GENERAL PROCEDURE FOR SYSTEMATIC QUALITATIVE ANALYSIS OF UNKNOWN ORGANIC COMPOUNDS
[Link] EXPERIMENT OBSERVATION INFERENCE
PRELIMINARY TESTS
1. Color of the compound Yellow Presence of aromatic nitro group
Presence of phenol and aromatic
Brown
amines
Red Presence of azo compounds
Presence of
Colorless
acids/esters/ketones/aldehydes
2. State of existence Solid May be presence of carbohydrates,
amides, thiourea
Liquid May be presence of alcohol, aldehyde,
amine
3. Odor of the compound Carbide/Phenol smell May be presence of phenol
Pleasant fruity smell May be presence of esters/ketones
Fishy smell May be presence of aromatic amines
Ammonium smell May be presence of amines
No characteristic smell May be presence of acid or amide
4. Solubility Soluble in water May be carbohydrates, diamides,
aliphatic aldehydes/ketones/acids
Soluble in sodium hydroxide May be phenols or aromatic carboxylic
acids
Soluble in hydrochloric acid May be aromatic amines
TESTS FOR SATURATION/UNSATURATION
5. Bayer’s Test Immediate decolorization Presence of unsaturated compound
To a small amount of the No decolorization or slow Presence of saturated compound
compound, water or alcohol is decolorization
added. To this solution, Bayer’s
reagent (dilute alkaline potassium
permanganate) was added drop by
drop with shaking
6. Bromine water test Immediate decolorization Presence of unsaturated compound
To a small amount of the No decolorization or slow Presence of saturated compound
compound, water or alcohol is decolorization
added. To this solution, bromine
water was added drop by drop with
shaking
KMnO4 OH
H2C CH2 HO
H2O
TESTS FOR ALIPHATIC/AROMATIC COMPOUND
7. Flame test Burns with smoky flame Presence of aromatic compound
A small amount of the substance
No smoky flame Presence of aliphatic compound
was taken in a spatula and
introduced into the flame
8. Nitration test Appearance of yellow color Presence of aromatic compound
A small quantity of the substance solution or precipitate
was taken in a test tube to which No yellow color solution or Presence of aliphatic compound
few drops of conc. sulphuric acid precipitate
and conc. nitric acid were added
and boiled on a water bath for few
minutes. The mixture was cooled
and carefully transferred into a
beaker containing 10 ml of water
NO2
+ HNO3
TESTS FOR SPECIAL ELEMENTS (NITROGEN, SULPHUR, HALOGENS)
Preparation of sodium fusion extract
A small amount of organic compound and sodium metal was taken in a fusion tube, heated gently and then strongly
in the flame until the bottom of the tube becomes red hot. The red-hot tube was then plunged into 10 ml of distilled
water taken in a mortar, grinded well with a pestle and filtered.
The filtrate was tested for the presence of special elements such as nitrogen, sulphur and halogens
Preparation of zinc and sodium carbonate extract (Alternate method)
A small amount of organic compound and mixture of sodium carbonate & zinc dust was taken in a fusion tube, heated
gently and then strongly in the flame until the bottom of the tube becomes red hot. The red-hot tube was then plunged
into 10 ml of distilled water taken in a mortar, grinded well with a pestle and filtered.
