Quest 1 chromatography, 2021

(50 points total)

Useful information:

tr −tm tr 2
Kpart = Cs / Cm k= N= 16 �w� N=L/H
tm

tB −tA t −t
H = A + B/u + Cu Rs = 1 α = tB−tm
(wA +wB ) A m
2

μe V 1 V
N= Rs = (μe1 − 𝜇𝜇e2 ) � �
2D 4√2 D(μ�e +μeo )

Multiple choice (4 points each). If there is more than one correct answer, mark all that are
correct.

1. Which of these liquid chromatography detectors gives the most structural

information about the analyte?

a. single stage mass spectrometric detector

b. uv/vis absorption detector
c. fluorescence detector
d. mass spectrometric detector

2. Which of these broadening effects would get worse by increasing the length of the
column?
a. eddy diffusion
b. longitudinal diffusion
c. mass transfer equilibrium
d. all of these
e. none of these

3. Which of these broadening effects would get worse by increasing the flow rate of
the mobile phase?
a. eddy diffusion
b. longitudinal diffusion
c. mass transfer equilibrium
d. all of these
e. none of these
4. (8 points) Sketch the major components of a reverse-phase HPLC instrument with
a fluorescence detector. A block diagram is fine.

sample
pump loop column F det.
Solv 1 Solv 2

injector

5. (5 points) Provide an example of a solvent gradient that might be used on the

reverse-phase HPLC instrument in question 5. A picture might help in your
discussion, but please label specifics of your gradient.

Starting solvent mixture should be very polar, say 95% H2O: 5% CH3OH/CH3CN
the polarity of the gradient show decrease either step-wise or continuously to a
mixture such as 50% H2O: 50% CH3OH/CH3CN. You can go all way to 5% H2O :
95% CH3OH/CH3CN, but that is not necessary. To get full credit, you must have
given me specific miscible polar solvents and the gradient should result in a decrease
in polarity

6. (5 points) What causes net migration of buffer to the cathode in capillary

electrophoresis? You may draw a picture to help your discussion. PLEASE be
brief!!!!!
Electro-osmotic flow is responsible for net migration of the buffer to the cathode.
This occurs because the walls of the capillary are negatively charged and cations
from the buffer solution for an electrical double layer. The cations from the double
layer are attracted to the negatively charged cathode and get pulled towards it
dragging the rest of the buffer and your analytes with them
7. Method choice (20 points). Consider the following titles of articles from the
Journal of Chromatography B from 2016. I have removed the method names
from the titles. Given the information in the titles and my comments, fill in a
method (and specific detector if I ask for it) that would be acceptable for the
study. There may be the possibility of more than one correct answer for each title.

a. “Simultaneous determination of major phytocannabinoids, their main metabolites,

and common synthetic cannabinoids in urine samples by [method and detector
removed].”
(method + detector)

The cannabinoids are small organic molecules that

are slightly polar.
GC or reverse-phase LC with an appropriate detector MS would be
appropriate for both, FID for GC, uv-vis for LC would also be appropriate.

b. “Use of [method removed] for simultaneous analysis of small ions in saliva

samples from wrestlers undergoing a weight training program.”
(method)

ion chromatography or CE

c. “Pre-study and in-study validation of a [method removed] method with different

detection modes for the analysis of monoclonal antibody aggregates.”
(method)

The proteins aggregates in the study ranged

from 14 -144 kDa.

size exclusion chromatography or gel electrophoresis

d. “Multi-residue analysis of 26 organochlorine pesticides in Alpinia oxyphylla by

[method removed] after solid phase extraction and acid cleanup.”
(method and detector)

All of these pesticides are volatile.

was fishing for GC-ECD, but FID or MS would also work.

other answers are possible. If you provided one that is not quite correct, I indicated
why.