HABIB GIRLS’ SCHOOL

HIGHER SECONDARY SECTION

XII
CHEMISTRY
LAB MANUAL
Compiled by:
Aisha Hareem

pg. 1
 Table of content
Stage I
Detection of acidic and basic radicals ------------------------------ 03
1. Some important definitions ------------------------------------------ 04
2. Acidic radicals --------------------------------------------------------- 05
(CO32-, SO32-, Cl–, Br–, I–,NO3–, SO42- )
3. Basic radicals ---------------------------------------------------------- 15
(Pb2+ , Cu2+ , Fe3+, Fe2+, Al3+ , Co2+, Ni2+, Zn2+ , Ba2+, Ca2+, Mg2+ , NH4+)
Stage II
Detection of functional group ---------------------------------------- 30
4. Phenolic group --------------------------------------------------------- 31
5. Aldehydic group ------------------------------------------------------- 32
6. Carboxlic group -------------------------------------------------------- 33
Stage III
Biochemistry ----------------------------------------------------------- 34
7. Identify glucose as reducing sugar in the glucose /urine sample
of a diabetic patient ----------------------------------------------------35
8. Observe the digestion of starch by salivary amylase ------------- 36
9. Observe the denaturation of protein by urea solution ------------ 37
Stage IV
Preparation of different compounds ---------------------------------38
10. Preparation of iodoform -----------------------------------------------39
11. Preparation of copper amine complex ------------------------------ 40
12. Preparation of azo dye from amine ----------------------------------43
Stage V
13. Detection of elements in an organic compounds ----------------- 45

pg. 2
 STAGE I
DETECTION OF
ACIDIC AND BASIC
RADICALS

pg. 3
SOME IMPORTANT DEFINATIONS
Salt Analysis:
It analysis involves the identification of the cation and anion of an inorganic salt. It is
also called systematic qualitative analysis.
Qualitative Analysis:
It is the analysis which involves the detection, identification and confirmation of the
radicals present in a salt.
Salt:
A salt is an ionic compound which is formed by the neutralization of an acid with a base
so it composed of a cation from a base and an anion from an acid.
Aqueous solution:
It is a solution of salt in water.
Original Solution:
It is a homogeneous solution of salt in a solvent (usually water). It is also called O.S. It
must be transparent.
Turbidity:
The suspended particles present in a solvent.
Precipitates:
The insoluble substance present in a solution mixture.
Dry Test:
When a test is performed with a dry salt is called dry test.
Wet Test:
When a test is performed with the solution of a salt , is called wet test.
Confirmatory Test:
A test which is performed to confirm the radical that has already been identified by dry
and wet test.

pg. 4
 ACIDIC
RADICALS
It is that part of the salt which is obtained from the
acidic part during the formation of salt. It is also
known as anion. It usually contains a
Non-metal.

pg. 5
A Guide to Common Anions in Salt Analysis:
Anions Group
CO32-, NO2–, SO32-, S2- Group 1

Cl–, Br–, I–, CH3COO–, NO3–, C2O42- Group 2

PO43-, SO42- Group 3

Preliminary Tests (Anions & Acid Radicals):

The salt analysis for anions involves carrying out preliminary tests and group-wise test
to find the salt’s anion. If the test yields
positive results, a confirmatory test needs to be carried out in order to confirm if the
anion is present or not in the salt.

Preliminary Tests (Group 1 Anions):

Procedure: Take a small amount of salt solution in a test tube and add some drops of
sulphuric acid (H2SO4)/ Hydrochloric acid (HCl) to it. If you observe no change, then
you can carry out preliminary tests for Group 2 anions.

Anions Results
Carbonate (CO32-) An odourless and colourless gas is released. It turns limewater milky.
Sulphite (SO32-) Releases a pungent-smelling, colourless gas
Sulphide (S2-) A colourless gas is released which smells of rotten eggs.

Nitrite (NO2–) A pungent-smelling gas is released, which is light brown in colour.

pg. 6
Preliminary Tests (Group 2 Anions):
Procedure: Take a small amount of the solid salt in the test tube and add a few drops of
concentrated sulfuric acid (H2SO4). If you notice no change, then you can carry out
preliminary tests for Group 3 anions.

Anions Results
A pungent-smelling gas is released, that is white in color. This further
–
Chloride (Cl ) increases in intensity when a glass rod, coated with ammonium
hydroxide is brought close to the brim of the test tube.
Bromide (Br–) Reddish-brown fumes are released.
Iodide (I–) Violet fumes are released.
Acetate
Fumes, smelling like vinegar, are released.
(CH3COO–)

Nitrate (NO3–) Pungent-smelling brown fumes are released.

A combination of carbon monoxide and carbon dioxide will be

Oxalate (C2O42–)
released in a bubbly effervescence.

