Difference between reversed phase and normal phase chromatography

The high-performance liquid chromatography is also known as high-pressure liquid chromatography. It is used to separate, identify, and quantify each component based on their polarity. The HPLC system consists of a column, injector, UV/VIS or PDA detector, pump, degasser, and reservoir, etc. There are different types of HPLC chromatography are available based on their composition and method of separation. It includes reverse phase chromatography, normal phase chromatography, size-exclusion chromatography, and ion-exchange chromatography is available.
In RP-HPLC the stationary phase is polar and the mobile phase is non-polar, whereas in NP-HPLC the stationary phase is non-polar and the mobile phase is polar. This is the key difference between reversed phase and normal phase HPLC.

Normal phase chromatography:

1. In the very non-polar environment, the hydrophilic analytes will interact with each other. In the mobile phase, the hydrophilic molecule will move towards adsorb outside and inside a particle if the surface is also hydrophilic. Increasing the polarity of the mobile phase will reduce adsorption and finally, the sample molecules will elute off the column. This occurrence is called a normal phase chromatography. This is a very potent technique and it is used mostly as an analytical technique in analytical chemistry.
2. Normal phase HPLC systems are parallel to the flash-column chromatography
3. A silica stationary phase is eluted with a non-polar solvent or a fairly non-polar solvent mixture. In RP-chromatography, only organic solvents are used.
4. In the normal phase, non-polar analytes elute rapidly as compared to polar analytes.

Reversed-Phase Chromatography:

1. The work of this technique is the opposite of the normal phase. Reducing the polarity of the mobile phase by adding more organic solvents decreases the hydrophobic interaction between solid support and solute, resulting in desorption. The molecules have more hydrophobic takes more time on the solid support and the more organic solvent concentrations necessary for the promotion of desorption.
2. The reversed phase is the works opposite of normal-phase chromatography.
3. With a polar solvent mixture, a non-polar stationary phase is eluted.
4. In the reverse phase, polar molecules elute rapidly as compared to non-polar analytes.

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