The filtrate was tested for the presence of special elements such as nitrogen, sulphur and halogens
9. Test for nitrogen
A few ml of the extract was treated
with freshly prepared ferrous
sulphate solution, to which sodium Prussian blue color was seen Presence of nitrogen
hydroxide solution was added and
boiled. The mixture was cooled, and
a few drops of dil. hydrochloric acid
was added
10. Test for sulphur
Violet or purple color was
A few ml of the extract was treated Presence of sulphur
with 2-3 drops of freshly prepared seen
sodium nitroprusside solution
11. A few ml of the extract was treated Presence of sulphur
with dil. acetic acid and 2-3 drops of Black precipitate was obtained
lead acetate solution
12. Test for nitrogen and sulphur
A few ml of the extract was treated Red or blood red color was
Presence of nitrogen and sulphur
with dilute hydrochloric acid and 2- obtained
3 drops of ferric chloride was added
3NaCNS + FeCl 3 Fe(CNS)3 + 3NaCl
Ferric thiocyanate (red)
13. Test for halogens Curdy white precipitate was Presence of chlorides
A few ml of the extract was formed
acidified with dilute nitric acid and Pale yellow precipitate was Presence of bromides
treated with 2-3 drops of silver obtained
nitrate solution Yellow precipitate insoluble in Presence of iodides
ammonium hydroxide solution
was obtained
TESTS FOR FUNCTIONAL GROUPS (PRELIMINARY)
14. CARBOXYLIC ACIDS
A small amount of sample solution Blue litmus turns red Presence of carboxylic acids
was tested with blue litmus paper
15. A small amount of sample solution Slow effervescence was Presence of carboxylic acids
was treated with sodium observed
bicarbonate solution
R - COOH + NaHCO3 RCOONa + H2O + CO2
effervescence
16. PHENOLS Violet/blue/green colour Presence of phenols
Neutral ferric chloride test solution
A small amount of sample solution
Flocculated white precipitate Presence of α-naphthol
was treated with a few drops of
was seen
neutral ferric chloride solution
6C6H5OH + FeCl3 → [Fe(C6H5O)6]3– (violet colour complex)+ 3HCl + 3H+
17. A small amount of sample solution Reddish brown color of Presence of phenols
was shaken with bromine water bromine water was discharged
18. AMIDES
Action of sodium hydroxide
A small quantity of the sample was
Red litmus turns blue due to
heated with few drops of sodium Presence of amides
the evolution of ammonia
hydroxide solution. A red litmus
paper is placed on the mouth of the
test tube to get exposed to the
vapors evolved
19. Hydroxamate test
A small quantity of the sample was
treated with hydroxylamine
hydrochloride and 10% sodium
hydroxide solution, gently boiled Red to purple color was Presence of aliphatic amides
for 1- 2 minutes. The mixture was obtained
cooled and acidified with dil.
hydrochloric acid. To this, a few
drops of ferric chloride solution
was added
20. Hydrogen peroxide test
A small quantity of the sample was
diluted with water. To this, 4-8
drops of hydrogen peroxide Blue or brown was obtained Presence of aromatic amides
solution was added, heated to
boiling, cooled and a few drops of
ferric chloride solution was added
21. THIOAMIDES
Action of sodium hydroxide
solution Ammoniacal smell evolves Presence of thioamides
A small quantity of the sample was
heated with sodium hydroxide
solution
22. ALDEHYDES AND KETONES
Borsche’s test
Yellow or orange or red Presence of aldehydes/ketones
A small quantity of the sample was precipitate was obtained
treated with 2,4- dinitrophenyl
hydrazine
23. Phenyl hydrazine test
A small quantity of the sample was Yellow precipitate was
treated with ethanol and phenyl Presence of aldehydes/ketones
obtained
hydrazine. The mixture was heated
on a boiling water bath for 10 min.
24. Sodium bisulphite test
A small quantity of the sample was White crystalline precipitate
treated with sodium bisulphite Presence of aldehydes/ketones
was obtained
solution and shaken vigorously for
few min.
25. Schiff’s test
A small quantity of the sample was Rose red solution was Presence of aldehydes
treated with Schiff’s reagent and obtained
shaken well for few min.
26. Sodium nitroprusside test
A small quantity of the sample was
treated with sodium nitroprusside Red or purple solution was Presence of ketones
solution and excess of sodium obtained
hydroxide solution was added and
shaken well
- -
[Fe(CN)5NO] 2 + CH3COCH3 [Fe(CN)5NO.CH3COCH3] 2
Red color
27. Legal’s test
A small quantity of the sample was Orange solution turning to
treated with water, 5 drops of Presence of ketones
purple was observed
sodium hydroxide solution and 5
drops of glacial acetic acid
28. ESTERS
A small quantity of the sample was
treated with sodium hydroxide White precipitate was Presence of esters
solution and boiled for 10 min., obtained
cooled and excess of dil. sulphuric
acid was added
29. Hydrolysis test
A small amount of the sample was
dissolved in alcohol. To this, dil.
