PRACTICAL ORGANIC CHEMISTRY

DETECTION OF ELEMENTS PRESENT:

The most commonly occurring elements in organic compounds are carbon, hydrogen, oxygen,
nitrogen, sulphur, halogen elements and less commonly phosphorous, mercury, arsenic etc. Carbon and hydrogen
are detected as follows. The organic compound is mixed with an equal amount of dry copper oxide and strongly
heated. The carbon present in the compound is oxidised to carbon dioxide (turns limewater milky) and hydrogen to
water (turns anhydrous copper sulphate blue).

C + 2CuO  CO2 + 2Cu

2H + CuO  H2O + Cu
CO2 + Ca(OH)2  CaCO3 + H2O
limewater white

CuSO4H2O + 4H2O  CuSO45H2O

white blue

Nitrogen, sulphur, phosphorous and halogen elements are identified by sodium fusion test (Lassaign’s test).

Lassaign’s test:
In this test the elements are converted into ionisable inorganic substances so that ionic tests of
inorganic analysis may be applied. This may be accomplished by fusing the organic compound with
metallic sodium. In this way NaCN, Na2S, NaX are formed when the respective elements are present.

ELEMENTS PRESENT IONIC COMPOUNDS FORMED UPON FUSION

WITH METALLIC SODIUM
C and N NaCN
S Na2S
C, N and S NaCNS
Halogens NaX
Phosphorous Na3P
It is essential to use excess sodium in order to get both Na2S and NaCN separately instead of
NaCNS.
NaCNS+2Na   NaCN+Na 2S
But even NaCNS may be identified by adding iron (III) ions which give red coloration due to the
formation of Fe(SCN)3.
3CNS- +Fe +3   Fe(SCN)3
red colouration
I. Identification of NaCN:
Cyanide ion, and hence nitrogen in the sample, may be detected by the Prussian Blue test.
The filtered alkaline solution, resulting from the action of water upon the sodium fusion, is treated
with iron (II) sulphate and thus forms sodium hexacyanoferrate (II). Upon boiling the alkaline
iron (II) salt solution, some iron (III) ions are inevitably formed by the action of air. These iron
(III) ions combine with [Fe(CN)6]-4 ions to form Prussian Blue.
FeSO4 +6NaCN   Na 4 [Fe(CN) 6 ]+Na 2SO 4
3Na 4[Fe(CN) 6 ]+2Fe2 (SO4 )3 
 Fe4 [Fe(CN) 6 ]3  +6Na 2SO4
Prussian Blue
Iron (III) chloride should not be added since its yellow colour makes the Prussian Blue appear
greenish. There is no necessity of adding it either. A sufficient concentration of iron (III) ions are
formed by the atmospheric oxidation.
If silphide ions are present, a black precipitate of FeS is obtained. To remove the sulphide
ion, boil the mixture for 30 seconds, and acidify with dil. H2SO4; the iron (II) sulphide dissolves
and the Prussian Blue colour persists.
FeS+H 2SO4   H2S  +Fe2 (SO 4 )3
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II. Identification of Na 2S:

Sulphur, as sulphide ion, may be detected by precipitation as black lead sulphide with lead
acetate solution.
S-2 +Pb(CH3COO)2   PbS  +2CH3COO-
black
It may also be identified by purple colour produced on addition of sodium
pentacyanonitrosoniumferrate(II).
2Na 2S+Na 2 [Fe(CN)5 NO]   Na 4 [Fe(CN)5 NOS]
Sodium nitroprusside purple
III. Identification of halides:
Halides are identified by the formation characterstic silver halides upon addition of
AgNO3/HNO3.
NaCl+AgNO3   AgCl  +NaNO3
White ppt which
is soluble in dil. NH 3 ,
NaCN and Na 2S2O3.

