PRACTICAL ORGANIC CHEMISTRY

POC: CLASSIFICATION TESTS FOR FUNCTIONAL GROUPS

Functional Groups Reagent Observation Reaction Note

1) Easily oxidized
compounds give a
positive test:
a) Most aldehydes
give a positive
test.
b) Formic acid and
Bayer’s reagent HO OH
KMnO4 its esters give a
Cold. dil. Pink colour disappears + MnO2 positive test.
alk.KMnO 4 2) Alcohols give a
positive test.
3) Phenols and aryl
C C C C
amines give a
positive test as they
get oxidized to
quinone.
HO
Reddish brown colour Br2 / H2O
Br2 / H2O
disappears
Br

Br
Reddish brown colour Br2 / CCl4
Br2 / CCl 4
disappears
Br

Cu2 Cl2 / NH3 Cu2Cl2 / NH3

Red precipitate R C C H R C C Cu
R C C H

AgNO3 / NH3 AgNO3 / NH3

White precipitate R C C H R C C Ag
 PRACTICAL ORGANIC CHEMISTRY
Na Alkyne should be a
Na Effervescence of H2 R C C H R C CNa liquid
Used to distinguish
ArH + H2SO4 (fuming)  ArSO3H + between saturated
Fuming H2 SO 4 Dissolves completely aliphatic and
H 2O
aromatic
ArH
hydrocarbons.

1) Can just be used to

identify thepresence
2ROH + 2Na  2RONa + H2 of acidic hydrogen
ROH, RCOOH, ArOH Na Effervescence of H2
2RCOOH + 2Na  RCOONa + H2 2) Can be used to
distinguish between
alcohols and ethers
Cloudiness appears Allyl and benzyl
3° ROH
immediately alcohols also give
ZnCl2
coloudiness
Cloudiness appears immediately
2° ROH Anhyd. ZnCl 2 ROH + HCl   RCl (cloudiness)
within 5 minutes
/Con.HCl + H 2O
(Lucas Reagent)
Cloudiness appears
1° ROH
only on heating
 PRACTICAL ORGANIC CHEMISTRY
Victor Mayer Test

Gives blue colouration

in step (iii) which turns
1° ROH i) I2 / Red P red on reaction with
ii) AgNO2 NaOH

iii) HNO 2 Gives blue colouration

2° ROH in step (iii) which does
iv) NaOH not react with NaOH

No reaction after step

3° ROH
(ii)

I2 /Red P AgNO2 HNO2 NaOH

H3 CH2 C OH H3 CH2 C I H3 CH2 C NO2 H3 C C NO2 H3 C C NO2

N N
OH ONa
nitrolic acid blue Red

CH3 CH3 CH3

I2 / Red P AgNO2 HNO2
H3 C C OH H3C C I H3 C C NO2
CH3 CH3 CH3
 PRACTICAL ORGANIC CHEMISTRY
OR

Ceric ammonium ROH +  NH4 2 Ce  NO3 6  NH4 2 Ce  NO3 5 + HNO3

blood red colouration
nitrate
In case of 1° and 2° alcohols the colour disappears after a while
due to oxidation to aldehdyes and ketones.


ROH + KOH  ROK + H 2 O
Xanthate Test
Yellow precipitate or ROK + S C S RO C SK
KOH  CS2
colouration
S
(yellow ppt.)

CrO (orange)
RCH2OH 
3 RCOOH +
Alcoholic group
K 2Cr2O 7 /
Orange colour changes Cr 3  (green) 3° alcohols do not give
Conc. H 2 SO 4 to green CrO (orange) this test
R 2CHOH 
3 RCOR +

Cr 3  (green)

Following groups can

give this test
I2/NaOH
RCH(OH) CH 3   CHI3 + H 3C CH , C CH 3
Iodoform Test
RCOONa OH O
I2 /NaOH Yellow precipitate X
H 3C CH , H C C
3

X X
 PRACTICAL ORGANIC CHEMISTRY

1) Compounds having active methylene group

First the hydrogen atoms of active methylene group are replaced and a diiodo compound is formed. It then
cleaves as follows to give iodoform.

