SIZE EXCLUSION CHROMATOGRAPHY principle| instrumentation|lab experiment|graph|Application

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Gel filtration chromatography (also known as size exclusion chromatography, molecular sieve chromatography, or gel permeation chromatography) is a chromatographic method in which molecules in solution are separated by their size, and in some cases molecular weight.
In gel filtration chromatography, the stationary phase consists of porous beads packed into a column. The mobile phase is the running buffer or other solvent. Sample components partition between the stationary and mobile phases based on their size-based accessibility to the pores of the matrix beads.
The smaller the size of the molecule or particle, the greater access it has to the pores of the matrix (or the accessible stationary phase), and the slower it moves through the column. Very large particles, which are completely excluded from the pores, elute first from the column at the void volume.
The separated components are visualized as a plot of the volume of the mobile phase eluted through the column versus the detector signal. This plot, called a chromatogram, shows the location of the individual peaks and the quality of resolution of these peaks.The location of the peaks, their heights, and widths convey a great deal of information about the sample components as well as the column.
In this video we have discussed complete laboratory procedure of size exclusion chromatography and also, how to calculate KD (distribution coefficient) of a particular component from chromatogram (Graph) and how KD valve can be use to determine the molecular weights of the sample components And the Application of size exclusion chromatography.

Please watch this video for more details.

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