NaBr+AgNO3 
 AgBr  +NaNO3
Light yellow ppt which
is soluble in conc. NH 3 ,
NaCN and Na 2S2O3 .
NaI+AgNO3 
 AgI  +NaNO3
Yellow ppt which
is soluble NaCN and Na 2S2 O3 .
Cyanide and sulphide ions both interfere with this test for halides by forming silver cyanide
(white) and silver sulphide (black) precipitates respectively. If nitrogen or sulphur has been
detected, therefore, the interfering ions must be removed. To remove cyanide and sulphide ions,
make 2-3 ml of fusion solution just acidic with dilute HNO3, and evaporate to half of the original
volume in order to expel hydrogen cyanide and/or hydrogen sulphide which may be present.
Alternately, add Ni(NO3)2 to the fusion solution, filter off the Ni(CN)2 and/or NiS, and proceed
for the test for halide ions (nickel halides are soluble in water).
The presence of halogens may be further confirmed by the Beilstein test. In this test the
compound is heated along with pure copper oxide in the Bunsen flame: the corresponding copper
halide is formed, which, being volatile, imparts an intense green or bluish-green colour to the
mantle of the flame.

III. Identification of phosphorous:

The presence of phosphorous may be indicated by a smell of phosphine during the sodium
fusion and the immediate production of a jet-black colour when a piece of filter paper moistened
with silver nitrate solution is placed over the mouth of the ignition tube after the sample has been
dropped in the hot sodium.
Alternately, treat 1 ml of the fusion solution with 3 ml of concentrated nitric acid/Na2O2
and boil for 1 minute. Cool and add an equal volume of ammonium molybdate reagent. Warm the
mixture and allow to stand. If phosphorous is present, a yellow crystalline precipitate is obtained.

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PO -34 + (NH 4 ) 3 (MoO4 )12 +H + 

 (NH 4 )3[P(Mo3O10 ) 4 ]  +H 2O
obtained from yellow
-3
oxidation of P
by HNO3 / Na 2O 2

TESTS TO DISTINGUISH/CHARACTERISE ORGANIC COMPOUNDS

Let us first study about the solubility behaviour of different organic compounds and then about chemical
reagents that can characterise different functional groups.

THE STUDY OF SOLUBILITY BEHAVIOUR

All solubilities discussed here under have the arbitrary standard: 0.1g solid or 0.2ml liquid in 3ml of the solvent.
Solvents discussed here are
a) water
b) ether
c) 5% NaOH
d) 5% NaHCO3
e) 5% HCl
f) conc. H2SO4
Solubility in H2O:
Water is a poor solvent for all hydrocarbons. Salts are usually water-soluble. Other compounds fall
between these two extremes: these include alcohols, esters, aldehydes, ketones, acids, ethers, amides, nitriles and
amines. Acids and amines are more soluble than neutral compounds.
For homologous series of mono-functional alcohols, esters, aldehydes, ketones, acids, ethers, amides,
nitriles, and amines, the upper limit of water solubility is found at about the member containing four carbon
atoms(remember the limit would be elsewhere for a different ratio of solute to solvent)
Solubility in ether
Non-polar and slightly polar compounds dissolve in ether. Salts are insoluble in ether. Many organic
compounds that are insoluble in water are soluble in ether. If a compound is soluble in both water & ether, it probably
i) is non-ionic ii) contains five or less carbon atoms iii) has a functional group that is polar (ex. –OH) and capable of
hydrogen bonding and iv) does not contain more than one strongly polar group.
If a compound dissolves in water but not ether, it may i) be ionic (a salt) or ii) contain two or more polar groups but
not more than four carbon atoms per polar group.
Solubility in dil. HCl
Most compounds that are soluble in dil. HCl contain a basic nitrogen atom. AMINES: Most aliphatic amines
(10,20,&30) form salts with HCl . Aryl groups reduce the basicity of nitrogen atom. Primary aromatic amines (e.g.
aniline), although more weakly basic than primary aliphatic amines, are soluble in dil.HCl. Secondary and tertiary
aromatic amines (e.g. diphenyl amine, triphenyl amine) are insoluble because their basic strengths are reduced to
such an extent that they do not form salts with dil. HCl. Aryl alkyl amines (containing no more than one aryl group)
and alicyclic amines, however, do dissolve. AMIDES: Amides which are insoluble in water are generally unaffected
by 5% HCl but may dissolve in higher concentration (10%-20%). Many substituted amides, that are water insoluble,
dissolve in 5% HCl.
Solubility in dil. NaOH & dil. NaHCO3
Carboxylic acids, sulfonic acids, phenols, thiophenols, thiols (but not alcohols), imides, aryl sulfonyl
derivatives of primary amines( Hinsberg test ), oximes, primary and secondary nitro compounds dissolve in dil.
NaOH. Enols of 1,3-diketones also dissolve in dil.NaOH. Carboxylic acids and sulfonic acids are soluble in
dil.NaHCO3 with evolution of CO2. Some phenols substituted with electron-withdrawing groups (e.g. picric acid, 2,4,6-
tribromophenol and 2,4-dinitrophenol) also dissolve in NaHCO3. Primary and secondary nitro compounds, imides,
and oximes are insoluble in NaHCO3.
Solubility in conc.H2SO4
The most important group of compounds to exhibit solubility in this reagent are those containing oxygen.
Reason: they form oxonium salts.
Unsatured hydrocarbons dissolve through formation of soluble alkyl hydrogen sulfates.
R-CH=CH-R + H2SO4   R-CH2-CH(SO3H)-R
Some aromatic hydrocarbons dissolve in conc. H2SO4. Reason: they undergo sulfonation.
ArH + 2H2SO4 
 ArSO3H + HSO4- +H3O+

