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Org. Chem. Manual, 2013 PDF
Org. Chem. Manual, 2013 PDF
College of Pharmacy
Department of Pharmaceutical Chemistry
A laboratory manual on
HO O OH
O
Azhar M. Jaism (M. Sc. Ph. Chem.)
Duraid H. Mohammad (M. Sc. Ph. Chem.)
Oct., 2012
Contents
ii
Tools and Glass Wares
Below are the mostly used tools and glass wares in organic chemistry laboratory
(courses II and III):
iii
litm
mus paperr spatula spatula
a
glass rod
r (stirreer) water bath
h Hood
iv
Org. Chem. Lab.
1
Qualitative Analysis of
Organic Compounds
H2O
insol. sol.
5% NaOH Ether
5% HCl 5% NaHCO3
insol. sol.
SI2
litmus unchanged
litmus blue
96%
litmus red
cold H2SO4 insol. sol.
insol. sol.
B
SI1
AI 2 AI 1
I N
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.
Org. Chem. Lab.
4
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 water-
soluble compounds only and no further solubility tests using the
remaining solvents are to be done.
5% HCl
If the compound is insoluble in water and sodium hydroxide
solution (and, hence, insoluble in sodium bicarbonate solution too), this
means that the compound is not an acid but, rather, is either a basic
compound or a neutral compound. 5% hydrochloric acid solution, which
can dissolve basic compounds such as amines (RNH2), is used for such a
compound. If the compound is soluble in this solvent, then it is given
class B. This class includes primary, secondary, and tertiary amines.
Identification of Alcohols
A
lcohols are organic compounds that may be considered as
derivatives of water in which one of the hydrogen atoms of
water molecule (H-O-H) has been replaced by an alkyl or
substituted alkyl group. Therefore, properties of alcohols may be related
to properties of both water and hydrocarbons. The alkyl group could be
primary, secondary, or tertiary, and may be open chain or cyclic.
Accordingly, alcohols may be defined as organic compounds that contain
hydroxyl groups attached to alkyl, substituted alkyl, or cyclic alkyl group.
H2
H2 C CH 3
R OH H 3C OH C CH
H3C
H3C OH
OH
the general methanol ethanol sec-butanol
f ormula
OH
C OH
H 3C CH3 C
H2
H 3C OH
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.
Org. Chem. Lab.
7
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.
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.
O
2[(NH 4)2Ce(NO 3)6] + RCH2OH RC H + 2[(NH4) 2Ce(NO 3)5] + 2HNO3
ceric ammonium
alcohol aldehyde
nitrate
OR
(NH 4)2Ce(NO3)6 + ROH (NH 4)2Ce(NO 3)5 + HNO 3
ceric ammonium
alcohol
nitrate
(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.
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.
ZnCl 2 + HCl ZnCl3- + H+
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.
Org. Chem. Lab.
11
A
ldehydes are compounds of the general formula RCHO;
ketones are compounds of the general formula RCO. The
groups R and may be aliphatic or aromatic, and in one
aldehyde, formaldehyde, R is hydrogen.
O
O O
C
C C H
R H H3C H
O
O O
C C
H C
H3C CH3 CH3
OH
O
O
C
C
C CH3
H2
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.
Org. Chem. Lab.
13
R H
C O+H Y R C OH
H
Y
NO 2 NO 2
R R
+
O2N NHNH2 + C O O2N N N C + H2O
R H R
2,4-dinitrophenyhydrazine 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.
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.
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).
4. Polymerization reaction
To 0.5 mL of formaldehyde or salicylaldehyde add 0.2 gm of
resorcinol and drop-by-drop concentrated sulfuric acid to get a red or
reddish violet color, or a white ring that changes to a reddish violet ring.
OH
OH
OH
O conc. H2SO4 HO HO OH
+ 2
C - H2O
OH H H OH
resorcinol formaldehyde
OH
polymer
5. Cannizzaro reaction
Benzaldehyde, salicylaldehyde, and formaldehyde can undergo
Cannizzaro reaction because they do not have an alpha hydrogen atom.
O O O
C C C
NaOH
H H OH ONa
+ +
heat
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.
Identification of Phenols
P
henols are organic compounds with a hydroxyl group
attached directly to benzene or substituted benzene. They
have the general formula Ar-OH. Examples of them include
phenol (also known as carbolic acid), hydroquinone, resorcinol, o-cresol,
m-cresol, p-cresol, -naphthol, and -naphthol.
