HPLC analysis is an interesting process utilizing a solid and stationary phase with a mobile or liquid phase. A glass or metal, or even plastic tube, called a column, is used to house the stationary phase, usually silica beads or a kind of porous solid laminated with specific compounds at different levels to attract the items needing separation. As the reactive solvent containing the compound moves through the column, separate molecules or even atoms can attach themselves to the stationary phase, thus separating the compound into its constituents.

We have always used so many techniques in the lab over the years but one of these included the synthesis of individual proteins. There was also a quite a bit of ion chromatography being done, which, as you are probably aware is a type of liquid chromatography.

This all begins with the purifying of amino acids and this can be achieved by using HPLC columns to separate the individual amino acids from different proteins. The column has a stationary phase that can separate the individual amino acids as the proteins pass through the column. Solvents are used to liquify and homogenize the proteins, and the solvent becomes a carrier, known as the mobile phase. The proteins are separated in an efficienct way as it is pumped through the system.

Particle size as well as pore size in the stationary process of an high performance liquid chromatography column is very important to the velocity at which the phase can separate the compound travelling through it. Silica is a common substance used in reverse-phase chromatography, which would be facilitated in a HPLC method system. Silica is not the only substance, and great care should be taken when using certain acidic solvents. Temperature is also important, as excessive heat can damage the silica. The benefit of silica is its electrical makeup, the pore sizes that can be found on the beads, and the surface area.