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Reminder: All aromatic hydrocarbons do not dissolve readily in conc. H2SO4. But they are readily soluble in
fuming sulphuric acid.
CHEMICAL TESTS FOR THE IDENTIFICATION OF FUNCTIONAL GROUPS
For a characterisation test, we select a reaction that is rapidly and conveniently carried out, and that gives
rise to an easily observable change.
ALKANES
An unknown compound is characterised as alkane on the basis of negative evidence. Upon quanlitative
analysis, an alkane gives negative tests for all elements except carbon and hydrogen.
An alkane is insoluble not only in water, dil.acid and dil.base but also in conc.H2SO4.
ALKENES
1. decolorize Br2/H2O(alkynes, phenols,and aldoses also do the same).
2. decolorize cold, dil. and neutral permanganate solution(Baeyer test) (this test is also responded to by
alkynes & aldehydes)
3. soluble in conc.H2SO4(some aromatic hydrocarbons and almost all compounds having oxygen are also
soluble in the same reagent).
ALKYNES
In their characterisation tests, alkynes resemble alkenes.
1. they decolorize Br2/CCl4
2. they also decolorize cold dil.KMnO4
Acidic alkynes (terminal alkynes) react with certain heavy metal ions, mainly Ag+& Cu+2, to form insoluble
acetylides.
With AgNO3 in ethanol or ammonia -white ppt.
With Cu2Cl2 in ammonia -red ppt.
AROMATIC HYDROCARBONS

1. because of very high carbon to hydrogen ratio aromatic hydrocarbons, on burning, produces thick
black smoke.
2. they are soluble in fuming H2SO4 . They are distinguished from alcohols and other oxygen containing
compounds by their failure to dissolve immediately in cold conc.H2SO4
3. aromatic hydrocarbons with side chains decolorize hot KMnO4(atleast one alpha hydrogen is
essential if the side chain is saturated).
ALKYL HALIDES
In characterisation tests alkyl halides resemble alkanes. They are readily distinguished, however, by
qualitatitve analysis, which shows the presence of halogen.
AgNO3 test: Almost all alkyl halides give ppt. with AgNO3. Reactivity toward alcoholic AgNO3 follows the
sequence RI>RBr>RCl. For a given halogen atom, reactivity decreases in the order 30>20>10. Allylic & benzylic
halides are highly reactive toward AgNO3.
Aryl, vinyl halides and bridge head halides do not react.
But bear in mind the fact that acid halides also respond to this reagent.
Beilstein’s Test: A green color is imparted to the flame if small amount of organic compound is taken on copper
wire.
ALCOHOLS
Alcohols are not oxidised by cold dil.KMnO4 but 10 & 20 alcohols are oxidised by it under hot conditions.
Alcohols are soluble in conc.H2SO4
1. alcohols release hydrogen gas on treatment with Na.
2. 10& 20 alcohols with CrO3/H2S04: within two seconds, the clear orange color turns blue-green and
becomes opaque(aldehydes also behave similarly).
3. ester formation: alcohols form esters most of which are sweet-smelling.
4. Cerric ammonium nitrate- alcohols give red color.
(NH 4 ) 2 [Ce(NO 3 ) 6 ]+2C2 H 5OH 
 [Ce(NO3 ) 4 (C 2H 5OH) 2 ]+2NH 4 NO 3
Distinguishing 10,20&30 alcohols among themselves.
(a) Lucas test:
1. Lucas reagent-HCl/ZnCl 2
2. 10 alcohols-no appreciable reaction, so no cloudiness.
3. 20 alcohols-cloudiness in 5 min.
4. 30 alcohols-cloudiness almost instantaneously.

