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Homework #5 (26-2, 26-6, 26-7, 26-11, 27-8, 27-19, 27-21, 27-22, 27-25, 28-4, 28-13,

28-15, 28-16, 28-17, 28-18)

26-2. Describe the general elution problem.

The general elution problem is the problem that arises when attempting to separate a
group of molecules which have very different retention factors. The problem is that no
single isocratic or isothermal separation will satisfactorily separate all of the molecules.
Conditions appropriate to weakly retained molecules (those with small retention factors)
will be inappropriate for the strongly retained molecules (those with large retention
factors), and the same is true in reverse. The solution is to use gradient elution or
temperature programmed conditions.

26-6. What variables are likely to affect the α value for a pair of analytes?

The selectivity factor is equal to the ratio of the two retention factors which will also be
equal to the ratio of the distribution coefficients. Anything that changes this ratio will
change the selectivity coefficient. Temperature will affect the value of the equilibrium
constants (freshman chem), but it will likely only weakly affect the ratio, depending upon
the enthalpies. Changing the composition of the stationary phase will affect the
distribution coefficients, and in LC changing composition of the mobile phase will also
have an impact on the equilibrium constant.

26-7. How can the retention factor for an solute be manipulated?

The retention factor can be manipulated by any of the above factors that affect the value
of the equilibrium constant (temperature, composition of stationary phase, composition of
mobile phase (in LC)), but you can also change the ratio of the volume of the stationary
phase to the mobile phase. This can be accomplished any number of ways, for example
by changing the thickness of the stationary phase on the packing material or by changing
the diameter of the capillary.

26-11. What is gradient elution?

Gradient elution is the process of varying the composition of the mobile phase during the

27-8. Describe the principle upon which each of the detectors listed in Problem 27-7 is
based. (a) TCD, (b) atomic emission, (c) thermionic, (d) electron capture, (e) flame
photometry, (f) photoionization.

(a) TCD operates on the principle of differences in thermal conductivity. Gases eluting
from the column are passed over a hot element (the temperature of which is closely
monitored). As the thermal conductivity of tha gases passing the element change, so does
the heat transport from the element, and its temperature changes. This detection method
is based on the average thermal conductivity and so will not be very sensitive, as the
analyte is diluted in the mobile phase.
(b) atomic emission operates by creating a plasma

27-19. The same polar compound is gas chromatographed on an SE-30 (very non-polar)
column and then on a carbowax 20M (very polar) column. How will K = cS/cM vary
between the two columns?
A polar compound will interact attractively with the polar stationary phase more than it
will interact with the nonpolar stationary phase. As a result, the Cs for the SE-30 will be
less than the Cs for the carbowax 20M, and therefore K20M > KSE-30.

27-21. A GLC column was operated under the following conditions:

column: 1.10m x 2.0mm, packed with Chromosorb P; weight of stationary liquid added,
1.40g, density of liquid 1.02g/mL
pressures: inlet 26.1psi above room; room, 748 torr
measured outlet flow: 25.3 mL/min
temperature: room, 21.1C; column, 102.0C
retention times: air, 18.0s; methyl acetate, 1.98min; methyl n-propionate, 4.16min;
methyl n-butyrate, 7.93min.
peak widths at base: 0.19, 0.39, 0.79 min, respectively
(a) the average flow rate through the column.
(b) the corrected retention volumes for air and the three esters.
(c) the specific retention volumes for the three components.
(d) the distribution coefficients for the three esters.
(e) the retention time and corrected retention volume for methylhexanoate.

a) the average flow rate through the column in mL/min is determined from equation 27-3
on page 702 F = Fm x Tc/T x (P-PH2O)/P
F= (25.3mL/min)((102.0+273.2)/(21.1+273.2))((748-18)/748) = 31.4mL/min
b) the retention volumes for air and the three esters are given by equations 27-1 and 2,
thus by simple multiplication of the retention times by the average flow rate.
Vair = ((18.0s/60s)min) 31.4mL/min = 9.44mL
Vma = 1.98min 31.4mL/min = 62.3mL
Vmp = 4.16min 31.4mL/min = 131.0mL
Vmb = 7.93min 31.4mL/min = 249.6mL
The corrected retention volumes include an additional pressure term j given by equation
27-5 j = (3/2)((Pi/P)2-1)/((Pi/P)3-1), where Pi is the inlet pressure
Pi = 748torr+ 26.1psi(760torr/14.7psi) = 1349torr and so Pi/P = 1349/748 = 1.80 and thus
j = 1.5(1.802-1)/(1.803-1) = 0.695
As a result the corrected retention volumes are
V0air = 0.695 9.44mL = 6.56mL
V0ma = 0.695 62.3mL = 43.3mL
V0mp = 0.695 131.0mL = 91.0mL
V0mb = 0.695 249.6mL = 173.5mL
c) the specific retention volumes are calculated via equation 27-6 on page 702
Vg = (273.2/Tcolumn)(V0R-V0m)/W and so for air this will be zero
Vg-ma = (273.2/(102.0+273.2)) (43.3mL-6.56mL)/1.40g = 19.1mL/g
Vg-mp = (273.2/(102.0+273.2)) (91.0mL-6.56mL)/1.40g = 43.9mL/g
Vg-mb = (273.2/(102.0+273.2)) (173.5mL-6.56mL)/1.40g = 86.8mL/g
d) the distribution coefficient depends upon the specific retention volume and the density
of the stationary phase and the temperature according to equation 27-7 on page 702
K = Vg ρs Tc/273.2 = Vg (1.02g/mL) (102+273.2)/273.2 = 1.40Vg and so for the esters
Kma = 1.40Vg = 1.40g/mL 19.1 mL/g= 26.7
Kmp = 1.40Vg = 1.40 g/mL 43.9 mL/g= 61.5
Kmb = 1.40Vg = 1.40 g/mL 86.8 mL/g= 121.5
(e) these three esters represent a homologous series, and as such you would expect that
the log of the corrected retention times would depend upon the carbon number in a linear
way. First make a table of the log corrected retention times and the carbon number and
then determine the slope and intercept.

