Experiment 9

AMINES
Arenas, E., Bacay, J.A., Siruno, A.
College of Science
University of the Philippines Baguio

Abstract

Amines are organic derivatives of ammonia in which one or more hydrogen atoms have
been replaced by an alkyl or aryl group. In this experiment, characterization of amine
was done in order to understand the behavior of amines through different reactions and
also to differentiate the types of amines. In the basicity of amines, three representatives
were tested. The experimental pH values obtained from these representatives were
closely similar to their theoretical pH values except for aniline. In the carbylamines test,
primary amines react with choloroform and KOH to form isocyanide that has a fishy odor.
In the third experiment, aniline was mixed with concentrated H2SO4 resulting to salt
formation. The last part of the experiment is the Hinsberg test which was done in order
to identify the primary, secondary and tertiary amine among the representative amines.
The primary amine will produce sodium salt of sulfonamide as product and is soluble to
aqueous sodium hydroxide solution, acidification of which results to the precipitate of
sulfonamide and this was observed in aniline and thus identified as primary amine.
Secondary amine will also form sulfonamide however it do not contain acidic amide
hydrogen therefore not soluble to aqueous sodium hydroxide solution, acidification of
sulfonamide product that are insoluble to aqueous sodium hydroxide solution will still not
make it soluble, this is observed in methyl aniline and thus identified as secondary amine.
Tertiary amines do not react in benzenesulfonyl chloride but reacts to sulfonyl chloride
to form insoluble salt of benzenesulfonic acid but then dissolved on acidification with HCl
since a water soluble amine hydrochloride salt is formed. Amines are widely used in the
industry from producing bulletproof vest for military purposes to biochemical uses. An
important amine in the body is the serotonin which is one of the primary neurotransmitters
for the brain.

I. Introduction

Amines are derivatives of ammonia, NH3, in which one or more hydrogen atoms of ammonia have been
replaced by an organic group. They are classified as primary, secondary, or tertiary, according to the
number of groups attached to the nitrogen. The structures of primary, secondary, and tertiary amines are
presented by Figure 1.

Figure 1. Primary, secondary and tertiary structures

of Amines

Retrieved from:
https://chem.libretexts.org/Bookshelves/Introductory_Chemistry/Book%3A_The_Basics_of_GOB_Che
mistry_%28Ball_et_al.%29/15%3A_Organic_Acids_and_Bases_and_Some_of_Their_Derivatives/15.1
0%3A_Amines_-_Structures_and_Names
The designation of amines as primary, secondary, and tertiary is different from the usage of these terms in
connection with alcohols and alkyl halides. In these latter two cases there is only one organic group (R), so
that the terms are used to designate the type of carbon to which the alcohol or halide function is attached.
(“Amines”, n.d.).

The lower aliphatic amines are gases with fishy odor. Primary amines with three or more carbon
atoms are liquid and still higher ones are solid. Aniline and other arylamines are usually colorless but get
colored on storage due to atmospheric oxidation. Lower aliphatic amines are soluble in water because they
can form hydrogen bonds with water molecules. However, solubility decreases with increase in molar mass
of amines due to increase in size of the hydrophobic alkyl part. Higher amines are essentially insoluble in
water. Considering the electronegativity of nitrogen of amine and oxygen of alcohol as 3.0 and 3.5
respectively, you can predict the pattern of solubility of amines and alcohols in water. (National Council of
Educational Research and Training, 2015).

Basicity of amines is related to their structure. Basic character of an amine depends upon the ease
of formation of the cation by accepting a proton from the acid. The more stable the cation is relative to the
amine, more basic is the amine.

There are different reactions that amines undergo. One of which is the carbylamine test. This test
is done to distinguish primary amines from secondary amines. The carbylamine test is also called the
isocyanide test due to the formation of isocyanide as a positive indicator of the test. Secondary and tertiary
amines will not react with chloroform (CHCl3) and alcoholic potassium hydroxide (KOH) but primary aliphatic
and aromatic will react with the reagents. (Tagala, n.d.)