Preliminary Tests (Group 3 Anions):

Significantly, Group 3 anions do not have any prominent preliminary test. These are the
phosphate and sulphate ion groups,
and if no positive test results are obtained, you must directly carry out the confirmatory
tests for these.

pg. 7
Carbonate CO32-

EXPERIMENT OBSERVATION INFERENCE EQUATIONS

(DRY TESTS)

Add HCl (dilute) or H2SO4 Brisk effervescence of CO2 Dilute acid group ( CO32- ,
(dilute) to the original salt. HCO3- S2- , NO2- , SO3 2- , Na2CO3 +H2SO4 → Na2SO4 +H2O +CO2
S2O3 2-) is present
Pass the gas through lime Lime water turns milky
water May be CO32- or HCO3- CO2 + Ca(OH)2 → CaCO3 +H2O

(WET TEST)

To the salt solution (original White ppt which is soluble CO3 2- is indicated
solution) add AgNO3 in dilute HNO3 with the 2AgNO3 +Na2CO3 → 2NaNO3 + Ag2CO3
solution. evolution of CO2

(CONFIRMATORY
TEST)
To the O.S add MgSO4 White ppt of MgCO3 in CO3 2- is confirmed.
solution. the cold state
Na2CO3 +MgSO4 → Na2SO4 + MgCO3

pg. 8
Sulphite SO32-

EXPERIMENT OBSERVATION INFERENCE EQUATIONS

(DRY TEST)
Add HCl (dilute) or H2SO4 SO2 gas comes out Dilute acid group(CO32-, Na2SO3 +H2SO4 → Na2SO4 +H2O +SO2
(dilute) to the original salt. which smells as HCO3 - , S2- , NO2- , SO32-
burning sulphur. , S2O3 2- may be present.
Pass the gas through K2Cr2O7 turns green. SO32- or S2O32- may be H2SO4 + 3SO2 + K2Cr2O4→ K2SO4 + Cr(SO4)3 + H2O
acidified K2Cr2O7 solution. present.

(WET TEST)
To the O.S add AgNO3 A white ppt soluble in SO32- is indicated.
solution. dilute HNO3 is
produced. SO3 is also
given out during the 2AgNO3 +Na2 SO3 → 2NaNO3 + Ag2SO3
addition of dilute
HNO3.

(CONFIRMATORY
TEST)
To the O.S add bromine The reddish brown SO32- is confirmed.
water. colour of bromine is Na2SO3 +H2O + Br2 → Na2SO4 +HBr
discharged.
OR
To O.S add acidified KMnO4 pink colour of KMnO4 SO32- is confirmed. Na2SO3 +3H2SO4 + KMnO4→ 5Na2SO4 +3H2O + K2SO4 +
solution discharge 2MnSO4

pg. 9
Chloride Cl -
EXPERIMENT OBSERVATION INFERENCE EQUATIONS

(DRY TEST)
Add HCl (dilute) to the No Reaction Dilute acid group ( CO32- , ----------
original salt HCO3- , S2- , NO2- , SO3 2- ,
S2O3 2-) is absent
Add H2SO4 (conc) to the Colorless gas with pungent Concentrated acid group 2NaCl + H2SO4 → NaHSO4 + HCl
original salt and warm. smell giving dense white (Cl -, Br -, I -, NO3-) may be
fumes with NH4OH. present.

(WET TEST)

To the salt solution (original White ppt which is soluble Cl - is indicated AgNO3 + NaCl → NaNO3 + AgCl
solution) add AgNO3 in excess of NH4OH but
solution. insoluble in dilute HNO3
formed.
(CONFIRMATORY
TEST)
Add K2Cr2O7 (solid) and Orange brown fumes of Cl - confirmed.
conc. H2SO4 to original salt chromyl chloride (CrO2Cl2) CrO2Cl2 + 2NaOH → Na2CrO4 + 2HCl
and heat. are produced.
Pass these orange brown A bright yellow ppt. of Cl - is indicated
vapors in NaOH sol. and lead chromate PbCrO4 is Na2CrO4 + Pb(CH3COO)2 → 2CH3COONa +PbCrO4
then add lead acetate sol. obtained.

pg. 10
Bromide Br-
EXPERIMENT OBSERVATION INFERENCE EQUATIONS
(DRY TEST)
Add HCl (dilute) No Reaction Dilute acid group ( CO32- , ----------------------------
HCO3- S2- , NO2- , SO3 2- , S2O3
2-
) is absent
Add H2SO4(conc) to the White fumes of HBr along Concentrated acid group (Cl -, 2NaBr + H2SO4 → NaHSO4 + HCl
original salt and warm. with reddish brown vapours Br -, I -, NO3- ) may be
of Br2 having pungent smell present.
are evolved.
(WET TEST)
To the O.S add AgNO3 A pale yellow ppt insoluble Br - is indicated AgNO3 + NaBr → NaNO3 + AgBr
solution. in dilute HNO3 but partially
soluble in NH4OH obtained.

