Chapter 25: Q32P (page 710)

Question: what are the general steps in developing an isocratic separation for reversed-phase chromatography?

Expert verified

General steps in developing an isocratic separation.

Choose a detector that runs a wide gradient to determine whether isocratic

Step by step solution

To create an isocratic separation for reversed-phase chromatography, we need to follow this steps.

Determine the analysis's goal.
choose a sample preparation method.
Choose a detector that runs a wide gradient to determine whether isocratic or gradient elution is appropriate.

Then,

for isocratic elution adjust the mobile phase composition to achieve retention of $0.5 \leq k \leq 20$ for all compounds.
Adjust relative retention by varying the solvent composition, column temperature, organic solvent type, and column type.
optimise column dimensions in order to increase plate number or decrease separation time.

Hence,.

Determine the analysis's goal. choose a sample preparation method. Choose a detector that runs a wide gradient to determine whether isocratic or gradient elution is appropriate. for isocratic elution adjust the mobile phase composition to achieve retention of $0.5 \leq k \leq 20$ for all compounds. Adjust relative retention by varying the solvent composition, column temperature, organic solvent type, and column type. optimise column dimensions in order to increase plate number or decrease separation time.