Pink color disappears Presence of esters
sodium hydroxide solution and
phenolphthalein was added and
heated gently
30. AMINES
Diazotization test
A small amount of the substance
was dissolved in dil. hydrochloric Scarlet red dye appears Presence of amines
acid and cooled in ice water. To
this, 1 ml of sodium nitrite solution
and β-naphthol in sodium
hydroxide solution were added
31. Action of conc. sulphuric acid
A small amount of the substance White precipitate was Presence of amines
was warmed with conc. sulphuric obtained
acid
32. Action of bleaching powder
A small amount of the substance Transient purple color turning
Presence of amines
was treated with water and a few brown was obtained
drops of bleaching powder solution
33. CARBOHYDRATES
Molisch’s test
A small amount of the substance Violet ring was formed at the
Presence of carbohydrates
was treated with Molisch reagent. junction of two layers
2-3 drops of conc. sulphuric acid
was added slowly through the sides
of the test tube
TESTS FOR FUNCTIONAL GROUPS (CONFIRMATORY)
1. CARBOXYLIC ACIDS
Esterification test
A small amount of the sample was Pleasant fruity smell was Presence of monocarboxylic acid is
heated with ethanol and conc. observed confirmed
sulphuric acid, cooled and the odor
was observed
2. Neutral ferric chloride test Presence of formic acid or acetic acid
Red solution was obtained
A small quantity of the substance (monocarboxylic acid)
was heated with dilute ammonia Light brown precipitate was Presence of succinic acid and phthalic
solution until the ammoniacal smell obtained acid (dicarboxylic acid)
disappears. To this mixture, 2-3 Violet color was obtained Presence of salicylic acid
drops of neutral ferric chloride (monocarboxylic acid)
solution was added Buff color precipitate was Presence of benzoic acid
obtained (monocarboxylic acid)
Presence of tartaric acid
(monocarboxylic acid), oxalic acid
No characteristic change
(dicarboxylic acid) or citric acid
(tricarboxylic acid)
3. Fluorescein test
A small amount of the sample was
heated with few quantities of
resorcinol and two drops of conc. Red solution with green Presence of dicarboxylic acid is
sulphuric acid. The mixture was fluorescence was obtained confirmed
cooled and poured into a beaker
containing 10 ml of dilute sodium
hydroxide solution
4. PHENOLS
Phthalein test
A small amount of sample and half
the quantity of phthalic anhydride,
Red/blue/green/yellow-green
2 drops of conc. sulphuric acid were Presence of phenol is confirmed
solution was obtained
taken in a test tube. The mixture
was heated and poured into a
beaker containing dil. sodium
hydroxide solution
5. Reimer-Tiemann reaction
A small quantity of the sample was
Red/deep blue/green/yellow
treated with dil. sodium hydroxide Presence of phenol is confirmed
solution was obtained
solution and chloroform. The
mixture was heated on a water bath
6. Liebermann nitroso test
A small quantity of the sample was
heated with sodium nitrite
Red color turning to deep blue
solution, cooled and a few drops of Presence of phenol is confirmed
was observed
conc. sulphuric acid was added,
followed by water and excess
sodium hydroxide solution
7. Diazotisation test
A small quantity of aniline in dil.
