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College of Pharmacy
Department of Pharmaceutical Chemistry
A laboratory manual on
for second year students
HO
OH
Contents
Tools and glass wares..................................................................................................... iii
Qualitative analysis of organic compounds ..................................................................... 1
Determination of solubility class ......................................................................................2
Identification of alcohols .................................................................................................6
Identification of aldehydes and ketones .........................................................................11
Identification of phenols ................................................................................................18
Identification of carboxylic acids ....................................................................................26
Identification of carboxylic acids salts ............................................................................37
Identification of alkyl and aryl halides ............................................................................39
Identification of amines .................................................................................................43
ii
reagent bottles
glass stoppers
dropper
Bunsen burner
washing bottle
beaker
test tubes
tripod stand
wire gauze
iii
litm
mus paperr
spatula
spatula
a
Peetri dish
graduaated cylin
nders
funnel
filteer papers
gradu
uated pipette
glass rod
r (stirreer)
water bath
h
Hood
iv
Qualitative Analysis of
Organic Compounds
H2O
insol.
sol.
5% NaOH
insol.
Ether
sol.
sol.
5% NaHCO3
5% HCl
insol.
sol.
sol.
SI1
AI 2
litmus red
sol.
SI2
litmus blue
insol.
litmus unchanged
96%
cold H2SO4
insol.
insol.
AI 1
Discussion on solvents
Water
Water is a polar solvent with a dielectric constant equals to 80. It
has the ability to form hydrogen bonding and can act either as an acid or a
base. Therefore it can dissolve:
Salts of ammonium ion (RNH4+) or organic acids salts with
alkali metal cations (RCOO-).
Ionic compounds.
Polar compounds like dissolves like.
Organic compounds with low molecular weight (less than 5
carbon atoms) such as alcohols, aldehydes, ketones, and
carboxylic acids.
Water is useful to determine the degree of acidity of a compound,
even if the compound is insoluble in water, using litmus paper (acidic,
basic, or neutral).
Water is the first solvent used to determine the solubility class of a
compound. If the compound is water soluble, the next step is to test its
solubility in ether.
Ether
Ether is a non-polar solvent having a dielectric constant of 4.3. It
cannot form hydrogen bonding (unassociated liquid). Therefore, it differs
from water in that it cannot dissolve ionic compounds such as salts. It
dissolves most water insoluble compounds; therefore, in the
determination of solubility class, the importance of ether is for watersoluble compounds only and no further solubility tests using the
remaining solvents are to be done.
Accordingly two probabilities are there:
1. Compounds soluble in both water and ether.
These compounds:
are non-ionic.
contain five or less carbon atoms.
contain an active group that is polar and can form hydrogen
bonding.
contain only one strong polar group.
This division of compounds is given S1class and includes, e.g., aldehydes,
ketenes, and aliphatic acids.
2. Compounds soluble in water only (but not in ether).
These compounds:
are ionic.
contain two or more polar groups with no more than four
carbon atoms per each polar group.
This group is classified as S2 class and includes ionic salts such as salts of
carboxylic acids and amines and compounds with more than one active
group such as poly hydroxylated compounds and carbohydrates.
Note that solubility in ether is tested only for water-soluble
compounds. For water insoluble compounds use the left side of the
solubility classification scheme, i.e. test solubility in sodium hydroxide
rather than ether.
5% NaOH & 5% NaHCO3
Water insoluble compounds must be tested first in 5% sodium
hydroxide solution which is a basic solvent. It reacts with water insoluble
compounds that are capable of donating protons such as strong and weak
acids. The stronger the acid, the weaker the base it can react with. Water
insoluble compounds that dissolve in 5% sodium hydroxide solution must
also be tested for solubility in 5% sodium bicarbonate solution.
Therefore, for water insoluble acidic compounds sodium hydroxide
solution is considered as a detecting solvent whereas sodium bicarbonate
solution is called as a sub classifying solvent since it can react with strong
acids only. That is, these two solvents give an idea about the acidity
degree of the compound. Note that testing solubility in 5% sodium
bicarbonate solution is not needed if the compound is insoluble in 5%
Identification of Alcohols
R OH
the general
f ormula
H 3C OH
H2
C
H2
C
H3C
methanol
OH
ethanol
H3C
CH 3
CH
OH
sec-butanol
OH
C
C
H2
OH
H 3C
CH3
H 3C
OH
tert-butanol
cyclohexanol
benzyl alcohol
(cyclic)
(aromatic)
Physical properties
Alcohols are colourless liquids with a special faint odour. Benzyl
alcohol and cyclohexanol have characteristic odours.
