Experiment 5

CHEM 613
Experimental Organic Chemistry
2013
Anastazija Ristovska

Post-Lab
Separation of Acidic, Basic & Neutral Compounds

The purpose of this experiment was to transform an organic acid, base and neutral
compound into water-soluble ionic forms and extract them in the water layer. The three
compounds were given to us by the TAs in a mixture dissolved in ether. Each student was given
a different combination of three compounds chosen from lists of acidic, neutral, and basic
compounds. Then based on the physical properties known for each compound their unknown
identity was determined. The list of possible acidic compounds was benzoic acid, 4chlorobenzoic acid, m-nitrobenzoic acid, and m-bromobenzoic acid. The possible basic
compounds were p-nitroaniline, 4-chloroaniline, p-toluidine, 2-aminobenzophenone, and 4aminoacetanilide. The possible neutral compounds list is: benzyl, fluorine, phenanthrene,
m-dinitrobenzene, and benzophenone.
The ether mixture is first extracted with acid in order to remove the base, separating the
aqueous layer that contains the organic acid and alkalizing it with a hydroxide a step that sets
free the organic base in a precipitate form. The remaining ether solution is extracted with a base
in order to isolate the organic base in the water layer; the water layer is then acidified with HCl in
order to set free the organic base in the form of a precipitate.
The neutral compound that is left in the organic layer is isolated by simply evaporating
the ether solvent which is very volatile anyways, and has a much lower boiling point than all the
neutral organic compounds on the list of possible unknown compound identities. Each time an
aqueous layer was separated a reverse extraction was performed on it with MBTE. The ether
used all throughout was methyl tert-butyl ether.
Melting points were used for the preliminary identification of the unknown compounds,
and their identities were confirmed using spectral analysis. The acidic compound was found to
be 4-chlorobenzoic acid with melting point 238-242C, the neutral compound was identified as
benzyl with m.p. 91-95C, and the basic compound was found to be p-toluidine with m.p. 4246C.

Acidic compound:

4-chlorobenzoic acid

m.p. 238-242C

Neutral compound:

Benzyl

m.p. 91-95C

Basic compound:

p-Toluidine

m.p. 42-46C

Initially the three unknown compounds mixture was given to me in the form of a dark
yellow slur that looked creamy, wet, and easily spreadable over surfaces. Its components being
organic substances, it was all easily dissolved in 6.25ml MBTE in a test tube. Using glassware
calibration on a precision scale, the exact weight of the slur administered to the test tube was
known to be 325.9mg. The test tube was closed with a cap and mixed vigorously until all the
yellow slur formed a uniform MBTE solution. 3ml 10% HCl was added to the test tube and the
tube was mixed vigorously with cap closed for about 1min. The solution was left to settle, and
two visible layers were obtained.
The aqueous layer was colorless and transparent on the bottom and the organic MBTE
layer was transparent yellow on the top portion of the test tube content. By adding the acid we
have separated the base by neutralizing it, isolating it in the aqueous bottom layer. Using a
pipette the bottom layer was extracted to another test tube. New 3ml 10% HCl were added to
the organic MBTE solution and the same process ending with pipette extraction was repeated all
over again three times.
It is important to note that the identity of the three compounds was still unknown, and
all students, each having different combinations of different compounds, were performing the
same procedure using the same organic solvent, and the same acid and base combination in same
concentrations. This means only one thing, that the procedure for separating most organic acids,
bases and neutral compounds should be same or very similar for each compound.
After 9ml of aqueous layer have been isolated in the new test tube, a reverse extraction
was performed on the using MBTE, in order to recover as much as possible of any acidic or
neutral compound that might have escaped into the aqueous layer in tiny MBTE droplets
because of pipette technique imprecision. This extract was returned to the original test tube.
After the reverse extraction a generous amount of 10% NaOH was added to the aqueous
extracts and the formation of a precipitate was observed. The test tube got heated, indicating an
exothermic reaction upon restoring the compound its basicity.
It seemed like the precipitate formation was highly sensitive to pH. When measuring the
pH of the solution after the first addition of NaOH, it was in the range 0-1. Still more hydroxide
needed to be added. When a very basic environment was achieved, the test tube was placed on a
water heat bath, which finally allowed for precipitate formation. The product was isolated using
vacuum filtration. The identity of the organic base was determined to be p-toluidine using MS,
IR, and melting point analysis.
2

Turning back to the original test tube now containing only the acidic and neutral
compounds dissolved in MBTE, 3-4 portions of 2ml 10% NaOH were added. All throughout
the organic layer maintained yellow color. After vigorously mixing and letting the two phases i.e.
two layers settle, the aqueous NaOH layer was transferred to another (third) test tube. A
generous amount of 10% HCl was added to this test tube until the formation of a white flaky
precipitate was observed. The test tube heated, indicating that the restoration of the acidity of
the organic acid was an exothermic reaction in this case.
This precipitate was the acidic compound. It was isolated using vacuum filtration, and
through MS, IR, and melting point analysis its identity was determined to be 4-chlorobenzoic
acid. The yielded amount of 4-chlorobenzoic acid was 97mg.
In order to isolate the neutral compound we needed to do more work with the MBTE
organic layer in the initial test tube that now was left containing only the neutral organic
compound but was still yellow. The solution was rinsed with 3ml water and the aqueous layer
was extracted, repeating the procedure three times.
All extraction and washing processes could have as well been performed using a
separation funnel; however due to the small solution volumes we were working with pipette
extraction was more appropriate. The test tube was then placed in a water heat bath, and the
ether layer was allowed to evaporate. When all the ether evaporated, only the neutral compound
was left in the test tube. Its identity was determined using MS, IR, and melting point, and was
found to be benzil. The yielded amount of benzil was 7.4mg.