(Benzyl and allyl alcohols behave similarly as 30 alcohols. But in the case of allyl alcohol no cloudiness is
found. The reason: allyl chloride is soluble in the reagent.)
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Reminder: this test is useful only for alcohols containing not more than 5 carbons. The reason: alcohols
containing more than 5 carbons are not soluble in the reagent.
(b) Iodoform test (NaOH/I2 or NaOI): alcohols having CH3-CH(OH)- skeleton can be characterised by iodoform
test.
(c) Victor Mayor Test: (also called as RBC test; Red, Blue, Colourless)
Procedure: treat the alcohol with PI3; treat the resultant alkyl iodide with AgNO2; dissolve the resultant nitro
compound in conc.NaOH and then add excess of NaNO3 solution. Upon cautiously acidifying with dil.H2SO4, the
following effects may be observed.
(i) Primary alcohols: intense red color, disappearing on acidification.
(ii) Secondary alcohols: dark blue or dark green color
(iii) Tertiary alcohols: no coloration.
Needless to say, the same test can also be used to distinguish 10,20,&30 nitro alkanes among themselves.
POLYHYDRIC ALCOHOLS
1. Boric acid test: Add a few drops of phenolphthalein to a 1 per cent solution of borax; a pink coloration
is produced. The addition of polyhydric alcohol causes the pink color to disappear. This reaction is due
to the combination of two cis- hydroxyl groups of the compound with boric acid to form reversibly a
much stronger acid.
2. Periodic acid test (for 1,2-glycols,  -hydroxy aldehydes,  -hydroxy ketones): Add periodic acid (or
para periodic acid, H5IO6) to the compound and then add AgNO3. A white ppt. of silver iodate (AgIO3)
is formed.
3. Molisch’s test: see carbohydrates.
ETHERS:
Ethers are soluble in conc.H2SO4. Ethers are also identified by cleavage of with hot conc.H2SO4 and
identification of the resultant products.
PHENOLS:
1. soluble in NaOH but not in NaHCO3.
2. Phenols form colored complexes (ranging from green through blue and violet to red) with FeCl 3.
Carbonyl compounds which form stable enols also respond to this test.
3. they decolorize bromine water.
4. Liebermann Test : See amines
ALDEHYDES AND KETONES:
1. 2,4-DNP: aldehydes and ketones give orange colored crystalline ppt.
2. NaHSO3: aldehydes and methyl ketones give crystalline ppt.

Chemical differentiation between aldehydes and ketones

1. Schiff’s test:
Schiff’s reagent (not Schiff’s base): Freshly prepared SO2 is sent into P-rosaniline hydrochloride solution
until its magenta color is disappeared. Aldehydes restore the magenta color, but ketones do not.
2. Tollen’s test:
Tollens’ reagent: Ammonical silver nitrate, [Ag(NH3)]+
Aldehydes reduce Ag+ to Ag (silver mirror), ketones do not
3. Fehling,s test:
Fehling’s solution: Mixture of copper (II) sulphate, NaOH and Rochelle salt (sodium potassium tartrate).
Aldehydes produce red ppt.(Cu2O), ketones do not.
Aromatic aldehydes and glyoxal do not give red precipitate with Fehling’s solution.
4. Benedict’s test:
Benedict’s reagent is similar to Fehling’s solution. The only difference is sodium potassium citrate in the
place of sodium potassium tartrate. Aldehydes produce red ppt.(Cu2O),ketones do not
Reminders:
1. formic acid ,like aldehydes , responds to Tollen’s and Fehling’s tests.
2. -hydroxy ketones behave similarly as aldehydes and respond to all the above tests.

