Problem

Set 6 11/15/2018

1. How does gas-liquid and gas-solid chromatography differ?

In gas-liquid chromatography, the stationery phase is a liquid that is immobilized on a solid. Retention of
sample constituents involves equilibria between a gaseous and a liquid phase. In gas-solid chromatography,
the stationery phase is a solid surface that retains analytes by physical adsorption. Here separation
involves adsorption equilibria.

2. What are the principal advantages and principal limitations of a FID detector?

Advantages of flame ionization detector (FID): high sensitivity, large linear range, low noise, ruggedness,
ease of use, and response that is largely independent of flow rate.
Disadvantage of FID: destructive

3. List the variables that lead to band broadening in gas-liquid chromatography.

Band broadening arises from:
o very high or very low flow rates
o large particles making up packing
o thick layers of stationery phase
o low temperature
o slow injection rates

4. The following data are for liquid chromatographic column
Length of the column 24.7 cm
Flow rate 0.313 mL/min
VM 1.37 mL
VS 0.164 mL
A chromatogram of a mixture of species A, B, C, and D provided the following data:
Retention Time, 𝑡𝑡" (min) Width of Peak Base (W), min
Nonretained 3.1 (𝑡𝑡$ ) -
A 5.4 0.41
B 13.3 1.07
C 14.1 1.16
D 21.6 1.72
Calculate the number of plates (N) and retention factor (k) for each peak.
Calculate the resolution for species B and C.
The length of column necessary to separate the species B and C with resolution of 1.5.

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 Problem Set 6 11/15/2018

𝑡𝑡𝑡𝑡𝑅𝑅𝑅𝑅 2 5.4 2
a) Number of plates for species A 𝑁𝑁𝑁𝑁𝐴𝐴𝐴𝐴 = 16 = 16 = 2775
𝑊𝑊𝑊𝑊 0.41
𝑡𝑡𝑡𝑡𝑅𝑅𝑅𝑅 −𝑡𝑡𝑡𝑡𝑀𝑀𝑀𝑀 5.4−3.1
Retention factor for species A 𝑘𝑘𝑘𝑘𝐴𝐴𝐴𝐴 = = = 0.74
𝑡𝑡𝑡𝑡𝑀𝑀𝑀𝑀 3.1
Similar calculations for species B, C and D
2 tR C − tR B 2 14.1−13.3
b) 𝑅𝑅𝑅𝑅s BC = WB +WC
=
1.07+1.16
= 0.72

𝑅𝑅𝑅𝑅S column 1 𝑁𝑁𝑁𝑁column 1 0.72 2418

c) 𝑅𝑅𝑅𝑅S column 2
=
𝑁𝑁𝑁𝑁column 2
=
1.5
=
𝑁𝑁𝑁𝑁column 2
⟹ 𝑁𝑁𝑁𝑁2 = 10495
𝐿𝐿𝐿𝐿column 1 24.7
plate height for column 1 𝐻𝐻𝐻𝐻column 1 = = = 0.0102 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐
𝑁𝑁𝑁𝑁𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐 1 2418
𝐿𝐿𝐿𝐿column 2 = 𝑁𝑁𝑁𝑁column 2 𝐻𝐻𝐻𝐻column 1 = 10495 × 0.0102 0.0097 = 107.2 𝑐𝑐𝑐𝑐𝑐𝑐𝑐𝑐

5. Define isocratic elution.

In an isocratic elution, the solvent composition is held constant throughout the elution.

6. Define reversed-phase packing.

In reversed-phase packing, the stationery phase is nonpolar and the mobile phase is quite polar.

7. Indicate the order in which the following compounds would be eluted from an HPLC column containing a
reversed-phase packing: benzene, diethyl ether, n-hexane

diethyl ether , benzene, n-hexane

In reversed-phase chromatography the stationery phase is nonpolar and the mobile phase is polar.
Therefore, the most polar component elutes first, and increasing the mobile phase polarity increases the
elution time. This is because nonpolar compounds will spend relatively more time in nonpolar stationery
phase as the mobile phase becomes more polar.