O O O O
I2 / NaOH NaOH
CH3COCHI2 + CH3COONa
H3 C CH3 H3 C CH3
I I I2 / NaOH

CH3I + CH3COONa

2)Molecules like CH3 COCH2COOEt , CH3 COCH2CN , CH3 COCH2NO2 etc do not give this test as they cleave to give
CH3 COONa and CHI2COOEt / CHI2CN / CHI2NO2 etc after initial substitution of active methylene hydrogen
atoms.
3)Sterically hindered methyl ketones like Ph 3CCOMe do not give this test as the hydrolysis proceeds through
sterically unfavoured tetrahedral intermediate.
Ph Ph Ph
I2 / NaOH OH OH
CH3 CI 3 CI 3
Ph Ph Ph
Ph Ph Ph
O O O
(highly unstable)

CH3 CH3
O O
NaOI NaOH
H3C H3C
CH3 CI3
CH3 CH3
 PRACTICAL ORGANIC CHEMISTRY

1) In case of naphthols
the reaction is
carried out in
alcoholic medium
otherwise a white
ppt is formed
2) o-nitrophenol does
not give neutral
FeCl3 test
Violet, blue, green or 3
Phenolic group FeCl 3 6 PhOH + FeCl 3   PhO 6 Fe  2) Carboxylate ions
red
give blood red
colouration
3) enols give violet
colouration
(compounds with
appreciable enol
content only, not
simple aldehydes
and ketones)

(PhOH-violet, m-cresol – violet; o- cresol - blue-violet; p-cresol – blue; catechol – green; resorcinol – blue-violet;
quinol – transient blue changes immediately to red-brown; -naphthol – Blue-violet; -naphthol – green)

Ceric ammonium Brown colour or Phenol gets oxidized to

nitrate test precipitate quinone

O OH O

Phthalein test
O +2 O
Phthalic anhydride
+ Conc. H2SO4 Pink colouration O

followed by few OH
drops of NaOH
HO
Phenolphthalein
 PRACTICAL ORGANIC CHEMISTRY
OH OH O

p-nitrosophenol is
NaNO2
yellow in colour
H2SO4
Libermann’s test Brown solid, giving red quinone monoxime is
colour in ethanol and NO NOH
green in colour
NaNO2 +
forms a blue solution in Over all Colour
Conc. H2SO4 PhOH
alkali. change:
HO N O Yellow → green →
brown→ red → blue
Indophenol (Red in ethanol)

OH OH Aniline also gives a

white ppt.
Br Br
NH2 NH2
Br2
Bromine water test Br Br
Br2
White precipitate H2O
Br2 /H2O H2O

Br
Br
white ppt.
white ppt.

Benzene diazonium Colour varies

chloride ( PhNH2 + depending on the
Azo dye test Bright colored dye
phenol and amines
NaNO2 + Conc.HCl)
selected.
 PRACTICAL ORGANIC CHEMISTRY

N
NHNH2 HN
Brady’s Test
NO2 NO2
2,4-Dinitrophenyl Crystalline yellow,
orange or red C O +
hydrazine (2,4-
Aldehydes DNP) in presence precipitate
andKetones of Conc. H2SO4 NO2 NO2
C H yellow, orange
(or) red ppt.
O
C R 1) MeCHO, PhCHO,
methylketones and
O OH sterically
NaHSO3 test
White precipitate C O + NaHSO3 unhindered cyclic
Saturated solution SO3Na ketones give this
(takes 5-10 min)
of NaHSO3 white ppt. test
2) Aromatic ketones
do not give this test.

Schiff’s Test
Para rosaniline Aromatic aldehydes
hydrochloride Violet-purple (pink) and acetone react
decolurised with colour slowly
SO2

RCHO O O OH
2 + H3N C NHSO2H Cl H2 N C NH S C H Cl + H2SO3
R H
aldehyde HO3 S 2 O R 2
Schiff's reagent (violet-purple
(colourless) solution)
 PRACTICAL ORGANIC CHEMISTRY

1) Easily oxidizable
compounds like
RNHOH and quinol
give precipitate with
Tollen’s test TR.
Ammoniacal 2) Formic acid gives
Shining silver mirror RCHO +  Ag  NH3 2  RCOO + Ag  s 
AgNO3 this test
3) -hydroxy ketones
give this test as they
get oxidized to
diketo compound or
-hydro acid

Fehling’s test 1) Formic acid gives

this test
Aq. CuSO4 + Reddish brown
sodium potassium precipitate 2) Aromatic aldehydes
RCHO + Cu2+  RCOO- + Cu2O(s) do not give this test
tartarate
(Rochelle’s salt) (reddish which is mainly
dissolved in NaOH brown) attributed to their
solution unreactivity.
[A complex cupric 3) Glyoxal does not
tartarate is formed give this test.
on mixing both the
reagents)
Benedict’s Test
CuSO4 + sodium
citrate in alkaline
solution
 PRACTICAL ORGANIC CHEMISTRY

Legal Test Benzophenone does

Sodium Wine red colouration not give this test.
RCOR nitroprusside test which changes to
yellow on standing
Na 2  Fe  CN 5 NO 
+ NaOH

1) Effervescence can’t
be seen if the
RCOOH + NaHCO 3 →RCOONa + CO2 + solution is warmed.
RCOOH NaHCO 3 Effervescence of CO2 2) di or tri
H2O
nitrophenol,
polyhalo phenols
give this test.