OH OH OH
OH
OH
phenol hydroquinone resorcinol
(quinol, hydroquinol)
OH
OH OH
CH 3
CH3
CH3
HO
OH
-naphthol -naphthol
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.
OH OC2H5
aq. NaOH
+ C2H5I + 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.
Org. Chem. Lab.
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,
violet or blue
resorcinol
o- and p-cresol greenish blue
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 OH
O OH
C
+ 2 + 2H2O
O O
C OH
O O
phthalic anhydride resorcinol fluorescein
(pale red with green fluorescence
in alkaline medium)
Org. Chem. Lab.
23
The fluorescence of resorcinol is due to the presence of an oxygen
linkage between the two phenolic nuclei (in basic medium).
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
o-cresol red-violet
p-cresol no change
pale red colour with green
resorcinol
fluorescence
hydroquinone violet colour
If the resultant colour is not so clear you can dilute with water.
4. Reimer-Tiemann reaction
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
p-cresol yellow
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
Org. Chem. Lab.
25
disappearance of the purple color and the formation of a brown
suspension, which is manganese (II) oxide, at the bottom of the test tube
is a positive result for the presence of phenols.
C
arboxylic acids are organic compounds that have a carboxyl
group attached to an alkyl group(RCOOH) or to an aryl
group (ArCOOH). The 'R' may be a hydrogen and the result
is formic acid. They may be mono carboxylated, multi carboxylated,
substituted (e. g., hydroxyl groups), or they may be aromatic
O O O
C OH H C OH H3C C OH
COOH CH2COOH
COOH CH2COOH
CH2COOH
COOH HO CHCOOH
HO CCOOH
HO C CH3 HO CHCOOH
H
CH2COOH
COOH COOH
OH
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.
Org. Chem. Lab.
27
Their boiling points are generally high due to the association
through hydrogen bonds: two molecules of the carboxylic acid are
held together by two hydrogen bonds rather than one.
O HO
R C C R
OH O
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.
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
Org. Chem. Lab.
28
solution neutral the excess ammonia should be removed by gently heating
the test tube in a water bath with shaking from time to time until both the
odour of ammonia and the pink colour disappears. (In case of oxalic,
tartaric, citric and lactic acids keep a portion of their neutral solution for
use in calcium chloride test). Cool the solution and then add few drops of
ferric chloride solution to get different colours (solutions or precipitates)
as follows:
acid result
As mentioned in the above table formic acid and acetic acid form a
red coloured solution in this test:
a) Iodoform test
Lactic acid can undergo iodoform formation reaction since it
contains a free methyl group and a hydrogen attached to the carbon
bearing the hydroxyl group:
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:
acid
FeCl3 test H2SO4 test CaCl2 test Fenton's test
citric
oxalic
tartaric
lactic
Org. Chem. Lab.
37
C
arboxylic acids salts are organic compounds with the
general formula (RCOOM) where (RCOO-) refers to the
carboxylic acid part and (M+) is the alkali part which, in this
experiment, may be either a metal cation (Na+ or K+) or ammonium
(NH4+). These salts are colourless or white crystalline solids and are
soluble in cold or hot water.
During ignition observe the colour of the flame. Sodium salts burn with a
golden yellow flame whereas potassium salts burn with a purple flame.
Org. Chem. Lab.
38
Identification of ammonium cation
Repeat the ignition procedure mentioned above and note that
ammonium salts don't leave any residual solid except the carbon coming
from decomposition of the organic part. After cooling, addition of few
drops of dilute hydrochloric acid does not result in any effervescence.
Ammonium cation can be detected as follows. Place few crystals of
the salt in a test tube and add 0.5 mL of 10% sodium hydroxide solution.
At this stage free ammonia is liberated and can be smelt easily:
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.
Physical properties
All alkyl halides and chlorobenzene are colourless liquids when
pure except iodoform, CHI3, which is a yellow crystalline solid with a
characteristic odour. Methyl iodide, ethyl iodide and bromide,
chloroform, and carbon tetrachloride have sweetish odours. Benzyl
chloride has a sharp irritating odour and is lachrymatory. Chlorobenzene
possesses aromatic odour.