Aliphatic alcohols burn with blue flame (without smoke) while
aromatic alcohols burn with yellow smoky flame.
Boiling points of alcohols are considerably high (being associated
liquids); they increase as the molecular weight (number of carbons)
increases.
Alcohols are miscible with water except benzyl alcohol,
cyclohexanol, and sec-butanol (which is very slightly soluble in
water.
Solubility classification
Alcohols are polar compounds because of the presence of the
hydroxyl group which is also responsible for their ability to form
hydrogen bonding. The degree of the polarity depends on the size
of the alkyl side chain; the polarity decreases as the size of the
alkyl side chain increases, or in another word, as the hydroxyl
group /hydrocarbon ratio of alcohols increases, their water
solubility increases and vice versa. Besides, low molecular weight
alcohols are soluble in water due to hydrogen bonding ability with
water molecules. Therefore, alcohols that are soluble in water and
ether are classified under class S1 such as ethanol and methanol.
Alcohols that are insoluble in water are related to class N such as
benzyl alcohol, sec-butanol, and cyclohexanol.
Which alcohol has solubility class S2, what are the
structural requirements present in this alcohol that
made it under this solubility class?
Chemical properties
Alcohols are neutral compounds that dont change the colour of
litmus paper.
All reactions of alcohols are related to its active hydroxyl group
and are of two types:
a) removal of the hydroxyl itself as in the reaction with hydrogen
halides to form alkyl halides or in the dehydration reaction to
form a double bond.
b) removal of the proton only from the active hydroxyl as in the
formation of esters or in the reaction with active metals such as
sodium.
1. General test (Ceric ammonium nitrate reagent)
Ceric ammonium nitrate (yellow solution) is an oxidizing agent
that reacts with alcohols to give a red complex and with phenols to give a
brown to greenish brown precipitate.
O
2[(NH 4)2Ce(NO 3)6] + RCH2OH
ceric ammonium
nitrate
alcohol
alcohol
(yellow)
(red)
Procedure
Water soluble (miscible) alcohols;
mix two drops of the alcohol with one drop of ceric ammonium
nitrate solution. A red complex indicates a positive test.
Water insoluble (immiscible) alcohols;
mix two drops of the alcohol with 0.5 ml dioxane, shake well, and
add one drop of the reagent to get a positive red complex.
This test gives positive results with primary, secondary, and
tertiary alcohols (up to 10 carbons), poly hydroxylated compounds such
as carbohydrates, and hydroxylated carboxylic acids, aldehydes and
ketones.
2. Specific tests
a) Iodoform (Haloform) test
This test is specific for alcohols which have a free methyl group
and a hydrogen attached to the carbon bearing the hydroxyl group such as
ethanol and sec-butanol.
OH
H 3C C
H
Procedure
(Note: read the procedure carefully and completely before starting)
Dissolve about 3 drops of the alcohol in about 2 mL of distilled
water (or 2 mL of dioxane for water insoluble compounds), add about 1
ml of 10% sodium hydroxide solution, then add iodine solution drop wise
with shaking until either a yellow iodoform precipitate is formed, in
which case the test is positive and is completed, or the dark colour of the
iodine solution is present. In the latter case allow the solution to stand for
3 minutes during which period check for the appearance of the yellow
precipitate at the bottom of the test tube. If there is no precipitate, warm
the solution in water bath (60 C) for about 3 minutes with shaking from
time to time and check for the yellow precipitate. During warming, if the
colour of iodine disappears, add few additional drops of iodine solution
with shaking until either the yellow precipitate is formed or the dark
iodine colour persists, and then complete warming. Then get rid of the
excess iodine by the addition of 10% sodium hydroxide solution drop
wise with shaking to obtain the yellow precipitate. If the precipitate is not
formed, allow the solution to stand for 10 minutes to get the positive
result. Finally if no precipitate is formed after the 10 minutes- standing
period, dilute the solution with an equal volume of distilled water to
obtain the iodoform precipitate. It is important to proceed through all
these steps so that only at the final step you can say that the test is
negative.
