Answer Posted / vinod kumar, nektar therapeuti

1.If molecular weight less than 2000. based on their
solubility follow as below
2.If water soluble and non ionic follow reverse phase.
3.If water soluble and ionic follow reverse phase ion pair
method.
4.If organic soluble follow normal phase.
5.If molecular weight is more than 2000.Based on their
solubility follow as below.
6.If organic soluble follow gel permeation chromatography
7.If water soluble ion exchange or gel filtration.
8.Once the technique is fixed optimise column.
9.Optimise buffers
10.optimise pH of buffers
11.optimise concentration of buffers.
12.optimise type of organic solvent.
13.optimise column temperature
14.optimise gradient programme to improve resolution and to
elute out late eluting peaks.

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