1878
1324

1557

2861

768
633

669
689

1096

1021

1121

718

1178

1450

2920
2963
3012
3024

3222
3339

3419

1264

1621
1517

812

Figure 1: p-Toluidine FTIR (KBr) Spectrum

The organic base spectral analysis is shown in figure 1; p-Toluidine has a methyl group
attached to a benzene ring, and an amino group attached at position 4 relative to the methyl
group on the benzene ring. The bands at 3339cm-1 and 3419cm-1 are indicative of the two N-H
bonds vibrations. The band at 3222cm-1 comes from the C-H aromatic vibrations. The stretches
between 2861cm-1 and 2963cm-1 come from the CH3 vibrations. The 3012cm-1 and the 3021cm-1
stretches are most likely due to the C-H aromatic vibrations. The peak at 1621cm-1 comes from
the double bond C=C vibrations of the benzene ring. The 1517cm-1 vibration is also most likely
due to the in-ring C-C vibrations (symmetric or asymmetric). The 1246cm-1 peak comes from the
C-N bond stretch. Between 669cm-1 and 812cm-1 there is a C-H aromatic oop stretch.

106

100

50
NH2

15

10
20
(mainlib) p-Aminotoluene

28 30
30

37 39 41 43
40

77 79

51 53 55
50

61 63 65 67
60

73 75
70

81
80

104

86 89 91 93
90

100

110

120

Figure 2: p-Aminotoluene Mass Spectrum

The molecular weight peak in the mass spectrum is given at m/z=107. The m/z=106
somes when one proton is separated from anywhere in the molecule, and we can only speculate
where this proton was cleaved off relative the molecule stability and the chance that after the
proton leaves the molecule will attain positive charge. Due to the fact that all the other peaks in
this mass spectrum are very short compared to the molecular peak and the full molecular ion
peak, it can be concluded that p-aminotoluene is a relatively stable molecule. The peak at
m/z=77 represents that molecular fragment left when both the methyl group as well as the
amino group are cleaved off from the benzene ring. The peak at m/z=79 results from the
cleavage of the molecule such that a -C-CH3 fragment is separated and what is left is five carbon
atoms of the benzene ring together with the amino group minus one proton. It can as well result
when the C-NH2 fragment of the molecule is cleaved off.

Figure 3: Benzil FTIR (KBr) Spectrum

The benzil compound mass spectrum is expected to have a carbonyl stretch in the
1660cm-1 to 1760cm-1 range, and indeed we do see two carbonyl stretches, supposedly due to the
two carbonyl groups, one at 1660cm-1 and the other one at 1677cm-1. The 3064cm-1 stretch is
representative of the aromatic C-H bond vibrations. The 1579cm-1 and 1594cm-1 stretches are
the aromatic C-C in-ring vibrations. Between 696cm-1 and 676cm-1 there is an aromatic C-H
oop stretch.
105

100

77
50

51

27

39 43

20
30
40
50
(mainlib) Ethanedione, diphenyl-

74

57 61
60

70

85
80

117
90

100

110

120

149
130

140

150

165
160

210

181
170

180

190

200

210

220

Figure 4: Benzil Mass Spectrum

The molecular weight peak is at m/z=210 and it is very weak, meaning that the molecule is not
stable under the molecular fragmentation technique used in the mass spectrometer. The m/z=77
peak comes from the phenyl fragments, i.e. just the benzene rings after they have been cleaved
off from the rest of the molecule. The strongest peak is at m/z=105 and comes from the
fragment obtained when the benzil molecule is cleaved exactly in half, i.e. the single bond
between the two carbonyl carbon atoms O=C-C=O is broken.

139

100

156

OH

50

111
75

141

158

50

Cl

113
38
0

14 17

27 31

10
20
30
(mainlib) Benzoic acid, 4-chloro-

40

45

55
50

61 65 69
60

70

85
80

92
90

128

99 103
100

110

120

130

140

150

160

170

Figure 5: 4-Chlorobenzoic Acid Mass Spectrum

The MS analysis of the organic acid isolated via our separation technique shows its molecular
weight peak at m/z=156. The m/z=158 peak is likewise a molecular peak, and it is due to the
less abundant chlorine isotope 37Cl with 24% natural abundance. The 35Cl isotope showing in the
peak at m/z=156 is found in nature with 76% abundance. This similar pattern and peak height
ratio can be seen for m/z=139 and m/z=141, and for m/z=111 and m/z=113, indicating that
the fragments represented by those peak pairs contain chlorine. The m/z=111 peak represents
the benzene ring with just chlorine attached to it, i.e. the carboxyl functional group has been
cleaved from the molecule for this fragment to form. The m/z=139 peak is due to the cleaving
of the OH group only, leaving the rest of the molecule intact. The m/z=75 represents the
benzene ring only, after both the carboxyl as well as halogen functional groups have been
cleaved from it.