(CONFIRMATORY
TEST)
To O.S add CS2 , few drops Orange red colouration in Br - confirmed. 10KBr + 2KMnO4 + 8H2SO4 → 6K2SO4
of acidified KMnO4 and CS2 layer is formed. +2MnSO4 + 8H2O + 5Br2
shake well.

pg. 11
Iodide I -

EXPERIMENT OBSERVATION INFERENCE EQUATIONS

(DRY TEST)

Add HCl (dilute) to the No Reaction Dilute acid group ( CO32- , ----------------------
original salt - -
HCO3 S2- , NO2 , SO3 2- , S2O3
2-
) is absent
Add H2SO4(conc) to the Violet fumes of I2 are Concentrated acid group ( Cl -,
original salt and warm. evolved which condenses to Br -, I -, NO3- ) may be present. NaI + H2SO4 → NaHSO4 + HI
form black mass.

(WET TEST)
To the salt solution A deep yellow ppt formed -
I is indicated
(original solution) add which is insoluble in AgNO3 + Nal → NaNO3 + Agl
AgNO3 solution. NH4OH& dilute HNO3

(CONFIRMATORY
TEST)
To O.S , add CS2 or CCl4 A Violet color is produced -
I is confirmed. 10KI + 2KMnO4 + 8H2SO4 → 6K2SO4 +2MnSO4
and few drops of KMnO4 in CS2 or CCl4
+ 8H2O + 5I2
and shake well.

pg. 12
Nitrate NO3 -

EXPERIMENT OBSERVATION INFERENCE EQUATIONS

(DRY TEST)
Add HCl (dilute) to the No Reaction Dilute acid group ( CO32- ,
original salt HCO3- S2- , NO2- , SO3 2- ,
S2O3 2-) is absent
Add H2SO4(conc) to the White fumes of HNO3 ocme Concentrated acid group ( Cl
original salt and warm. out which have pungent -
, Br -, I -, NO3- ) may be NaNO3 + H2SO4 → Na2SO4 + HNO3
odour. present.

(WET TEST)
To the salt solution (original No ppt -
NO3 is indicated.
solution) add AgNO3
solution.
(CONFIRMATORY
TEST)

RING TEST:
To the O.S.add FeSO4 sol. Brown ring -
NO3 is confirmed. FeSO4 +3NO2 +H2O → 2HNO3 + [Fe (NO)SO4]
freshly prepared & conc.
H2SO4 carefully along the
sides of the test tube.

pg. 13
Sulphate SO42-
EXPERIMENT OBSERVATION INFERENCE EQUATION

(DRY TEST)
Add HCl (dilute) No Reaction dilute acid group ( CO32- ,
HCO3- S2- , NO2- , SO3 2- ,
S2O3 2-) is absent

Add H2SO4(conc) to the No Reaction Concentrated acid group ( Cl

original salt and warm - - - -
, Br , I , NO3 ) is absent.
Add BaCl2(soln) to the White ppt May be SO42- or PO43-
original salt solution. Na2SO4 +2 BaCl2 → 2NaCl +BaSO4

Add HCl (dilute) to White ppt insoluble. SO42- confirmed.

white ppt.

(CONFIRMATORY
TEST)
Add lead acetate to O.S White ppt SO42- confirmed.
Na2SO4 + Pb(CH3COO)2 → 2CH3COONa + PbSO4

pg. 14
 BASIC
RADICALS
It is that part of the salt which is obtained from the basic part
during the formation of salt. It is also known as cation. It usually
contains a metal

pg. 15
A GUIDE TO COMMON ANIONS IN SALT ANALYSIS:
Always check the colour of the salt first. Certain cations have coloured salts. If the salt is coloured,
you can directly conduct a preliminary test for the cation that forms salts of that colour. The cations
that form coloured salts are tabulated below:

Colour of the Inorganic Salt Cation

Blue Cu2+

Deep blue Co2+ (anhydrous salt)

Greenish-blue Cu2+ (hydrated salt)

Green Ni2+ (hydrated salt)

Light green Fe2+

Yellow, brown, or yellowish-brown Fe3+

Pale pink Mn2+

Rose-red Co2+ or mercuric iodide (HgI2)

Purple or dark green Cr3+

pg. 16
COMMON CATIONS (BASIC RADICALS) FOR SALT ANALYSIS

Group Group Reagents Cations

Group 1 O.S. + dil. HCl Pb2+

Group 2 O.S. + dil. HCl, + H2S Cu2+

Group 3 O.S. + NH4Cl + NH4OH Fe3+, Fe2+, Al3+

Group 4 O.S. + NH4Cl + NH4OH + H2S Co2+, Mn2+, Ni2+, Zn2+

O.S. + NH4Cl + NH4OH +

Group 5 Ba2+, Ca2+, Sr2+
(NH4)CO3

O.S. + NH4Cl + NH4OH +

Group 6 Mg2+ , NH4+
Na2HPO4

pg. 17
Lead Pb+2

DRY TEST

Add HCl (dil) to salt solution. white ppt. obtained. May be Pb+2 Ag+2, Hg+ Pb(NO3)2 + 2HCl → 2HNO3 +PbCl2