Alkyl and aryl halides (RX, ArX) have boiling points higher
than the parent hydrocarbon because of the heavier molecular weight.
Accordingly, for a given compound, iodides have the higher boiling point
than bromides and chlorides.
In spite of their polarity alkyl halides are insoluble in water due to
their inability to form hydrogen bonds. They are soluble in most organic
solvents.
Iodo-, bromo-, and polychloro- compounds are denser than water.
CH 2Cl Cl
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).
Org. Chem. Lab.
40
C2 H5 OH
RX + AgNO 3 AgX + RONO2
Ag+ + +
R X R X Ag AgX + R+
NO3-
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:
R CH2X R H Br
R3CCl RI C C C C H2C
H H H CH2X CH2Br
R H R X
ArX HCCl3 CCl4 C C C C
H X H H
acetone
RBr + NaI RI + NaBr
I-
R X I C X 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
Org. Chem. Lab.
42
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.
Identification of Amines
A
mines are basic organic compounds that are considered as
derivatives of ammonia. They are classified as primary,
secondary, or tertiary according to the number of groups
attached to the nitrogen atom: RNH2, R2NH, or R3N respectively where R
is any alkyl or aryl group.
Physical properties
Like ammonia, amines are polar compounds and all of them can
form intermolecular hydrogen bonds except tertiary amines.
H CH 3 H
H 3C N H N H N CH 3
H H
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.
Org. Chem. Lab.
45
Modified Ramini test
To 1 mL of the modified sodium nitroprusside reagent add 1 mL of
saturated aqueous zinc chloride solution, 5 drops of acetone, and about 30
mg of the amine. Primary and secondary aromatic amines produce
orange-red to red-brown colours within a period of few seconds to 5
minutes. Tertiary aromatic amines give a colour that changes from
orange-red to green over a period of about 5 minutes.
Modified Simon test
Follow the above procedure exactly but use 5 drops of 2.5 M
acetaldehyde solution instead of acetone. Primary aromatic amines give
an orange-red to red-brown colour within 5 minutes; secondary aromatic
amines give a colour changing from red to purple within 5 minutes;
tertiary aromatic amines give a colour that changes from orange-red to
green over a period of about 5 minutes.
Examples are outlined in the following table:
Modified Modified
Amine Ramini test Simon test
Ramini test Simon test
violet with
dicyclohexylamine deep red
precipitate
turbid orange,
aniline changes to brown
henna colour
turbid red turbid henna
o-phenylenediamine
brown colour
light brown light brown
p,p-diaminodiphenylmethane
precipitate precipitate
brown brown changes
N,N-dimethylaniline changes to to green (rapid
green change)
orange- red to
diphenylamine pale orange
red brown
Org. Chem. Lab.
46
2. Nitrous acid test.
The reaction of amines with nitrous acid (HNO2) is another test
that classifies the amine not only as primary, secondary, or tertiary, but
also as aliphatic or aromatic.
Primary aromatic and aliphatic amines react with nitrous acid to
form an intermediate diazonium salt. The aliphatic diazonium salts
decompose spontaneously by rapid loss of nitrogen, particularly when the
original amino group is attached to a secondary or tertiary carbon, while
most aromatic diazonium salts are stable at 0C but lose nitrogen slowly
on warming to room temperature.
R
R2NH + HONO N N O + H 2O
R
2 amine nitrous N -nitrosoamine
acid (yellow oil or solid)
Tertiary aliphatic amines do not react with nitrous acid, but they
form a soluble salt.
R3N + H + R3NH+
3 aliphatic
soluble
amine
Tertiary aromatic amines react with nitrous acid to form the orange-
coloured hydrochloride salt of the C-nitrosoamine. Treating the solution
with base liberates the blue or green C-nitrosoamine.
Org. Chem. Lab.
47
NaOH
C -nitrosoamine
(green)
Procedure
Nitrous acid is prepared instantaneously by the reaction of sodium
nitrite and hydrochloric acid:
N Ar
N
ONa ONa
ArN2Cl + + NaOH + NaCl + H2O
diazonium salt
azo dye
of primary sodium-2-naphthol (red- orange)
aromatic amines
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.
S
C
R2NH + CS2 + NH 4OH R2N SNH 2 + H2O
carbon disulfide
dialkyldithiocarbamate
NiCl2
S
C
R 2N S 2Ni
Org. Chem. Lab.
49
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.