What are the limitations of using reverse HPLC?

Expert Solution

HPLC (High performance Liquid chromatography) is a type of column chromatography which pumps an analyte (solution mixture) in a solvent (also called the mobile phase) at some high pressue, through a column with a packing material (stationery phase).

In Normal phase chromatography the stationery phase (hydrophillic) is polar and the mobile phase is non-polar. In revese HPLC also called reverse phase chromatography, stationery phase (hydrophobic) and non-polar and mobile phase is polar. Since the polarity of stationery and mobile phase is inverted hence, it is reverse phase.

Although the technique is very useful and there are only a handful of limitations for this technique. Thus I would like to mention a few of them.

Limitations of using reverse HPLC

1. Reverse phase HPLC cannot be sometimes used for the seperation of proteins since the organic solvents used in the reverse phase HPLC can denature many proteins.

2. Column Chromatography in general is a time consumind process.

3. More amounts of Mobile phase (solvents) is essential.

4. The technique is automatic and this makes the technique more complicated and expensive as well.

queen_honey_blossom answered 2 years ago

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