Add NH4OH excess to white White ppt. insoluble in

May be Pb+2
ppt. excess

WET TEST

To the white ppt. obtained White ppt. remains

Pb+2 is indicated
add dil HCl unaffected

CONFIRMATORY TEST

Add KI sol. to O.S Yellow ppt. Pb+2 is confirmed. Pb(NO3)2 +2KI → 2KNO3 +PbI2

Add K2CrO4 solution to O.S Yellow ppt. Pb+2 confirmed. Pb(NO3)2 +K2CrO4 → 2KNO3 + PbCrO4

pg. 18
Copper Cu+2
TEST FOR EXPERIMENT OBSERVATION INFERENCE EQUATIONS
+2 +2 +
Add HCl (dil) to salt No reaction Group 1 (Pb Ag , Hg
solution. is absent)
GROUP 1

WET TEST

Add HCl (dil) and Brownish black ppt is May be Cu+2 , Pb+2 , Bi+3 , CuSO4 +H2S → H2SO4 + CuS
H2S gas or solution formed Hg+2
to salt solution

To the above ppt add Blue solution is formed Cu+2 indicated

1:1 HNO3 and boil
GROUP 2

CONFIRMATORY
TEST

Add potassium ferro- Choclate brown ppt. Cu+2 confirmed 2CuSO4 + K4[Fe(CN)6] → 2K2SO4 +
cyanide sol. to O.S obtained. Cu2[Fe(CN)6]

Add NaOH soln. to light blue ppt which turns Cu+2 confirmed CuSO4 + 2NaOH → Na2SO4 +
the salt solution black on heating is formed Cu(OH)2

pg. 19
Aluminium Al+3
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)
Group Add HCl (dil) to salt No Reaction Group 1 ( Pb+2 Ag+2, Hg+ )
1 solution. is absent

Group Add HCl (dil) and H2S No Reaction Group 2( Cu+2 , Pb+2 , Bi+3 ,
2 gas or solution to salt Hg+2 Cd+2 ) is absent.
solution
Group Add NH4Cl (solid) and White gelatinous ppt. May be Al+3
3 NH4OH excess to salt obtained.

(CONFIRMATORY
TEST)
Lake Test :

Add NaOH soln. to salt White ppt. soluble in Al+3 confirmed AlCl3 + NaOH → 3NaCl + Al(OH)3
solution. excess.
OR
To O.S add litmus sol, A blue ppt, is seen Al+3 confirmed AlCl3 + 3NH4OH → 3NH4Cl + Al(OH)3
then add HCl dil. and floating in solution
NH4OH

pg. 20
Ferrous Fe+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)

Group Add HCl (dil) to salt No Reaction Group 1 ( Pb+2 Ag+2,

1 solution. Hg+ ) is absent

Group Add HCl (dil) and H2S No Reaction Group 2( Cu+2 , Pb+2 ,
2 gas or solution to salt Bi+3 , Hg+2 Cd+2 ) is
solution absent.
Group Add NH4Cl (solid) and Green ppt. obtained May be Fe+2or Cr+3
3 NH4OH excess to salt

(CONFIRMATORY
TEST)
Add potassium ferro- Bluish white ppt. Fe+2 confirmed FeSO4 + 2K4 [Fe(CN)6]3 → Fe3[ Fe(CN)6] + 3K2SO4
cyanide solution to the obtained
O.S
To O.S add NaOH Greenish white ppt. Fe+2 confirmed FeSO4 + NaOH → Fe(OH)2 + Na2SO4
solution

pg. 21
Ferric Fe+3
TEST EXPERIMENT OBSERVATION INFERENCE
EQUATIONS
FOR
(WET TEST)
Group Add HCl (dil) to salt No Reaction Group 1 ( Pb+2 Ag+2,
1 solution. Hg+ ) is absent

Group Add HCl (dil) and H2S No Reaction Group 2( Cu+2 , Pb+2 ,
2 gas or solution to salt Bi+3 , Hg+2 Cd+2 ) is
solution absent.
Group Add NH4Cl (solid) and Reddish Brown ppt. May be Fe+3
3 NH4OH excess to salt obtained.

(CONFIRMATORY
TEST)
Add potassium ferro- Blue ppt. obtained Fe+3 confirmed 4FeCl3 + 3K4 [Fe(CN)6]3 → Fe4[ Fe(CN)6]3 + 12KCl
cyanide to the O.S

OR

To O.S add NaOH Reddish-brown ppt Fe+3 confirmed FeCl3 + 3NaOH→Fe(OH)3 + 3NaCl
solution insoluble in excess

pg. 22
Zinc Zn+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)
GROUP Add HCl (dil) to salt Group 1 (Pb+2 Ag+2, Hg+
1 solution. is absent)
No reaction
GROUP Add HCl (dil) and H2S gas No reaction Group 2 (Cu+2 , Pb+2 ,
2 or solution to salt solution Bi+3 , Hg+2 )is absent

