Chapter 28 Liquid Chromatography

1. List the types of substances to which each of the following chromatographic

methods is most applicable:
(a) gas-liquid

(b) liquid adsorption
(c) liquid-liquid partition
(d) reversed-phase partition
(e) ion exchange

(f) gel permeation

(g) gas-solid

(h) gel filtration

(i) ion-pair

2. Describe three general methods for improving resolution in partition

chromatography.

3. Describe a way to manipulate the retention factor of a solute in partition

chromatography.

4. How can the selectivity factor be manipulated in (a) gas chromatography and (b)
LC? 


5. In preparing a hexane-acetone gradient for an alumina HPLC column, is it des able

to increase or decrease the proportion of hexane as the column is eluted? 


6. What is meant by the linear-response range of a detector?

7. Define

(a) isocratic elution. 

(b) gradient elution.

(c) stop-flow injection.

(d) reversed-phase packing.

(e) normal-phase packing.

(f) ion-pairing chromatography.
(g) ion chromatography.

(h) bulk property detector.

 (i) solute property detector.

(j) sparging. 


8. What is a guard column in partition chromatography? 


9. In what way are normal-phase partition chromatography and adsorption

chromatography similar?

10. What is the order in which the following compounds would be eluted from an HPLC
column containing a reversed-phase packing?

(a) benzene, diethyl ether, n-hexane

(b) acetone, dichloroethane, acetamide 


11. What is the order of elution of the following compounds from a normal-phase
packed HPLC column?
(a) ethyl acetate, acetic acid, dimethylamine

(b) propylene, hexane, benzene, dichlorobenzene

12. Describe the fundamental difference between adsorption and partition

chromatography.

13. Describe the fundamental difference between ion-exchange and size-exclusion

chromatography.

14. What types of species can be separated by HPLC but not by GC?

15. Describe the various kinds of pumps used in HPLC. What are the advantages and
disadvantages of each?

16. Describe the differences between single-column and suppressor-column ion

chromatography.

17. Mass spectrometry is an extremely versatile detection system for gas

chromatography. However, interfacing an HPLC system to a mass spectrometer is a
much more difficult task. Describe the major reasons why it is more difficult to
combine HPLC with mass spectrometry than it is to combine GC with mass
spectrometry.

18. Which of the GC detectors in Table 27-1 are suitable for HPLC? Why are some of the
detectors listed unsuitable for HPLC?
19. Although temperature does not have nearly the effect on HPLC separations that it
has on GC separations, it nonetheless can play an important role. Discuss how and
why temperature might or might not influence the following separations:

(a) a reversed-phase chromatographic separation of a steroid mixture.
(b) an adsorption chromatographic separation of a mixture of closely related
isomers.

20. Two components in an HPLC separation have retention times that differ by 15 s. The
first peak elutes in 9.0 min and the peak widths are approximately equal.
The
dead time tM was 65 s. Use a spreadsheet to find the minimal number of theoretical
plates needed to achieve the following resolution R, values: 0.50, 0.75,
0.90,1.0,1.10,1.25,1.50,1.75,2.0,2.5. How would the results change if peak 2 were
twice as broad as peak 1?

21. An HPLC method was developed for the separation and determination of
ibuprofen in rat plasma samples as part of a study of the time course of the
drug in laboratory animals. Several standards were chromatographed and the
following results obtained:

Next, a 10 mg/kg sample of ibuprofen was administered orally to a laboratory

rat. Blood samples were drawn at various times after administration of the
drug and subjected to HPLC analysis. The following results were obtained:
 Find the concentration of ibuprofen in the blood plasma for each of the times
given above and plot the concentration versus time. On a percentage basis,
during what half-hour period (first, second, third, etc.) is most of the ibuprofen
lost?

22. In a normal-phase partition column, a solute was found to have a retention time of
29.1 min, and an unretained sample had a retention time of 1.05 min when
the
mobile phase was 50% by volume chloroform and 50% n-hexane. Calculate (il) k for
the solute and (b) a solvent composition that would bring k down to a value of
about 10.