Advantages and disadvantages of size exclusion chromatography

The size exclusion chromatography or gel filtration chromatography is one of the high-performance liquid chromatography (HPLC) separation types. This technique is used to separate particles based on size, larger molecules travel faster throughout the column as compared to smaller molecules, as they penetrate deep into the holes. The gel permeation chromatography (GPC) and gel filtration chromatography (GFC) are two basic types of size exclusion chromatography (SEC). GPC uses a hydrophobic stationary phase and a non-aqueous mobile phase and GFC uses a hydrophilic stationary phase and an aqueous mobile phase for the separation of components.
The advantages of size exclusion chromatography are as follows.

• Good separation of large components from the small sample components with a less amount of solvent is the major advantage of size exclusion chromatography.

• The separation in gel filtration chromatography doesn't rely on the buffer composition, pH, temperature and ionic strength.

• Size exclusion chromatography provides a well-defined separation of analytes.

• This technique requires a short analysis time.

• There is no sample loss in this technique.

• The flow rate of the system can be set as per requirement.

• It gives a narrow band and good sensitivity.

• The lower amount of mobile phase required for isolation compared to other techniques.

The disadvantages of size exclusion chromatography are as follows.

• Mobile phase filtering is necessary to prevent the columns and interfere with detectors.

• The major disadvantage of size exclusion chromatography is that the limited number of peaks can be resolved since the run time is short.

• It has poor selectivity compared to other chromatographic techniques.

• The load capacity of the sample is another major disadvantage of gel filtration chromatography.

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