The History and Development of Reversed-Phase Chromatography

Reversed-phase chromatography (RPC) contains any chromatographic method using a non-polar stationary phase. The word “reverse” has a historical background. In the 1970s, most liquid chromatography was performed on unmodified silica or alumina. Their surface chemistry is hydrophilic and has a stronger affinity for polar compounds.

It is also called “positive phase (Normal) chromatography”. If an alkyl chain is covalently bonded to the support surface, the elution sequence is reversed. In reversed-phase chromatography, polar compounds are eluted first, while non-polar compounds are retained because they are an affinity for the reversed surface.

All mathematical and experimental studies used in other chromatographic methods are also applicable in reversed phase chromatography (e.g. chromatographic resolution is proportional to column length).

Today, reversed-phase column chromatography accounts for the vast majority of analytical liquid chromatography.

For example, in biochemical analysis, it can be used

1. Analysis of amino acids and peptides;
2. Separation of proteins;
3. Analysis of bases, nucleic acids, and nucleases;
4. Analysis of steroidal compounds;
5. And other separations such as catecholamines, histamine, sugar, and vitamins.