3 Classification

Detection of Fnnctional Groups

I. Metallic derivatives.

II. Carbohydrates and related compounds. Aldehydes and

ketones, acetals; polyfunctional carbonyl compounds.
Quinones. Carboxylic acids. Phenols and enols. Esters (of
carboxylic acids), lactones and acid anhydrides. Alcohols.
Ethers. Hydrocarbons.

III. Amines and polyfunctional amino-compounds (amino-

acids etc.). Salts of amines.

IV. Ammonium salts of carboxylic acids, aldehyde-ammonias,

amides, imides, nitriles. Acyl derivatives of bases (N-
substituted amides). Hydrazines, semicarbazide, hydroxyl-
amine, their salts and carbonyl derivatives. Condensation
products of aldehydes with ammonia or amines.

v. Nitro-, nitroso-, azoxy-, azo-, and hydrazo-compounds.

VI. Compounds containing halogen (but not nitrogen or sul-

phur). (Halogen-substituted acids, aldehydes, ketones,
alcohols, ethers, esters and hydrocarbons; acid halides.)

VII. Compounds containing nitrogen and halogen (but not

sulphur). (Hydro halides of bases; acid halides of nitrogen-
containing acids; halogenated bases, their hydrohalides and
acyl derivatives; halogenated nitro-compounds; nitro-
genous derivatives of halogeno-acids; N-halogeno-com-
pounds.)

B. Haynes, Qualitative Organic Analysis

© B. Haynes 1966
VIII. Compounds containing sulphur (with or without nitrogen
and/or halogen). Ammonium salts of simple and substituted
sulphonic acids. Sulphonic acids and their metallic salts;
sulphonamides. Amino-sulphonic acids. Sulphates of bases.
Thiourea, substituted thioureas, thioamides. Sulphonyl
halides. N-Halogeno-sulphonamides. Aldehyde and ketone
bisulphite compounds. Thiols. Thio-ethers. Thio-acids.
Esters of sulphonic acids.
 Classification

DETECTION OF FUNCTIONAL GROUPS

THE CLASSIFICATION tests given on the following pages have been

chosen for their reliability and general usefulness. In the main,
more than one test for a particular functional group has been
provided, since restrictions on the universal applicability of any
one test may arise through low solubility of the unknown substance
in the solvent used in the reaction, or interference by the molecular
environment of the functional group, leading to little or no reac-
tion.
Where important variations of the structure of the typical
functional group are likely to be encountered, the General Tests
for the type are followed by other tests designed to assist in a more
exact classification of the type variant. For example, the General
Tests for alcohols are followed by distinguishing tests for primary
secondary and tertiary alcohols. Naturally, the latter tests need
not be applied unless positive results have been obtained in the
General Tests.
Usually, positive results from two tests for a particular func-
tional group are required in order that the presence of the group
can be considered proved.
If the preliminary tests have yielded little helpful information
except a knowledge of the elements present, the compound should
be subjected to tests for those functional groups which are con-
sistent with the results of the elements test. Thus, if the compound
contains carbon, hydrogen and possibly oxygen only, the class
tests for carbohydrates, carbonyl compounds, acids, etc., are
performed in that order, as given in the following pages. If a
positive result cannot be obtained in these tests, it is likely that
elements actually present have not been detected, and the sodium
fusion should be repeated using a smaller amount of the substance
than previously.
43
QuoJitame Organic Analysis
The possibility that the compound under examination contains
more than one functional group must be borne in mind. The
preliminary examination may have furnished evidence to this
effect, and if so, after one group has been proved to be present,
further classification tests should be applied in order to confirm or
deny the presence of other groups. The additional tests to be
applied will be determined by the results of the preliminary
examination.
The results of the preliminary and classification tests must agree.

SECTION I

Metallic derivatives

The metallic derivatives of organic compounds that are most likely

to be encountered in the course of practice analyses are: salts of
carboxylic acids, sulphonic acids, phenols and enols, alkyl hydro-
gen sulphates; metallic derivatives of amides, imides and sulpho-
namides; sodium bisulpbite adducts of carbonyl compounds.
The presence of a metal is shown by a residue when the com-
pound is ignited in air (preliminary tests, pages 18, 19). This residue
should be identified by the normal methods of inorganic analysis.
The identification of the acid or pseudo-acid does not necessarily
depend on the isolation of the acidic substance, especially if the
metal present is sodium or potassium. Thus, the alkali metal salts
of carboxylic and sulphonic acids may be characterized by direct
conversion into the S-benzyltbiuronium salts, amides or anilides.
Carboxylic salts may be also used to prepare the p-nitrobenzyl and
p-bromophenacyl esters. Metallic derivatives of phenols may be
converted into aryloxyacetic acids.
The isolation of the acid or pseudo-acid may be achieved by
treating the original substance with a small excess of dilute hydro-
chloric acid and removing a sparingly soluble acid by filtration
(beware of the possible precipitation of a metallic chloride or
oxychloride), a volatile acid by distillation, and a non-volatile
water-soluble acid by extraction with ether. The free acid or
pseudo-acid is then analysed in the usual manner. Isolation of an
acid which is non-volatile, water-soluble and sparingly soluble in
ether, such as oxalic, tartaric or citric, is not easily accomplished
44
 Classification
by the above methods. In this event the tests described on pages
53-55 should be applied to the original substance, unless the latter
contains an interfering metal, which must first be removed, e.g. by
the procedure below.
If the original substance contains a metal other than an alkali
metal (salts of such metals are often sparingly soluble in water) it
may be necessary to remove the metal by boiling the substance
with saturated sodium carbonate solution for 10 minutes. Cool,
filter and treat the filtrate with a slight excess of dilute hydrochloric
acid. Add a small excess of ammonium hydroxide solution and
boil off the excess of ammonia. This neutral solution of the
sodium salt of the organic acid may be used for the tests on
pages 53-55, and (after concentration) for the preparation of
derivatives.
An aromatic amino-acid may be liberated from a solution of one
of its salts by treatment with a slight excess of acetic acid.
The analysis of sodium bisulphite compounds of aldehydes and
ketones is discussed on page 103.

SECTION II

Carbohydrates and glycosides

Aldehydes, ketones and related compounds
Quinones Carboxylic acids Phenols and enols
Esters, lactones and acid anhydrides of carboxylic acids
Alcohols Ethers Hydrocarbons

All of these compounds in their simple unsubstituted forms con-

tain carbon, hydrogen and oxygen only, with the exception of
hydrocarbons (hydrogen and carbon only).
The compounds are considered in the order given above, and
where appropriate, for a given class of compound, brief reference
is made to the chemistry of important related polyfunctional com-
pounds. These polyfunctional substances may contain elements
(other than sulphur) in addition to carbon, hydrogen and possibly
oxygen, but in those cases chosen for mention here the more
characteristic reactions remain due to the oxygen function.
45
Qualitative Organic Analysis
CARBOHYDRATES AND GLYCOSIDES

All the common members of these classes are solids. Mono- and
disaccharides are soluble in water and sparingly soluble in ether.
Many polysaccharides dissolve in water to give colloidal solutions.

General test (Molisch's test)

Dissolve or suspend a small quantity (ca 50 mg) of the substance in
2 m1 of water, add two drops of an alcoholic solution of a-naphthol
(10%) and shake the mixture well. Carefully pour 1 ml of concen-
trated sulphuric acid down the side of the test-tube and allow to
stand. Within a minute or two a violet or red colour should have
formed at the junction of the sulphuric acid and the aqueous layer,
and on shaking the whole should assume a violet hue, with forma-
tion of a blue precipitate. A positive result indicates that the
substance is a carbohydrate or a glycoside. This is a very sensitive
test: traces of filter-paper will give a positive result. Other com-
pounds, e.g. benzilic acid, yield reddish colours.
Test for glycosides. Treat a small quantity of the substance with
concentrated sulphuric acid. A definite colour should be produced
within a few minutes, e.g. amygdalin-cannine; salicin-crimson;
saponin-red-brown.

Tests for mono- and disaccharides

(a) Fehling's solution (test for reducing sugars). If Molisch's test is
positive but the glycoside test is negative, heat on a boiling water-
bath for one minute, 1 ml of an aqueous solution of the substance
and 1 ml each of Fehling's solutions A and B. A red precipitate of
cuprous oxide indicates that a reducing sugar is present. Other
compounds, e.g. aldehydes, also reduce Fehling's solution.
(b) Barfoed's test (for reagent preparation, see page 221). Heat
2 m1 of Barfoed's reagent with 2 ml of an aqueous solution of the
substance for 2 minutes on a boiling water-bath. A red precipitate
of cuprous oxide indicates the presence of a monosaccharide.
If test (a) is positive but test (b) is negative, the substance must
be a reducing disaccharide, the commonest examples being maltose
and lactose. Iftests (a) and (b) are both negative, the compound is
probably a non-reducing disaccharide, the only common example
being sucrose. In either case proceed to tests (e) and (f).
46
 Classification
(c) Pentose test. If test (b) is positive, dissolve a small quantity of
the substance in dilute hydrochloric acid and add about O· 1 g of
phloroglucinol. Boil for one minute. The formation of a red colour
indicates a pentose. (Other sugars may give yellow or brown
colours.)
(d) Seliwanoff's ketose test. Dissolve 50 mg of resorcinol in a mix-
ture of 4 m1 of water and 2 m1 of concentrated hydrochloric acid.
Add 5 ml of a 5 %solution of the sugar and heat gently to boiling.
Ketoses cause the formation of a red colour within two minutes.
Prolonged heating should be avoided, as aldoses will then give a
positive result.
(e) The 'rapid furfural' test. Careful observation is essential in this
test.
Heat to boiling a mixture of 1 m1 of an aqueous solution of the
sugar, 1 m1 of a 1 % solution of ex-naphthol in ethanol, and 8 m1
of concentrated hydrochloric acid. A violet colour appears in the
following cases: colour immediately boiling-fructose, sucrose;
colour after from half to one minute-maltose, glucose, lactose.
(f) Sucrose. To 1 ml of a saturated solution of nickel ammonium
sulphate add a small quantity of sucrose and O· 5 m1 of concen-
trated hydrochloric acid. Boil gently. A green colour, changing to
yellow and then red, is a positive test.

Starch. When treated with iodine solution, starch gives a blue

colour, discharged by heating but restored on cooling.
Identification. Sugars: see page 119. Starch: hydrolyse with
boiling dilute hydrochloric acid (20 minutes), and show the
presence of monosaccharide (glucose) in the neutralized residual
liquid.

ALDEHYDES AND KETONES

General test (Brady's reagent, * p. 222, gives reagent preparation).

Shake 3 m1 of a stock solution of 2,4-dinitrophenylhydrazine
sulphate in methanol with 0·1 ml of the substance, or if solid, a
* The formation of 2,4-dinitrophenylhydrazones may also be
carried out in 'diglyme',-see page 121.
47
Qualitative Organic AlUllysis
solution of 0·1 g of it in methanol. A reddish or yellow precipitate
indicates a compound of this class. Occasionally precipitation is
slow and may be hastened by warming on a water-bath for not
more than 5 minutes. An oily precipitate usually crystallizes on
standing.
If a precipitate is not obtained within 15 minutes, dilute the
solution very carefully with 2N sulphuric acid. It is advisable to
collect and wash the derivative and determine its melting point to
ensure that it is the 2,4-dinitrophenylhydrazone of the test sub-
stance, and not 2,4-dinitrophenylhydrazine (m.p. 194°) or its
sulphate (m.p. 185j. (Dinitrophenylhydrazine sulphate is stable
only in the presence of excess of sulphuric acid; injudicious dilu-
tion of the reagent or addition of a basic compound promotes
hydrolysis and the precipitation of 2,4-dinitrophenylhydrazine.)
Differentiating tests
Aldehydes. These compounds are more reactive than ketones, and
may usually be detected by the following tests.
(a) Schiff's reagent. Add one drop of the substance or 50 mg
dissolved in pure (aldehyde-free) alcohol to 1 m1 of reagent.
DO NOT HEAT. The production of a strong pink colour almost
immediately is a positive test. If the result is positive, add three
drops of concentrated hydrochloric acid; a blue colour indicates
formaldehyde, most other aldehydes yielding an almost colourless
mixture. (A few aromatic aldehydes give a negative Schiff's test;
some ketones produce a pale pink colour. Owing to the sensitivity
of the test, aldehydic impurities may give a positive result. Solu-
tions with an alkaline reaction to litmus interfere with the test
reaction.)
(b) Tollen's reagent (see preliminary tests for its preparation). Add
about 0·1 g of the substance or a solution of it in alcohol to 2 m1 of
the reagent contained in a clean test-tube. Aldehydes reduce the
reagent to silver which forms a black precipitate or a silver mirror.
Water-insoluble aldehydes reduce slowly and it may be necessary
to warm gently on a water-bath, with occasional shaking. [The
following also react: formates, tartrates, lactates, ex-diketones,
ex-hydroxy ketones, some quinones, some phenols, and a few simple
ketones (usually slowly).]
Ketones. These compounds do not usually give positive results in
Schiff's and Tollen's tests (with the reservations stated above).
48
 Classification
Additional tests
Ketones, and aldehydes (e.g. acetaldehyde) containing the group
CH3CO- respond to the following tests, which are often useful in
effecting a more precise classification of the substance under
examination.

Nitroprusside test. Treat the compound with 1 ml of freshly pre-

pared sodium nitroprusside solution, and add excess of sodium
hydroxide solution, when a red colour develops.

Iodoform test. To 1 ml of the carbonyl compound or its solution in

water or dioxan, add 5 ml of sodium hydroxide solution followed
by iodine/potassium iodide solution* until the iodine colour
remains on shaking. If no yellow precipitate of iodoform appears
after 3 minutes, warm to 60°, adding iodine solution if the colour
fades. Add drops of 2N sodium hydroxide solution to remove the
excess of iodine, add water to dilute the solution, and allow to
stand for 15 minutes. Yellow crystals of iodoform (m.p. 120j
separate. (Dioxan may contain alcoholic impurities; perform a
blank test.)
The iodoform test is also positive for compounds of some
other classes, e.g. certain alcohols containing the grouping
[Link](OH).CH3 which in the early stages of the reaction are
oxidized to the methyl ketone.
Acetals and Polymerized Aldehydes will often give some of the
above tests directly, but more generally only after heating with
dilute sulphuric acid, when the aldehyde is produced. Acetals
should be hydrolysed thus to an aldehyde and an alcohol, and both
products identified (see Table 3).

