Molecular Weight and Molecular Size by GPC-SEC

Who wants to measure molecular weights andmeasurement of size based on how it moves,
sizes?which means that it is much more useful in
The answer is a surprisingly broad range ofindustrial or practical settings.
people and companies. Molecular weight and size isSize exclusion chromatography
most often used in identification of proteins, and inSize exclusion chromatography is the method by
characterization of polymers.which scientists determine the molecular size (not
Molecular Weight is commonly measured inthe weight!), of a particular substance. Molecules
industries such as::are separated in columns packed with porous
* Pharmaceutical industrysubstances, which might include glass beads,
* Biotech researchpolystyrene gels, silica gel, etc. Larger molecules
* Medical analysiselute more quickly through the columns, since the
* The food industrymolecules cannot fit into as many spaces. A
* Petroleum and polymer industriesconcentration detector is placed at the bottom of
Radius of gyration and Hydrodynamic radiusthe GPC columns to determine the amount of
Molecules are not simple shapes - unlike the fairlymaterial of each size fraction. In traditional SEC
compliant, mathematically-uncomplicated atom,GPC systems, operators need to pass known
which is spherical, molecules are difficult to definestandards through the columns before the sample.
as a simple shape. Hence two measurementsBy creating a calibration curve of size versus
have become industry standards, and analyticalelution time, particle size of unknown samples can
instruments are geared towards obtaining these;be calculated.
the radius of gyration, and the hydrodynamicIn more recent times, addition of other detection
radius.techniques such as Static Light Scattering and the
Radius of gyrationIntrinsic Viscosity detector provide direct
Scientific instruments can be used to identify themeasurement of molecular weight and size so
center of mass and dimensions of a molecule; thisthat traditional calibration techniques are not
is its radius of gyration. It is measured directlyrequired.
using static light scattering; however this methodGel permeation chromatography
has limitations at sizes lower than 10-15nm, andSEC is also commonly referred to by other
also for large molecules like polysaccharides.names. When an aqueous solution is used to carry
Viscometers are the tool which is most oftenthe sample through the column, the technique is
used for determining radius of gyration, using theoften called "Gel Filtration Chromatography", and
Flory-Fox equation.the name "Gel Permeation Chromatography" is
Hydrodynamic radiusoften used when solvents are used to carry the
This is a behavioral property of a substance - asubstance in question through the packed columns.