Both ethanol and sec-butanol give positive iodoform test and they
can be differentiated only by testing their solubility in water; sec-butanol
is less soluble in water than ethanol.
b) Lucas test
This test often provides classification informations on alcohols and
is used to distinguish between the different types of alcohols (primary,
secondary, or tertiary). It depends on the formation of alkyl chloride as a
second liquid phase.
Lucas reagent is prepared from anhydrous zinc chloride and
concentrated hydrochloric acid. Zinc chloride is added to increase the
ionization of hydrochloric acid.
ZnCl3- + H+
ZnCl 2 + HCl
R
OH + HCl
R
tertiary alcohol
ZnCl 2
Cl
R
tertiary alkyl halide
insoluble in water
+ H2O
Procedure
Mix 2-4 drops of the alcohol with few drops of Lucas reagent and
observe the results:
benzyl alcohol gives immediate result as shown by the appearance
of two phases.
tertiary alcohols give two phases that separate within 2-3 minutes.
secondary alcohols give two phases that separate after 15-20
minutes (giving a cloudy solution).
in primary alcohols one layer appears.
10
11
H3C
f ormaldehyde
benzaldehyde
O
C
acetaldehyde
O
H3C
CH3
O
C
CH3
OH
acetone
salicylaldehyde
acetophenon
O
O
C
C
H2
C
CH3
benzophenone
Physical properties
All aldehydes and ketones are liquids except formaldehyde, which
is gas (boiling point -21C), and benzophenone, which is solid
(melting point 48C). Formaldehyde is handled either as an
aqueous solution (formalin, an aqueous solution of 40%
formaldehyde and 15% methanol.) or as one of its solid polymers:
paraformaldehyde, (CH2O)n, or trioxane, (CH2O)3.
Low molecular weight aldehydes and ketones (less than 5 carbons)
are appreciably soluble in water, although they do not have the
ability to form hydrogen bonds (unlike alcohols), aromatic ones are
insoluble in water, and all of them are soluble in organic solvents.
They are colorless except benzaldehyde, which has a pale yellow
colour (due to oxidation) with a characteristic odour.
The boiling points of aldehydes and ketones are lower than those of
the alcohols from which they are derived; isopropyl alcohol boils at
82.5C while its oxidation product, acetone, boils at 56C, ethanol
boils at 78C while its oxidation product, acetaldehyde, boils at
21C.
Aliphatic aldehydes and ketones burn with a blue flame (without
smoke) while aromatic ones burn with a yellow smoky flame.
Solubility classification
Aldehydes and ketones that are soluble in water are soluble in ether
too and are classified under class S1(e.g., formaldehyde and
acetone).Aldehydes and ketones that are insoluble in water are
classified under class N such as benzaldehyde and benzophenone.
Chemical properties
All reactions of aldehydes and ketones are related to the carbonyl
group (the active group).
Aldehydes contain a hydrogen atom attached to its carbonyl while
ketones dont. This difference in the chemical structure affects their
chemical properties in two ways:
a) aldehydes are easily oxidized to the corresponding acids and
have reducing properties while ketones are not oxidized under
similar conditions and do not show reducing properties.
b) aldehydes are usually more reactive than ketones towards
nucleophilic addition, the characteristic reaction of carbonyl
group.
12
13
H
R
C
O+H
OH
NO 2
R
O2N
NHNH2 +
+
C O
R
O2N
R
2,4-dinitrophenyhydrazine
+ H2O
N N C
R
H
2,4-dinitrophenyhydrazone
(imine)
yellow-orange ppt.
Procedure
To 2 drops of the compound add 3 drops of the reagent, a yellow or
orange precipitate will be formed. If the compound is insoluble in water,
dissolve it in 1 mL of methanol and then add the reagent.
2. Differentiation between aldehydes and ketones
Differentiation between aldehydes and ketones is achieved by taking
the advantage of the fact that aldehydes can be easily oxidized while
ketones cannot (they need stronger oxidizing agents).Two reagents can be
used for this purpose, Tollens reagent or Fehlings reagent. Only
aldehydes give positive results with these two reagents.
a) Tollen's test (Reduction of ammoniacal silver nitrate)
Tollens reagent is the combination of silver nitrate solution with
ammonium hydroxide in the presence of sodium hydroxide solution.
