Practical Organic Chemistry

[1] Sodium Metal test :-

Compounds which have acidic hydrogen or active hydrogen liberate H2 gas with sodium
metal.
Na Metal 1
Z–H ZNa + H
2 2

Acidic Hydrogen :- Hydrogen of electronegative atom (O, N, X, S, CSP)

Na Metal 1
R–C CH R–C CNa + H
2 2

Na Metal 1
+ H
2 2
-
O

O O O O
Na Metal 1
CH3–C–CH2–C–CH3 CH3–C–CH–C–CH3 + H
-
O 2 2
[2] Test for Unsaturation:-
[a] Bromine water test :-
Reagent :- Br2 / H2O
 Reddish brown colour of bromine water disappear with alkenes and alkynes.
 Benzene does not give this test, whereas aniline and phenol give this test.

[b] Baeyer’s reagent test :-

Reagent :- Cold dil. KMnO4 or 1% alkaline KMnO4
 Pink colour of Baeyer’s reagent disappear with alkenes and alkynes.

Br2/H2O Br2/H2O
CH3–CH2–CH3 –ve –ve

Br2/H2O
CH3–C C –CH3 +ve
Br2/H2O
Br2/H2O –ve
CH2 CH2 +ve
[3] Test for terminal alkynes :-
[a] Tollen’s test :-
Reagent :- AgNO3 / NH4OH or [Ag(NH3)2]+
 Terminal alkynes give white precipitate with Tollen’s reagent.
AgNO3/NH4OH
R–C CH R–C CAg + NH4NO3 + H2O
(White ppt.)
[b] Ammoniacal cuprous chloride test :-
Reagent :- Cu2Cl2 / NH4OH
 Terminal alkynes give red precipitate with ammoniacal cuprous chloride.
Cu2Cl2/NH4OH
R–C CH R–C CCu + NH4Cl + H2O
(Red ppt.)
 Internal alkynes do not give Tollen’s test and ammoniacal cuprous chloride test.

AgNO3/NH4OH AgNO3/NH4OH
CH3–C C–CH3 –ve CH2 CH2 –ve

Cu2Cl2/NH4OH Cu2Cl2/NH4OH
CH3–C C–CH3 –ve CH3–C CH +ve
[4] Test for Alkyl halides:-
[a] Beilstein test :-
 A copper wire is heated in a Bunsen burner flame to form a coating of copper(II)
oxide. It is then dipped in the halogen containing compound and once again heated
in Bunsen burner flame.
 Copper halide (volatile) is formed, which imparts an intense green or bluish green
colour.
 This test is not given by fluorides since copper fluoride is not volatile.
[b] With AgNO3 :-
 If alkyl halides are capable of carbocation formation, then they will give precipitate
with AgNO3.
Cl Cl O
R3C–Cl R–C–Cl
[5] Test for Alcohols :-
[a] Ceric ammonium nitrate test :-
Reagent :- (NH4)2[Ce(NO3)6]
 Alcohols give characteristic red colour with ceric ammonium nitrate.
[b] Lucas test :-
Reagent :- Conc. HCl / Anhyd. ZnCl2
ZnCl2
R–OH + HCl R–Cl + H2O
(White ppt.)
(Turbidity)
(Cloudiness)

 Lucas reagent is used to distinguish 1 alcohol, 2 alcohol and 3 alcohol.

3 alcohol - Instant turbidity
2 alcohol - Turbidity appears in 5 min.
1 alcohol - Turbidity appears in 30 min. or does not give appreciable reaction.
 Allylic alcohols and benzylic alcohols also give instant turbidity.
CH2 CH–CH2–OH Ph–CH2–OH
[c] Victor Meyer test :-
Reagent :- 1. P + I2 2. AgNO2 3. HNO2 4. Base
P + I2 AgNO2 HNO2 NaOH
R–CH2–OH R–CH2–I R–CH2–NO2 R–C–NO2 Red colour
N–OH
P + I2 AgNO2 HNO2 NaOH
R2CH–OH R2CH–I R2CH–NO2 R2C–NO2 Blue colour
N O
P + I2 AgNO2 HNO2 NaOH
R3C–OH R3C–I R3C–NO2 No reaction Colourless

[d] Dichromate test :-

Reagent :- K2Cr2O7 / H+
 1 alcohols and 2 alcohols are oxidized by K2Cr2O7/H+.
H+
R–CH2–OH + K2Cr2O7 R–COOH + Cr+3
(Orange) (Green)

R2CH–OH + K2Cr2O7 H+ R2C O + Cr+3

(Orange) (Green)
[6] Test for Phenols :-
[a] Litmus paper test :- Phenol turns blue litmus into red.
[b] Neutral FeCl3 test :-
Reagent :- Neutral FeCl3
Ph–OH + FeCl3 [Fe(Ph–O)6] -3
(Violet blue complex)
 Phenol and enol give violet blue complex with neutral FeCl3.