GROUP Add NH4Cl (solid) and Group 3 (Al+3,Fe+3, Fe+2

3 NH4OH excess to salt or Cr+3) is absent
solution. No reaction
May be Ni+2, Cr+3, Zn+2
GROUP
4
Add NH4Cl (solid)
NH4OH excess and H2S to
Greyish white ppt.
Mn+2 ZnCl2 +H2S → 2HCl +ZnS
salt solution
CONFIRMATORY
TEST
Zn+2 confirmed
Add NaOH to salt solution. a white gelatinous ppt
which is soluble in ZnCl2 +2NaOH → 2NaCl +Zn(OH) 2

excess of NaOH
Zn+2 confirmed
To O.S add NH4OH White ppt. soluble in
excess NH4OH ZnCl2 +2NH4OH→ 2NH Cl +Zn(OH)
4 2

pg. 23
Nickle Ni+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
WET TEST

GROUP Add HCl (dil) to salt No reaction Group 1 (Pb+2 Ag+2, Hg+ is
1 solution. absent)
GROUP Add HCl (dil) and H2S gas or No reaction Group 2 (Cu+2 , Pb+2 , Bi+3 ,
2 solution to salt solution Hg+2 )is absent
GROUP Add NH4Cl (solid) and No reaction Group 3 (Al+3,Fe+3, Fe+2 or
3 NH4OH excess to salt Cr+3) is absent
solution.

May be Ni+2, Cr+3, Zn+2 Mn+2

GROUP Add NH4Cl (solid) NH4OH
4 excess and H2S to salt
Black ppt.
NiCl2 + H2S → 2HCl + NiS
solution
CONFIRMATORY TEST

Ni+2 confirmed.
Add NaOH to salt solution. Green ppt.
NiCl2 + 2NaOH → 2NaCl +
Ni(OH)2
Ni+2 confirmed.
Add Na2CO3 sol. to the O.S Green ppt.
NiCl2 + Na2CO3 → 2NaCl +
NiCO3

pg. 24
Cobalt Co+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR

WET TEST
Group 1 Add HCl (dil) to salt No reaction Group 1 (Pb+2 Ag+2, Hg+
solution. is absent)
Group 2 Add HCl (dil) and H2S gas No reaction Group 2 (Cu+2 , Pb+2 ,
or solution to salt solution Bi+3 , Hg+2 )is absent

Group 3 Add NH4Cl (solid) and No reaction Group 3 (Al+3,Fe+3, Fe+2

NH4OH excess to salt or Cr+3) is absent
solution.
May be Ni+2, Cr+3, Zn+2
Group 4 Add NH4Cl (solid)
NH4OH excess and H2S to
Black ppt. original salt is
deep pink. Mn+2 CoCl2 + H2S → 2HCl + CoS
salt solution
CONFIRMATORY
TEST
Co+2 confirmed
Add NaOH to salt
solution.
Blue or violet ppt.
CoCl2 + 2NaOH → 2NaCl + Co(OH) 2

Co+2 confirmed
Add Na2CO3 sol. to the
O.S
Blue or violet ppt.
CoCl2 + Na2CO3 → 2NaCl + CoCO3

pg. 25
Barium Ba+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)
Group Add HCl (dil) to salt solution. No Reaction Group 1 ( Pb+2 Ag+2, Hg+ )
1 is absent
Group Add HCl (dil) and H2S gas or No Reaction Group 2( Cu+2 , Pb+2 , Bi+3 ,
2 solution to salt solution Hg+2 Cd+2 ) is absent.
Group Add NH4Cl (solid) and No Reaction Group 3 ( Al+3, Fe+2 or Cr+3 )
3 NH4OH excess to salt is absent
Group Add NH4Cl (solid) NH4OH No Reaction Group 4( Ni+2, Cr+3 Co+2
4 excess and H2S to salt solution Zn+2 Mn+2) is absent

Group Add NH4Cl (solid) , NH4OH White ppt. May be Ba+2, Sr+2 Ca+2
5 excess and ammonium Ag+2, Hg+ ) is absent
carbonate to the salt solution
(CONFIRMATORY
TEST)
Ba+2 confirmed
O.S +NH4Cl (solid) , NH4OH
excess and ammonium
Yellow ppt.
BaCl2 + K2CrO4 →2KCl + BaCrO 4

carbonate add acetic acid then

add potassium chromate
solution
OR

Ba+2 confirmed
To O.S add H2SO4 White ppt. insoluble in
conc. HCl BaCl2 + H2 SO4 → 2HCl + BaSO4

pg. 26
Calcium Ca+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)