N tr(s) tr’(s) Log(tr’)

2 118.8 100.8 2.003
3 249.6 231.6 2.365
4 475.8 457.8 2.661
6 2150 2132 3.329
Using the slope is 0.3286 and the intercept is 1.357 (from excel) you can obtain the last
line of values. With a retention time of 2150s, the corrected retention volume will be
V0mh = jFtr-mh = (0.695) (31.4mL/min) (2150s/60s) min = 782mL

27-22. From the data in problem 27-21, calculate:

(a) k’ for each compound.
(b) α values for each adjacent pair of compounds.
(c) the average number of theoretical plates and plate height for the column.
(d) the resolution for each adjacent pair of compounds.

(a) The k’ can be readily determined from the retention times. By using equation 26-8 on
page 680. k’ = tr-tm/tm values from this calculation are shown in the attached table.
(b) The α values for each adjacent pair of compounds can be readily determined by
simply plugging the retention times into equation 26-10 on page 680 a = k’A/k’B. Values
for these are shown in the attached table below. The selectivity coefficient in the table
represents the selectivity between that compound and the previous compound in the table.
(c) The number of theoretical plates is measured for each of the peaks and is shown in the
table along with the average value. These values were computed using the equation 26-17
on page 683 N = 16 (tr/Wb)2. the average number of theoretical plates and plate height for
the column.
(d) The resolution for each adjacent pair of compounds was determined by using equation
Wb Wb tr tr tr' k' ! N Rs
min sec min sec sec
water 0.3 18 0 0
methylacetate 0.19 11.4 1.98 119 101 5.6 1738
methylpropionate0.39 23.4 4.16 250 232 12.87 2.2976 1820 7.52
methylbutyrate 0.79 47.4 7.93 476 458 25.43 1.9767 1612 6.39

average 1723
second equation on page 689. It is not labeled but it reads as Rs = (tr)B – (tr)A / W. In my
calculation I used the average of the peak widths as the width. The values next to a
compound represent the resolution between that compound and the previous compound
on the list.

27-25. What would be the effect of the following on the plate height of a column?
(a) increasing the weight of the stationary phase relative to the packing weight.
(b) decreasing the rate of sample injection.
(c) increasing the injection port temperature.
(d) increasing the flow rate.
(e) reducing the particle size of the packing.
(f) decreasing the column temperature.

(a) Increasing the weight of the stationary phase relative to the packing weight.
This is essentially the same as making the stationary phase film thickness greater,
which will increase the plate height due to increasing the stationary phase
component of the mass transfer term.
(b) Decreasing the rate of sample injection.
Decreasing the rate of sample injection will cause all of the peaks to become
broadened, while this has nothing to do with the column, and therefore does not
represent a modification of the plate height (as described by the vanDeemter
equation) it will nonetheless lower the efficiency and increase the calculated plate
(c) Increasing the injection port temperature.
Increasing the injection port temperature (assuming no degradation of the analytes
or solvent) will enhance evaporation. This produces the opposite effect of slow
injections, because it makes the sample vaporization as quick as possible, and thus
makes a tighter more confined plug of sample at the start of the column. So
increasing the injector temperature will improve efficiency and decrease the plate
height. Again, you should note, as in the previous case this improvement is
unrelated to the column and has nothing to do with the van Deemter equation.
(d) Increasing the flow rate.
Increasing the flow rate from zero will first improve decrease the plate height, and
then above the optimum flow rate (minimum plate height) it will increase the
plate height.
(e) Reducing the particle size of the packing.
Reducing the packing particle size decreases the distance that the analyte needs to
travel in the mobile phase to gain access to the stationary phase. This decreases
the mobile phase mass transfer contribution to the plate height.
(f) Decreasing the column temperature.
Decreasing the column temperature will decrease the extent of longitudinal
diffusion, and this will improve efficiency leading to smaller plate heights.
However, decreasing diffusion will also reduce the mass transfer of the analyte
between the mobile and stationary phases, and this will reduce efficiency by
increasing both of the mass transfer components to the plate height. The total
impact of column temperature will depend on the relative sizes of each of these
contributions (B smaller with lower temp) and (C gets larger with lower temp) to
the plate height.

