Reversed Phase Chromatography Principle
The stationary phase covalently bound to the support in reverse phase chromatography is some linear hydrocarbon such as n-octyl. Extraction and purification by hydrophobic extraction, that is, when non-polar solutes are present in water, the interaction between solute molecules, the interaction between solute molecules and water molecules is much smaller than the interaction between water molecules, so solute molecules are from water Was “squeezed out”. It can be seen that the more hydrophobic the compound in reversed phase chromatography, the easier it is to extrude from the mobile phase, and the longer the residence time in the column, so the different compounds in reversed phase chromatography are separated according to their hydrophobic properties.
Reversed phase chromatography also separates compounds with groups of different polarities. The retention behavior of the solute molecules and the mobile phase can be altered by changing the solvent of the mobile phase and its composition and pH. In addition, the proportion of the mobile phase in the reverse phase chromatography is very large, and can be/from 0-100%, so that reverse phase chromatography can be used for the separation of water-soluble and fat-soluble compounds.