Aldehydes show positive result with this reagent because the reaction
between them involves the oxidation of the aldehyde to the corresponding
carboxylic acid with an accompanying reduction of the silver ions from
this reagent to silver element in the form of silver mirror on the inner side
of the test tube.
Procedure
Preparation of Tollens reagent
To 3mL of silver nitrate solution add 2-3 drops of 10% sodium
hydroxide solution, and then add drop wise very dilute ammonia
solution with continuous shaking until all the brown precipitate of
silver oxide is dissolved. This reagent should be freshly prepared
prior before use.
Add 2-3 drops of the compound to 2-3 mL of Tollens reagent, a
silver mirror will be formed. If no reaction occurs, warm the test
tube in water bath for few minutes (note that excessive heating will
cause the appearance of a false positive test by decomposition of
the reagent).
The formed silver mirror can be washed using dilute nitric acid. If
the test tube is not very clean, silver metal forms merely as a granular
gray or black precipitate. False-negative tests are common with water
insoluble aldehydes. A negative result indicates that the compound is a
ketone.
b) Reduction of Fehling's reagent
This test, like Tollens test, is used to distinguish aldehydes from
ketones. Only aldehydes can reduce Fehlings reagent (a deep blue
solution) to give a red cuprous oxide precipitate.
14
Procedure
Preparation of Fehling's reagent
Fehlings reagent is prepared by mixing exactly equal volumes of
Fehlings A and Fehlings B solution in a 1:1 ratio immediately
before use (usually 1 mL of each).Fehlings A solution is an
aqueous solution of copper sulfate pentahydrate (CuSO4.5H2O)
with few drops of concentrated sulfuric acid whereas Fehlings B
solution is an aqueous solution of potassium sodium tartrate
(C4H4KNaO6,4H2O) and sodium hydroxide.
Add 5 drops of the compound to 1 mL of Fehlings solution, and
then heat in water bath for 5 minutes (with shaking for water
insoluble compounds).
Aldehydes change the color of Fehlings solution from blue to green,
orange precipitate, and then red precipitate or copper mirror. Ketones
dont change the color of this reagent. On the other hand, this test does
not give a sharp result with aromatic aldehydes.
3. Special tests for aldehydes and ketones containing a terminal
methyl group
These compounds include acetaldehyde, acetone, acetophenone, and
benzyl methyl ketone. All of them have a methyl group attached to the
carbonyl group:
15
16
OH
O
+
OH
resorcinol
conc. H2SO4
C
H
HO
HO
OH
- H2O
OH
formaldehyde
OH
polymer
5. Cannizzaro reaction
Benzaldehyde, salicylaldehyde, and formaldehyde can undergo
Cannizzaro reaction because they do not have an alpha hydrogen atom.
In this type of reactions the aldehyde undergoes a self oxidationreduction in the presence of a strong basic medium to yield a mixture of
the corresponding alcohol and the salt of the corresponding carboxylic
acid (or the acid itself). Therefore, one molecule of the aldehyde serves as
the oxidizing agent while the other serves as the reducing agent.
O
C
benzaldehyde
O
H
C
+
O
H
NaOH
OH
heat
benzaldehyde
benzyl alcohol
C
+
ONa
sodium benzoate
Procedure
To few drops of benzaldehyde (or the other aldehydes) add 0.5 mL
of 30% sodium hydroxide solution and heat gently on a water bath with
shaking for five minutes. A precipitate of sodium benzoate is produced.
Dissolve this precipitate by adding few drops of distilled water, and then
add drops of concentrated hydrochloric acid to liberate benzoic acid as a
white precipitate.
As mentioned earlier formaldehyde can undergo this reaction
; however, this reaction can't be relied on for testing formaldehyde since
the acid produced, formic acid, is liquid that can't be observed separately
as compared to the solid benzoic acid resulted from benzaldehyde.
17
18
Identification of Phenols
OH
OH
OH
OH
phenol
hydroquinone
(quinol, hydroquinol)
resorcinol
OH
OH
OH
CH 3
CH3
o-cresol
m-cresol
CH3
p-cresol
HO
OH
-naphthol
-naphthol
19
Physical properties
Phenols are liquids (e. g., o- and m-cresol) or solid crystalline
compounds(e. g., phenol and resorcinol).