Group Add HCl (dil) to salt No Reaction Group 1 ( Pb+2 Ag+2, Hg+ )
1 solution. is absent
Group Add HCl (dil) and H2S No Reaction Group 2( Cu+2 , Pb+2 , Bi+3 ,
2 gas or solution to salt Hg+2 Cd+2 ) is absent.
solution
Group Add NH4Cl (solid) and No Reaction Group 3 ( Al+3 Fe+2or Cr+3 )
3 NH4OH excess to salt is absent
Group Add NH4Cl (solid) No Reaction Group 4( Ni+2, Cr+3 Co+2
4 NH4OH excess and H2S Zn+2 Mn+2) is absent
to salt solution
May be Ba+2, Sr+2 Ca+2
Group Add NH4Cl (solid) ,
5 NH4OH excess and
White ppt.
CaCl2 + (NH4)CO3 → 2NH Cl+ CaCO
4 3

ammonium carbonate to
the salt solution
(CONFIRMATORY
TEST)
Ca+2 confirmed
To the O.S add
(NH4)2C2O4
White ppt.
CaCl2 + (NH4)2C2O4 → 2NH Cl+4
CaC2O4
OR
To O.S add H2SO4 No white ppt. Ca+2 confirmed

pg. 27
Ammonium NH4+
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR (DRY TEST)
Group Add HCl (dil) to salt solution. No Reaction Group 1 ( Pb+2 Ag+2, Hg+ )
1 is absent
Group Add HCl (dil) and H2S gas or No Reaction Group 2( Cu+2 , Pb+2 , Bi+3 ,
2 solution to salt solution Hg+2 Cd+2 ) is absent.
Group Add NH4Cl (solid) and No Reaction Group 3 ( Al+3 Fe+2or Cr+3 )
3 NH4OH excess to salt is absent
Group Add NH4Cl (solid) NH4OH No Reaction Group 4( Ni+2, Cr+3 Co+2
4 excess and H2S to salt Zn+2 Mn+2) is absent
solution
Group Add NH4Cl (solid) , NH4OH No Reaction Group 5 ( Ba+2, Sr+2 Ca+2 )
5 excess and ammonium is absent
carbonate to the salt solution
Group Add NH4Cl (solid) , NH4OH No ppt. May be NH4+ K+ or Na+
6 excess and ammonium
hydrogen phosphate to salt
solution.
Add NaOH to solid salt and Smell of NH3 gas which NH4+ is indicated. NH4Cl+ NaOH → NaCL +H2O +NH3
warm gives white dense fumes
with the rod dippes in
conc. HCl
(CONFIRMATORY TEST)

Add Nessler's soln. & NaOH Brown ppt. NH4+ confirmed NH4OH + 2K2HgI4 +NaOH→
to O.S NH2Hg2I3+4KI+NaI+2H2O
OR
Add freshly prepared sodium Yellow ppt. NH4+ confirmed 3NH4Cl+Na3[Co(NO2)6] →3NaCl
cobaltnitrite to conc. O.S. +(NH4)3 Co(NO2)6]

pg. 28
Magnesium Mg+2
TEST EXPERIMENT OBSERVATION INFERENCE EQUATIONS
FOR
(WET TEST)
Group Add HCl (dil) to salt solution. No Reaction Group 1 ( Pb+2 Ag+2,
1 Hg+ ) is absent
Group Add HCl (dil) and H2S gas or No Reaction Group 2( Cu+2 , Pb+2 ,
2 solution to salt solution Bi+3 , Hg+2 Cd+2 ) is
absent
Group Add NH4Cl (solid) and No Reaction Group 3 ( Al+3 Fe+2 or
3 NH4OH excess to salt Cr+3 ) is absent
Group Add NH4Cl (solid) NH4OH No Reaction Group 4( Ni+2, Cr+3 Co+2
4 excess and H2S to salt solution Zn+2 Mn+2) is absent
Group Add NH4Cl (solid) , NH4OH No Reaction Group 5 ( Ba+2, Sr+2
5 excess and ammonium Ca+2 ) is absent
carbonate to the salt
Group Add NH4Cl (solid) , NH4OH White ppt. May be Mg+2 3MgSO4 +2(NH4)2HPO → (NH4)2SO4 +
6 excess and ammonium Mg3(PO4)2
hydrogen phosphate to salt
solution.
Add NaOH sol. to salt solution White ppt. May be Mg+2

(CONFIRMATORY
TEST)
Add NH4OH sol. to the salt white ppt. Mg+2 confirmed MgSO4 + 2NaOH → Na2SO4 +
solution Mg(OH)2

pg. 29
 STAGE II
DETECTION OF
FUNCTIONAL GROUP

pg. 30
 PHENOLIC GROUP
EXPERIMENT OBSERVATION INFERENCE EQUATIONS
To the sample solution, No CO2 gas evolved Phenolic group is
add freshly prepared indicated
NaHCO3 sol.

Treated the compound Blackening without Phenolic group is

with conc. H2SO4 effervances indicated
(CONFIRMATORY
TEST)
To the sample solution, Violet color is Phenol is 3ArOH +FeCl3 → Fe(OAr)3 + 3HCl
add freshly prepared produced. confirmed.
FeCl3 solution

pg. 31
 ALDEHYDIC GROUP
EXPERIMENT OBSERVATION INFERENCE EQUATIONS

To the organic De-colourization of Aldehydic group

compound, add KMnO4 takes place is indicated
alkaline KMnO4
solution

To the sample No CO2 gas evolved Aldehyde is

solution, add indicated
freshly prepared
NaHCO3 solution.