[7] Test for Carbonyl compounds :-

[a] 2,4-DNP test :-
Reagent :- 2,4-Dinitrophenyl hydrazine or Brady’s reagent or Borche’s reagent

O2N O2N
R R
H+
O + NH2–NH NO2 N–NH NO2
R R
(Yellow orange)
[8] Test for Aldehydes :-
[a] Tollen’s test :-
Reagent :- AgNO3 / NH4OH or [Ag(NH3)2]+
 Aldehydes give black precipitate or silver mirror with Tollen’s reagent.

R–COO-NH4+
NH4OH
R–CHO + AgNO3 + Ag
(Silver mirror)
(Black ppt.)
[b] Fehling solution test :-
Reagent :- Fehling solution A - CuSO4.5H2O
Fehling solution B - Sodium potassium tartrate or Rochelle salt
[c] Benedict solution test :-
Reagent :- Benedict solution A - CuSO4.5H2O
Benedict solution B - Sodium citrate
R–CHO + Cu++ R–COO- + Cu2O
(Red ppt.)
  Aliphatic aldehydes give red precipitate with Fehling solution and Benedict solution.
 Aromatic aldehydes do not give Fehling solution test and Benedict solution test.
 Formic acid, a- hydroxy ketone and hemiacetal also give Tollen’s test, Fehling solution
test and Benedict solution test.
R OR
a- hydroxy ketone –C–CH– Hemiacetal C
O OH H OH
[d] Schiff reagent test :-
 Magenta dye or p- Rosaniline hydrochloride is pink coloured. Its pink colour is
discharged by passing SO2 gas and the colourless solution is called Schiff reagent.
 Aldehydes react with this reagent to restore the pink colour.
[e] Mercuric chloride test :-
Reagent :- HgCl2
R–CHO + HgCl2 + H2O R–COOH + HCl + Hg2Cl2
(White ppt.)
R–CHO + Hg2Cl2 + H2O R–COOH + HCl + Hg
(Grey or
black ppt.)
[9] Iodoform test :-
Reagent :- I2 / NaOH or NaOI

 R–C–CH3 and R–CH–CH3 give yellow precipitate of iodoform with I2 / NaOH.

O OH
 R can be alkyl, aryl or hydrogen .
R–C–CH3 I2 / NaOH
R–COO-Na+ + CHI3
O (Yellow ppt.)

R–C–CH3 R–C–CH2I R–C–CHI2 R–C–CI3 R–COO-Na+ + CHI3

O O O O

Qus. Which of the following can give iodoform test.

O O
(A) CH3–CH2–OH (B) CHI2–CHO (C) CH3–CH2–CHO (D) Ph–C–CH2–C–Ph

Ans. A, B, D
[10] Test for Carboxylic acids :-
[a] Litmus paper test :-
 Carboxylic acids turn blue litmus into red.
[b] Sodium bicarbonate test :-
 Carboxylic acids liberate CO2 gas with NaHCO3.
R–COOH + NaHCO3 R–COO-Na+ + H2O + CO2
 All acids which are stronger than H2CO3 give CO2 with NaHCO3.
 R–SO3H, 2,4- dinitrophenol, picric acid, barbituric acid, squaric acid also liberate
CO2 gas with NaHCO3.

R–SO3H + NaHCO3 R–SO3-Na+ + H2O + CO2

OH

+ NaHCO3 No reaction
[11] Test for Amines :-
[a] Carbyl amine test :-
Reagent :- CHCl3 / alc. KOH
 Primary aliphatic amines and primary aromatic amines give unpleasant smell
of isocyanide.

R–NH2 + CHCl3 + KOH R–NC + KCl + H2O

(Alkyl isocyanide)

[b] Hoffman mustard oil test :-

Reagent :- (i) CS2 (ii) HgCl2

 Primary aliphatic amines and primary aromatic amines give isothiocyanate,

which has smell like mustard oil.
(i) CS2
R–NH2 R–N=C=S + HCl + HgS
(ii) HgCl2
(Alkyl isothiocyanate)
[c] Hinsberg’s test :-
Reagent :- (i) PhSO2Cl (ii) Aq. KOH
 Hinsberg’s reagent is used to distinguish 1 amine, 2 amine and 3 amine.

(i) 1Amine :-
O O
Pyridine KOH Soluble in base
R–NH2 + Cl–S–Ph R–NH–S–Ph
- HCl
O O
(ii) 2Amine :-
O O
Pyridine KOH
R2NH + Cl–S–Ph R2N–S–Ph Insoluble in base
- HCl
O O
(iii) 3Amine :-
O
Pyridine
R3N + Cl–S–Ph No reaction
O
[c] With NaNO2 / HCl or HNO2 :-
 It is used to distinguish primary aliphatic amine and primary aromatic amine.