RCONH2 Con.NaOH, heat Smell of ammonia RCONH2 + NaOH → RCOONa + NH 3

Carbylamine test CHCl /KOH

Nauseating smell or RNH2 
3

 RNC (offensive
CHCl 3 , alcoholic
Offensive smell smell)
KOH

Amines NaNO /HCl Aromatic amines

HNO2 (NaNO2+HCl) Effervescence of N2 RNH2 
2
ROH + N2 give diazonium salts
1° - RNH2
Hinsberg’s test
White precipitate RNH2 + PhSO2Cl → PhSO2NHR (white)
i) PhSO2Cl
soluble in NaOH
ii) NaOH
 PRACTICAL ORGANIC CHEMISTRY
2° amines
R2NH Yellow oily liquid RNH2 + HNO2  R2N – NO
HNO2  NaNO 2  HCl  (nitrosoamine)Yellow oily liquid

Hinsberg’s test
i) PhSO2Cl White precipitate RNH2 + PhSO2Cl  PhSO2NR2

ii) NaOH Insoluble in NaOH

Hoffman Mustard oil (Amines 1° - RNH2

test
Smell of mustard oil RNH2 + CS2  RNHC(SH)=S
i) CS2 and a black precipitate HgCl

2
 RNCS + HgS(s) + 2HCl
of HgS
ii) HgCl2 Smell of mustard oil

Aromatic amines
HNO2 ( NaNO2 form p-
3° amines Dissolves R 3N + HNO2  R 3N NO Cl  (salt)
+HCl)   nitrosoamines
(blue/green)

Hinsberg’s test No reaction

Mulliken-Baker test
i) Zn /NH4Cl Zn/NH Cl Ag 
Nitro compounds Black Precipitate RNO2  
4
 RNHOH  Ag(s) + RNO
ii) AgNO3 /NH3
 PRACTICAL ORGANIC CHEMISTRY
O O O
R
OH
+ H 2N C COOH N  Blue 
Amino Acids Ninhydrin Blue colour OH
H
O O HO
 CO2  RCHO  H2O

Amino acids having benzene ring get

Xanthoproteic test nitrated
Proteins Yellow colour
Conc.HNO3 (Tyrosine, tryptophan and
phenylalanine)

Peptide bonds complex with Cu2 just

like Biuret  NH2CONHCONH2 

Biuret Test H H
Violet colouration O R
O N
CuSO4 solution
Cu2
N O
R O
H H

Millon’s Test Tyrosine gets nitrated which then

Red
HNO3 + Hg(I) or complexes with Hg(I) or Hg(II) to give
precipitate/colouration
Hg(II) red coloration/precipitate

Sodium Cysteine complexes with sodium

Violet colouratioon
nitroprusside test nitroprusside with its –SH group.
 PRACTICAL ORGANIC CHEMISTRY
R

F R HN COOH
NO2 NO2
Sanger’s test H 2N COOH

2,4-FDNB Yellow precipitate

NO2 NO2

Yellow precipitate

Except trioses and tetroses all carbohydrates including

Molisch test polysaccharides give this test.
Carbohydrates -naphthol + Pentoses and hexoses get dehydrated to give furfural and 5-
Purple colouration
Con.H2SO4 (hydroxymethyl)furfural which gives purple colouration with -
naphthol

HO
O
H3O
HO HO O
 3 H2O
HO OH OH O
5-(hydroxymethyl) furfural
D-glucose
 PRACTICAL ORGANIC CHEMISTRY

OH OH
OH

HO HO
H3O O , H3O
HO O + 2 O O
 H2O
O  H,  2e
5-(hydroxymethyl) furfural -naphthol

OH OH
purple-colored dye

Iodine test Blue-black colouration Starch forms blue-black complex with iodine

cherry red precipitate Ketoses(pentoses and hexoses) get dehydrated by HCl, the
by ketoses within dehydrated product condenses with resorcinol to give cherry red
Seliwanoff’s test 2 minutes precipitate
Resorcinol + 6M HCl blue-green or peach Aldoses(pentoses and hexoses) get dehydrated by HCl, the
colour precipitate by dehydrated product condenses with resorcinol to give blue-green
aldoses after 2 minutes precipitate which may change to peach colour.

Ketoese get dehydrated faster and hence they give the test faster. Aldoses react very slowly and give faint colours.
 PRACTICAL ORGANIC CHEMISTRY
CH 2OH CH 2OH
O CH 2OH O CHO HO O O
H H, 0,5 O2
HO
OH  3 H2O OH
3 H2O
OH
O
2
OH
CH 2 OH

ELEMENTAL ANALYSIS
Element Test/Reagents Observation Reaction(s) Note

Na + C + N NaCN 1. Diazonium salts do

not give this test as
FeSO4 +6NaCN Na 4  Fe  CN 6  + they decompose to
give off nitrogen gas
Na 2SO4 during fusion with
sodium metal.