They are coloured due to air oxidation and have a special odour.
Pure compounds are colourless.
They have high boiling points because of intermolecular hydrogen
bonding.
Phenol itself is soluble in water due to its ability to form hydrogen
bonding with water. For other phenols the solubility in water
decreases by increasing the molecular weight. (cresols and
naphthols are insoluble in water).
Phenols burn with a yellow smoky flame due to the presence of
aromatic ring.
Chemical properties
Phenols are weak acidic compounds, so they are soluble in strong
alkaline solutions only (e. g., sodium hydroxide solution). For this reason
they fall into solubility class A2. Presence of electron withdrawing group
at the phenyl ring strengthens the acidity of the phenol making it of
solubility class A1 (e. g, nitrophenol). However, phenol itself is of
solubility class S1since it is water soluble.
Types of phenols reactions
1. Reactions at the phenolic hydroxyl group (-OH); e.g., ether
formation and salt formation:
phenol reacts with sodium hydroxide to form sodium
phenoxide.
OC2H5
+ C2H5I
phenol
aq. NaOH
+ HI
heat
Chemical reactions
1. Reaction with ferric chloride
Phenols react with ferric chloride to give coloured compounds due to
the presence of the enol group. Actually this reaction is considered as a
test for any compound with enol group.
20
21
Procedure
To a very dilute aqueous solution of the phenol (30-50 mg in 1-2 mL
water) or to a few crystals of the solid phenol (50-100 mg) dissolved in
water add 1 drop of ferric chloride solution and observe the resulting
colour:
compound
colour
phenol, m-cresol,
resorcinol
violet or blue
o- and p-cresol
greenish blue
hydroquinone
deep green
- and -naphthol
no special colour
22
Procedure
To a concentrated aqueous solution of the phenol or to the phenol
itself, add bromine water gradually. At first the bromine is decolourized
and then, on adding an excess, a white or yellowish-white precipitate of a
poly bromo-derivative is produced with all except hydroquinone and and -naphthol. On gradually adding bromine water to a solution of
hydroquinone, a deep red coloration is produced, followed by the
separation of deep green crystals which then dissolve giving a yellow
solution. The naphthols decolourize bromine water, but usually no
precipitate of the bromo compound can be obtained.
This test is not very satisfactory with those phenols which are
insoluble in water, owing to the difficulty of distinguishing the bromo
compound from the original phenol.
3. Phthalein test
Many phenols yield phthaleins which give special colours
(sometimes with fluorescence) in alkaline solutions when reacted with
phthalic anhydride and a little amount of concentrated sulphuric acid.
Phenol and resorcinol are examples.
HO
O
C
O
C
O
phthalic anhydride
OH
OH
+ 2
O
OH
O
resorcinol
fluorescein
(pale red with green fluorescence
in alkaline medium)
+ 2H2O
23
Procedure
In a dry test tube put about 0.1 g of the phenol and an equal amount
of phthalic anhydride or phthalic acid, mix well, and add 1-2 drops of
concentrated sulphuric acid. Heat gently on a direct flame for 1 minute
until the crystals of the mixture melt and fuse. Then cool the test tube and
add excess of 10% sodium hydroxide solution. Results should be as
follows:
compound
colour
-naphthol
green colour
-naphthol
phenol
red to pink
o-cresol
red-violet
m-cresol
blue to pink
p-cresol
no change
resorcinol
hydroquinone
violet colour
If the resultant colour is not so clear you can dilute with water.
