Question

1.Calculate the resolution of three isocratic (60% MeOH: 40% H2O) reverse phase chromatographic peaks having the following qualities:

•Peak 1: retention time 3.45 minutes FWHM: 0.13 minutes

•Peak 2: retention time 3.54 minutes FWHM: 0.16 minutes

•Peak 3: retention time 3.97 minutes FWHM: 0.29 minutes

•How can the experimental conditions be altered to enhance separation? (Give 2 options)

Answer 1

Resolution - 1.18 (the tri FWHM, & FW HM Betucen peak 1 and 2 = 1.18 (5.54 – 3.45) - 0-366 Between peak 2 and 3 = 1188 77 3785. Dos 1.18 (3.91-3.34) = 3.54) 0.29 +0.16 tot Between peak land 3 = 1.18(3.97-3.45) - 1.461 0.29 + 0.13 /

Alterations to enhance separation

Increase in theoretical plates increases resolution. This can be achieved by following ways

a) Changing mobile phase composition.

b) Changing column temperature