Another is the Hinsberg Test wherein another electrophilic reagent, benzenesulfonyl chloride,
reacts with amines in a way that provides a useful test for distinguishing primary, secondary and tertiary
amines. This test is conducted in aqueous base (NaOH or KOH), and the benzenesulfonyl chloride reagent
is present as an insoluble oil. Because of the heterogeneous nature of this system, the rate at which the
sulfonyl chloride reagent is hydrolyzed to its sulfonate salt in the absence of amines is relatively slow. The
amine dissolves in the reagent phase, and immediately reacts (if it is 1º or 2º), with the resulting HCl being
neutralized by the base. The sulfonamide derivative from 2º amines is usually an insoluble solid. However,
the sulfonamide derivative from 1º amines are acidic and dissolves in the aqueous base. Acidification of
this solution then precipitates the sulfonamide of the 1º amine. (Reusch, 2013).

In this experiment, the physical and chemical properties of amines are determined through different
reactions which are discussed in the following paragraphs.

II. Results and Discussion

A. Basicity of Amines
In the first part of the experiment, the basicity of aniline, benzylamine and methyl aniline were tested using
pH paper. A drop of each were placed in each pH paper. The pH of each were recorded based on the color
each of them produced. Table 1 is the recorded pH of the three compounds.

Table 1. Basicity of amines

Amine pH
Aniline 7
Benzylamine 11
Methylaniline 8

Theoretically, the pH of aniline is 8.1 (O'Neil, M.J. (ed.) 2013), while benzylamine has the pH of 11.6
(Heuer L.,2008 ;John Wiley & Sons,2006) and Methylaniline has the pH of 7.6 . Comparing the pH recorded
from the experiment and the actual pH value of the compounds the recorded values are close to the actual
values except for aniline, perhaps the color patterns produced mistook for the pH of 7 instead of 8 since
consecutive color patterns are nearly the same it just varies in darkness and intensity of bright colors. There
are five factors that affect the basicity of amines. First the basicity increases as negative charge on nitrogen
also increases, thus, the more lone pairs the nitrogen gets the more basic the amine is. Figure 2 below
illustrates this factor.

Figure
Figure2.3 The
The effect
effect of
of charge
charge in
in the
the basicity
basicity of
of amines
amine
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amines/

The second factor that affects the basicity of the amine is the resonance or conjugated vs. non-conjugated
amines. The more resonance structures or more lone pairs of electrons that can be delocalized into larger
pi system the lower the basicity. The higher the pKaH value is, the stronger base the compound will be.
Figures 3 and 4 illustrates the process of the second factor.
 Figure 3. Effects of resonance in basicity of amines

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amines/

Figure 4. Resonance-stabilized amines exhibit less basicity

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amines/
The test compounds used in the experiment, aniline has the pKaH of 4.63 while benzylamine has the pKaH
of 9.33 while methyl aniline has the pKaH of 4.85 so the basicity order is benzylamine is the most basic.
Next is methyl aniline and last is the aniline. Below are the structures of the three compounds.
 Aniline Benzylamine Methyl aniline

Images retrieved from:

https://pubchem.ncbi.nlm.nih.gov/compound/aniline/benzylamine/methylaniline#section=2D-Structure

The third factor that affects the basicity of amines is the inductive effects; the more electronegative the
substituent in comparison to the nitrogen itself, for instance oxygen or other halogens, the less basic the
amine will be. It is because, in such cases the nitrogen acts as “pi donor” instead of “pi acceptor”.
Simultaneously the presence of these more electronegative atoms than nitrogen decreases the pKaH value,
which further strengthens the lessened basicity of amines in such conditions. Figure 5 demonstrates the
process.

