Reversed phase chromatography (RP) is the most widely used technique in high-performance liquid chromatography (HPLC) up to now, mainly because it is suitable for the analysis of most of them. Nonpolar substances and many ionizable and ionic compounds. Most stationary phases used in reversed-phase chromatography are natural hydrophobic substances.
Therefore, the analytes are separated according to their hydrophobic interaction with stationary phases. The mechanism of hydrophobic separation can also be separated in the same way.
Users must be aware of the specific properties of the surface of the stationary phase they are using and the possible interaction between the surface of the stationary phase of the substance so that the possible matrix interaction can be taken into account when they use the inverse phase method.
For example, very hydrophobic sample substrates such as corn oil, highly aromatic substances, and wax can adhere to the stationary phase filling surface and change their properties. Biofluids containing proteins can also be adsorbed on the filling surface. Despite the best efforts of analysts to protect HPLC columns, matrix contamination of some analytes can adversely affect stationary phases.