what is hplc

Different types of Column Chromatography

The column chromatography is a type of chromatography it is one of the most effective separation techniques. Stationary phase (adsorbent) either silica gel or alumina is used and filled from the top of the column. The mobile phase is either solvent or a combination of solvents, passed along the sample through the stationary phase. The separation of the components depends on their polarity, as the polar compounds travel slowly through the column. This means that polar compounds absorb more strongly than non-polar.
Based on the stationary phase, there are four types of column chromatography available, and they are working on a different mechanism.
There are different types of column chromatography as follows.
1. Adsorption chromatography.
2. Ion-exchange chromatography
3. Partition chromatography
4. Gel chromatography

1. Adsorption chromatography is based on this interaction based on active sites and polarity of solid molecules and analytes at the stationary phase. Since the polarity of the stationary phase than the mobile phase is more polar, then the high polar compounds will remain firm until the stationary phase, while the low polar compounds will be tied less than the stationary phase. Compared with firmly bound compounds, the less-bound compounds will be eluted by the first mobile stage, so this technique proves that polar like polar.
2. Ion-exchange chromatography is a method that permits ions and polar molecules to be separated on the basis of their charge. It can be used for about any type of charge molecule contains large amino acids and proteins. It is regularly used in quality control, water analysis, and protein purification.
3. Partition chromatography, components are distinguished between the stationary phase and mobile, In the Partition chromatography, the stationary phase and the mobile phase are both are liquid.
4. The gel chromatography column is packed with gel in which the separation takes place, and the stationary phase is a solvent.


Go up