3
2. If the compound
Lassaigne’s Test Na 4  Fe  CN 6  + Fe  contains both
Prussian blue sulphur and Carbon,
Nitrogen Sodium Fusion 
colouration/precipitate NaFe  Fe  CN 6  + 3Na NaSCN is formed
Extract + FeSO4 which gives blood red
Blue colouration colouration with
FeCl3 solution

3 3. A compound
3 Na 4  Fe  CN 6  + 4 Fe  containing both S and
N when fused with
Fe4 Fe  CN6  +12Na  excess Na gives NaCN
3
and Na2S as excess
 PRACTICAL ORGANIC CHEMISTRY
Blue precipitate sodium decomposes
NaSCN formed.

Preparation of Sodium fusion extract(SFE)

The organic compound is fused with sodium metal in a fusion tube to give water soluble NaCN, NaSCN, Na2S, NaCl, NaBr & NaI. The
fused mixture is plunged into water when the tube is red hot. It is then boiled and filtered.

SFE + dil.AcOH + Black Precipitate

Pb2 + S2  → PbS (s)
Pb(OAc)2
Black

SFE + Pink colouration

Na2S+ Na 2  Fe  CN 5 NO  →
Na2[Fe(CN)5NO]
Na 4  Fe  CN 5 NOS 
Sulphur Pink colouration

White ppt
Oxidation test 3 KNO3  3KNO2  3 O
Fuse Compd with Na2CO3 + S + 3[O] Na2CO4 + CO2
KNO3 + Na2CO3. BaCl2  aq  + Na 2SO4  aq   BaSO4  +
Extract in water and 2NaCl(aq)
add BaCl2 solution

Cl  : Curdy white ppt

SFE + Dil.HNO3 +
Halogens Br  : Pale yellow ppt Ag   X   AgX
AgNO3 solution
I : Dark yellow ppt
H3PO4 +Magnesia mixture 
Compound + HNO3 +
Mg  NH4  P2O7  H2O 
Phosphorous heat and add White precipitate 
2MgNH4PO4  Mg 2P2O7  + 2NH3 +
Magnesia mixture
H2O
 PRACTICAL ORGANIC CHEMISTRY

Separation of binary mixtures of organic compounds.

Themixture of organic compounds can be separated by using appropriate solvent.
Most of the aromatic compounds are water insoluble due to large hydrophobic group of six carbon atom.
Aromatic acids are insoluble in water but are soluble in aqueous NaHCO3 solution or NaOH solution, due to salt
formation.
Phenols are water insoluble but are soluble in aqueous NaOH solution due to salt formation.
Phenols with two or more nitrogroups are soluble in NaHCO3 as well as NaOH.
Aromatic amines (Aniline 1º, 2º, 3º) are water insoluble but are soluble in aqueous HCl solution due to salt
formation.

Amides of the type RCONR 2 are basic and are soluble in 5 % HCl while RONHR and RCONH2 are neutral and are

insoluble in 5% HCl.
 PRACTICAL ORGANIC CHEMISTRY

SEPARATION OF A MIXTURE CONTAINING ACIDIC, PHENOLIC, BASIC AND NEUTRAL ORGANIC COMPOUNDS
The mixture is dissolved in organic solvent
organic layer

R COOH Ar OH R NH2 R H
carboxylic acid phenol amine neutral

extract with HCl

organic layer aqueous layer

R COOH Ar OH R H R NH3Cl

extract with NaHCO3 add NaOH

R NH2

organic layer aqueous layer collect

precipitate
Ar OH R H R COONa

extract with NaOH add HCl

R COOH

organic layer aqueous layer collect

precipitate
R H Ar ONa
evaporate
add HCl
solvent
Ar OH

collect
precipitate
 PRACTICAL ORGANIC CHEMISTRY

Ex: Separation of the organic compounds from a mixture containing Cinnamic acid, p-toluidine and anisole.
O
NH2 O
OH

Cinnamic acid p-toluidine anisole

dissolve mixture in CH2Cl2

extract with HCl (aq)
first aqueous layer first CH2Cl2 layer

NH3Cl O
O
OH

p-toluidinium
chloride (4)
add NaOH(s) extract with
extract with CH2Cl2 NaOH (aq)
NH2 second aqueous second CH2Cl2
layer
layer
O
O

ONa

purify by
crystallisation sodium cinnamate (5)
add conc. HCl
O purify by
distillation

OH

purify by crystallisation