4. Reimer-Tiemann reaction
Treatment of phenol with chloroform and aqueous sodium hydroxide
solution introduces an aldehyde group (-CHO) into the aromatic ring at
the ortho- or para-positions:
24
Procedure
To about 0.2 g of the phenol add 1mL of 30% sodium hydroxide
solution and 1 mL of chloroform, heat on water bath, and observe the
colour of the aqueous layer:
compound
colour
phenol
yellow or no colour
resorcinol
-naphthol
dark green
-naphthol
o-cresol
deep orange
m-cresol
pale orange
p-cresol
yellow
hydroquinone
deep brown
Procedure
Add 0.1 g or 0.2 mL (3-4 drops) of the compound to 2 mL of water
or ethanol. Add 2% aqueous potassium permanganate solution drop by
drop with shaking until the purple colour of the permanganate persists. If
the permanganate color is not changed in 0.5-1 minutes, allow the
mixture to stand for 5 minutes with occasional vigorous shaking. The
25
26
OH
carboxyl group
C
H
OH
H3C
formic acid
CH2COOH
COOH
CH2COOH
oxalic acid
succinic acid
CH3
HO
CHCOOH
HO
CHCOOH
lactic acid
CH2COOH
HO CCOOH
CH2COOH
tartaric acid
COOH
OH
acetic acid
COOH
COOH
HO
citric acid
COOH
OH
benzoic acid
salicylic acid
Physical properties
Only formic acid, acetic acid, and lactic acid are liquids at room
temperature. The others are solids.
Low molecular weight carboxylic acids are soluble in water and,
therefore, lie under class S1. Water insoluble acids dissolve in both
sodium hydroxide solution and sodium bicarbonate solution, being
classified under class A1. When they react with sodium
bicarbonate, they evolve carbon dioxide gas. This is considered as
a good simple indication of them.
27
HO
C
C
OH
Chemical properties
The acidic properties of carboxylic acids are attributed to the proton
of the carboxyl group. Mono carboxylic acids are weak acids except
formic acid, which is the strongest. The tendency of the alkyl group to
release electrons weakens the acid; thus formic acid is the strongest. On
the other hand presence of electron withdrawing groups (such as
halogens) especially on the alpha carbon increases the acidity.
Reactions of carboxylic acids are related to:
the proton as in salt formation reactions.
removal of the hydroxyl group as in conversion to derivatives such
as esters, amides, or acid chlorides.
substitution either in the alpha position of aliphatic acids or in the
meta position of aromatic ones.
Give an example of a carboxylic acid with halogenation (name and chemical structure).
Which parameter will you look for to compare
its acidity with other acids?
Give the general formula of esters, amides, and
acid chlorides.
Chemical reactions
1. General test (Ferric chloride test)
The acid solution should be made neutral before performing the test
with ferric chloride solution. This is achieved by adding very dilute
ammonia solution drop by drop with shaking to a solution of about 0.5 g
of the solid acid or 2 drops of the liquid acid in 1 mL water until the
medium becomes basic as indicated by changing the colour of litmus
paper to blue or changing the colour of phenolphthalein indicator from
colorless to pink, in which case the characteristic odour of ammonia is
predominant. At this stage the solution is slightly basic. To make the
28
acid
result
red solution
salicylic
violet solution
no special change
As mentioned in the above table formic acid and acetic acid form a
red coloured solution in this test:
29
30
31
conc. H2SO4
CH3COOC2H5 + H2O
ethyl acetate
(ester)
32
33
34
35
36
Methyl salicylate, also known as wintergreen oil, is used widely in pharmaceutical topical
preparations, give its main use with the name of a popular topical preparation.
Both ethyl acetate and methyl salicylate separate as an organic phase during ester
formation test, how can you detect the organic layer practically and theoretically?
Fill the following table:
citric
oxalic
tartaric
lactic
FeCl3 test
H2SO4 test
CaCl2 test
Fenton's test
During ignition observe the colour of the flame. Sodium salts burn with a
golden yellow flame whereas potassium salts burn with a purple flame.
37
Place a small filter paper over the top of the tube and fold it down around
the tube. Add 2 drops of 10% copper sulphate solution on the filter paper
covering the mouth of the test tube. Heat the test tube mildly on a flame
to boil the mixture. The liberated ammonia will react with the copper ions
present on the filter paper resulting in a blue colour.
What is the proper technique for smelling chemicals?
38
39
C2H 5 X
CHCl 3
metheyl halide
ethyl halide
chlorof orm
Cl
CH 2Cl
benzyl chloride
CCl4
carbon tetrachloride
chlorobenzene
Chemical reactions
1. Reaction with alcoholic silver nitrate.
Alcoholic silver nitrate reagent is useful in classifying halogen
compounds. Many halogen containing compounds react with silver
nitrate to give an insoluble silver halide (AgX), and the rate of this
reaction indicates the degree of reactivity of the halogen atom in the
compound. Besides, the identity of the halogen can sometimes be
determined from the colour of the silver halide produced; silver chloride
is white (turns to purple on exposure to light), silver bromide is pale
yellow, and silver iodide is yellow. These should, of course, be consistent
with results from elemental analysis (sodium fusion for detection of
halogens).