Figure 5. Inductive effect of electron withdrawing group to the basicity of amines

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amines/

The fourth factor is the hybridization, which implies that the less “s” character the amine have the more
basic it is. Figure 6 illustrates this process.
 Figure 6. The effect of hybridization in the basicity of amines
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amines/

The last and the fifth factor is the aromaticity regardless of tautomerism. It implies that in some cases
between amines with rings, aromatic amines are more basic than other amines that are with rings but not
aromatic. Figure 7 shows an example of this factor.

Figure 7. The effect of aromaticity to the basicity of amines

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amines/

B. Carbylamine Test

In the carbylamine Test, a primary amine is reacted to chloroform (CHCl3) and alcoholic potassium
hydroxide (KOH) to form isocyanide. In table 2, the gathered data from the experiment is stated.

Table 2. Carbylamine Test

Action taken Observations
Aniline Brown liquid
+ chloroform Heterogenous mixture
+ KOH Fishy odor was produced
*after heating: persistent foul odor
*+ 1.00mL of concentrated HCl: white ppt. and foul odor less persistent
The carbylamines test or also known as Hofmann isocyanide synthesis is a test to determine the presence
of a primary amine. These primary amines can be aliphatic or aromatic. (Chang, 2010) In this reaction, a
primary amine, chloroform and a base to synthesize isocyanides. The mechanism of this reaction includes
a formation of a dichlorocarbene intermediate through the process of dehydrohalogenation of chloroform
with the addition of amine. The formation of the dihalocarbene intermediate is vital in the process of
conversion since it serves as the electrophile wherein the lone pair electrons of the nitrogen atom will donate
to. The positive result of this test is indicated by the production of a fishy smell due to the formation of
isocyanide or carbylamines. In figure 8, the mechanism of the carbylamine reaction is illustrated.

Figure 8. Mechanism of the carbylamine test

The first step of the reaction is the dehydrohalogenation of chloroform (CHCl3) to have a dichlorocarbene
intermediate. In the dehydrohalogenation, there is a removal of a hydrogen halide in the substrate. The
dichlorocarbene intermediate is highly reactive due to its charge. Second step would be the attack of the
electrophilic dichlorocarbene to the nucleophilic nitrogen of the primary amine. In the last step the
elimination of the hydrochloric acid will then form the isonitrile product or isocyanide.

C. Salt Formation

In the third part of the experiment, 3 drops of aniline and 4 drops of concentrated sulfuric acid produces
sulfate or bisulfate of the anilinium ion. The data from the experiment is summarized in table 3 and reaction
mechanism is shown in figure 9.

Table 3. Salt Formation

Action taken Observations
Aniline Brown liquid
+ concentrated H2SO4 1st drop: evolution of gas
After 4 drops: formation of solid salt crystals
 Figure 9. Mechanism of the formation of anilinium ion

Image created on www.chemdoodle.com

Sulfuric acid is a strong acid thus the reaction was hasten and the formation of bisulfate was fast. Upon the
attack of the nitrogen on one of the hydrogen of sulfuric acid the bond between the hydrogen and oxygen
in the sulfuric acid was then break and the electron went back to the oxygen atom. The product produced
where in NH3 was obtained as a meta-director and it pulls electrons from the ring causing the slow
attachment of SO3H-.

D. Hinsberg Test

The Hinsberg test is used to distinguish a primary amine from secondary amine and tertiary amine.
Table 4 below summarizes the results in using two reagents, namely, benzenesulfonyl chloride and KOH.