POLYFUNCTIONAL COMPOUNDS
CONTAINING AN ACTIVE CARBONYL GROUP

The types of compounds in this category that are most likely to be

encountered and their more important characteristics are:

* For preparation of this solution, see page 223.

D 49
Qualitative Organic Analysis
1. a-Hydroxyketones (acyloins), e.g. benzoin, furoin, acetoin.
These compounds reduce Tollen's reagent and Fehling's solution.
They are oxidized rapidly by bismuth oxide to the corresponding
a-diketones, and this reaction may be used as a test as follows.
Shake o· I g of the substance with 3 m1 of glacial acetic acid and
heat with o· I g of bismuth oxide. A black precipitate of finely
divided bismuth is produced almost at once.
a-Hydroxyketones react normally with the reagents used to
characterize carbonyl compounds.

2. a-Diketones, e.g. diacetyl, benzil, furil. These substances are

yellow or orange in colour and are generally soluble in ether but
insoluble in water. They react with o-phenylenediamine to produce
quinoxalines (see Quinones, below). With 2,4-dinitrophenylhydra-
zine they yield bis-2,4-dinitrophenylhydrazones.

3. ~- Diketones, e.g. acetylacetone. These keto-enolic compounds

are soluble in aqueous sodium hydroxide but not in aqueous
sodium bicarbonate. They give a red colour with aqueous ferric
chloride. When treated (in alcohol) with a solution of o-phenylene-
diamine in 2N hydrochloric acid, they give the intense violet colour
of the salt of a heptazine. They react with 2,4-dinitrophenylhydra-
zine to yield pyrazoles.

4. y-Diketones, e.g. acetonylacetone. Most of these compounds

react normally with 2,4-dinitrophenylhydrazine.

5. Keto-acids, keto-esters. Normal reactions are obtained with

2,4-dinitrophenylhydrazine and semicarbazide.
Keto-acids are strongly acidic and may be characterized via the
carboxyl group (see carboxylic acids).
~- Keto-esters are enolic compounds, soluble in alkalis but
insoluble in aqueous sodium bicarbonate, and give a red colour
with aqueous ferric chloride. With phenylhydrazine they yield
pyrazolones.

6. Aliphatic halogenated aldehydes and ketones. In general these

compounds react normally with the usual reagents for the carbonyl
group (but see also pages 93, 94).
SO
 Classification

7. Aromatic aldehydes and ketones with nuclear substituents. The

commonest nuclear substituents are: nitro-, halogeno-, alkoxy-,
hydroxy-. In most instances it is advisable to prepare derivatives
via the carbonyl group rather than to attempt to reduce a nitro-
aldehyde to the amino-aldehyde, for example.
Phenolic and nitro-carbonyl-compounds often dissolve In
alkalis, the latter slowly and with some decomposition.

Derivatives. See page 121.

QUINONES

All are yellow to red solids. Most are soluble in ether but not in
water. Many of them dissolve in solutions of alkalis; these solu-
tions darken rapidly in air. They vary widely in reactivity.
Some react abnormally with reagents for the carbonyl group,
e.g. 2,4-dinitrophenylhydrazine may give azo-compounds.
p-Benzoquinone is noted for its reactivity, e.g. it reduces cold
ammoniacal silver nitrate, liberates iodine from acidified potas-
sium iodide solution, and gives the iodoform reaction. It is volatile
in steam and has a characteristic irritating smell.
By contrast, anthraquinone is very stable, having no reducing
properties. When heated with zinc dust and alkali it gives a bright
red colour, due to anthrahydroquinone, which vanishes on shaking
in the presence of air owing to regeneration of anthraquinone.

Derivatives (melting points are given in Table 2).

(a) Semicarbazone.
(b) In some cases specific reactions may be utilized, e.g. ortho-
quinones such as phenanthraquinone are converted to the quinox-
aline derivative thus: take equal weights of the quinone and
o-phenylenediamine and dissolve them separately in glacial acetic
acid so as to obtain hot saturated solutions. Mix the solutions and
cool the mixture; if a precipitate does not form at once, add a little
water. Recrystallize from acetic acid.
(c) Diacetate of the hydro quinone (see footnote, page 165).
51
Qualitatil'e Organic Analysis

CARBOXYLIC ACIDS

Most simple aliphatic acids are soluble in water. Simple aromatic

acids are sparingly soluble in cold water (more soluble in hot
water) and soluble in ether. Many aliphatic dibasic and hydroxy-
acids are readily soluble in water but sparingly soluble in ether.

General tests
(a) Dissolve 0·1 g of the substance (or three drops if it is a liquid)
in water or neutral alcohol, add two drops of phenolphthalein
solution and O· 1 N sodium hydroxide solution drop by drop with
shaking until a permanent pink colour is obtained. If the number
of drops of sodium hydroxide solution required to produce a pink
colour is greater than about four, and if the colour change is sharp,
the substance is probably a carboxylic acid. *
(b) Add a small quantity of the substance to O' 5 m1 of saturated
sodium bicarbonate solution contained in a narrow test-tube.
Effervescence, due to evolution of carbon dioxide, indicates the
presence of a carboxylic acid. *

Unsaturation. Consult the preliminary tests to ensure that the

compound does not contain a centre of unsaturation. If necessary,
repeat the bromine and potassium permanganate tests. Note that
in a few instances bromine water and bromine in carbon tetra-
chloride are not decolourized at room temperature despite the
olefinic character of the acid, e.g. maleic acid.

Differentiating tests for Fatty Acids

1. Ferric chloride test. Refer to the results obtained in this test
(preliminary examination). Propionic acid, butyric acids and
valeric acids may be distinguished from acetic and formic acids by
carrying out the test in the presence of amyl alcohol. If, when the
mixture is vigorously shaken, the red colour passes into the

* Positive results in both tests may be given by some nitro-

and polyhalogenophenols, some simple acid anhydrides, and some
very readily hydrolysable esters (e.g. methyl and ethyl esters of
oxalic and formic acid; methyl oxalate is a white crystalline solid).
For distinguishing test for esters and anhydrides, see pages 58, 59.
52
 Classification
alcohol layer, the presence of propionic or one of the higher fatty
acids is indicated.
Propionic acid may be distinguished from butyric and higher
acids by performing the ferric chloride test in the presence of
diethyl ether. The ether layer remains colourless in the case of
propionic acid, but if an acid higher than this is present, the ether
layer becomes coloured.

2. Acetic acid may be distinguished from formic acid by the ability

of the latter to reduce potassium permanganate solution, Tollen's
reagent and mercuric chloride solution. In the latter case a white or
grey precipitate of mercurous chloride (insoluble in dil. HCI) or
mercury is produced when the mixture is warmed.

3. Lanthanum nitrate test (for acetic and propionic acids). Mix

equal volumes of 5 % aqueous lanthanum nitrate, iodine solution
and the solution of the substance. Add a few drops of dilute
ammonium hydroxide solution, and heat slowly to boiling. The
presence of acetic or propionic acid is indicated by the appearance
of a blue colour. Sulphates interfere with the test.

Tests for Individual Acids

Certain polybasic and hydroxy-acids such as tartaric, citric, oxalic,
succinic, lactic, present difficulty in their identification when
present in dilute aqueous solution as their sodium salts, as for
example following the hydrolysis of an ester or an amide. In such
circumstances, the following qualitative tests may be found useful.
They should be applied to the solution of the sodium (or potas-
sium) salt, neutralized if necessary by treatment with dilute
mineral acid. (The mineral acid to be used will depend on the test
to be applied.)
Colour tests given here for some other acids should be used only
as confirmatory reactions.

Tartaric acid. (a) Make a saturated aqueous solution of ferrous

sulphate (do not heat). Add one drop of it to the neutral solution
of the compound, followed by two drops of 10 volume hydrogen
peroxide and an excess of sodium hydroxide solution. An intense
violet colour due to the ferric salt of dihydroxyfumaric acid, is a
positive test (Fenton's test).
53
 Qualitative Organic Analysis
(b) Treat the neutral solution of the compound with 5 %aqueous
ammonium vanadate, and acidify with dilute acetic acid. An
orange colour is produced.
Citric acid. (a) Add Deniges' solution to the neutral solutionofthe
compound and heat to boiling. Add drops of dilute potassium
permanganate solution with shaking. Discharge of the purple
colour of the permanganate followed by the rapid formation of a
heavy white precipitate indicates a citrate. (Chloride ions interfere.)
(b) Add calcium chloride solution to the neutral solution of the
substance and bring to the boil. A white precipitate forms slowly.
Oxalic acid. (a) Acidify the solution of the compound with dilute
acetic acid and add aqueous calcium chloride. A white precipitate
indicates an oxalate. Confirm by filtering and warming the washed
precipitate with dilute sulphuric acid and potassium permanganate
solution. The purple colour is discharged.
(b) To 1 ml of the copper-benzidine reagent (see page 221) add
drops of the neutral solution of the substance. The appearance of a
brown crystalline precipitate within about 3 minutes indicates the
presence of oxalate. (Sulphites react similarly. Some aromatic
acids give deep green precipitates.)
(c) Treat a very small amount of the solid acid or one of its salts
with a crystal of diphenylamine and four drops of syrupy phos-
phoric acid. Warm the mixture over a small flame. A blue colour
develops which vanishes on cooling but reappears when the
mixture is dissolved in alcohol.
Succinic acid (fluorescein test). Obtain the solid acid or metallic
salt and to a small quantity add 2 parts of resorcinol and 2 drops
of concentrated sulphuric acid. Gently warm the mixture, allow to
cool and pour into water. Addition of an excess of alkali produces
a red solution with an intense green fluorescence. (Similar results
are obtained with phthalic, glutaric and adipic acid; citric acid
gives a dirty brown solution with a blue fluorescence. The test
should be regarded as being positive only if a strong fluorescence
is produced, for a feebly positive result is sometimes obtained
when resorcinol is heated with sulphuric acid.)
Lactic acid. (a) Treat the acid or one of its salts with concentrated
sulphuric acid. Warm gently until the solution is yellow, allow to
cool and add two drops of a 5 % solution of guaiacol in alcohol.
54
 Cltulijicatio"
An intense red colour indicates the presence of a lactate. (Glycollic
acid gives a violet colour.)
(b) A positive result is obtained in the iodoform test; see page 49.

Phthalic acid (a) In a dry test-tube warm a little of the free acid or
one of its salts with an equal weight of phenol and 2 drops of
concentrated sulphuric acid. Cool the tube carefully, dissolve the
contents in water, and add an excess of dilute aqueous sodium
hydroxide to the solution. A bright red colour indicates the forma-
tion of a sodium salt of phenolphthalein.
(b) Phthalic acid gives a positive result in the fluorescein test
(see Succinic acid, above).

Salicylic acid. Gently warm a mixture of 2 m1 of alcohol (or

methanol), concentrated sulphuric acid (a few drops), and about
0·5 g of the acid (or one of its salts) for two minutes. Cool and
pour into water in a test-tube. A characteristic odour ('winter-
green') can be detected.

Pyru,ic acid (an «-keto-acid). (a) Pyruvic acid decolourizes

bromine water and potassium permanganate solution. It reduces
ammoniacal silver nitrate.
(b) Treat the solution of the acid in excess of dilute ammonia
solution with a dilute aqueous solution of sodium nitroprusside.
A violet colour, changed to red by an excess of alkali and to blue
by acetic acid indicates pyruvic acid.
(c) Pyruvic acid gives a positive result in the iodoform reaction
(see page 49).

Malonic acid. This acid is soluble in water and ether, and when
heated just above its melting point decarboxylates to produce
acetic acid.

ACIDS CONTAINING OTHER FUNCTIONAL

GROUPS

With some exceptions, acids containing functional groups in

addition to one or more carboxyl groups, may be characterized
in the same way as the simple acids. Those most likely to be
encountered are:
5S
Qualitative Organic Analysis
Aromatic nitro-acids (yellow in colour).
Aliphatic and aromatic halogeno-acids.
Aliphatic and aromatic amino-acids (amphoteric).
Aliphatic and aromatic hydroxy-acids (including phenolic
acids).
Keto-acids (see page 50).
For discussion of amino-acids, see page 74. Phenolic acids often
but not always give colours with aqueous ferric chloride. Aliphatic
halogeno-acids are considered on page 94.
Note. It is of course necessary to show the presence of the addi-
tional group(s) (nitro- etc.), wherever possible, by applying the
appropriate classification tests.
Derivatives. See page 123.

PHENOLS AND ENOLS

General tests
(a) Ferric chloride. Dissolve 0·03 g of the substance in 1 m1 of
chloroform and to this solution add I m1 of a solution of I g of
anhydrous ferric chloride in 100 m1 of chloroform. Shake the
mixture well and add one drop of pyridine. Phenols and keto-
enolic compounds give a blue, purple, red or greenish colour.
Many phenols do not give colours with ferric chloride when in
aqueous or alcoholic solution, but these will respond to the test
when it is performed as described above.
To distinguish between phenols and keto-enolic substances, add
a small amount of the substance to a solution of mercurous nitrate
in dilute nitric acid. Keto-enolic compounds give an immediate
grey precipitate of metallic mercury; phenols do not.

(b) Solubility. Simple phenols and enols are soluble In dilute

sodium hydroxide solution, but insoluble in sodium bicarbonate
solution. Nitro-, halogeno- and some polyhydric phenols are
sufficiently acidic to dissolve in sodium bicarbonate solution with
evolution of carbon dioxide.
Aminophenols are amphoteric substances, which behave
56
 Classification
normally as amines towards nitrous acid; their solutions in
alkali rapidly darken in air.

(c) Reducing powers. Phenols containing more than one hydroxyl

group possess reducing powers and some will reduce ammoniacal
silver nitrate solution in the cold. Aminophenols are also good
reducing agents.

Additional tests
1. The phtludein reaction. This test is satisfactory only if the
phenol has the position para to a hydroxyl group unsubstituted.
Heat 0·5 g of the compound and 0·5 g of phthalic anhydride
with two drops of concentrated sulphuric acid, for one minute.
Allow the mixture to cool and pour it into an excess of dilute
sodium hydroxide solution. Colours that may be produced are
indicated below:
Red: phenol, o-cresol.
Blue: m-cresol, catechol (slow).
Green: ex-naphthol.
Faint green with some fluorescence: p-naphthol.
Red solution with strong green fluorescence: resorcinol.

2. Millon's test. This test applies only to phenols having one

hydroxyl group, and at least one ortho position free.
Add 0·1 g of the substance to 2 ml of Millon's reagent (see
page 222) and heat to boiling in a beaker of boiling water. A red
colour is a positive result.

