weights that can be separated.
If an analyte is either too large or too small it will be either not retained or
completely retained respectively. Analytes that are not retained are eluted
with the free volume outside of the particles (Vo), while analytes that are
completely retained are eluted with volume of solvent held in the pores (V i).
The total volume can be considered by the following equation, where V g is
the volume of the polymer gel and V t is the total volume.
As can be inferred, there is a limited range of molecular weights that can be

separated by each column and therefore the size of the pores for the
packing should be chosen according to the range of molecular weight of
analytes to be separated. For polymer separations the pore sizes should be
on the order of the polymers being analyzed. If a sample has a broad
molecular weight range it may be necessary to use several GPC columns
in tandem with one another to fully resolve the sample.
APPLICATIONS;
GPC is often used to determine the relative molecular weight of polymer

samples as well as the distribution of molecular weights. What GPC truly
measures is the molecular volume and shape function as defined by
the intrinsic viscosity. If comparable standards are used, this relative data
can be used to determine molecular weights within 5%
accuracy. Polystyrene standards with PDI of less than 1.2 are typically
used to calibrate the GPC. Polystyrene (PS) is
asynthetic aromatic polymer made from the monomer styrene
Unfortunately, polystyrene tends to be a very linear polymer and therefore

as a standard it is only useful to compare it to other polymers that are
known to be linear and of relatively the same size.
MATERIAL AND METHODS;

Gel permeation chromatography is conducted almost exclusively
in chromatography columns. The experimental design is not much different
from other techniques of liquid chromatography. Samples are dissolved in
an appropriate solvent, in the case of GPC these tend to be organic
solvents and after filtering the solution it is injected onto a column. The
separation of multi-component mixture takes place in the column. The
constant supply of fresh eluent to the column is accomplished by the use of
a pump. Since most analytes are not visible to the naked eye a detector is
needed. Often multiple detectors are used to gain additional information
about the polymer sample. The availability of a detector makes the
fractionation convenient and accurate.
Gel
Gels are used as stationary phase for GPC. The pore size of a gel must be
carefully controlled in order to be able to apply the gel to a given
separation. Other desirable properties of the gel forming agent are the
absence of ionizing groups and, in a given solvent, low affinity for the
substances to be separated. Commercial gels like PLgel, Sephadex, BioGel (cross-linked polyacrylamide), agarose gel and Styragel are often used
based on different separation requirements.
Eluent
The eluent (mobile phase) should be a good solvent for the polymer, should
permit high detector response from the polymer and should wet the packing
surface. The most common eluents in for polymers that dissolve at room
temperature GPC are tetrahydrofuran (THF), o-
dichlorobenzene and trichlorobenzene at 130150 C for

crystalline polyalkynes and m-cresol and o-chlorophenol at 90 C for
crystalline condensation polymers such as polyamides and polyesters.
Pump
There are two types of pumps available for uniform delivery of relatively
small liquid volumes for GPC: piston or peristaltic pumps .
Detector
In GPC, the concentration by weight of polymer in the eluting solvent may
be monitored continuously with a detector. There are many detector types
available and they can be divided into two main categories. The first is
concentration sensitive detectors which include UV absorption, differential
refractometer (DRI) or refractive index (RI) detectors, infrared (IR)
absorption and density detectors. Molecular weight sensitive detectors
include low angle light scattering detectors (LALLS), multi angle light
scattering (MALLS).The resulting chromatogram is therefore a weight
distribution of the polymer as a function of retention volume.
The most sensitive detector is the differential UV photometer and the most
common detector is the differential refractometer (DRI). When
characterizing copolymer, it is necessary to have two detectors in
series. For accurate determinations of copolymer composition at least two
of those detectors should be concentration detectors. The determination of
most copolymer compositions is done using UV and RI detectors, although
other combinations can be used.
ADVANTAGES OF GPC;
As a separation technique GPC has many advantages. First of all, it has a
well-defined separation time due to the fact that there is a final elution
volume for all unretained analytes. Additionally, GPC can provide narrow
bands, although this aspect of GPC is more difficult for polymer samples
that have broad ranges of molecular weights present.
Finally, since the analytes do not interact chemically or physically with the
column, there is a lower chance for analyte loss to occur. For investigating
the properties of polymer samples in particular, GPC can be very
advantageous.
GPC provides a more convenient method of determining the molecular
weights of polymers. In fact most samples can be thoroughly analyzed in
an hour or less.
Other methods used in the past were fractional extraction and fractional
precipitation. As these processes were quite labor-intensive molecular
weights and mass distributions typically were not analyzed.
Therefore, GPC has allowed for the quick and relatively easy estimation of
molecular weights and distribution for polymer samples.
DISADVANTAGES OF GPC;
There are disadvantages to GPC, however. First, there is a limited number
of peaks that can be resolved within the short time scale of the GPC run.
Also, as a technique GPC requires around at least a 10% difference in
molecular weight for a reasonable resolution of peaks to occur.
In regards to polymers, the molecular masses of most of the chains will be
too close for the GPC separation to show anything more than broad peaks.
Another disadvantage of GPC for polymers is that filtrations must be
performed before using the instrument to prevent dust and other
particulates from ruining the columns and interfering with the detectors.
Although useful for protecting the instrument, the pre-filtration of the sample
has the possibility of removing higher molecular weight sample before it
can be loaded on the column. Another possibility to overcome these issues
is the separation by Field-flow fractionation (FFF).

Gel Permetion Chromatography 1

Uploaded by

Copyright:

Available Formats

Gel Permetion Chromatography 1

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Gel Permetion Chromatography 1

Uploaded by

Copyright:

Available Formats

GEL PERMETION CHROMATOGRAPHY:

Gel permeation chromatography (GPC) is a type of size exclusion

HOW GPC WORKS?

As can be inferred, there is a limited range of molecular weights that can be

GPC is often used to determine the relative molecular weight of polymer

Unfortunately, polystyrene tends to be a very linear polymer and therefore

MATERIAL AND METHODS;

dichlorobenzene and trichlorobenzene at 130150 C for

You might also like