(CONFIRMATORY
TEST)
(SILVER MIRROR
TEST )
To the organic A mirror is formed Aldehyde is R-CHO +2 Ag(NH3)2OH → R-COONH4
compound, add confirmed + 2Ag +3NH3 +H2O
Ammonical AgNO3
(Tollen's Reagent)
and heat on water
bath

pg. 32
 CARBOXLIC GROUP
EXPERIMENT OBSERVATION INFERENCE

To the organic compound, add No De-colourization of KMnO4 Carboxylic group is indicated

alkaline KMnO4 solution. takes place

To the organic compound, add Effervesces takes place and Carboxylic group is indicated
freshly prepared NaHCO3 sol. CO2 gas evolved

(CONFIRMATORY TEST)

To the sample solution, add freshly

Buff coloured ppt. is obtained Carboxylic group is Confirmed
prepared FeCl3 solution

pg. 33
 STAGE III
BIOCHEMISTRY

pg. 34
 GLUCOSE AS REDUCING SUGAR

OBJECT : Identify glucose as reducing sugar in the glucose /urine sample of a diabetic patient
CHEMICALS : Fehling solution A ,Fehling solution B, Benedict solution
THEORY :
Reducing sugar is any sugar that contain aldehyde group that are oxidized to carboxylic acid .any sugar that
either has no aldehydic group is capable of forming its solution through isomerism. Glucose is most important
carboxylic fuel in the body.it is monosaccharide as it cannot be broken down further by hydrolysis. It is
classified as hexose because of its 6 carbon skeleton. It is the strong reducing property of sugar which make it
relatively easy to measure.

RESULT
EXPERIMENT OBSERVATION

To the sample solution, add Brown /Orange or Red ppt are

Benedict solution and then heat. formed.
Glucose as reducing sugar is
confirmed.
To the sample solution, add Fehling Brown /Orange or Red ppt are
solution A and Fehling Solution B formed.
and then heat.

pg. 35
 DIGESTION OF STARCH
OBJECT : Observe the digestion of starch by salivary amylase
CHEMICALS : Amylase , starch solution, iodine solution
THEORY :
The digestion of a carbohydrate such as starch begins in the mouth, where is it mixed with saliva containing the
enzyme salivary amylase. Starch, a long chain of repeating glucose molecules, is hydrolyzed (cut) by amylase into
shorter polysaccharide chains and eventually into the disaccharide maltose (known as a reducing sugar), which
consists of two glucose subunits.

PROCEDURE:
 Take four test tubes and add 1 ml of iodine solution in each test tube.
 Prepare a mixture solution by mixing 5ml starch solution and 2ml saliva solution in a test tube.
 After one minute take out 2 drops of mixture solution and transfer it in to another test tube containing about 1
ml of iodine solution and note the color produced.
 Repeat the experiment after every 5 minutes till the test shows no blue color.

OBSERVATIONS:
Time 1 min. 5 min. 10 min. 15 min.

Color Intensity Deep Blue Blue Light Blue No Blue

Absences of blue color on addition to iodine solution means absences of starch in the mixture solution.

RESULT:
Starch gests hydrolysed by salivary amylase.

pg. 36
 PROTEIN UREA DENATURATION
OBJECT : Observe the denaturation of protein by urea solution

CHEMICALS : Egg White, Urea Solution (8%)

THEORY :
Denaturation can be brought about in various ways. Proteins are denatured by treatment with alkaline or acid,
oxidizing or reducing agents, and certain organic solvents. Interesting among denaturing agents are those that
affect the secondary and tertiary structure without affecting the primary structure. The agent most frequently used
for this purpose is urea. These molecules, because of their high affinity for peptide bonds, break the hydrogen
bonds and the salt bridges between positive and negative side chains, thereby abolishing the tertiary structure of
the peptide chain. When denaturing agents are removed from a protein solution, the native protein re-forms in
many cases.

EXPERIMENT OBSERVATION RESULT

To the egg white, add 2 ml of urea Coagulation occur on heating Protein is denatured
solution and heat

pg. 37
 STAGE IV
SYNTHESIS OF DIFFERENT
COMPOUNDS

pg. 38
OBJECT:
Prepare Iodoform
THEORY:
When halogen reacts with alcohol or carbonyl compound in the presence of a base,
multiple halogenation occurs at the carbon of methyl group and form haloform.
This reaction is called haloform reaction.
C2H5OH +4 X2 +6NaOH  CHX3 +HCOONa + 5 NaX + 5 H2O
Iodoform is an organoiodine compound with the formula CHI3 (also called
triiodomethane, a yellow, crystalline solid belonging to the family of organic
halogen compounds, used as an antiseptic component of medications for minor skin
diseases and have a tetrahedral molecular geometry. It is a relatively water-
insoluble yellow solid that is chemically reactive in free-radical reactions.
Iodoform has also been found in dental paste and root canal filling materials.
Iodoform is soluble in fatty acids and decomposes releasing iodine in nascent state
(96,7% of iodine) when in contact with secretions or endodontic infections.
CHEMICALS REQUIRED:
 Powdered Iodine (10 g)
 Ethanol (20 ml)
 NaOH
STRUCTURE AND EQUATION:
C2H5OH +4 I2 +6NaOH  CHI3 +HCOONa + 5 NaI + 5 H2O