3. Liebermann's reaction. Most phenols that have a position para

to a hydroxyl group free, respond to this test. Heat a crystal of the
phenol with a crystal of sodium nitrite in a dry test-tube for 20 sec.
Cool and add I ml of concentrated sulphuric acid. An intense
green-blue colour is produced, which changes to red when the
mixture is poured carefully into an excess of water, and becomes
blue when the solution is made alkaline.

4. Azo-dye formation. Most phenols and keto-enolic compounds

couple with diazotized primary aromatic amines to give highly
coloured azo-dyes (see pages 66 et seq.).
57
QlIIlIitative Organic Analysis
Derivatives:
Simple phenols: aryloxyacetic acid; ex-naphthyl carbamate;
p-toluenesulphonate ('p-tosyl' derivative).
Nitrophenols: a-naphthyl carbamate; benzoate.
Halogenophenols: aryloxyacetic acid; a-naphthyl carbamate.
Polyhydric phenols: benzoate; 3,5-dinitrobenzoate.
Aminophenols: as for primary aromatic amines, page 138 et
seq.
Enols: as for carbonyl compounds, page 50 (items 3 and 5).

ESTERS, LACTONES AND ANHYDRIDES

(OF CARBOXYLIC ACIDS)
General tests
(a) Hydroxamic acid test. This test depends on the conversion of
the ester or anhydride into a hydroxamic acid, (RCONHOH),
which is readily detected by the high colour of the ferric salt.
To 0·1 g of the compound add 1 m1 of a 5 %solution of hydro-
xylamine hydrochloride in methanol. Add drops of saturated
methanolic potassium hydroxide until the mixture is alkaline to
litmus. Boil for up to 1 minute, cool, and acidify with dilute
hydrochloric acid. Add two drops of ferric chloride solution. The
appearance of a violet or wine colour indicates the presence of a
compound of the above classes. (Urethanes, and esters of carbonic
and chloroformic acids often give negative results.)
Note. Anhydrides and acid halides will give a positive result
without the addition of the methanolic potassium hydroxide. Some
amides also give a positive result (page 81). Acyl halides and
amides should have betrayed their presence in the preliminary
tests. Trihalo compounds, e.g. CHCI3, give positive results.
If an anhydride is suspected, but the test gives a negative result,
heat O· 5 g of the compound with 0·5 m1 of n-butyl alcohol, and
examine for an ester as described above.
It is necessary to establish that esters are absent before using the
hydroxamic acid test for any other class of compound.

(b) Hydrolysis test. Dissolve about O· 5 g of the substance in 2 m1

of neutral alcohol and add one drop of approximately normal
58
 Classification
alcoholic potassium hydroxide. Add one drop of neutral phenol-
phthalein and warm the mixture in boiling water, together with a
test-tube containing a 'blank' mixture, i.e. lacking the original
substance. Fading of the pink colour in the test solution compared
with the blank, indicates the presence of an ester, lactone or
anhydride. (The latter often causes fading before the mixture has
become hot.)
Note. Readily hydrolysable esters such as those derived from low
molecular weight aliphatic alcohols and formic and oxalic acids
give instant decolourization in test (b), and so might be mistaken
for carboxylic acids. However, the esters will give test (a) while
carboxylic acids do not.

Test for anhydrides. Dissolve a little of the substance in the mini-

mum of benzene or chloroform and add a small amount of aniline.
Warm gently for a minute or two, and cool. A precipitate indicates
the presence of an anhydride. The precipitate is the anilide (if from
the anhydride of a monobasic acid) or the anilic acid (if from the
cyclic anhydride of a dibasic acid, e.g. phthalic, succinic), and
forms a useful derivative.

ESTERS CONTAINING OTHER FUNCTIONAL

GROUPS

The nature of groups (other than the ester function) present may
vary widely, depending on the substituents present in the parent
alcohol (or phenol) and carboxylic acid. (For sulphonic esters see
page 105.)
In most cases the substituted esters show the properties of the
individual groups present. Classification tests for such groups are
usually satisfactory and should be applied when appropriate.
For the present purpose it is possible to limit the number of
types of ester. The following are those most likely to be encoun-
tered, and unless a statement is made to the contrary, identification
by hydrolysis may be carried out normally.

1. Hydroxy-esters (including esters with free phenolic groups, e.g.

methyl salicylate), and keto-enolic esters (e.g. ethyl acetoacetate).
59
Qualitative Organic Analysis
An ester with a free phenolic group and a ,B-keto-enolic ester will
give a red colour with ferric chloride (pages 34-36). Keto-esters
may be conveniently characterized through the keto-group (see
page 50). ,B-Keto-esters may be hydrolysed with hot dilute sul-
phuric acid ('ketonic hydrolysis') to a ketone, which should then be
identified.

2. Halogenated esters. If the halogen is directly attached to an

aromatic nucleus, it will be found in either the acid or the alcohol
(or phenol) formed by normal hydrolysis. If the halogen forms part
of an alkyl function, it may well be released as halide ion during
alkaline hydrolysis; in this case it will not be possible to recover
the halogenated parent compound (see page 94).

3. Nitro-esters. These are usually almost white, or yellow. The

nitro-group is most likely to be a nuclear substituent of an aromatic
parent acid or alcohol (or phenol). Apply classification test for the
nitro-group.

4. Amino-esters, e.g. methyl anthranilate. Classification tests for

both the amino- and ester functions are positive. These compounds
are most conveniently characterized through the amino-group.

5. Cyano-esters. The most common example is cyanoacetic ester

(ethyl cyanoacetate). It is a pseudo-acidic liquid substance,
insoluble in water but soluble in alkalis. By virtue of its activated
methylene group it undergoes the Knoevenagel reaction. Warm
2 ml each of benzaldehyde and the ester with two drops of mor-
pholine (or piperidine) under reflux for 10 minutes. Cool, break up
any lumps, wash with a little alcohol and recrystallize the product
from alcohol (ethyl oc-cyanocinnamate, m.p. 51°).

Identification. Esters. see page 127. Anhydrides: see page

132.

ALCOHOLS

Simple alcohols have characteristic odours. Those with up to three

carbon atoms per molecule are completely miscible with water.
The higher alcohols are less soluble in water. Polyhydric alcohols
60
 Classification
are soluble in water but not in ether, and range in appearance from
viscous liquids to crystalline solids. By far the most common
alcohols are those containing only one type of functional group
(hydroxyl).
Note. A few less common alcohols cannot be detected by the ceric
ammonium nitrate test because of interference, e.g. by the amino-
group in an amino-alcoho1.

General tests
(a) Ceric ammonium nitrate test (see page 221). Add 2 m1 of water
to 1 m1 of reagent and treat with 2 drops of the substance or 2 drops
of its saturated aqueous solution. A red colour denotes an alcohol.
If the substance is insoluble in water, use the minimum of dioxan*
as solvent and diluent. (The test is satisfactory only if the number
of carbon atoms per molecule is less than ten. Phenols and arom-
atic amines give various colours.)

(b) Xanthate test. Carefully heat O· 5 m1 of the alcohol with 2

pellets of potassium hydroxide until the latter have dissolved. Cool,
add 1 m1 of ether and shake well. Add drops of carbon disulphide
to a limit of 0·5 ml. A yellow precipitate of the xanthate indicates
an alcohol. If no precipitate forms even on standing, but a pale
yellow colour develops, apply the confirmatory test below to the
solution.
Confirm by separating the precipitate and adding a little of it to
weak ammonium molybdate solution followed by an excess of
dilute hydrochloric acid. A colour ranging from red or plum to
blue, confirms an alcohol.
Simple primary and secondary alcohols give satisfactory results
provided they are not heavily contaminated with water. Tertiary
and polyhydric alcohols are in general not readily detectable by the
test. Esters, because of hydrolysis, and some ketones containing
the group CH3COCH2- give positive results.

(c) Vanadium oxinate test (see page 223). Treat 1 ml ofthe reagent
with 0·1 ml of the alcohol (or o· 1 g dissolved in the minimum of
xylene and glacial acetic acid (1 :1)). Shake well and allow to stand
* As dioxan may contain alcoholic impurities, a blank test is
advisable.
61
Qualitatille Organic Analysis
if necessary. Most alcohols cause a change of colour from the deep
blue of the reagent to pinkish-red. Tertiary alcohols react slowly.
Owing to low solubility in the solvent mixture, some polyhydric
alcohols do not respond very satisfactorily. Water causes de-
colourization. Aldehydes, amines and a few aromatic alcohols give
a yellow colouration.

(d) Acetyl chloride test (not suitable for tertiary alcohols). In a

small dry test-tube treat 0·5 ml of the dry compound with 0·3 m1
of acetyl chloride. Alcohols and other compounds having an easily
replaceable hydrogen atom give a vigorous reaction. Carefully add
3 m1 of water drop by drop, neutralize the aqueous layer with solid
sodium bicarbonate and examine the product. If the original is an
alcohol, an ester will have been formed, the odour of which may
be distinguishable from that of the original substance. A few drops
of the upper layer should be withdrawn and tested for an ester by
the hydroxamic acid test (see page 58).

Distinguishing tests
1. Iodoform test. Dissolve the original compound in water or
dioxan, add 1 m1 of 10% aqueous sodium hydroxide and then
drops of saturated iodine solution with vigorous shaking until a
deep iodine colour persists. If no yellow precipitate of iodoform
appears on standing, warm to 60° on a water-bath, adding more
iodine solution if necessary. Remove the excess of iodine with 10 %
sodium hydroxide solution, add water, separate the iodoform and
wash it with water (iodoform m.p. 120°).
The test is positive for compounds containing the group
CH3COR or CH3CH(OH)R, the most common alcohols in this
category being ethyl, isopropyl, s-butyl and s-amyl alcohol.
Methyl alcohol of commercial quality often gives a positive result.

2. Test for methyl alcohol. Add a crystal of resorcinol to 2 ml of

the substance and plunge a red-hot copper spiral into it. Repeat
twice. The formaldehyde formed combines with the resorcinol.
The product when cooled and treated with drops of concentrated
sulphuric acid gives a white turbid mixture which turns red and
finally produces a flocculent white precipitate. Tertiary butyl
alcohol gives a similar result, while ethyl alcohol gives a yellow
colour.
62
 Classification
3. Differentiation between primary, secollllary turd tertiary aIcohola.
(i) Lucas test (applies only to aliphatic and cyclo-aliphatic alcohols
having not more than six carbon atoms per molecule. Glycols
react anomalously).
To 1 m1 of the substance in a test-tube add 8 m1 of Lucas' reagent
(a saturated solution of anhydrous zinc chloride in concentrated
hydrochloric acid). Close the tube with a cork and after vigorous
shaking, allow to stand. The formation of an alkyl halide immis-
cible with water, may be observed as follows:
Primary alcohols: no reaction is usually visible, though impuri-
ties may cause a darkening or a slight cloudiness.
Secondary alcohols: a distinct upper layer should be visible in
from 10 to 60 minutes. (Isopropyl alcohol does not give a positive
result; allyl alcohol reacts in this test although it is a primary
alcohol and yields an upper layer of alkyl halide in from 6 to 10
minutes.)
Tertiary alcohols: two layers are apparent almost at once owing
to the great ease of formation of alkyl halides from these alcohols.
To decide if a cloudiness in the above test is due to a secondary
or a tertiary alcohol, treat 1 m1 of the substance with 6 m1 of
concentrated hydrochloric acid, shake well, and allow to stand.
Tertiary alcohols give an upper layer of an alkyl halide in a few
minutes; secondary (and primary) alcohols give a clear solution.
(ii) Oxidation. Distil about I g of the substance with a mixture
of 6 m1 of water, I g of chromic anhydride and 1 m1 of concen-
trated sulphuric acid in a small flask having a side-arm, the latter to
act as a condenser. Allow the arm to dip into 5 m1 of water con-
tained in a test-tube. Test the distillate for aldehyde (from a
primary alcohol) and ketone (from a secondary alcohol). By
delivering the distillate into a solution of 2,4-dinitrophenylhydra-
zine in methanolic sulphuric acid (Brady's reagent), crystalline
derivatives of the carbonyl compound can be obtained (see Alde-
hydes and Ketones, page 121).

4. Periodic acid test (for polyhydric alcohols). This test is based

on Malaprade's reaction, in which an alcohol having two hydroxyl
groups situated on adjacent carbon atoms is oxidized by periodic
acid, the latter being reduced to iodic acid which is detected in the
test as its silver salt.
2 m1 of a 0·5 %solution of periodic acid in water is treated with
one drop of concentrated nitric acid and the mixture is shaken
63
Qualitative Organic Analysis
thoroughly. A small crystal (or two drops) of the substance is
added and the whole shaken once more. Mter about 30 seconds,
two drops of dilute (3 %) silver nitrate solution are added. The
rapid formation of a white precipitate (of silver iodate) is a positive
test. A brown precipitate which dissolves on shaking is of no
importance.
Note. IX-Hydroxy-aldehydes (e.g. simple ~ugars), IX-hydroxy-
ketones, IX-hydroxy-acids and IX-amino-alcohols also give positive
results. The preliminary tests should have given evidence of the
presence of the non-alcoholic functional group in these com-
pounds.
Derivatives. See page 133.

ETHERS

All common ethers are sparingly soluble in water. They dissolve

in concentrated sulphuric acid (cooled in an ice-bath) but are
regenerated when the solution is poured into water, except for
some aromatic ethers which may be partially sulphonated and so
may be incompletely recovered.
Many ethers possess strong odours; those of simple aromatic
ethers such as anisole are sometimes confused with the odours of
the simple aromatic hydrocarbons.
Ethers, like hydrocarbons, are somewhat unreactive substances.
Consequently there is a lack of really convenient and reliable tests
for distinguishing between the two types of compound, and great
care is needed when attempting to assign a substance to one of
these two classes.

Tests
(a) The following test must NOT be used unless the ester group
has been tested for and found to be absent from the original
substance.
Make a mixture of 1 m1 of the substance and 4 m1 of glacial
acetic acid, and carefully add 1 m1 of concentrated sulphuric acid.
Heat the mixture so formed under reflux for 5 minutes, cool well,
and extract with benzene. Use the extract to test for an ester of
acetic acid formed by the fission of the ether (hydroxamic acid test,
64
 Classification
page 58). A positive result is indicated by the appearance of a
purple colour.
(b) Make a light-purple solution of iodine in carbon disulphide
and to 1 m1 of this solution add O' 5 ml of the original substance.
Shake the mixture well. A change of colour from purple to tan in
the carbon disulphide layer indicates an ether. It is best to compare
the test solution with a 'blank', i.e. a portion of the iodine solution
used in the test diluted with carbon disulphide to the same volume
as the test solution.
Too much reliance must not be placed on this test; some alipha-
tic hydrocarbons respond to it.
(c) If the substance is a liquid, treat it with a very small piece of
freshly cut sodium. Provided it is quite pure and dry, an ether
does not react (a pure hydrocarbon is also without action).
Derivatives. See page 135.

