Question

Answer the following questions

1) Which compound would you expect to have a shorter retention time on a reverse-phase HPLC column with a water/acetonitrile mixture adjusted to pH 3 with acetic acid — benzylalcohol or benzoic acid?

2) Draw a hypothetical chromatogram of a mixture containing compounds A and B in approximately equal amounts. Label the axes of the chromatogram, the peaks and any other pertinent information. If the void volume of the column is 2.5 min and the retention times of A and B are 5.0 and 6.0 min, respectively, what are the column capacity ratios of the two compounds? Are these peaks completely resolved?

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1.
The by reversed phase chromatography is used for the molecules which has some amt of hydrophobic character the process gives excellent recovery and resolution. The actual separation process involves the hydrophobic binding interaction between the solute molecule in the mobile phase and the immobilised hydrophobic ligand, i.e. the stationary phase in simpler words reversible adsorption and desorption of solute molecules with varying degrees of hydrophobicity to a hydrophobic stationary phase....

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