Friday, 5 October 2012


Reversed Phase HPLC is a chromatographic technique also know as Reversed-Phase Chromatography (RPC) or hydrophobic Chromatography. So let us clear what is reverse phase HPLC and discuss some of the information about reverse phase.

Basic Principle involve in this reverse phase chromatography process are as follows : 

"When the polar mobile phase is used  and non polar stationary phase is used for the separation of an analyte or a mixture in the solution which is given with the help of chromatography, then such kind of separation or the process is known as reversed phase chromatography."

Reversed- Phase chromatography or RP-HPLC separates all the molecules on the basis of differences in their hpdrophobicity. 

Reversed phase chromatography (RPC) is also one of the widely used or most popular technique for the separation at analytical scale.

Some of the reasons for its popularity are as follows:

  1. The columns used in RPC are stable and efficient.
  2. RPC is considered a robust technique.
  3. RPC allows precise control of various variables  such as ph, temperature, concentration and organic solvent type.
  4. RPC is applicable to a wide variety or wide range of molecules which includes charged and polar molecules.

Uses of reverse phase HPLC or (RP-HPLC)

  • This technique is widely used in pharmaceutical as well as clinical work to separate hydrophobic compounds.
  • It is possible to apply serum, urine directly to column.
  • It can separate charged molecule.

Thank you