HYDROCARBONS
1. Unsaturated Hydrocarbons
(a) Mix some of the substance with an equal volume of cold
concentrated sulphuric acid (CARE!). Unsaturated aliphatic and
alicyclic hydrocarbons, aromatic hydrocarbons with unsaturated
side-chains, and highly alkylated hydrocarbons, dissolve.
(b) Tests for unsaturation.
(i) The bromine test (see preliminary tests). Bromine in carbon
tetrachloride instantly decolourized indicates unsaturation, though
in some cases decolourization may only occur on warming.
(ii) Treat 0·2 g of the substance in 2 ml of acetone with dilute
potassium permanganate solution added drop by drop. A brown
colour or precipitate is a positive test, though not if less than
3 drops of permanganate are decolourized.

2. Saturated Hydrocarbons
Place 2 m1 of20% fuming sulphuric acid (CARE!) in a DRY test-
tube, add O· 5 ml of the compound and shake gently. Aromatic
hydrocarbons dissolve; saturated aliphatic and alicyclic com-
pounds are insoluble.
Identification. See page 136 and Tables 12 and 13.
E 65
 Classification

SECTION III

Amines. Salts of Amines. Polyfuncti01lll1 Compoumls which contain

one or more Amino-Groups.
(Amino-compounds containing sulphur are considered in Section
VIII.)

AMINES

The majority of the simple aliphatic amines and aromatic side-

chain amines (e.g. benzyl amine) are liquids with powerful odours
resembling but not identical with that of ammonia. They dissolve
in water yielding alkaline solutions. Simple heterocyclic amines are
also soluble in water. All compounds of these types dissolve readily
in dilute mineral acids.
Except for some polyfunctional aromatic amino-compounds
(e.g. aminophenols, some amino-acids, phenylenediamines) aro-
matic amines are not soluble in water. Most dissolve in dilute
hydrochloric acid, but some weak bases, e.g. nitroanilines,
diphenylamines and polyhalogeno-anilines dissolve only in con-
centrated acids. A few aromatic amines (e.g. naphthylamines)
dissolve in concentrated hydrochloric acid and then slowly pre-
cipitate as their sparingly soluble hydrochlorides. In all cases of
doubt as to the solubility of a suspected amine in hydrochloric acid,
a little of the filtered solution should be treated with an excess of
alkali, when the amine (if insoluble in water) should be regenerated
as an oil or solid. Amino-phenols and amino-acids are amphoteric
substances (see pages 74-76).

General test-the action of nitrous acid; see diagram opposite.

Limitations of the test. Under the conditions used for the test, a
clear distinction between the various types of amine is not always
obtained. For example, some primary aliphatic amines (e.g.
methylamine) do not react normally with nitrous acid to yield the
corresponding alcohol and nitrogen, and in any case unless the
test is carried out very carefully a steady evolution of nitrous fumes
from the reaction mixture may be erroneously reported as nitrogen.
Again, some common secondary aliphatic amines give water-
66
 AMINE
I
Treat with
nitrous acid
I
Oear solution ..
TurbI·d·Ity, orI precIpItate
obtained (A) of an oil or solid (B)
I I
I 1 I I
Vigorous evolution When poured into Original compound Turbid S~IUtion or Deep red-brown
of nitrogen an excess of alkaline liberated when separation of an solution, possibly with
,B-naphthol solution is made oily or solid a brown or yellow
solution, an azo alkaline nitrosamine precipitate of the
I dye is produced (yellow to brown) hydrochloride of a
p-nitroso-compound
I I I I I
Primary Primary Tertiary Secondary Tertiary
aliphatic amine aromatic amine aliphatic or amine B(i) aromatic amine
A(i) A(ii) heterocyclic B(ii)
amines A(iii)

Some members of Various substances Some secondary o-Diamines and m-Diamines also
this class cannot be may give misleading amines are also arylhydrazines give give brown precipitates
detected in this way results here regenerated products of similar
unchanged appearance. Some
secondary amines
do not readily give
nitrosamines
~
Qualitatipe Organic Analysis
soluble nitrosamines, and others react only very slowly. The latter
may be liberated unchanged on making the solution alkaline (cf.
tertiary aliphatic and heterocyclic amines).
For the above reasons, it is recommended that, for aliphatic
amines, greater importance should be attached to the additional
tests listed for the appropriate type of amine (page 71).
The test, if properly conducted, is satisfactory for the detection
of primary, secondary and tertiary aromatic amines.

Precautions. The following points of technique are important.

1. The large excess of acid used to dissolve the amine and react
with the sodium nitrite is necessary to avoid undesirable by-
products being formed. Do not use less than the volume stated.
2. The temperature of the reaction solution should initially be as
near 0° as possible; it must not be allowed to rise above 10° at any
time.
3. Slow addition of the sodium nitrite solution, efficient stirring
and external (ice-bath) cooling are essential, otherwise high local
solution temperatures may be developed.
4. Nitrous acid reacts rapidly but not instantaneously with
amines. Consequently. in testing for completion of the reaction
(presence of unreacted nitrous acid), the starch-iodide test for free
nitrous acid should not be applied until 5 minutes after the addition
of a portion of the sodium nitrite solution.
5. Use only fresh starch-iodide paper as external indicator; test
it with a drop of dilute hydrochloric acid, when it should not show
a blue colour at once. (Starch-iodide papers, when wet, become
blue in the laboratory atmosphere in a relatively short time due to
airborne oxidants.)

Procedure. Dissolve 0·5 g (or O· 5 ml) of the amine in a mixture of

3 ml of concentrated hydrochloric acid and 2 ml of water. Cool the
solution to 5° in an ice-bath. Prepare an ice-cold solution of 0·4 g
of good quality sodium nitrite in 4 ml of water and add it slowly
in small portions, stirring continuously, to the amine solution.
Keep the solution below 10° during this operation. Leave the
solution in the ice-bath for 5-10 minutes and then place a drop of
the solution on starch-iodide paper. It should show an immediate
blue colour, indicating the presence of an excess of nitrous acid.
If this is not so, add a further small volume of sodium nitrite
68
 Classification
solution, allow the mixture to stand in ice for 5 minutes, and test
with starch-iodide paper again.
(If the amine is not very soluble in hydrochloric acid, dissolve it
in a mixture of water (3 ml) and concentrated sulphuric acid (4 ml),
cool as above, and treat it with sodium nitrite dissolved in sul-
phuric acid of the same concentration as used for the amine.)
The following observations and tests are made on the reaction
mixture. (Keep the mixture in ice when not being tested.)

A. H the solution remains clear

(i) A sustained and vigorous evolution of gas (nitrogen) indicates
that the original substance was an aliphatic primary amine or an
aromatic side-chain primary amine. Additional tests for these
compounds are given on page 71.
(ii) Pour a few drops of the reaction mixture into a solution of
0·5 g of ,a-naphthol in 5 ml of dilute (2N) sodium hydroxide
solution. A precipitate or solution of a bright red or orange azo-
dye indicates a primary aromatic amine. After purification the azo-
dye is sometimes useful as a derivative of the amine (for melting
points of a small number of azo-dyes see Table 24, page 209).
Notes. (a) Traces of primary amine in technical samples of secon-
dary or tertiary aromatic amines may give a positive result.
(b) It is important that an excess of the diazonium solution shall
not be added to the ,a-naphthol solution, otherwise cx-nitroso-,a-
naphthol may be precipitated (yellow to light brown). The test
solution must be alkaline at the end of the reaction.
(c) If the azo-dye is appreciably soluble, suspect the presence of
hydroxyl or sulphonic groups in the original amine.
(tl) In the above test, anilides sometimes give positive results due
to the initial formation of nitroso-amides, (CtiHsN(NO)COR),
which with alkali form azo-compounds, (C6HSN:NONa). These
then couple with the ,a-naphthol to form red dyes. Anilides how-
ever do not give the dinitrochlorobenzene test (page 71).
(e) In this test, o-diamines do not react normally; they give
azimides (yellowish colours or precipitates). These should be
differentiated from the visually similar products obtained from
secondary amines (nitrosamines) and from some aromatic tertiary
amines (p-nitroso-compounds).
m-Diamines give intense dark brown azo-products with nitrous
acid, e.g. Bismarck Brown from m-phel1ylenediamine.
69
Qualitative Organic Analysis
(f) Some primary arylhydrazines (ArNHNHV may give mis-
leading results in this test. They may be distinguished from arom-
atic primary amines by the ease with which they reduce Fehling's
solution.
(iii) Carefully add an excess of dilute sodium hydroxide solution
to a portion of the reaction solution. Tertiary aliphatic and
heterocyclic amines, (and some aliphatic secondary amines) which
do not react with nitrous acid, are liberated unchanged.

B. H the mixture is turbid, is red-brown, and/or contains an on or

solid precipitate
(i) The formation of a yellowish emulsion or precipitate may
indicate the presence of the N-nitrosamine of a secondary amine.
(o-Diamines (see A(ii) above) produce yellowish colours or
precipitates; arylhydrazines (see page 85) give turbid solutions, but
may be distinguished by the ease with which they reduce Fehling's
solution.)
To confirm the presence of a nitrosamine, apply Liebermann's
test:
Place a portion of the reaction liquor in a small separatory
funnel, add a little urea to destroy excess of nitrous acid, and shake
with ether to extract the nitrosamine. Separate the ether layer,
wash it with dilute alkali and then with water. Evaporate the ether
on a previously heated water-bath. To two drops of the resulting
oil (or a few mg if a solid) add a drop of concentrated sulphuric
acid. A blue colour here indicates the presence of nitrosodiphenyl-
amine. H there is no blue colour, treat two drops of the oil (or a
few mg if a solid) with a small crystal of phenol and warm gently
for 20 seconds. Cool and add I m1 of concentrated sulphuric acid.
An intense green-blue colour is produced, changing to red when
poured into 30 m1 of water, and becoming greenish-blue when an
excess of sodium hydroxide solution is added.
(ii) A deep red-brown solution which may deposit a yellow or
brown precipitate on standing in ice probably indicates the
presence of the hydrochloride of a p-nitroso-compound derived
from a tertiary aromatic amine. The hydrochloride will be in-
soluble in ether (contrast nitrosamines).
Treat the reaction mixture with a slight excess of dilute sodium
hydroxide solution. A precipitate of the free p-nitroso-compound,
which is characteristically green or bluish-green, shows the original
70
 Classification
substance to be an aromatic tertiary amine. Extract the derivative
with ether, dry the extract with anhydrous sodium sulphate, and
evaporate the ether. Recrystallize from light petroleum and deter-
mine the m. p. (Table 17). (A dark brown precipitate on making the
reaction mixture alkaline may indicate the original substance to be
a m-diamine, e.g. Bismarck Brown from m-phenylenediamine.)

Additional tests
1. Aliphatic (and aromatic side-chain) primary amines
(a) Rimini's test. Make a solution or suspension of the amine in
water (one drop to 3 mI) and to it add 1 mI of acetone and one drop
of a 1 %solution of sodium nitroprusside in water. A red or violet
colour develops within about 2 minutes. It is important that the
acetone used is free from acetaldehyde. (Some secondary amines
of commercial quality give a positive result due to impurities.)

(b) 2,4-Dinitrochlorobenzene test. Moisten a strip of filter paper

with a saturated alcoholic solution of 2,4-dinitrochlorobenzene.
To it add a drop of an aqueous or alcoholic solution of the amine,
or if a salt, warm the latter with dilute sodium hydroxide solution
and hold the filter paper in the vapours evolved. An intense yellow
colour indicates a primary amine. Some secondary amines give a
paler colour, more slowly.

(c) Carbylamine test. This test is very sensitive and may give a
positive result with a secondary or a tertiary amine containing a
small amount of primary amine as impurity. Some anilides may
give weakly positive results owing to slow hydrolysis to primary
amines.
Add 3 drops of chloroform to about 0·1 g of the substance,
followed by 2 mI of alcoholic potassium hydroxide, and gently
warm the mixture. The foul smell of a carbylamine indicates a
primary amine. As soon as the odour has been detected, add an
excess of concentrated hydrochloric acid and wait until the smell
has gone before pouring the mixture away.
Note. Owing to the toxic nature of carbylamines (isocyanides) this
test should be carried out in a fume cupboard.

(d) Hinsberg's reaction. Primary amines may be sometimes dis-

tinguished from secondary amines by the fact that their p-toluene-
71
Qlllllitative Organic Analysis
sulphonyl derivatives (Schotten-Baumann method, page 139) are
acidic and soluble in alkali whereas those of secondary amines are
insoluble in alkali. This distinction is not completely reliable
because some primary amines give non-acidic disulphonamides,
and others form sparingly soluble sodium salts.

2. Aromatic primary amines

See tests l(b, c, tl) above.

3. Aliphatic and aromatic secondary amines

(a) Simon's test. A few secondary amines (e.g. diethylamine, mor-
pholine, piperidine, piperazine) do not readily give nitrosamines.
These and most other secondary amines give Simon's test, which
works best only with freshly-prepared acetaldehyde solution.·
Suspend or dissolve one drop (or a few mg) of the compound in
3 m1 of water and add 1 m1 offresh acetaldehyde solution, followed
by two drops of sodium nitroprusside solution. A deep blue colour
is produced within five minutes, fading slowly to blue-green and
then yellow.

(b) Carbon disulphide reagent (for preparation, see page 222). A

secondary amine reacts with carbon disulphide to form a di-
substituted dithiocarbamate, which readily yields an insoluble
nickel salt. The test is more satisfactory with aliphatic amines, but
some aromatic amines give a positive result, e.g. N-ethylaniline,
N-methylaniline.
To 5 m1 of water add two drops (or a few mg) of the amine, and
if the latter is insoluble in water, two drops of concentrated
hydrochloric acid. Place 1 m1 of the nickel-carbon disulphide
reagent in a test-tube, add 1 m1 of concentrated ammonia solution
and 1 m1 of the amine solution. A precipitate indicates a secondary
amine, while a slight turbidity may be due to the presence of a
secondary amine as impurity.