(i) Primary aliphatic amine :-

NaNO2 / HCl
R–NH2 R–OH + H2O + N2

(ii) Primary aromatic amine :-

NH2 N2+Cl-

NaNO2 / HCl b- Naphthol

Orange-red dye
pH = 8 - 10

(iii) Secondary amine :-

NaNO2 / HCl
R2NH R2N–N=O
(Yellow oily liquid)
[11] Test for Amides :-
 Amides give smell of ammonia when heated with alkali.
Reagent :- Conc. NaOH

R–C–NH2 + NaOH R–COO-Na+ + NH3

O

 Amides also liberate N2 gas with NaNO2 / HCl.

NaNO2 / HCl
R–C–NH2 R–COOH + H2O + N2
O
[12] Test for Esters :-
 Esters have sweet smell or fruity smell.
 When esters is heated with NaOH and Phenolphthalein then pink colour
disappears .
R–C–OR + NaOH + Phenolphthalein R–COO-Na+ + R–OH
O (colourless solution)
[13] Test for Nitro :-
[a] Mulliken-Barker test :-
Reagent :- (i) Zn / NH4Cl
(ii) AgNO3 / NH4OH
Zn/NH4Cl AgNO3/NH4OH
R–NO2 R–NH–OH R–N=O + Ag
(Black ppt.)

 Nitro alkane and nitro benzene give black precipitate on reaction with Zn / NH4Cl
followed by Tollen’s reagent.
AgNO3/NH4OH
R–NH–OH Black ppt.

AgNO3/NH4OH
R–C CH White ppt.

AgNO3/NH4OH
R–CHO Black ppt.

AgNO3/NH4OH
HCOOH Black ppt.
[14] Test for Carbohydrate :-
[a] Molisch test :-
 It is used for detection of all type of carbohydrate (monosaccharide, disaccharide
and polysaccharide).
Reagent :- (i) 1% Alcoholic solution of a- naphthol.
(ii) Conc. H2SO4
 Molisch reagent is added to aqueous solution of carbohydrate followed by conc.
H2SO4 along the side of test tube. A violet ring is formed at the junction of the
two layers.
Pentose Conc. H2SO4 (Furfural)
CHO
O

Hexose Conc. H2SO4

HO–CH2 CHO
O

Furfural + a- naphthol Violet ring

[b] Barfoed test :-
Reagent :- (CH3COO)2Cu
 It is used for detecting the presence of monosaccharides. Monosaccharides react
with copper(II) acetate and give brick red precipitate of Cu2O.

[c] Iodine test :- For starch

Reagent :- Iodine solution (KI + I2 or KI3)
 When iodine solution (orange or yellow) comes in contact with starch it turns
dark black-blue.
[d] When carbohydrate is heated in dry test tube, it melts turn brown and finally black,
giving a characteristic smell of burning sugar.
[e] Conc. H2SO4 is added to sugar, reducing it to black substance.
Conc. H2SO4
C12H22O11 12 C + 11 H2O
(Black)
Test to distinguish between Glucose and Fructose :-
[a] Seliwanoff’s test :-
Reagent :- Resorcinol & Conc. HCl
Fructose - Red colour is formed rapidly
Glucose - Light pink colour is formed slowly
 Fructose and Sucrose are two common sugars which give a positive test.

[b] Bromine water test :-

Reagent :- Br2 / H2O
Glucose - Bromine water oxidise glucose to gluconic acid, hence decolourise the solution.
Fructose - Negative test
 Aldose and Ketose are distinguished by bromine water test and Seliwanoff’s test.
[14] Test for Proteins or Amino acids :-
[a] Biuret test :-
 It is used for detecting presence of peptide bond.
Reagent :- NaOH solution & 1% CuSO4 solution
 Blue solution turns violet on contact with protein or substance having peptide bond.
 Biuret test is confirmatory test for urea.
180 ℃
NH2–C–NH2 NH2–C–NH–C–NH2 + NH3
D
O O O
[b] Ninhydrin test :-
 It is used for amino acid and protein with free –NH2 group.
O
Reagent :- Proline does not
OH
OH
give ninhydrin test. COOH
NH
O
 All amino acid in which a- amino group is primary give violet colour (Ruhemann’s
purple) with ninhydrin.
[c] Xanthoproteic test :-
 Aromatic amino acid (Tyrosine and Tryptophan) give yellow colour when heated
with conc. HNO3.
Qus. The correct observation in the following reaction is :
Glycosidic bond cleavage Seliwanoff’s reagent
Sucrose
Hydrolysis
A + B ?

(A) Formation of blue colour (B) Formation of violet colour

(C) Formation of red colour (D) Gives no colour

Ans. (C)

Sol. Sucrose Glycosidic bond cleavage

Hydrolysis Glucose + Fructose

Glucose and Fructose are distinguished by Seliwanoff’s test. Red colour is formed
rapidly with fructose.
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