Size Exclusion Chromatography

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Size Exclusion Chromatography

Size Exclusion Chromatography (SEC) is the separation technique based on the molecular size of the components.

Size exclusion chromatography is a kind of method to separate different size of molecules that put in solution. It was first discovered by two scientists who named Grant Henry Lathe and Colin R Ruthven. Both of them received the John Scott Award for this fabulous invention.

There are various applications for Size exclusion chromatography such as biochemical aspect and polymer synthesis. For application in biochemical aspect, this technique can find out the quaternary structure of purified proteins which possess slow exchange times, since it can be carried out under native solution conditions and preserve macromolecular interactions. The reason why we use this technique for purification is Size exclusion chromatography is a low resolution chromatography method as it does not identify similar species very well. It can also test the tertiary structure of protein as it measures the hydrodynamic volume, allowing folded and unfolded versions of the same protein to be distinguished. Besides using in biochemical research, it is able to find the distribution of the sizes of polymer molecules like if a solvent is chose and run, we can create a calibration curve to determine the sizes of polymer molecules in it.

It is better to introduce the mobile phase and stationary phase first. Stationary phase is the solid absorbent or the pore(SEC) with solid support that allow sample across through it while the mobile phase is the sample percolate through or along to the stationary phase.

In SEC, separation is achieved by the differential exclusion from the pores of the packing material, of the sample molecules(mobile phase) as they pass through a bed of porous particles(stationary phase).

For the principle of the SEC, molecules of different sizes can be separated by this technique because of differential time spent inside a...