• Freshly prepared acetaldehyde may be obtained by heating a

small coil of copper wire to redness, and plunging it repeatedly
into 5 m1 of 50 %alcohol in a heat-resisting glass test-tube. Repeat
the heating and immersion in the alcohol several times, cooling
the test-tube under the tap. Use the final cold solution for Simon's
test.
72
 ClIIssifieation
4. Tertiary amines
Purely aromatic tertiary amines, e.g. triphenylamine, do not dis-
solve in water or in dilute acids, but dissolve in concentrated
sulphuric acid. They are almost devoid of basic properties and so
do not give quaternary ammonium iodides (see below). They are
not often encountered, the common tertiary amines being of the
classes:
Aliphatic, e.g. triethylamine
Mixed aliphatic-aromatic, e.g. N,N-dimethylaniline
Tertiary heterocyclic bases, e.g. pyridine
Tertiary amines do not give the carbylamine reaction. With the
exception of mixed aliphatic-aromatic amines (which yield p-
nitro so-compounds), members of these classes do not react with
nitrous acid. The following tests are often useful.

(a) Methiodide formation. * Add 5 drops of methyl iodide to 5 drops

of the amine in a dry test-tube and stand for 5 minutes. Warm for a
further 5 minutes on a water-bath, cool in ice and scratch the sides
of the tube with a glass rod. A precipitate or complete solidifica-
tion, indicates a tertiary amine. To confirm, dissolve the quater-
nary ammonium salt in water and shake with a slight excess of
silver oxide. Filter and test the filtrate with litmus, which should
show a strongly alkaline reaction owing to the presence of the
quaternary ammonium hydroxide.
(b) Nessler's reagent (for preparation see page 222). Dissolve the
base in dilute hydrochloric acid and add Nessler's solution. A white
precipitate is formed. (This test should be used with caution: some
compounds other than tertiary amines may give positive results,
e.g. anilides, various amino-compounds.)
(e) To distinguish pyridine from quinoline. Dissolve one drop of the
base in 1 ml of dilute hydrochloric acid and add a few drops of
saturated aqueous potassium dichromate solution. A copious

• Primary and secondary amines, when treated with excess of

methyl iodide, yield similar products, so that this test does not
necessarily give unambiguous proof of the presence of a tertiary
amine. Avoidance of excess of methyl iodide is obviously impor-
tant.
73
Qualitati,e Organic Analysis
orange-yellow precipitate of the dichromate salt is formed with
quinoline, but pyridine does not give a precipitate.
Deri,ati,es. See pages 138-142.

POLYFUNCTIONAL AMINO-COMPOUNDS

The commonest are:

1. Diamines 6. Amino-azo compounds
2. Amino-acids 7. Amino-esters
3. Aminophenols 8. Amino-anilides
4. Amino-alcohols 9. Nitro-amines
5. Aminoketones 10. Halogeno-amines
Except where indicated below, the functional groups present in
such compounds show the normal properties, and may be detected
by the appropriate classification tests.

1. Phenylenediamines
These compounds are soluble in water, and their solutions give
colours with ferric chloride solution (see preliminary tests). 0- and
m-Phenylenediamines do not react normally in the nitrous acid
test (q.v.). o-Phenylenediamine combines with oc-diketones or
o-quinones (q.v.) to give quinoxalines.

2. Amino-carboxylic acids
(i) Aliphatic amino-acids, +NH3-CH-COO-
I
R
Only the cx-amino-acids, which are the commonest, will be con-
sidered. Many are soluble in water, but not in ether, and are almost
neutral. They dissolve in alkalis and acids, (amphoteric). They
decarboxylate in the soda-lime test, evolving amines.
(a) Sodium bicarbonate. Carbon dioxide is liberated only slowly,
and may not be detected until after a lapse of several minutes.
(b) Ferric chloride. Aqueous or alcoholic solutions of amino-
acids give red colours with this reagent.
(c) Sorensen's reaction. Dissolve about 0·2 g of the substance in
74
 ClassijiCfltion
water or alcohol and neutralize with dilute sodium hydroxide to
phenolphthalein. Add a few ml of neutralized (phenolphthalein)
formaldehyde. The immediate disappearance of the pink colour
indicates the presence of an amino-acid.
(d) The ninhydrin test.* Heat an aqueous solution of the sub-
stance with a few drops of a 0·2 %solution of ninhydrin reagent.
A blue colour is given by IX-aminO acids.
(e) Copper sulphate. Add a few drops of aqueous copper sulphate
(Fehling's solution A) to an aqueous solution of the substance.
Many aliphatic amino-acids give a deep blue colour.
(f) Ice-cold nitrous acid solution causes evolution of nitrogen as
with aliphatic amines (q.v.).
(g) In the carbylamine test, the characteristic odour is usually not
readily detected, owing to the fact that the isocyanide here is
acidic and forms a metallic salt which renders it non-volatile.

(ii) Aromatic amino-acids. These are generally sparingly soluble in

cold water but are more soluble in hot water. Unlike the aliphatic
amino-acids, aromatic amino-acids usually cause a vigorous
evolution of carbon dioxide from sodium bicarbonate solution.
The tests and comments in (b) and (g) above, are applicable to
these acids. With nitrous acid, aromatic amino-acids give diazo-
nium salts, which couple normally with alkaline ,8-naphthol.

(iii) N-Acylamino-acids (e.g. N-benzoylglycine). Compounds of

this type show strongly acidic character. On hydrolysis with
moderately concentrated sulphuric acid they yield an amino-acid
and a carboxylic acid (e.g. glycine and benzoic acid from N-
benzoylglycine).

Derivatives. (i) Aliphatic amino-acids. (a) Prepare the

benzoyl derivative by the method described on page 139
(Schotten-Baumann), but using sodium carbonate solution
instead of sodium hydroxide. The derivative must be
precipitated from the mother liquor by final acidification.
(b) Prepare the a-naphthyl isocyanate derivative thus:
Add 1 g of the isocyanate to a solution of 0·5 g of the

* Positive results are also obtained with ammonium salts, some

amines, and proteins.
75
Qualitative Organic Analysis
amino-acid in 25 ml of N/2 alkali. Shake the mixture for
5 minutes and allow to stand for a further 25 minutes.
Filter to remove any diarylurea, acidify the filtrate with
hydrochloric acid, collect the precipitate* and wash it with
cold water. Recrystallize the product from water or
alcohol.
(ii) Aromatic amino-acids. Prepare the formyl, acetyl (see
pages 141, 138), or benzoyl derivative (see above). p-Nitro-
benzyl esters of these acids may be prepared as described
on page 124.

3. AminophenoIs
These compounds are colourless when pure but are frequently
encountered as brown solids. They are soluble in alkalis and acids
and some are soluble in water. The preliminary tests will have
shown the reducing properties of a substance of this class. Amino-
phenols react normally with nitrous acid.
Derivatives. As for primary aromatic amines. If preparing
a benzoyl derivative, use an excess of the reagent, to
convert to the dibenzoyl compound.

4. Amino-alcohols
These are oily liquids, soluble in water, insoluble in ether, and
strongly alkaline. The amino-group interferes with the ceric
ammonium nitrate test for the alcohol function.
Derivative. Picrate.

5. Aminoketones (e.g. p-amino-acetophenone)

The amino-group may interfere with the 2,4-dinitrophenylhydra-
zine test for the carbonyl group.
Derivatives. As for amines.

6. Amino-esters (e.g. methyl anthranilate)

In aromatic compounds of this type, both functional groups may
be detected normally.
Derivatives. As for amines.
* It may be necessary to cool for an hour to obtain complete
precipitation.
76
 Classification
7. Amino-anilides
The common examples of this type of compound are mono-acyl
derivatives of aromatic diamines, such as p-amino-acetanilide,
H2NC6H4NHCOCH3'
On hydrolysis, (see classification tests for anilides), a diamine is
liberated, and it should be noted that 0- and m-diamines do not
give clear diazonium solutions when treated with nitrous acid
(page 69).
If the original substance is the mono-acyl derivative of an 0-
diamine, the action of heat causes cyclization, with formation of an
alkylbenziminazole (e.g. o-amino-acetanilide gives methylben-
ziminazole, m.p. 175°).
Derivatives. As for amines; the products of hydrolysis of
the original substance obviously give alternative routes to
identification.

8. Amino-azo-compounds
On reduction with tin and hydrochloric acid, these compounds
give two amines, at least one being a diamine.
Derivatives. As for amines.

9. Nitro-amines (e.g. nitro-anilines)

Nitro-amines are yellow or orange crystalline solids, sparingly
soluble in water, and not very soluble in dilute hydrochloric acid.
They dissolve in concentrated hydrochloric acid (except poly-nitro-
anilines) and may be diazotized and coupled with ,B-naphthol in the
usual manner.
In the soda-lime test, 0- and p-nitro-amines evolve ammonia.
The nitro-group(s) may be detected by the usual classification
tests, though it must be remembered that when reduced with tin
and hydrochloric acid, nitro-amines give diamines (or poly-
amines), and these may not react normally with nitrous acid (see
page 69).
Derivatives. Acetyl, formyl or benzoyl derivatives.

10. Halogeno-amines
These compounds are weaker bases than the corresponding simple
amines, and dissolve readily only in fairly concentrated acid. The
77
Qualitative Organic Analysis
only important compounds are those in which the halogen is
attached to an aromatic nucleus, and in this position it is not
'mobile' as determined by the preliminary tests.
Derivatives. As for simple amines.

AMINE SALTS

Salts of amines with mineral acids are usually sufficiently soluble

in water to give strongly acid solutions. The salts with organic
acids are also generally acid to litmus.
Treat the solution of the substance with a slight excess of
sodium hydroxide solution, * and extract the liberated amine with
ether. Identify the amine. The aqueous residue, after extraction,
contains the sodium salt of the acid, which should be identified by
the usual inorganic tests, e.g. excess of nitric acid and silver nitrate
solution for halide, excess of hydrochloric acid and barium chloride
solution for sulphate, or by the tests on pages 52-55, if the acid is
organic.
Note. Aniline sulphate is sometimes confused with sulphanilic
acid. The latter, unlike the former, does not give the test for
sulphate ion with hydrochloric acid and barium chloride.

* If the original compound 'is a salt of an aminophenol, the latter

may be precipitated when the solution is just neutral but will
dissolve when alkali is present in excess. Salts of amino-acids
behave in this manner also; the amino-acid may be removed by
the addition of acetic acid to the neutral solution.
78
 Classification

SECTION IV

Ammonium Salts of Carboxylic Acids Aldehyde-ammonias

Amides lmides Nitrites
Acyl Derivatives of Bases
Hydrazines, Semicarbazide, Hydroxylamine,
their salts and carbonyl derivatives
Condensation Products of Aldehydes with Ammonia or Amines
With the exception of some of the free bases all of these com-
pounds can be hydrolysed to yield ammonia and/or an organic
base.
They are considered in the order given above.

AMMONIUM SALTS, ALDEHYDE-AMMONIAS,

AMIDES, IMIDES, NITRILES

All compounds of the above types liberate ammonia with varying

ease when treated with sodium hydroxide solution.
Ammonium salts, aldehyde-ammonias, amides (except forma-
mide) and imides are crystalline solids. Ammonium salts, aldehyde-
ammonias, simple amides and nitriles, and some imides are soluble
in water; the solutions of aldehyde-ammonias are alkaline, those of
imides (and some amides) are feebly acid.
Nitriles are liquids or low melting solids. Alkyl cyanides usually
have moderate water-solubility, while aryl cyanides are sparingly
soluble in water and some smell of almonds.
Nitriles liberate ammonia much more slowly than amides, and
in the soda-lime test distil partly unchanged.

AMMONIUM SALTS OF ACIDS;

ALDEHYDE-AMMONIAS

General test (which differentiates these substances from simple

amides and imides)
Grind a small quantity of the substance with magnesia in a small
79
Qualitative Organic Analysis
mortar, and just moisten with water. Evolution of ammonia is a
positive result.
Test for ammonium salts. To 1 ml of a 4 % solution of phenol in
water, add 1 ml of 5 %sodium hypochlorite solution and a crystal
(or a few drops of aqueous solution) of the original substance.
Shake the mixture and warm gently. A blue colour confirms an
ammonium salt.
Note. If the substance is suspected to be an ammonium salt of a
nitrogen-containing acid, heat it with sodium hydroxide solution
until all ammonia has been driven off. Acidify the solution,
evaporate it carefully to dryness, and apply the Middleton test for
nitrogen. A positive result will confirm the presence of nitrogen in
the free acid.
Derivatives. Boil the substance with alkali to remove
ammonia, acidify, and isolate the acid. Prepare derivatives
of the latter (page 123). (Some colour tests for carboxylic
acids may be made directly on the ammonium salt.)

Tests for aldehyde-ammonias. (a) Boil a small quantity of the

substance with a few m1 of dilute sulphuric acid, collecting a little
of the distillate. Examine the distillate for the presence of aldehyde.
If the test is negative, the aldehyde may be non-volatile and so must
be removed from the mother liquor by ether extraction and sub-
sequent evaporation of the solvent, followed by tests for aldehyde.
Identify the aldehyde in the usual manner.
(b) Treat 3 m1 of the 2,4-dinitrophenylhydrazine reagent with not
more than 0·1 g of the substance in aqueous methanol. Aldehyde-
ammonias give the 2,4-dinitrophenylhydrazone of the parent
aldehyde, which serves to identify the latter.

AMIDES AND IMIDES

(including amides and ester-amides of carbonic acid)

Unlike simple amides (RCONH2), simple imides (RCONHCOR')

contain an acidic hydrogen atom, and they behave similarly to
weak acids. Consequently an imide, if water-insoluble, can be
distinguished from an amide by its solubility in dilute alkali.
80
 Classification
Imides are rather more resistant to alkaline hydrolysis than are
amides.

General tests
(a) Heat the substance with 30 % sodium hydroxide solution.
Amides and imides evolve ammonia. Cool the solution and care-
fully add dilute hydrochloric acid: amide derivatives of carbonic
acid (urea, mono-substituted ureas, urethanes, guanidine) evolve
carbon dioxide from the acidified solution. If the amide or imtde
is derived from a sparingly soluble acid, the latter will precipitate
from the acidified solution and should be removed for identifica-
tion.
Note. With hot alkali mono-substituted ureas give both ammonia
and an amine, e.g. phenylurea, (C6H sNHCONHv yields ammonia
and aniline. Careful acidification of the cooled mixture then causes
evolution of carbon dioxide.
(b) Provided the test is carefully carried out, treatment of the
substance with nitrous acid (see Amines) can be shown to cause
evolution of nitrogen.

