The Essentials to note about Chromatography

November 20, 2020

Chromatography is the collective term for a set of techniques used to separate mixtures. These techniques include gas chromatography GC, thin layer chromatography TLC, Size exclusion Chromatography SEC, and higher performance liquid chromatography HPLC.

Both Phases

Chromatography involves passing a mix dissolved in mobile phase via stationary phase. The mobile phase is generally a liquid or a gas that transfers the mixture to be separated via a column or horizontal sheet that has a solid stationary phase.

Liquid Chromatography

Liquid chromatography LC is a separation technique in which the mobile phase is a liquid. It can be carried out in either a column or a plane. LC is particularly helpful for the separation of electrons or ions that are dissolved in a solvent. Simple liquid chromatography includes a column using a fritted bottom that holds a stationary phase in equilibrium with a solvent. Commonly used stationary phases include solids, ionic groups on a resin, liquids on an inert solid support and porous inert particles. The mixture to be separated is loaded on the surface of the column followed by more solvent. The various components in the mixture pass through the column at different rates due to the variations from the partitioning behavior between the mobile liquid and stationary phases. Liquid chromatography is more widely used than other methods like gasĀ chromatogram since the samples analyzed do not have to be vaporized. Additionally, the variations in temperature have a slight impact in liquid chromatography, unlike in other kinds of chromatography.

High Performance Liquid Chromatography HPLC

Present day liquid Chromatography that normally utilizes tiny packing particles and a rather higher pressure is referred to as HPLC. It is basically an extremely improved form of column chromatography frequently used by biochemists to different amino acids and proteins because of their different behavior in solvents regarding the amount of electronic charge of every one. Instead being allowed to trickle through a column under gravity, the solvent is forced through under high pressures up to 400 atmospheres, which makes the process much quicker. Since smaller particles are used, with their dimensions being determined by a particle size analyzer, there’s greater surface area for connections between the stationary phase and the molecules flowing past it. This then allows for much greater separation of these components in the mix.

There are many advantages of HPLC. For one, it is an automated process which only requires a couple minutes to produce effects. This is a huge step up from liquid chromatography, which uses gravity rather than a high-speed pump to force components through the thickly packed tubing. HPLC produces results which are of a high resolution and are simple to read. Additionally, the tests can easily be reproduced via the automated procedure.