40
RX + AgNO 3
R X
Ag+
C2 H5 OH
AgX + RONO2
Ag
R+
NO3-
AgX + R+
R ONO 2
Procedure
Add one drop or a couple of crystals of the unknown to 2 mL of
2% ethanolic silver nitrate solution. If no immediate reaction is observed,
stand for 5 minutes at room temperature and observe the result. If no
reaction takes place, warm the mixture in water bath for 30 seconds and
observe any change. If there is any precipitate (AgX) add several drops of
1 M nitric acid solution to it; silver halides are insoluble in this acid.
tert- chlorides, methyl and ethyl iodides, allylic chlorides, and ethyl
bromides give fast result at room temperature:
CH2X
R
R3CCl
RI
C C
C C
H
Br
H2C
CH2X
CH2Br
pri- and sec- chlorides, benzyl chloride, and 1-chloro-2,4dinitrobenzene give result only on warming:
R
ArX
HCCl3 CCl4
C C
H
R
C C
41
RCl + NaI
acetone
RBr + NaI
RI + NaCl
RI + NaBr
I-
I R
Procedure
To 1 mL of the sodium iodide-acetone reagent in a test tube add
two drops of the compound. If the compound is a solid, dissolve about 0.1
g in the smallest possible volume of acetone, and add the solution to the
reagent. Shake the test tube, and allow the solution to stand at room
temperature for 3 min. Note whether a precipitate is formed and also
whether the solution turns reddish brown, because of the liberation of free
iodine. If no change occurs at room temperature, place the test tube in
water bath at 50C.Excessive heating causes loss of acetone and
precipitation of sodium iodide, which can lead to false-positive results. At
the end of 6 min, cool to room temperature and note whether a reaction
has occurred. Occasionally, a precipitate forms immediately after
combination of the reagents; this represents a positive test only if the
precipitate remains after the mixture is shaken and allowed to stand for 3
minutes.
3. Differentiation between alkyl and aryl halides
(Formaldehyde- sulfuric acid test)
With this test aryl halides (chlorobenzene) produce pink, red, or
bluish red colour whereas alkyl halides produce yellow, amber, or brown
colour.
Procedure
This reagent is prepared at the time of use by adding 1 drop of
formaldehyde to a test tube containing 1 mL concentrated sulfuric acid. In
another test tube add 1 drop of the compound to be tested to 1 mL of
hexane. From this solution take 1-2 drops and add them to 1 mL of the
reagent. Shake well and observe the colour.
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Identification of Amines
Physical properties
Like ammonia, amines are polar compounds and all of them can
form intermolecular hydrogen bonds except tertiary amines.
H
H 3C N H
CH 3
N H
N CH 3
compounds. Methylamine is gas while o-phenylenediamine and p,pdiaminodiphenylmethane are solids. The others are liquids.
All amines are capable of forming hydrogen bonds with water, thus
those with six carbon atoms or less are quite soluble in water. They are
soluble in organic solvents as ether, alcohol and benzene.
All of them have fish like odour except the methylamines and
ethylamines which smell just like ammonia.
Aromatic amines are colourless when pure, but they are easily
oxidized by air becoming coloured. They are generally very toxic and can
be absorbed through the skin.
Chemical reactions
All classes of amines (primary, secondary, and tertiary) have an
unshared pair of electrons on the nitrogen atom, just like ammonia. That
is why they are similar to ammonia in their chemical behavior (mainly
basicity and neocleophilicity).
1. Ramini and Simon tests (Sodium nitroprusside tests).
(conventional Ramini and Simon tests)
In Ramini test the amine reacts with acetone and the product
interacts with sodium nitroprusside (Na2[Fe(NO)(CN)5].2H2O) that is
dissolved in 50 % aqueous methanolic solution to produce a coloured
complex. In Simon test acetone is replaced by 2.5 M acetaldehyde
solution. These two tests distinguish between primary and secondary
aliphatic amines.