Test for imitles. Treat a saturated solution of the substance in

methanol with a saturated methanolic solution of potassium
hydroxide. Many imides yield white precipitates of their potassium
salts.

Test for aliphatic amities. Aliphatic amides, but not aromatic

amides, (with the exception of salicylamide, which contains a free
phenolic group and reacts directly with ferric chloride solution)
give the hYdroxamic acid test under the following conditions:
Add O· 1 g of the compound to 1 m1 of 5 %methanolic hydro-
xylamine hydrochloride reagent and boil for three minutes. When
a drop of ferric chloride solution is added to the cooled solution, a
wine-red or purple colour is produced.

The biuret test. This test is a general one for compounds containing
two -CONH- groups attached to one another directly, or attached
to the same carbon or nitrogen atom. It is therefore given by
proteins and peptides. Of the simpler compounds responding to the
test, urea, malonamide and oxamide are important. For malona-
mide and oxamide the initial fusion may be omitted.
F 81
Qualitative Organic Analysis
Gently heat a small amount of the substance so that it just melts
and evolves ammonia. When the mixture solidifies owing to the
formation of biuret, usually after about one minute, allow the
solid to cool, dissolve it in warm dilute sodium hydroxide solution,
cool, and add one drop of very dilute copper sulphate solution.
A purple or blue colour develops.
Xanthydrol test. Dissolve the compound in 50 % acetic acid and
add to the solution 1 m1 of 5 %methanolic xanthydrol reagent. An
immediate precipitate of the xanthyl derivative is given by urea,
urea salts and mono-substituted ureas. After recrystallization from
aqueous dioxan, the product can be retained as a derivative.
Urease test (specific for urea). Add a few drops of phenolphthalein
to a solution ofO' 2 g of the substance in 5 m1 of water, followed by
a pinch of jack bean meal or a crushed urease tablet. Shake the
mixture, which slowly becomes pink due to the hydrolysis of urea
to ammonium carbonate (alkaline).
Tests for urethanes (esters of carbamic acid, H 2NCOOH).
(a) Gently warm a little of the compound with a solution of
potassium hydroxide in ethyl alcohol. Crystalline potassium
cyanate is precipitated. Filter and dissolve the solid in water. To
this solution add a fairly concentrated solution of aniline hydro-
chloride in water, when phenylurea, m.p. 147°, is precipitated.
(b) If test (a) is positive, hydrolyse the compound by refluxing it
with dilute sodium hydroxide solution for five minutes, dilute the
cooled liquor, and distil off about one third of the liquid. Apply
tests to this distillate for alcohols.
(c) Urethanes do not usually give a positive result in the hydrox-
amic acid test for esters (page 58).

Tests for guanidine. Guanidine is usually encountered in the form

of its salts which are stable in aqueous solution.
(a) Warm the compound with dilute alkaline potassium per-
manganate solution. Guanidine and its simple derivatives effect
reduction of the reagent in a short time.
(b) Treat the compound with a concentrated solution of sodium
hypochlorite; a red to orange colour develops.
(c) Gently heat the substance in a small tesHube so that it just
melts. Maintain this condition until a brownish residue is obtained,
82
 Classification
when strong heat is applied and a strip of filter paper, previously
treated with drops of solutions of 8-hydroxyquinoline in ethanol
and potassium cyanide in water (CAUTION), is held at the mouth
of the tube. Within about 2 minutes, a scarlet stain appears on the
paper. This test is given by urea (but only after prolonged pyroly-
sis) and by thiourea.
Derivatives. See page 142.

NITRILES

These do not evolve ammonia with cold alkali, and only slowly on
heating with alkali.

General tests
(a) Boil the compound (1 g) under reflux with 10 ml of 50% (v/v)
sulphuric acid for 10 minutes. Cool and treat a portion of the
hydrolysate with an excess of aqueous sodium hydroxide. Ammo-
nia is evolved. If the nitrile gives rise to a sparingly soluble acid, the
latter may precipitate from the bulk of the hydrolysate, and should
be isolated with a view to its identification.
(b) Reduce the substance to the corresponding primary amine by
dissolving 0·1 g of it in 5 ml of ethanol and adding a small piece of
sodium. When the sodium has dissolved, detect the primary amine
by adding a few drops of chloroform and warming gently. The
repulsive odour of carbylamine indicates the presence of a cyanide.
(Aliphatic nitriles give fairly good results in this test, but generally
aromatic nitriles do not react appreciably.) Perform the test in a
fume-cupboard, and destroy the carbylamine by adding excess of
concentrated hydrochloric acid and allowing to stand.
Derivatives. See page 144.

ACYL DERIVATIVES OF BASES

In this group are included N-substituted imides, (RCONR'COR");

and N-substituted amides, (e.g. anilides, PhNHCOR; and
RR'NCOR'').
These compounds are crystalline solids, insoluble in cold water
(except for some aliphatic members), but soluble in hot water.
Some are soluble in concentrated mineral acids.
83
Qualitative Organic Analysis
Compounds containing an acyl group derived from a ,8-keto-
acid, e.g. acetoacetanilide, possess keto-enolic properties, dissolv-
ing in alkalis, and giving red colours with neutral ferric chloride
solution (see preliminary tests). Anilides derived from phenolic
acids may react similarly with these reagents.
Some acylated arylamines (e.g. anilides) give misleading results
in the nitrous acid test for amines (page 69).
The soda-lime test (preliminary tests) decomposes acylated bases
into the base and the parent acid, though the latter may then be
decarboxylated to a hydrocarbon. Hydrolysis of these compounds
by alkali is often slow, and the following method is preferable.

General test
Hydrolyse the substance by heating together in a small reflux
apparatus, I m1 of diethylene glycol, I ml of concentrated hydro-
chloric acid, I m1 of syrupy phosphoric acid, and 0·5 g of the
compound. Boil the mixture for about two minutes, allow it to
cool, and decant a small portion. To this add an excess of aqueous
sodium hydroxide solution-evolution of a volatile amine indicates
that the original substance is derived from a simple aliphatic amine.
(Ammonia may be liberated from a mono-substituted urea, e.g,
phenylurea.)
If an amine has not been detected, add 5 m1 of dilute hydro-
chloric acid to the bulk of the hydrolysate, filter if necessary (the
residue may be a sparingly soluble acid and/or a sparingly soluble
hydrochloride of the base), cool the solution in ice, and treat it
with 5 ml of a 2 %ice-cold sodium nitrite solution. The separation
of a yellow oil or solid on standing indicates that the original is an
acylated secondary amine, the yellow product being the corres-
ponding nitrosamine. * (If in doubt, apply Liebermann's test
(page 70).)

* Note that an acyl derivative of an o-diamine releases the latter

on hydrolysis; these diamines give yellow azimides with nitrous
acid, and such products may be confused with nitrosamines. If a
m-diamine is set free during the hydrolysis, it will produce a deep
brown precipitate when treated with nitrous acid (see page 69).
A liberated arylhydrazine may simulate a secondary amine when
treated with nitrous acid (page 70), and may yield a red dye with
alkaline ,8-naphthol (pages 69-70).
84
 Classification
If no products of reaction have been detected, pour a few drops
of the solution into a solution of 0·5 g of ,8-n~phthol in 5 ml of
aqueous sodium hydroxide (2N). The formation of a red or orange
azo-dye indicates that the original compound is an acylated
primary aromatic amine.
Acyl derivatives of amino-acids, e.g. benzoylglycine (hippuric
acid) hydrolyse normally in the above test to yield two acids. Thus
benzoic acid and glycine are obtained from hippuric acid. Because
it is soluble in water, acids and alkalis, the amino-acid will be
difficult to isolate.

Test for anilides, toluidides, etc. In a dry test-tube place 3 ml of

concentrated sulphuric acid and to it add 0·1 g of the compound.
Shake until most of it has dissolved. Add O· 1 g of finely powdered
potassium dichromate and shake vigorously at once. A deep rose,
plum or purple colour, which may be transient, indicates a
compound of the type ArNHCOR.
For simple acylated primary arylamines the test is satisfactory.
Thiocarbanilide and some nuclear-substituted anilides (e.g. p-
chloro-, p-bromo-, p-nitro- and p-ethoxy-acetanilide) give negative
results.
Derivatives. See page 144.

HYDRAZINES, SEMI CARBAZIDE,

HYDROXYLAMINE, THEIR SALTS
AND CARBONYL DERIVATIVES

1. The free bases. The bases most likely to be encountered are the
arylhydrazines and semicarbazide. They are reducing agents
towards ammoniacal silver nitrate, potassium permanganate and
Fehling's solution, and dissolve with varying ease in dilute mineral
acid solutions, some substituted phenylhydrazines requiring
moderately concentrated acids for dissolution. The arylhydrazines
respond to some of the tests for primary and secondary aromatic
amines (pages 69-70).
Derivative. Prepare a derivative as described under Alde-
hydes and Ketones, page 121, using a known pure carbonyl
compound. (See also Table 21.)
85
Qualitative Organic Analysis
2. The salts with mineral acids. These compounds when treated
with a slight excess of sodium hydroxide solution become conver-
ted to the bases, which are usually precipitated as solids or heavy
oils, except in the cases of semicarbazide and hydroxylamine,
these latter compounds being water-soluble.

Derivative. Prepare a derivative as indicated in (1) above,

using either the free base or the salt, and a known pure
carbonyl compound.

3. The carbonyl derivatives. The majority of these compounds are

insoluble in water. Arylhydrazones are usually yellow to red in
colour.

Tests
(a) Warm about 0·1 g of the compound in the minimum quantity
of methanol with 3 ml of a methanolic solution of 2,4-dinitro-
phenylhydrazine sulphate (Brady's reagent). Most compounds of
this class yield a precipitate of the 2,4-dinitrophenylhydrazone of
the parent aldehyde or ketone, and the latter can thus be identified.
(b) Hydrolyse the compound by boiling it with concentrated
hydrochloric acid for about 30 seconds. Cool the mixture, make
alkaline with sodium hydroxide solution, and add 2 m1 of Fehling's
solution. A red precipitate of cuprous oxide obtained when the
mixture is warmed shows the presence of an oxime, semicarbazone
or an arylhydrazone in the original.
If the above tests are positive, hydrolyse 2 g of the substance by
heating it under reflux with 10 m1 of concentrated hydrochloric
acid for 30 minutes. Cool the hydrolysate and remove the liberated
carbonyl compound by ether extraction, filtration or distillation.
Prepare from it a suitable derivative (see page 121).
Make the residual liquid alkaline and remove any precipitated
base in order to prepare from it a suitable derivative (see (1 ) above).
If a base has not been detected, evaporate the solution with a
slight excess of concentrated hydrochloric acid on a water-bath,
dissolve the residue in the minimum quantity of water and divide
the resulting solution into 2 parts. Use one portion for each of the
following tests:
(i) To 1 portion add 1 drop of benzoyl chloride, make the
mixture alkaline by careful addition of alcoholic potassium
86
 Classification
hydroxide solution, and heat to boiling. Cool at once, acidify
with dilute hydrochloric acid, and add 2 drops of aqueous ferric
chloride solution. A red to purple colour indicates that the
solution contains hydroxylamine, and hence shows that the
original substance is an oxime.
(ii) Use the second portion to confirm the presence of a
powerful reducing agent, namely semicarbazide, hydroxylamine,
or less likely, hydrazine.
For melting-points of semicarbazides and some oximes, see
Table 2; for melting-points of some hydrazines, see Table 21.

CONDENSATION PRODUcrS OF ALDEHYDES

WITH AMMONIA OR AMINES

These compounds when boiled with dilute hydrochloric acid yield

the aldehyde and either ammonium chloride or the hydrochloride
of the parent amine. The aldehyde may be removed for further
examination by ether extraction or distillation. When the residual
liquid is made alkaline and warmed, ammonia or the parent amine
will be liberated.
The amine and/or the aldehyde should be identified in the usual
way.

SECTION V

NITRO-, NITROSO·, AZOXY·, AZO·, AND

HYDRAZO-COMPOUNDS

As normally encountered, these compounds are coloured as

follows:
Nitro-compounds-yellow, depending on the number of
nitro-groups.
Nitroso-compounds-colourless or yellow (nitroso-dialkylani-
lines are green).
Azoxy-compounds-yellow.
87
Qualitative Organic Analysis
Hydrazo-compounds-colourless when pure, but usually
coloured.
Azo-compounds-highly coloured, usually red or yellow.
Most are insoluble in water, and unless other groups such as
-S03Na, -COOH, or phenolic -OH are present they are insoluble
in alkali (except for primary and secondary nitro-paraffins,
RCH2N02 and RR'CHN02 which dissolve in alkalis).
In the soda-lime test (preliminary examination) nitro-com-
pounds may be reduced to a small extent to the corresponding
amine.
Hydrazo-compounds reduce ammoniacal silver nitrate and
Fehling's solution, and this fact should be borne in mind with
respect to Mulliken's test (page 89).

General tests
(a) Titanous chloride. To a solution of 0·1 g of the substance in
acetone (about 5 ml) add 5 ml of a 5 %solution of titanous chloride
in dilute hydrochloric acid. Warm the mixture on a water-bath,
adding more acetone if necessary to produce a homogeneous
solution. Discharge of the mauve colour of the reagent within
2 minutes constitutes a positive result. (Compare with a blank.)
The reagent also reduces quinones and hydroxylamines.
(b) Reduction with tin and acid. Mix O· 5 g of the substance with
5 ml of concentrated hydrochloric acid and 2 ml of alcohol. Add
about 3 small pieces of granulated tin and cool if necessary to
moderate the reaction. Warm under reflux on a water-bath until
the original compound has passed into solution (about 20 minutes).
Decant the mixture from unchanged tin, and withdraw 1 ml of the
solution. To this add 2 ml of water, cool in ice to 5°, and dropwise
add 1 ml of a 10 %aqueous sodium nitrite solution. Transfer a few
drops of the resulting solution to 5 ml of an ice-cold solution of
,8-naphthol in 2N sodium hydroxide.
With the exception of aliphatic nitro-compounds, all the above
substances produce red, orange or red-brown azo-dyes.
(If required, the bulk of the reduction mixture can slowly be
made alkaline until the first-formed precipitate of stannous
hydroxide redissolves, the mixture cooled, and the amine removed
by ether extraction or steam-distillation. It may then be examined
as described on pages 68-71.)
88
 Classification
Note. In this particular test N-substituted amides such as anilides
are hydrolysed by the acid, and if primary aromatic amines are
thereby formed, will yield azo-dyes.
0- and m-Nitro-amines and nitro-anilides will form 0- and m-
diamines when treated with tin and acid. These diamines do not
diazotize normally with nitrous acid (see pages 69, 70).