CARBOXYLIC ACIDS:
1. they dissolve in aq.NaOH & aq.NaHCO3 (sulfonic acids are even more acidic than carboxylic acids,
but they contain sulphur, which can be detected by elemental analysis).
2. ester formation: add some amount of ethanol to the sample. If acid is present, ester is formed. In
most cases, esters are sweet-smelling.

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3. also useful, in identifying a particular carboxylic acid, is neutralisation equivalent; equivalent weight
of the acid as determined by titration with a standard base.
ACID HALIDES:
Most acid halides undergo ready hydrolysis as they are highly reactive to give an acidic solution and the
halide ion produced can be detected by adding AgNO3 sol.
AMINES:
Amines are identified chiefly through their basicity. A water-insoluble compound that dissolves in cold
dil.HCl-or a water-soluble compound (not a salt) whose aqueous solution turns litmus blue- must almost certainly
be an amine.
Distinguishing 10,20,&30 among themselves:
Hinsberg test:
Hinsberg reagent: benzene sulfonyl chloride.
10 amines: add Hinsberg reagent; a ppt. is formed; then add NaOH; the ppt. disappears; the ppt. reappears
on acidification.
20 amines: add Hinsberg reagent; a ppt. is formed; the ppt. does not disappear on adding base.
30 amines: add Hinsberg reagent; a ppt. is formed (not because of any reaction like in the case of 10 or 20
amines); ppt. disappears on acidification.
Nitrous acid test:
Add sodium nitrite and HCl to the sample. After standing for 3-4 minutes if a clear solution is obtained with
a continuous evolution of N2 gas, the substance is a primary aliphatic or aralkyl amine. If there is apparently no
evolution of N2 from the clear solution, add one-half of the solution to a cold solution of 2-naphthol in NaOH
solution. The formation of a colored azo-dye indicates the presence of a primary aromatic amine; in which case
warm the other half of the diazotised solution and note the evolution of N2. If a colorless solution is obtained which
gives an immediate and sustained positive test with starch-iodide paper when only a little sodium nitrite solution
has been added, the compound is a tertiary aliphatic amine. The formation of N-nitrosoamines which usually
separate as orange-yellow oils or low melting solids indicates the presence of a secondary amine. You can confirm
the formation of nitrosoamine by the Liebermann nitroso reaction. This consists in warming the nitrosoamine
with phenol and conc. H2SO4. First red color appears which turns blue by the addition of base.
H2SO4 PhOH PhOH
R2NNO R2NH + HONO HO NO HO N O

indophenol (red)

NaOH

NaO N O

blue
Carbylamine reaction: 10 amines (both aliphatic and aromatic) respond to this test.
KOH  RNC + KCl + H O
RNH 2 + CHCl3  2

foul smelling
compound
Hoffmann mustard oil reaction: 10 amines give black ppt. (HgS) when treated with CS2 followed by HgCl 2.
Diazo Test: This test is for aromatic 10 amines. For example, aniline is first treated with NaNO2/HCl and then with
any highly activated benzene ring. The resultant products are coloured compounds (dyes).

NH2 N2Cl

NaNO2
HCl

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1.

N
N 2Cl N
OH OH
+

-Naphthol
red

2. N2 Cl OH OH

-Naphthol
NO2

red brown

3. NMe2

-O3S N2Cl + -O3S N=N NMe2

Yellow

H+

+
-O3S NH=N NMe2

Methyl orange

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CARBOHYDRATES
Molisch’s test:
This is a general test for carbohydrates. Take the sample and add to it 2-naphthol in ethnol or chloroform.
Allow conc. H2SO4 to flow down the side of the inclined tube. If a carbohydrate is present, a red ring appears at the
common surface of the liquids. The color quickly changes to purple on standing or shaking. This test is also given
by all polyhydroxy compounds.
Br2/H2O: aldoses decolorise bromine water.

PROTEINS:
(a) Biuret Test: alkaline solution of protein treated with a drop of aq. CuSO4; bluish color is obtained.
This test is also given by urea.
(b) Ninhydrin Test: Proteins treated with a puridine solution of ninhydrin give color ranging from deep
blue to violet color.

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