To distinguish between aromatic amines (primary, secondary and
tertiary) the modified Ramini and Simon tests are applied. These tests
use the same reagents and procedure of the conventional tests but the
modifications are the replacement of the methanolic solution of sodium
nitroprusside by a solution of sodium nitroprusside in dimethylsulfoxide
(modified sodium nitroprusside reagent) and the use of a saturated
aqueous solution of zinc chloride instead of water.
Procedure
Ramini test
To 1 mL of methanolic sodium nitroprusside solution add 1 mL of
distilled water, 5 drops of acetone, and about 30 mg of the amine. In most
cases the characteristic colour appears in a few seconds, although in some
cases 2 minutes may be necessary.
Simon test
Follow the above procedure exactly but use 5 drops of 2.5 M
acetaldehyde solution instead of acetone. Up to 2 minutes may be needed
for the colour to develop.
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Modified
Ramini test
Modified
Simon test
aniline
turbid orange,
changes to
henna colour
brown
o-phenylenediamine
turbid red
brown
turbid henna
colour
p,p-diaminodiphenylmethane
light brown
precipitate
light brown
precipitate
N,N-dimethylaniline
brown
changes to
green
brown changes
to green (rapid
change)
diphenylamine
orange- red to
red brown
pale orange
Amine
Ramini test
Simon test
tert-butylamine
deep red
red-brown
dicyclohexylamine
deep red
violet with
precipitate
diethylamine
red-brown
deep blue
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Secondary amines undergo a reaction with nitrous acid to form Nnitrosoamines, which are usually yellow oils or solids. These are
carcinogenic compounds; therefore, students are advised not to perform
nitrous acid test for secondary amines.
R
R2NH + HONO
2 amine
nitrous
acid
N N O + H 2O
N -nitrosoamine
(yellow oil or solid)
Tertiary aliphatic amines do not react with nitrous acid, but they
form a soluble salt.
R3N + H +
3 aliphatic
amine
R3NH+
soluble
Tertiary aromatic amines react with nitrous acid to form the orangecoloured hydrochloride salt of the C-nitrosoamine. Treating the solution
with base liberates the blue or green C-nitrosoamine.
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O N
NHR2Cl + H2O
NaOH
O N
C -nitrosoamine
(green)
Procedure
Nitrous acid is prepared instantaneously by the reaction of sodium
nitrite and hydrochloric acid:
NaNO 2 + HCl
sodium nitrite
HONO + NaCl
nitrous acid
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N Ar
ONa
ArN2Cl +
diazonium salt
of primary
aromatic amines
ONa
+ NaOH
+ NaCl + H2O
azo dye
(red- orange)
sodium-2-naphthol
Procedure
Dissolve 0.1 g of 2-naphthol in a mixture of 2 mL of 10 % sodium
hydroxide solution and 5 mL distilled water. Add 2 mL of the cold
diazonium solution and observe the result. The formation of a red- orange
dye (red precipitate in case of phenol) with evolution of gas only upon
warming indicates that the compound is a primary aromatic amine.
4. Carbon disulfide reagent test (for secondary aliphatic amines).
S
R2NH + CS2 + NH 4OH
carbon disulfide
R2N
SNH 2 + H2O
dialkyldithiocarbamate
NiCl2
S
R 2N
2Ni
Procedure
In a test tube dissolve 50 mg (1-2 drops) of the amine in 5 mL
distilled water (or 1-2 drops of concentrated hydrochloric acid if
necessary). In another test tube mix o.5- 1 mL of concentrated ammonia
solution with 1 mL of nickel chloride in carbon disulfide reagent
(NiCl2/CS2). Add 0.5- 1 mL from the first test tube to the second one. A
definite precipitation indicates that the unknown is a secondary amine. A
slight turbidity is an indication of a trace of a secondary amine as an
impurity.
5. Lignin test (for primary and secondary aromatic amines).
This test depends on the action of lignin in the newsprint paper.
Procedure
Dissolve 10- 20 mg of the amine in a few drops of ethanol and
moisten a small area of newsprint paper with this solution. Place 2 drops
of 6 N hydrochloric acid on the moistened spot. The immediate
development of a yellow or an orange colour is a positive test for a
primary or secondary aromatic amine.
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