(c) Mulliken's test. Dissolve 0·1 g of the compound in a mixture of

1 m1 of alcohol and 1 m1 of water by warming, add 1 m1 of ammo-
nium chloride solution and a pinch of zinc dust. Boil the mixture
briefly, and filter directly into ammoniacal silver nitrate solution
(or Tollen's reagent-see preliminary tests).
A black precipitate of silver indicates a nitro-, nitroso-, azo- or
azoxy-compound.
The original substance should be tested to ensure that it does not
itself reduce ammoniacal silver nitrate.
Note. This test is useful for detecting one of the above functional
groups in the presence of a primary aromatic amino-group (e.g.
in nitro-amines). It will also detect the nitro-group in nitro-sub-
stituted anilides.

Distinguishing tests
1. Test for m-dinitro-compounds (Janowsky's test). Dissolve the
compound in acetone and add 2N sodium hydroxide solution.
m-Dinitro-compounds yield a red or purple colour. The test must
be interpreted with caution: a variety of polynitro compounds give
positive results. Amino and phenolic groups interfere.

2. Hydrazo-compounds (RNHNHR'). These compounds reduce

Fehling's and Tollen's solutions.

3. Nitroso-compounds. (a) C-Nitroso-compounds. These are

generally found as nitrosophenols or p-nitroso-derivatives of
dialkylanilines. Their colour is usually green (p-nitrosodialkylani-
lines) or yellow (salts of p-nitrosodialkylanilines, nitrosophenols).
(b) N-Nitroso-compounds, (nitrosamines). These compounds
are usually yellow. Compounds of this type (and p-nitrosophenol)
generally respond to the Liebermann reaction (page 70).
89
QlUIlitative Organic Analysis
4. 'Nitroparaffins. Shake the compound (3 drops) with a solution
of 0·1 g of sodium nitrite in 1 m1 of 30 %aqueous sodium hydrox-
ide. Primary and secondary nitroparaffins dissolve. Dilute the
solution with 3 m1 of water and add dropwise 2N sulphuric
acid. Primary nitroparaffins give a red colour which is destroyed
by excess of acid, secondary nitro-compounds a blue or green
colour soluble in chloroform, whereas tertiary nitro-compounds
(RR'R"CNO.v do not react.

s. Azoxy-compounds (RN(O)NR'). Heat a very small quantity

(ca 1 mg) of the substance with 1 m1 of concentrated sulphuric
acid at 90° for 30 seconds. Azoxy-compounds are converted to
hydroxy-azo-compounds, which possess rich orange or red-brown
colours.

Identification. (a) The most general method used for identi-

fying compounds of this section involves reduction to the
corresponding primary amines and characterization of the
latter.
(Unsymmetrical azo- or azoxy-compounds on reduction
with tin and hydrochloric acid produce a mixture of two
amines, while hydrazo-compounds may yield amine(s)
formed by normal reduction of the hydrazo-link and a
diamine produced by a benzidine rearrangement. Nitro-
samines, on reduction with tin and acid, give the secondary
amines from which they were derived.)
Identification by reduction is invalidated by the
presence of other reducible groups such as carbonyl,
cyano-, etc.
(b) Aromatic nitro-hydrocarbons may be characterized
by conversion to polynitro-compounds. This can be a
hazardous operation, and should be conducted with
caution.
(c) The nitro-derivatives of mono-alkylbenzenes may be
oxidized to the corresponding nitrobenzoic acids.
(d) Some aromatic dinitro-compounds may be reduced
to the corresponding nitro-amines by treatment with
ammonia and hydrogen sulphide.
For experimental details of these methods, see page
146.
90
 Classification
POLYFUNCfIONAL NITRO-COMPOUNDS
The most commonly occurring compounds of this type are listed
below. Except where a statement to the contrary is made, they
show the usual properties of the individual groups present.

1. Nitro-acids.
Derivatives. As for carboxylic acids (page 123).
2. Nitro-aldehydes and nitro-ketones. These compounds give the
reactions of both carbonyl compounds and nitro-compounds.
Derivatives. As for carbonyl compounds (page 121).
3. Aromatic nitro-ethers.
Derivatives. As for aromatic nitro-hydrocarbons (page 146).
4. Nitrophenols. These compounds dissolve in sodium hydroxide
solution, giving yellow or red solutions. Many are soluble in
sodium bicarbonate solution, a fact which distinguishes them from
simple phenols.
Derivatives. As for phenols (page 126).
5. Nitro-amines. These are normally encountered as yellow to red
solids. Their salts are colourless. Reduction gives the correspond-
ing diamines, but this procedure is not recommended as a means
of preparing derivatives.
Derivatives. Formyl; acetyl; benzoyl (pages 138-141).
6. Nitro-esters. Compounds of this class should be hydrolysed as
described on pages 127, 128 and the products of hydrolysis iden-
tified in the usual manner (see nitro-acids, above). See also Tables
5 and 8.
7. Nitro-alcohols. See Table 23.
8. Nitro-amitIes. These compounds should be converted to the
parent nitro-acids in the usual manner.
9. Acylated nitro-amines (e.g. P-N02C6H4NHCOCH3). The nitro-
group in such compounds may be detected by Mulliken's test
(page 89). Tin and hydrochloric acid reduction causes simultaneous
91
Qualitative Organic Analysis
hydrolysis at the amide group, with formation of a diamine.
(0- and m-Primary diamines do not diazotize normally when
treated with nitrous acid (see page 69).) For this reason it is
strongly advised that if the compound under investigation is sus-
pected to be an anilide containing a nitro-group, the tests for the
substituted amide function should be made before an examination
for the nitro-group.

10. Nitro-halogeno-hydrocarbons. These compounds are neutral,

and insoluble in water. The determination of the degree of
mobility of the halogen in the compound affords a means of
allocating the substance to its class.
(a) Gently warm the compound with 2N nitric acid and silver
nitrate solution. If a precipitate of a silver halide is formed, the
compound is probably a dinitro- or a trinitro-derivative in which
the nitro-groups are para and/or ortho to the halogen atom(s).
Derivative. As the halogen in these compounds is mobile,
boiling with sodium hydroxide solution for 15 minutes
causes its replacement by a phenolic group. The phenol so
formed may be precipitated from the liquor by the addition
of an excess of concentrated hydrochloric acid, and may be
recrystallized from dilute hydrochloric acid. Determine its
melting-point.

(b) If test (a) is negative, boil the substance for 15 minutes with
2N alcoholic potassium hydroxide, cool, acidify with 2N nitric
acid, and test the solution for halide ions with aqueous silver
nitrate. A precipitate of silver halide indicates that the compound
is most probably either (i) a nitro-substituted aromatic halide with
the halogen present in a side-chain, e.g. a nitro benzyl halide, or
(ii) a mono-nitro-halogeno-benzene with the halogen ortho or para
to the nitro-group. To distinguish, the compound is boiled with
concentrated sodium hydroxide solution. An intense yellow colour
which pales on cooling and adding hydrochloric acid indicates that
the compound is of type (ii).
(c) If tests (a) and (b) are both negative, the compound is
probably ofthe halogeno-m-nitro- type, e.g. m-nitrochlorobenzene.
Derivatives. See page 148 and Table 23.
92
 ClIIssification
11. Acid halides of nitro-acids.
Derivatives. As for simple acid halides (page 149).

SECTION VI

COMPOUNDS CONTAINING HALOGEN

(BUT NOT NITROGEN OR SULPHUR)

Alkyl Halides Aryl Halides

Halogen-substituted Aliphatic Acids, Akohola, Aldehydes, Ketones,
Esters and Ethers
, Aliphatic Polyhalides Acid Halides

In compounds of this type, the preliminary tests should have

yielded valuable information, for a knowledge of the degree of
mobility of the halogen they contain is vitally important.

ALKYL HALIDES

These compounds do not dissolve in cold concentrated sulphuric

acid. With the exception of the alkyl iodides, substances of this
type do not readily react with aqueous silver nitrate solution.
Aromatic side-chain halides such as benzyl chloride behave like
simple aliphatic halides in many respects.
Derivatives. Aliphatic mono-halides, page 148. Aromatic
side-chain halides, page 147.

ARYL HALIDES

In this category are included all aromatic compounds containing

halogen linked directly to the nucleus, except those having in
addition a more reactive group such as hydroxyl or carboxyl, etc.,
for compounds containing these more sensitive groups will already
93
Qualitative Organic Analysis
have been classified as phenols, acids etc., rather than through the
(usually) inert halogen function.
Aryl halides do not usually give a positive test for halogen when
warmed with alcoholic silver nitrate solution, but certain activating
groups may modify the normal reaction, e.g. see nitro-halogeno-
hydrocarbons, (page 92).
Derivatives. See page 147.

HALOGEN-SUBSTITUTED ALIPHATIC ACIDS,

ALCOHOLS, ALDEHYDES, ESTERS, KETONES,
AND ETHERS

In general, compounds of these classes react as do the correspond-

ing non-halogenated compounds, and identification proceeds by
the same route as for the latter compounds, except as indicated
below.

Esters of aliphatic halogeno-acids (other than chloroformates).

Some members of this class, particularlY'ihe ex-bromo esters, have
very irritating odours.
When applying the classification tests for esters, it must be
remembered that many aliphatic compounds containing halogen,
though not esters of organic acids, yet will give a positive result in
the hydrolysis test (test (b), page 58), owing to hydrolysis at the
halogeno-alkyl group, with consequent consumption of alkali.
Derivatives. (a) Prepare the amide by shaking the ester
with concentrated ammonia solution, and recrystallize the
separated amide from dilute alcohol. (For melting-points,
see Table 5.)
(b) Hydrolysis of the ester will cause breakdown of the
acid from which the ester is derived, so that this process
can only give useful information about the alcoholic part
of the ester molecule.
Esters of chloroformic acid. Compounds of this class react with
warm water to give fumes of hydrogen chloride, and carbon
dioxide. They possess sharp odours and are lachrymatory. They
may not respond to the hydroxamic acid test for esters (page 58).
94
 Classification
Derivatives. (a) Prepare the amide (urethane) by the method
described for acid chlorides (page 149), finally adding a
layer of ether, in which the product dissolves. Evaporate
the ether and recrystallize the residue from alcohol.
(b) The chloroformic ester may be converted into the
carbanilate by the method described for preparing anilides
from acid chlorides (page 149). Melting points are in
Table 28.

a.-Halogeno-ethers. Such a compound decomposes in warm water,

giving halogen hydracid, an alcohol and an aldehyde. The alde-
hyde produced may be detected and identified by treating the warm
solution with Brady's reagent, when its 2,4-dinitrophenylhydra-
zone is precipitated.
The more common compounds of this type are:
a.-chloroethyl ether, b.p. 98° - with water gives ethanol,
acetaldehyde.
a.a.'-dichloroethyl ether, b.p. 116° - with water gives acetalde-
hyde.
cx,8-dichloroethyl ether, b.p. 140° - with water gives ethanol,
chloroacetaldehyde.

ALIPHATIC POL YHALIDES

Compounds of this class are generally rather inert, and there are
few satisfactory derivatives (see Table 27). Useful tests are:

Test for chloroform ami bromoform. Warm gently a pellet of

potassium hydroxide with one drop of aniline, 2 m1 of ethyl
alcohol and a few drops of the original substance. The evil odour
of carbylamine indicates that the original substance is one of the
above compounds.

Test for chloroform, bromoform, iodoform. Boil a small quantity of

the substance with a solution of a few crystals of resorcinol in 2 m1
of dilute aqueous sodium hydroxide. A red colour in the solution
is a positive test.
95
Qualitative Organic [Link]

ACID HALIDES

Simple aliphatic acid halides fume in moist air, react vigorously

with water, and give immediate precipitation of silver halide when
treated with nitric acid and silver nitrate solution. Their aqueous
solutions are strongly acid.
Aromatic and high molecular weight aliphatic acid halides
behave similarly but less vigorously.
Derivatives. See page 149.

SECTION VII
COMPOUNDS CONTAINING HALOGEN
AND NITROGEN (BUT NOT SULPHUR)

Most of the compounds likely to be encountered containing these

elements, (in addition to carbon, hydrogen and possibly oxygen),
will have been classified by tests for the nitrogen-containing func-
tions given on previous pages. For comparison, they are listed
here, together with notes on compounds not yet considered.

Hydrohalides of bases (e.g. PhNH2HC1, PhNHNH2.HC1), see

pages 78, 86.

[Link] halides of nitrogen-containing acids (e.g. 3,5-dinitrobenzoyl

chloride), see pages 93, 149.

Halogenated bases, their hydrohalides and acyl derivatives (e.g.

chloroanilines, etc.). Examine as for the non-halogenated com-
pounds; see pages 66 et seq., 78, 83.

Halogenated nitro-compounds. See page 92.

Nitrogenous derivatives of halogeno-acids (ammonium saits,

amides, anilides).
(a) Ammonium salts. Treat the compound with 2N sulphuric
acid and isolate the organic acid. In test (a) for carboxylic acids
(page 52), an aliphatic halogeno-acid may give a fading end-point,
96
 Cltusijiclltio"
due to hydrolysis at the halogeno-a1kyl group and consequent
consumption of alkali. In such a case, identification of the acid by
determining its equivalent weight is clearly not possible. For
derivatives, see page 123.
(b) Amides and anilides. Hydrolyse in the usual way (pages 80 et
seq., 83 respectively) and identify the base and/or the halogeno-
acid. An aliphatic halogeno-acid may lose its halogen during the
hydrolysis, more especially if an alkaline hydrolysing mixture is
used.
N-Iudoge"o-compounds (e.g. N-bromosuccinimide). These com-
pounds liberate halogen very readily, and this fact will have been
established in the preliminary examination. They liberate iodine
from acidified potassium iodide solution (page 27).
Identification is by hydrolysis to the free acid (page 143), and
characterization of the latter.

SECTION VIII

COMPOUNDS CONTAINING SULPHUR

(WITH OR WITHOUT NITROGEN AND/OR
HALOGEN)

Ammonium salts 0/ sulpho"ic acids

Sulphonic acids Sulphonamides
Amino-sulpho"ic acids Sulphates 0/ bases
Thiourea, substituted thioureas, thioamides
Sulpho"yl halides N-HalosulpholUlmides
Aldehyde and ketone bisulphite compounds
Thiols Thio-ethers Thio-acids
Esters 0/ sulphonic acids

Compounds containing sulphur are rather less likely to be en-

countered by the student than compounds having nitrogenous
functions as their principal groups. In addition, sulphur is readily
and reliably detected in the sodium fusion test for elements. For
these reasons it is convenient to group together all common com-
pounds containing sulphur, irrespective of the nature of other
elements that may be present, (in addition to carbon, hydrogen and
possibly oxygen).
G 97
Qualitative Organic Analysis

AMMONIUM SALTS OF SIMPLE AND

SUBSTITUTED SULPHONIC ACIDS

Treat the substance with dilute aqueous sodium hydroxide.