hydrochloric acid was cooled in ice
bath. To this, sodium nitrite
Orange to scarlet red dye was
solution was added. The cold Presence of phenol is confirmed
observed
solution was added into another
test tube containing sample
dissolved in sodium hydroxide
solution
8. AMIDES
Biuret test
A small quantity of the sample was
heated in a dry test tube until
Purple or violet color solution
ammonia gets evolved. To the white Presence of amides is confirmed
was obtained
solid obtained on cooling, few drops
of sodium hydroxide solution and 2-
3 drops of copper sulphate solution
was added
2NH2CONH2 NH2CONHCONH2 + NH3
Urea Biuret Ammonia
9. Urea nitrate test
A small quantity of the sample was White crystalline precipitate
Presence of amides is confirmed
treated with water and conc. nitric was obtained
acid solution
10. Urea oxalate test
A small quantity of the sample was White crystalline precipitate
Presence of amides is confirmed
treated with water and oxalic acid was obtained
solution
11. THIOAMIDES
Lead acetate test
A small quantity of the sample was
Black color solution was
heated with 2 ml of potassium Presence of thioamides is confirmed
obtained
hydroxide solution, cooled and a
few drops of lead acetate solution
was added
RNHCSNHR + 4KOH 2RNH2 + K2CO3 + K2S + H2O
K 2S + (CH3COO)2Pb PbS + 2CH3COOK
12. Ferric chloride test
A small quantity of the sample was
Blood red color solution was
dissolved in water and a few drops Presence of thioamides is confirmed
obtained
of ferric chloride solution was
added
13. ALDEHYDES
Tollen’s test
Black precipitate or silver
A small quantity of the sample and Presence of aldehydes is confirmed
mirror was obtained
few drops of tollen’s reagent were
boiled on a water bath for 5-10 min.
RCHO + 2[Ag(NH3)2]OH RCOONH4 + 2Ag + 3NH3 + H2O
14. Fehling’s test
A small quantity of the sample and
few drops of Fehling’s solution A & Red precipitate was obtained Presence of aldehydes is confirmed
B were boiled on a water bath for 5-
10 min.
RCHO + 2CuO RCOOH + Cu2O
15. Semicarbazide test White crystalline precipitate
Presence of aldehydes is confirmed
A small quantity of the sample & was obtained
semicarbazide hydrochloride and
sodium acetate were boiled on a
water bath for 5-10 min.
16. Benedict’s test
A small quantity of the sample and
Red precipitate was obtained Presence of aldehydes is confirmed
1 ml of benedict’s reagent were
boiled on a water bath for 5-10 min.
17. KETONES
Iodoform test
To a small quantity of the sample, Yellow precipitate was
Presence of ketones is confirmed
iodine solution and excess of obtained
sodium hydroxide solution were
added and shaken well
18. ESTERS
Hydroxamic acid test
To a small quantity of the sample, 2
to 3 drops of alcohol, 0.2 gm of
hydroxylamine hydrochloride, 5 ml
of sodium hydroxide solution were Violet/reddish brown solution
Presence of esters is confirmed
added and boiled gently. The was obtained
mixture was cooled and 1-2 drops of
neutral ferric chloride solution and
excess of dil. hydrochloric acid,
again a few drops of ferric chloride
solution were added
19. AMINES
Rimini test
About 2 drops of amine, 1 ml of Presence of primary aliphatic amine is
Violet red color was obtained
acetone, 3 ml of water and 2 drops confirmed
of 1% sodium nitroprusside
solution was added and shaken well
20. About 2 drops of amine, 2 drops of
Presence of secondary aliphatic amine
1% sodium nitroprusside solution Blue color was obtained
is confirmed
were added and shaken well
21. Hinsberg test Presence of primary amine is
Yellow color was obtained
To the sample, 2 ml of pyridine, 2 ml confirmed
of freshly prepared 2% sodium Presence of secondary amine is
Orange color was obtained
hydroxide solution were added and confirmed
shaken well. To this, 2 drops of
Deep orange red color was
benzene sulphonyl chloride was Presence of tertiary amine is confirmed
obtained
added
22. CARBOHYDRATES
Fehling’s test
To a small amount of the substance, Presence of carbohydrates is
Red precipitate was obtained
1 ml of each of Fehling’s solution A confirmed
& B were added and heated on
boiling water bath for 5-10 min.
23. Benedict’s test
To a small amount of the substance,
Presence of carbohydrates is
1 ml of Benedict’s reagent was Red precipitate was obtained
confirmed
added and heated on boiling water
bath for 5-10 min.
24. Barfoed’s test
To a small amount of the substance,
Presence of carbohydrates is
1 ml of Barfoed’s reagent was Red precipitate was obtained
confirmed
added and heated on boiling water
bath for 5-10 min.
25. Tollen’s test
To a small amount of the
Black precipitate or silver mirror Presence of carbohydrates is
substance, 1 ml of Tollen’s reagent
was obtained confirmed
was added and heated on boiling
water bath for 5-10 min.