28-4.How can the selectivity factor be manipulated in (a) gas chromatography; (b) liquid
The selectivity factor can be manipulated in much the same way as the retention factor in
the previous problem, because something that changes the retention factor MAY affect
the ratio of two retention factors, aka the selectivity factor.
a) In the case of GC the mobile phase composition is not as important as the column
temperature. Changing the column temp can change the equilibria for both analytes, if it
changes one more than the other then there will be a change in the selectivity. This could
simply be via a different temp isothermal run or it could involve temperature
programming. Alternatively you could change the composition of the stationary phase,
thereby enhancing the retention of one analyte over the other, really obvious in chiral
b) In the case of HPLC, the temp can also be programmed or changed, but more
significant impact will likely come from variation of the mobile phase composition, via
different isocratic runs, or gradient elution. Of course altering the composition of the
stationary phase is also a solution for LC. One last possibility, which is similar to
changing the mobile phase composition, is to add an additive to the mobile phase that
complexes or interacts with one of the analytes.

28-13. In a normal phase partition column, a solute was found to have a retention time of
29.1min, while 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 (a) k’ for the solute
and a (b) solvent composition that would bring k’ down to a value of about 10.
Using table 28-2 on page 743 and the equation for normal phase partition
chromatography on page 742 (28-3), but first we need to determine the initial value for
the retention factor k’i = tr-tm / tm = 29.1-1.05/1.05 = 26.71 (quite large). The initial value
for the polarity index was Pi = 0.5Phexane + 0.5Pchloroform = 0.5(0.1+4.1) = 2.1. To reduce
the value to 10 then 10/26.71 = 10(Pi-Pf)/2 thus Pf = 2.1 – 2 log(10/26.71) = 2.95. to get this
polarity index 2.95 = 0.1x + 4.1 - 4.1x and thus x = 0.288. So 28.8% hexane with 71.2&

28-15. Suggest a type of liquid chromatography that would be suitable for the separation

(a) and
(b) CH3CH2OH and CH3CH2CH2OH
(c) Ba2+ and Sr2+
(d) C4H9COOH and C5H11COOH
(e) high-molecular-weight glucosides
a) These extremely nonpolar polyaromatic hydrocarbons are well suited to adsorption
b) These small polar (essentially non-ionizable) molecules will be well suited to normal
phase partition chromatography
c) Atomic ions such as these are best separated with ion exchange chromatography,
although ion pair would also be a possibility.
d) These short chain ionizable highly polar molecules are well suited for normal phase
and possibly ion pair chromatography (in fact these were also described in the text as an
example for ion exclusion chromatography)
e) Of the techniques introduced so far, high molecular weight glucosides will be best
separated with size exclusion chromatography.

28-16. What is a suppressor column and why is it employed?

The suppressor column is an ion exchange column that is opposite to the kind of ion
column used for the analytical separation. That is, if you are separating anions with the
analytical column, then the suppressor would be a cation exchanger. The primary
function of the ion suppressor column is to decrease the ionic conductivity of the mobile
phase that often contains a high fraction of supporting electrolyte (in the form of H+ or

28-17. On a silica gel column, a compound was found to have a retention time of 28.0min
when the mobile phase was toluene. Which solvent, carbon tetrachloride or chloroform,
would be more likely to shorten the retention time?
From table 28-2 on page 743 you can find the polarity indices for these three solvents
toluene is 2.4, CCl4 is 1.6 and CHCl3 is 4.1. Recalling that this is normal phase you
would use equation 28-3 on page 742, which tells you that smaller retention indices and
thus smaller retention times will result when the polarity index for the mobile phase is
larger. So to shorten the retention time you should use a solvent that has a higher polarity
index and in this set of solvents that is chloroform.

28-18. For a normal phase separation, predict the order of elution of

(a) n-hexane, n-hexanol, benzene
(b) ethyl acetate, diethyl ether and nitrobutane.
For part (a) the elution order would be first n-hexane, benzene and then n-hexanol, and
for part (b) first diethyl ether, ethyl acetate and then nitrobutane
In normal phase partition chromatography, the stationary phase is more polar than the
mobile phase, and thus the more polar compounds will be retained longer than the less
polar compounds. For molecules with similar polarities, elution will depend on the
molecular polarizability.