Table 4. Hinsberg Test

Observations
Test Compounds
Benzenesulfonyl chloride +KOH
Test compound is soluble with
aniline Formation of residue
pH 14
Formation of brown solid layer
methyl aniline Formation of residue
with pH 14
Formation of two distinct layers
triethyl amine No formation of residue
with pH 14

Theoretically, the following reactions occurred during the experiment. For the primary amine (aniline), which
reacted with benzenesulfonyl chloride (A) to produce sulfonamides (B) which contains an acidic amide
hydrogen that reacts with KOH to form the salt of the sulfonamide (C). The salt is soluble in the aqueous
KOH solution. The acidification of the solution produces a precipitate of the sulfonamide (B). The reaction
for primary amines are presented in Figure 10.
 KOH

Figure 10. General reaction of primary amines in Hinsberg Test with KOH

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amines

For the secondary amine (methyl aniline), it reacts with benzenesulfonyl chloride to form a sulfonamide (D)
that does not contain an acidic amide hydrogen, hence, it is not soluble in the aqueous KOH. Acidification
of the mixture dissolve the insoluble sulfonamide. Figure 11 presents the reaction of secondary amines in
the Hinsberg Test.

Figure 11. General reaction of primary amines in Hinsberg Test with KOH

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amines

Lastly for the tertiary amine (triethyl amine), does not react with benzenesulfonyl chloride. However, KOH
eventually reacts with the sulfonyl chloride to form salt of benzenesulfonic acid (E). Triethyl amine does not
dissolve in the initial reaction mixture, unless it is a low molecular weight soluble amine, and dissolves on
acidification with HCl since a water soluble amine hydrochloride salt (F) is formed. Figure 12 presents the
general reaction of tertiary amines in the Hinsberg Test.

Figure 12. General reaction of tertiary amines in Hinsberg test with KOH

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amines
 III. Summary and Conclusion

The basicity of amines is affected by five factors, inductive effect, hybridization, aromaticity, electron
pairs on the nitrogen atom and resonance structures. The higher the pKa of the amine the more basic it
becomes. The basicity of amines also depends on the number of pair electrons the nitrogen has, so the
primary amines are the most basic, next is the secondary amines and the least basic is the tertiary amines.
Among the compounds tested on the laboratory benzylamine is the most basic, next is the methyl aniline
and the least basic is the aniline.

The Carbylamine test is used to determine primary aliphatic or aromatic amines. Dehydrohalogenation
of the chloroform and the amine forms a dichlorocarbene intermediate. This test produces isocyanide
indicated by a production of a foul fish odor. Only primary amines can undergo this test due to the free
electron pair of nitrogen.

In the salt formation, sulfuric acid as a strong acid and aniline having NH 2 which is a ortho-para director
enabled the fast reaction resulting to formation of salt or bisulfate of anilium ion. Upon the attack of nitrogen
atom on one of the hydrogen atoms of the sulfuric acid cleaving then the bond between the hydrogen and
oxygen results to a salt having NH3 and SO3H- wherein NH3 is meta-director deactivator causing slow
attachment of SO3H- to the ring.

For primary amines, when added to Hinsberg test solution, gave a clear solution (indicating that it is
soluble), which, on acidification with HCl, formed a precipitate (residue). In secondary amines, on the other
hand, reacted with the Hinsberg test solution in which it exhibited a suspension of an insoluble solid or oil
(forming two distinct layers) which did not dissolve on acidification with HCl. And lastly, on tertiary amines,
in which there was no reaction that occurred with benzenesulfonyl chloride in the test solution, hence it
initially provided an insoluble solid or oil, which, on acidification with HCl, dissolved to give a clear solution
of the salt amine.

IV. References

Chang, R. (2010). Chemistry 10th Edition. New York: McGraw-Hill.

Factors of basicity of amines. Retrieved on April 19, 2019 from

https://www.masterorganicchemistry.com/2017/04/26/5-factors-that-affect-basicity-of-amines/

National Council of Educational Research and Training. (2015). Amines. Retrieved from http://ncert.nic.in/

Reusch, W. (2013). Amine Reactivity. Retrieved from https://www2.chemistry.msu.edu/faculty/reusch/

virttxtjml/amine1.htm

Tagala, J. (n.d.). Amines II and III. Retrieved from https://www.academia.edu/22538807/Amines

_II_and_III