pg. 39
PROCEDURE:
 Dissolve powdered iodine in ethanol (violet color disappear, brown sol.
obtained)
 Add NaOH solution to the above mixture till yellow crystals of
iodoform obtained.
 Filter and wash the crystals with cold water and dry.
 M.P of pure CHI3 crystals = 119° C

pg. 40
OBJECT:
Preparation of Copper Ammine Complex

THEORY:
Tetraamminecupric sulfate is the salt with the formula [Cu(NH3)4]SO4·H2O. This
dark blue to purple solid is a salt of the metal complex [Cu(NH3)4(H2O)]2+. It is
closely used for the production of cellulose fibers in the production of rayon. It is
used to print fabrics, used as a pesticide and to make other copper compounds like
copper nano-powder.

CHEMICALS:
 CuSO4 (5 g)
 H2SO4 (1-2 ml)
 Ammonia Solution
 Ethanol

STRUCTURE:

SYNTHESIS:
This compound can be prepared by adding concentrated solution of ammonia to a
saturated aqueous solution of copper sulfate pentahydrate followed by precipitation
of the product with ethanol or isopropanol.
4NH3 + CuSO4·5H2O → [Cu(NH3)4]SO4·H2O + 4 H2O
The deep blue crystalline solid tends to hydrolyse and evolve (release) ammonia
upon standing in air. It is fairly soluble in water. The brilliant dark blue-violet color
of tetraamminecopper(II) sulfate solution is due to presence of [Cu(NH3)4]2+.
Often, the dark blue-violet color is used as a positive test to verify the presence of
Cu2+ in a solution.

pg. 41
PROCEDURE

 Dissolve CuSO4 in 20 ml water, light blue

solution is obtained.
 Add H2SO4 to get clear solution
 Add ammonia solution till the deep blue
solution appeared.
 Add ethanol to the deep blue solution till
needle like blue crystals are obtained.
 Filter and wash with ethanol and dry.

pg. 42
OBJECT: Prepare azo dye from amine

THEORY:
Azo dyes are organic compounds bearing the functional group R−N=N−R′, in which
R and R′ are usually aryl. They are a commercially important family of azo
compounds, i.e. compounds containing the linkage C-N=N-C.

CHEMICALS REQUIRED:
 Primary aromatic amine
 HCl
 2% NaNO3
 β-Naphthol
 NaoH

AZO-DYE TEST:
This test is performed in order to confirm the presence of aromatic amines. Aromatic
amines forms a diazonium salts when treated with HNO 2. These diazo compounds
couple with phenols like β-naphthols forms an orange azo dye which is insoluble in
water.

pg. 43
PROCEDURE:
 Take three test tubes and labeled it as A, B and C.
 Test tube A: dissolved the organic compound with HCl and dilute it with water.
 Test tube B: add sodium nitrite solution.
 Test tube C: Dissolve β-naphthol with NaOH.
 Place all the three test tubes in an ice bath. Now add the contents in test tube B to
the mixture in test tube A.
 The resulting mixture is added to the contents in test tube C.
 Red-orange dye is formed.

USES:
Azo dyes are the most important synthetic colorants which have been widely used
in textile, printing, paper manufacturing, etc.

pg. 44
 STAGE V
DETECTION OF ELEMENTS
IN ORGANIC COMPOUND

pg. 45
OBJECT: Detection of nitrogen(N), Sulphur(S) and Halogens (Cl, Br ,I) in organic compound
THEORY: Besides carbon, hydrogen, oxygen, the most commonly occurring elements in organic
compounds are nitrogen, sulphur and halogens. Since organic compounds are organic in nature and
there is no direct method to detect these element, we can use the sodium fusion test (Lassaigne’s test)
in which the organic compound is converted in to ionic form by treating with sodium metal.
Na + C + N → NaCN
2Na + S → Na2S
Na + X → NaX ( X= Cl, Br, or I)
LASSAIGNE’S TEST:

pg. 46
S.R
Experiment Observation Inference
No.
1 Test for Nitrogen, N: Lassaign’s Prussian Blue Nitrogen is present.
solution 2 ml + 2 drops NaOH + 1-2 Coloration.
ml FeSO4, boil + dil. H2SO4
2 Test for Sulphur,S: Lassaign’s Violet Coloration. Sulphur is present.
solution 1- 2 ml + 1 drops sodium
nitroprusside. Black ppt of lead
Lassaign’s solution 1- 2 ml + acetic sulphide is formed.
acid (few drops) + lead acetate
3 Test for Halogens,X: Lassaign’s a) White ppt of AgCl3 Chlorine is present.
solution 1 ml + AgNO3 and few soluble in NH4OH.
drops of conc HNO3 b) Pale yellow ppt of Bromine is present.
AgBr partially
soluble in NH4OH.
c) Yellow ppt of AgI Iodine is present.
insoluble in
NH4OH.

pg. 47