Centeno, Carl Jerome N.

Date Performed: April 26, 2018

2015-09151 Date Submitted: May 10, 2018

CHEM 140: Organic Analysis

Exercise 5: Polyfunctional Mixture

Unknown Samples:
4-bromoacetophenone 4-nitrobenzaldehyde

Postlab Discussion:

1. During the selection of appropriate solvent system, take photograph of the TLC plate after iodine vapor
visualization and paste them into your laboratory notebook

1 2 3 4 5 6 7
\
a. What happens
2 when you increase the polarity of the solvent system?

The solvent system constituents optimized in the exercise were hexane and ethyl acetate. Based
from their chemical structures (Figure 1), hexane is the non-polar part of the solvent system
since it does not contain any electronegative elements, as compared to ethyl acetate, which alter
the dipole moment of the molecule. Changing the polarity of the solvent system depends on
what type of chromatography was employed for sample separation. In this exercise, a mixture
of two compounds were separated using thin layer chromatography (TLC).
 A B
Figure 1. Chemical structures of (A) hexane, and (B) ethyl acetate.

TLC is a type of partition chromatography where a thin film of the liquid stationary phase can
form on the surface of a solid support. The stationary phase is a nonvolatile polar liquid held
as a thin layer on the surface of an inert solid. The mixture to be separated is usually carried by
a non-polar liquid or gas mobile phase. The more soluble component in the mobile phase will
be the fastest to reach the ends of the chromatographic plate after the solute disseminate
themselves between the stationary and mobile phases.

When the polarity of the solvent system is increased, the polar molecules present in the sample
will have a greater interaction with the polar stationary phase thereby allowing the sample to
be retarded on the plate while the non-polar part of the sample will be easily dissolved by the
mobile phase causing it to move higher up the plate. In addition, increasing the polarity of the
solvent system will cause an overall increase in the retention factor (Rf) values of all the
samples since it interacts with the polar stationary phase causing the samples to move higher
up the plate. However, changing the polarity of the mobile phase will normally not result in the
reversal of the Rf values.

The Rf of the sample is dependent on the interactions of the polar stationary phase and the
solvent system used. If the solvent-sample interaction is greater than the silica-sample
interaction, then the sample analyzed shall move higher up the plate since it spends less time
interacting with the stationary phase.

b. What is the most appropriate solvent system for separating the components of the mixture?
discuss your answer.

To determine the optimal solvent system for the unknown mixture, the concentrations of the
polar and non-polar moieties of the solvent system were evaluated. As seen from table 1,
different solvent systems with increasing polarity were formulated to determine the optimal
solvent system for the 4-bromoacetophenone and 4-nitrobenzaldehyde mixture. Solvent
systems with pure hexane and ethyl acetate were also prepared. Based from the ΔRf values
presented (Figure 2), solvent 6 (3 Hexane: 7 EtOAc) was the most appropriate solvent to
separate the samples. Solvent 6 yielded a ΔRf value of 0.25 and two distinct spots at 2.8cm and
4.2cm. It is also worth noting that solvent 2 (Pure EtOAc) yielded the highest ΔRf value of
0.267857143 however, the solvent system was not used since its is highly polar and can
increase the individual Rf values of the sample causing decreased separation.
Table 1. Data on the effect of increasing solvent system polarity on the Rf values of the unknown mixture.
Solvent Distance Travelled (cm) Rf Values
Solvent System Solvent Front (cm) ΔRf
No. Spot 1 Spot 2 Spot 1 Spot 2
1 Pure Hexane 6.3 0.3 0.8 0.047619048 0.126984127 0.079365079
2 Pure EtOAc 5.6 3.4 4.9 0.607142857 0.875 0.267857143
3 9 Hexane: 1 EtOAc 6 1.7 2.6 0.283333333 0.433333333 0.15
4 7 Hexane: 3 EtOAc 6.1 2.7 3.4 0.442622951 0.557377049 0.114754098
5 1 Hexane: 1 EtOAc 6.4 2.6 3.6 0.40625 0.5625 0.15625
6 3 Hexane: 7 EtOAc 5.6 2.8 4.2 0.5 0.75 0.25
7 1 Hexane: 9 EtOAc 5.5 4.2 4.4 0.763636364 0.8 0.036363636

ΔRf Values
0.3

0.25

0.2
ΔRf values

0.15

0.1

0.05

0
0 1 2 3 4 5 6 7 8
Solvent Number

Figure 2. ΔRf Values of solvent system at different polar-non-polar proportions.

2. After performing column chromatography, were you able to separate the two compounds? Which
compound eluted first? Draw the structures of the two compounds and discuss why that compound
eluted first using the chosen solvent system. If you were not able to separate the compounds, what
recommendations can you give to improve the experiment?

The two-compound mixture were separated using the 3 Hexane: 7 EtOAc solvent system, the TLC
chromatograms for 4-bromoacetophenone and 4-nitrobenzaldehyde were presented on figure 3.
 A B
Figure 3. TLC chromatograms of (A) 4-bromoacetophenone, and (B) 4-nitrobenzaldehyde.

After solvent optimization, the sample mixture was subjected to column chromatography to
separate its individual components. Thirty (30) test tubes with 1mL eluent were collected and
subjected into two chemical tests: (1) Jones oxidation, and (2) 2,4-DNP test, to differentiate the
eluted compounds. Results have shown that the first component, 4-nitrobenzaldehyde (Figure 4),
was eluted at 11mL since the eluent tested positive on jones oxidation (Figure 5) and negative on
2,4-DNP test. 4-nitrobenzaldehyde yielded a retention factor (Rf) value of 0.116666667. On the
other hand, the second component, 4-bromoacetophenone (Figure 4), was eluted at 14mL based
from its positive result on the 2,4-DNP test (Figure 5) and its negative result on jones oxidation. 4-
bromoacetophenone yielded a retention factor (Rf) value of 0.754385965.

A B
Figure 5. The chemical structure of (A) 4-bromoacetophenone and (B) 4-nitrobenzaldehyde.
 A B
Figure 6. Positive result for (A) Jones oxidation, and (B) 2,4-DNP test.

4-nitrobenzaldehyde eluted first on the column chromatography due to its high affinity towards the
polar solvent. However, its movement was retarded during TLC since the solvent-sample
interaction is less than the present silica-sample interaction causing it to interact more with the
stationary phase. On the other hand, 4-bromoacetopenone is less polar compared to 4-
nitrobenzaldehyde since it contains less electronegative groups thereby causing it to be eluted last.