Ammonium salts evolve ammonia in the cold. Boil the solution to
drive off all ammonia, cool, add a slight excess of dilute sulphuric
acid, and very carefully evaporate to dryness. Examine the residue
for nitrogen by Lassaigne's test. (Part of the residue may be sub-
mitted to the test for sulphonic acids, given below.)
If nitrogen is found to be present, the parent acid probably
contains an amino- (see amino-sulphonic acids, page 100) or a
nitro-group. The latter may be detected in the original compound
by applying the tests on page 88.
If neither nitrogen nor halogen have been found to be present in
the acid, the original compound is probably the ammonium salt
of a simple sulphonic acid.
Derivatives. See page 149. (The ammonium salt may be used
directly.)

SULPHONIC ACIDS (AND THEIR METALLIC

SALTS); SULPHONAMIDES

Simple aliphatic sulphonic acids are hygroscopic liquids; aromatic

sulphonic acids are hygroscopic solids. Sulphonic acids are soluble
in water giving strongly acid solutions. Sulphonic acids unlike
sulphates do not yield white precipitates when treated with dilute
hydrochloric acid and barium chloride solution.
Sulphonamides are crystalline solids. Those derived from
ammonia or a primary amine are soluble in alkalis (see below).
In the soda-lime test (preliminary examination) sulphonamides
evolve ammonia or an amine depending on whether they are
simple (RS02NH2) or substituted (RS02NHR', RS02NR'R'')
respectively. Sulphanilamides give ammonia and an amine.
The presence of a nitro-group may be detected by applying the
appropriate classification tests (page 88) to the original substance.
A nuclear halogen atom will not in general be mobile (see pages
25,26).
98
 Classification
General test
Fuse about 0·5 g of the substance with 2 g of potassium hydroxide
in a hard-glass test-tube, or better, in a nickel crucible for 10
minutes. A simple sulphonamide evolves ammonia, while a sub-
stituted sulphonamide liberates an amine, * the vapour of which
may be detected by means of filter-paper moistened with saturated
ethanolic 2,4-dinitrochlorobenzene solution; a strong yellow
colour is a positive test.
Allow the melt to cool, and treat it with a little water so as to
dissolve as much of the residue as possible. Acidify the solution and
test for evolved sulphur dioxide and/or hydrogen suphide by using
filter strips impregnated with acidified potassium dichromate solu-
tion and lead acetate solution respectively. All compounds of this
class should evolve sulphurous vapours at this stage. It is feasible in
some cases to extract the acidified solution with chloroform and to
test the extract for the presence of a phenol by means of the
modified ferric chloride test (page 36); a positive result confirms
the presence of a sulphonate or sulphonamide in the original
substance.
Note. A few compounds such as thiourea and its mono-substitu-
tion products give a positive result in the above test, even though
they are not of the class now being considered. They can be dis-
tinguished readily by differences in solubility and other tests (see
pages 101, 102).
Hydrolysis test. Hydrolyse the compound (0·5 g) by heating with
5 m1 of 80 % sulphuric acid (1 vol. water and 3 vols. conc.
H 2S04) at 1500 for about 15 minutes. Allow to cool, and with care
pour a portion of the hydrolysate into a moderate excess of dilute
sodium hydroxide solution. If ammonia is not evolved (indicating
original compound was not a simple sulphonamide), examine the
rest of the hydrolysate for amine by treating with sodium nitrite
solution as described in the General Test for acylated bases (page
83).

Identification
Sulphonic acids and their salts. (a) Convert to the sulphonyl
chloride using phosphorus pentachloride, and hence prepare the
amide or anilide (see page 149).
* A sulphanilamide gives ammonia and an amine.
99
Qualitative Organic Analysis
(b) Prepare the S-benzylthiuronium salt (see page 124).

Sulphonamides-simple (RS02NH:z). These compounds are soluble

in aqueous alkalis.
(a) Prepare the benzoyl derivative (see page 151).
(b) Prepare the xanthyl derivative (see page 150).
(c) Prepare the acetyl derivative (see page 150).
(d) Hydrolyse to the free acid and identify the latter (see page
151).

Sulphonamides-primary substituted (RS0 2NHR'). These com-

pounds are soluble in aqueous alkalis.
(a) Prepare the methyl derivative (see page 151).
(b) Prepare the acetyl derivative (see page 151).
(c) Hydrolyse to the free acid and amine and identify these pro-
ducts (see page 152).

Sulphonamides-secondary substituted (RS02NR'R"). These com-

pounds are not soluble in aqueous alkalis.
Identification is by hydrolysis to the free acid and amine, and
characterization of the products. It must be remembered that the
amine that is produced from a sulphonamide of this type will be a
secondary amine (see page 152).

AMINO-SULPHONIC ACIDS
The more common members are soluble in alkalis but not in acids
owing to suppression of the basic function.
Distillation with soda-lime yields a primary amine (sulphanilic
acid gives aniline, while metanilic acid gives m-aminophenol).
With bromine water, sulphanilic acid yields tribromoaniline;
metanilic acid does not.
Sulphanilic acid and naphthionic acid yield p-benzoquinone and
oc-naphthoquinone respectively when each is heated with man-
ganese dioxide and concentrated sulphuric acid. The pungent
odours of the products are readily detected.
100
 CltusijicGtio"
General tests
(a) Members of this class respond to the fusion test for sulphonic
acids (q.v.).
(b) Dissolve the substance in sodium carbonate solution, cool in
ice, and treat with an ice-cold solution of sodium nitrite. Add
dilute hydrochloric acid slowly until the solution is slightly acid
and then pour it into a solution of ,a-naphthol in sodium hydroxide
solution, keeping the mixture alkaline. The production of a red
soluble azo-dye is a positive test.
Deri,flti,es. S-Benzylthiuronium salt; determination of
equivalent weight (both as for carboxylic acids, pages 123,
124).

SULPHATES OF BASES

The base should be isolated as described in the general section

dealing with amine-salts (page 78). The free base should then be
examined by the methods described on pages 66 et seq., 85.

THIOUREA; SUBSTITUTED THIOUREAS;

THIOAMIDES (RCSNH2 )

All common compounds in this category are solids. Thiourea is

moderately soluble in cold water, while substituted thioureas are
generally sparingly soluble.
Thioamides and substituted thioureas dissolve in dilute sodium
hydroxide solution. When boiled with this reagent, thiourea and
simple thioamides evolve ammonia, while substituted thioureas
slowly liberate ammonia and/or an amine.

General test
Make a suspension of yellow mercuric oxide in alcohol and add to
it a very small amount of the original substance. The production of
black mercuric sulphide when the mixture is gently warmed con-
stitutes a positive result.
101
Qualitative Organic Analysis
Test for thiourea. Heat a little of the compound until it just melts.
Cool, dissolve the product in water and add aqueous ferric chloride
solution. The appearance of a blood-red colour (indicative of the
presence of ammonium thiocyanate) shows the original substance
to be thiourea.
Test for substituted thioureas. Heat the compound under reflux
with 30 %aqueous sodium hydroxide. Mono-substituted thioureas
yield both ammonia and an amine; di-substituted thioureas of the
type RNHCSNHR yield the amine RNH2, while di-substituted
thioureas of the type RNHCSNHR' give a mixture of the amines
RNH2 and R'NH2. Isolate the base that has been formed and
identify it in the usual way (page 66 et seq.).
Test for simple thioamides. Heat the substance under reflux with
dilute sulphuric acid. The thioamide is thus hydrolysed to the
corresponding carboxylic acid, which should be isolated by filtra-
tion or distillation and identified (page 52).
Derivatives. Miscellaneous; see Table 19.

SULPHONYL HALIDES

These compounds are hydrolysed by hot water (see below). If the

original substance contains a nuclear halogen substituent, e.g.
p-bromobenzenesulphonyl chloride, only the chlorine is labile. The
presence of a nitro-group may be determined by the usual tests
(page 88).

General test
Boil a little of the compound with distilled water for 5 minutes.
Cool, filter if necessary, acidify the filtrate with 2N nitric acid, and
add silver nitrate solution. A white or yellowish precipitate of silver
halide is formed.

Reduction. Most sulphonyl halides respond to the following test.

Boil 0·2 g of the original compound with 5 ml of dilute hydro-
chloric acid and a pinch of zinc dust. The characteristic odour of a
thiophenol indicates the presence of a sulphonyl halide.
Derivatives. Sulphonamide and sulphonanilide (page 152).
102
 Classification

N-HALOSULPHONAMIDES (ArS02N <g! (or Na»)

These compounds liberate iodine from potassium iodide solution
which has been acidified with dilute sulphuric acid. (See page 27.)
The commonest compounds are chloramine T, which contains
sodium, and dichloramine T, which does not.
Derivative. Boil the compound (0· 5 g) with 10 m1 of 2N
aqueous sodium hydroxide and 10 m1 of 20 volume hydro-
gen peroxide, for 15 minutes. Cool, acidify with concen-
trated hydrochloric acid, boil until the solution is clear, and
cool in ice. Filter off the solid product and recrystallize it
from water or from dilute alcohol. p-Toluenesulphonamide
(m.p. 1371 is thus formed from both chloramine T and
dichloramine T.
Chloramine T has no definite melting-point. Dichloramine T
melts at 81°.

ALDEHYDE AND KETONE BISULPHITE

COMPOUNDS

Compounds of this class are readily identified by treating with

dilute sulphuric acid and testing for sulphur dioxide. When this
gas has been detected, heat a fresh sample of the original substance
with dilute sodium carbonate solution in a small distillation
apparatus, and pass the condensate into methanolic 2,4-dinitro-
phenylhydrazine sulphate. The formation of an orange precipitate
indicates the presence of an aldehyde or ketone. The precipitate
may be collected, purified and used as a derivative.
Derivatives. See Table 2.

THIOLS (RSH)

This class includes the thioalcohols and the thiophenols, the latter
being less common. They possess penetrating, unpleasant odours
103
[Link] Orgflllic A1IIIIysis
and are sparingly soluble in water, but dissolve in alkalis forming
salts. They decolourize potassium permanganate solution.

Tests
(a) To mercuric chloride solution add a small amount of the
original substance. The precipitation of the mercuric mercapto-
chloride (white) and the liberation of hydrochloric acid in the
solution (litmus paper) is indicative of the presence of a thioI.
(b) The following test is given by thioalcohols only. Dissolve a
few drops of the mercaptan in ethanol and add this solution to
2 m1 of a I % solution of isatin in concentrated sulphuric acid.
A green colour develops.
(c) Most mercaptans (thioalcohols) and some thiophenols give a
purple colouration (which may be transient) when their solutions
in dilute aqueous ammonium hydroxide are treated with sodium
nitroprusside solution.
[Link]. See page 152.

THIO-ETHERS (RSR')

These are neutral compounds, relatively resistant to the action of

alkali. They may be characterized by oxidation to the correspond-
ing sulphone (see page 153).
Note. Thio-ethers are often contaminated by traces of thiols.

THIO-ACIDS

This term is often used for true thio-acids such as thioacetic acid
(CH3COSH) and thiobenzoic acid (C6HSCOSH) as well as for
thiol-acids such as thiosalicylic acid (4I4(SH)COOH) and
thioglycollic acid (HSCH2COOH).
All compounds of these types have powerful odours, and they
are fairly readily decomposed by hot water or aqueous alkalis.
104
 Cltusijicatio"
General test
Boil the substance with a small excess of aqueous sodium hydr-
oxide, cool, and filter if necessary. Add 1 m1 of 20 volume hydrogen
peroxide and boil to oxidize sulphide. Cool, neutralize the solution,
and test the solution for the appropriate simple carboxylic acids
(acetic, benzoic, glycollic, etc.) by treatment with aqueous ferric
chloride, and by other tests given on pages 52-55.
Derivatives. (a) Thiosalicyclic acid. Prepare the acetyl
derivative.
(b) Thioacetic and thiobenzoic acids. Gently warm I g
of the acid with 1 m1 of aniline so that hydrogen sulphide
is evolved. Cool, pour into water wash the precipitate with
dilute hydrochloric acid and sodium carbonate solution.
Recrystallize the acetanilide or benzanilide from aqueous
alcohol.
(c) Thioglycollic acid. Prepare the benzylidene deriva-
tive by mixing 1· 5 g of the acid with 1 m1 of benzaldehyde
and allowing to stand until the mixture solidifies. Wash the
product with light petroleum and recrystallize it from
aqueous acetone. It has a melting point of 124°.

ESTERS OF SULPHONIC ACIDS (RS02 0R')

The number of compounds of this type that are likely to be

encountered is fairly small. They may be hydrolysed by hot
aqueous alkali as described for carboxylic esters (page 127), and
the alcoholic (or phenolic) component identified in the usual way.
The sulphonic acid should be isolated as its sodium (or potassium)
salt by making the hydrolysate (after removal of the alcoholic or
phenolic component) only just alkaline by the careful addition of
dilute sulphuric acid, and then evaporating to dryness. The solid
residue is then subjected to tests for sulphonic salts, and may be
used to prepare a derivative of the sulphonic acid.
An alternative method of isolating the sulphonic acid as its
sodium salt is to saturate the hydrolysate with common salt
(sodium hydroxide having been used in the hydrolysis procedure).
The melting points of some of the simpler sulphonic esters are
as follows:
105
Qualitative Organic Analysis
Methyl p-toluenesulphonate, 28°
Ethyl p-toluenesulphonate, 33°
Phenyl benzenesulphonate, 35°
p-Cresyl p-toluenesulphonate, 70°
ex-Naphthyl p-toluenesulphonate, 88°
Phenyl p-toluenesulphonate, 96°
,B-Naphthyl p-toluenesulphonate, 125°.
The detailed consideration of other compounds containing
sulphur, e.g. sulphoxides (RSOR,), sulphones (RS02R ,), esters
of sulphuric acid (ROS0 20H and ROS0 20R)" sulphites
(ROSOOR'), sulphonium salts (R3S +X-), disulphides (RSSR,),
sulphinic compounds, and compounds containing phosphorus,
is beyond the scope of this book.

106