CHARACTERIZATION OF HYDROCARBONS

TEST
Nitration

REAGENT
H2SO4, HNO3
(1:1)
Br in DCM

PRODUCT
Yellow globule
Oily layer of nitrobenzene
Decoloration of red solution

REACTION
Substitution of aromatic ring

NOTE
benzene

Addition of bromine

Permanganate/
Baeyers
(basic)

Aqueous KMNO4, NaOH

Decoloration of violet solution

Formation of brown pptMnO2

Oxidation

DNPH

2,4- dinitrophenylhydrazine

Yellow orange to red ppt

Addition,
Elimination

Dichromate
(acidic)

K2Cr2O7, H2SO4

Blue green soln of Cr2(SO4)3

Oxidation of 1, 2, 3 aldehyde

Tollens

Substitution

3-OH

NH2OH, HCl, FeCl3

Base-catalyzed substitution
reaction
Substitution of ester
Complexation of hydroxamic acid
with ferric ion

2 OH

Hydroxamic

Formation of silver ppt

(silver mirror)
Immiscible mixture of alkyl Cl
and H2O
Formation of yellow ppt =
methyl carbonyl
Formation of wine red soln

Oxidation of aldehyde

Iodoform

Ag(NH3OH) or
AgNO3/NaOH/NH3
Concentrated HCl, anhydrous
ZnCl2
NaClO, KI

cyclohexane
alkene
Oxidizable:
cyclohexan
e
aldehyde
2 OH
alkene
Carbonyl:
aldehyde
ketone
Oxidizable:
cyclohexan
e
aldehyde
2 OH
alkene
aldehyde

Bromine

Lucas

*aqueous KMNO4 violet

solution

ester

STRUCTURES
Hexane

Aldehyde (formalin)
n-propanol

Cyclohexane
2-propanol
Ketone (acetone)

Toluene

Tertbutatnol

Naphthalene

Ester (ethyl acetate)

9. CHARACTERIZATION OF FATS & OILS

TEST
Acrolein

FOR
glycerol

Bromine

unsaturation
(double bond)

Saponificatio
n

break ester
bond

REAGENT
KHSO4 (oxidizing
agent)
Br2 water solution

Alcoholic KOH
not all reacts
(excess)
O.5M HCl excess
reacts

PRODUCT
Burnt fat odor

REACTION
Dehydration, oxidation

NOTE
No color reaction

Red color

Decolorization of Br2 H20

solution

Iodine number gram of iodine

absorbed by fatty acid in 100g of
fat/oil
Test indicates molecular weight
Saponification # - mgKOH that
will complete saponify 1 gram of
fat sample
High SN = low molecular weight

Neutralization of HCl and

the excess KOH after the
basic hydrolysis

High SN, low molecular weight:

coconut oil, fish oil
Low SN, high mol. weight: olive oil,
palm oil
Ester = C bond

8. CHARACTERIZATION OF AMINO ACIDS

TEST
Biuret

FOR
peptide

Ninhydrin

alpha-amino acid

Xanthoproteic

REAGENT
NAOH, Cu2SO4

PRODUCT
Rose pink to purple,
violet
Triketohydrindine hydrate Blue to blue-violet
Yellow proline,
hydroxypoline
Yellow solution

activated aromatic
except
phenylalanine
+ tyrosine,
tryptohan
indole
(tryptophan)

HNO3, conc. NAOH

Mg, conc. H2SO4, oxalic

acid

Violet interphase

Sakaguchi

guanido grp
(arginine)

Alpha-napthol, NaOH,
NaOBr

Cherry red

Millons

phenol tyrosine,
hydrobenzene

Hg, HNO3

Flesh to red ppt

Lead Acetate

amino acid
cysteine

NaOH, Pb (CH3COO)2

Brown to black ppt

Hopkins Cole

x- methionine

REACTION
Complexation of Cu2with amide
Oxidative deamination,
decarboxylation, and
reduction of Ninhydrin

NOTE
More peptide = more
intense color
More free alpha-amino
acid = more intense
(Darker color)

Electrophilic aromatic
substitution, nitration of
aromatic ring

Reduction of oxalic acid

to glycoxylic acid
Condensation of an
indole ring with aldehyde
Base catalyzed
condensation of alphanapthol with the
guanidine grp of adenine
Mercuration/complexatio
n with Hg2+ of
nitrohydroxyphenal ring
Degradation, reduction of
sulfur to sulfide
precipitation

Glycoxylic acid
(aldehyde) condense
Mg reducing agent

Color- Hg salt of
nitrated tyrosine

11. CHARACTERIZATION OF LIPIDS

TEST
Acrolein
Salkowski

FOR
glycerol
sterols
(cholesterol)

REAGENT
KHSO4
Conc. H2SO4, CHCl3

PRODUCT
Burnt fat odor
Cherry red
interphase

LiebermannBurchard

sterols
(cholesterol)

Conc. H2SO4, acetic

anhydride

Emerald green
solution

Phosphate

ammonium molybdate,
HNO3

Yellow ppt

Krauts

choline (lecithin,
sphingomyelin)
carbohydrate
moiety

KI, bismuth subnitrate

Dark orange ppt

Alpha-napthol,
95% ethanol, conc.
H2SO4
Ninhydrin

Violet ring interphase Condensation,

dehydration, hydrolysis

Molisch

Ninhydrin

alpha-amino acid
group, cephalin

Blue violet solution

REACTION
Dehydration, oxidation
Addition followed by
condensation to form
steroid
Esterification of OH at
C-3 with acetic
anhydride
Epimerization of double
bond at C-5
Decomposition and
precipitation

NOTE

When attached to
organic cpd.
Gray/white substance
= ash
H OH (phosphate +
NaO)
P phosphate
inorganic phosphorus
CO2

Complexation reaction

Oxidative deamination
followed by
condensation

Sulfuric acid
dehydration

12. CHARACTERIZATION OF NUCLEIC ACIDS

TEST
Dische

FOR
DEOXYRIBOSE
(DNA)

REAGENT
Diphenylamine, H2SO4

PRODUCT
Blue solution

Murexide

PURINE

HNO3, NH4OH/KOH

Purple red
residue

Br2, water, Ba(OH)2

Purple red
precipitate

ammonium molybdate,
HNO3, fusion mixture (KNO3,
NA3CO2)

Yellow ppt

4Ns

WheelerJohnson
Phosphate

PYRIMIDINE
2Ns
P

REACTION
Dehydration of deoxyribose
forming hydroxyl
levulinaldehyde
Complexation reaction of
levulinaldehyde with
diphenylamine
Ocidation of purine forming
dialuric acid and alloxan
Condensation reaction forming
alloxantin
Neutralization producing the
purple red murexide (ammonium
purpurate)
Oxidation of dialuric acid
Neutralization to form purple
barium salt of dialuric acid
Decomposition organic to
inorganic phosphate
Precipitation of inorganic
phosphate

NOTE

When attached to
organic cpd.
Gray/white
substance = ash
H OH
(phosphate + NaO)
P phosphate
inorganic
phosphorus
CO2