8. Indicate the order of elution of the following compounds from a normal-phase packed HPLC column:
ethyl acetate, acetic acid, dimethylamine

ethyl acetate , dimethylamine , acetic acid

In normal-phase chromatography the stationery phase is polar and the mobile phase is nonpolar.
Therefore, the least polar component elutes first, and increasing the mobile phase polarity decreases the

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 Problem Set 6 11/15/2018

elution time. The more polar the solvent (mobile phase), the greater the eluent strength on a polar column
and the shorter the retention time.

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

Nonvolatile and thermally unstable compounds can be separated by HPLC by not GC.

10. 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. Find minimum number of
theoretical plates needed to achieve the resolution, RS, values of 1.25 and 1.50.

(𝑡𝑡" )& = 9 min
15
(𝑡𝑡" )- = 9 + = 9.25 min
60
65
𝑡𝑡$ = = 1.031579 min
60
Selectivity factor
(𝑡𝑡" )- − 𝑡𝑡$ 9.25 − 1.08333
𝛼𝛼 = = = 1.031579
(𝑡𝑡" )& − 𝑡𝑡$ 9.00 − 1.08333
retention factor for the second peak
(𝑡𝑡" )- − 𝑡𝑡$ 9.25 − 1.031579
𝑘𝑘- = = = 7.538462
𝑡𝑡$ 1.031579
Number of theoretical plates for resolution of 1.25
𝛼𝛼 ? 1 + 𝑘𝑘- ? 1.031579 ? 1 + 7.538462 ?
𝑁𝑁 = 16𝑅𝑅>? @ A B ?
C = 16 ∙ 1.25 B C B C = 34225
𝛼𝛼 − 1 𝑘𝑘- 1.031579 − 1 7.538462
Number of theoretical plates for resolution of 1.50
𝛼𝛼 ? 1 + 𝑘𝑘- ? 1.031579 ? 1 + 7.538462 ?
𝑁𝑁 = 16𝑅𝑅>? @ A B C = 16 ∙ 1.50? B C B C = 49284
𝛼𝛼 − 1 𝑘𝑘- 1.031579 − 1 7.538462

11. The two chromatograms shown below were recorded using reverse phase HPLC. Chromatogram B was
recorded on the same column and with the same sample as chromatogram A, but the separation is very
different. This is because the mobile phase was changed between the two separations. Explain how the

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 Problem Set 6 11/15/2018

mobile phase was changed to affect the separation so dramatically.

a. Chromatogram B was recorded using a mobile phase with higher eluent strength than
chromatogram A.
b. Chromatogram B was recorded using mobile phase with lower temperature.
c. Chromatogram B was recorded using a mobile phase with lower eluent strength than
chromatogram A.
d. The two chromatograms are identical.

GFH
12. Suggest a plausible formula of the type CxHyNz if the nominal mass of molecular ion M F = 79 and I =
G

5.9%?
a. C3HN3
b. C6H7
c. C4H3N2
d. C5H5N.

13. Of the following compounds, which would you expect to elute first from a nonpolar gas chromatographic
column?
a. Methanol (CH3OH)
b. Ethanol (CH3CH2OH)
c. n-Propanol (CH3CH2CH2OH)
d. n-Butanol (CH3CH2CH2CH2OH)
e. n-Pentanol (CH3CH2CH2CH2CH2OH)

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 Problem Set 6 11/15/2018

14. What useful information can be found from a Van Deemter plot?
a. The selectivity factor.
b. Optimum mobile phase flow rate.
c. Optimum column temperature.
d. Optimum column length.

15. What is eluent?

a. Eluent is the solvent or gas coming out of the column.
b. Eluent is the solvent in liquid chromatography.
c. Eluent is the solvent or gas going into a column.
d. Eluent is the solution injected on to a chromatographic column.

16. Which column gives narrower peaks in chromatographic separation: smaller plate height or larger plate
height?
a. Larger plate height gives less band spreading.
b. Plate height does not affect band spreading.
c. I do not know.
d. Smaller plate height gives less band spreading.