DECCAN EDUCATION SOCIETY’s

FERGUSSON COLLEGE (AUTONOMOUS) PUNE-411028

DEPARTMENT OF CHEMISTRY

F.Y. B. Sc. PRACTICAL LABORATORY MANUAL

SEMESTER I (2019-2020)

S.N CONTENTS
I LABORATORY SAFETY RULES
II GENERAL FORMAT OF INORGANIC QULITATIVE ANALYSIS
III FORMAT FOR INORGANIC QUALITATIVE ANALYSIS
IV INORGANIC QUALITATIVE ANALYSIS OF KNOWN COMPOUND
V GENERAL FORMAT OF ORGANIC QULITATIVE ANALYSIS
VI FORMAT FOR ORGANIC QUALITATIVE ANALYSIS
VII ORGANIC QUALITATIVE ANALYSIS OF KNOWN COMPOUND
VIII DETERMINATION OF PHYSICAL CONSTANTS
IX PURIFICATION BY CRYSTALLISATION
X PURIFICATION BY SUBLIMATION
XI THIN LAYER CHROMATOGRAPHY

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 1

 LABORATORY SAFETY RULES

Aim: To understand the correct ways of handling Chemicals, Reagents, Equipments and
performing experiments in the lab.
Chemistry laboratories can be hazardous and dangerous if the rules are not followed. In a
chemistry lab, a student may handle materials which are carcinogenic, flammable, and explosive.
Some of these materials and equipment may also cause severe burns, cuts, if handled improperly
or carelessly. Most accidents that occur in the chemistry laboratory are a result of carelessness,
impatience, improper or unauthorized experimentation, and disregard for safety rules or proper
operating procedures. In order to minimize the chances of an accident, certain rules and regulations
must be obeyed, while performing experiments in a chemical laboratory. Therefore, it is not
advisable for anyone to work in a laboratory without proper knowledge of the hazards involved.
The student is expected to exercise proper judgements and precautions at all times when working
in the laboratory. Instructors will discuss specific safety precautions relevant to each experiment
during laboratory sessions.
Safety and Laboratory rules!
1. Unauthorized experiments or work should not be performed in the laboratory.
2. Approved eye protection must be worn at all times in the laboratory. In case of any injury
eyes must be rinsed immediately with large quantities of water.
3. Long hair must be tied and loose clothing must be avoided while working in a laboratory.
4. Appropriate clothing must be worn at all times while in the laboratory. Legs must be
completely covered below the knee by proper choice of clothing.
5. Closed shoes with socks must be worn at ALL times – open-toed shoes, backless shoes,
sling backs, and sandals are not permitted.
6. Awareness of the location and proper use of first aid kits, fire extinguishers, fire blankets,
safety showers, eye wash devices is required.
7. The labels of chemicals must be read carefully.
8. Eating, smoking, and drinking to be avoided in a chemistry laboratory.
9. Hands must be thoroughly washed before leaving the laboratory.
10. Fume hoods must be used for tests that involve toxic or irritating vapors.
11. Mouth suction is never used to fill a pipette.
12. The open end of test tube must never be pointed towards oneself or anyone else.
13. Water should never be poured into concentrated acid.
14. Proper procedure for igniting and operating a laboratory burner must be learnt. Flame
must be extinguished when the burner is not being used. No flammable reagents to be kept
nearby before lighting the burner.
15. Liquid and solid waste containers must be properly used at all times.
16. Chemicals should never be placed directly on the balance pan. Always proper weighing
container must be used, when using a balance to weigh a chemical.
17. Unused chemicals should not be returned to their original container (unless directed to do
so by the instructor).
18. The lids, caps, and stoppers must be securely replaced, after removing reagents from
containers.
19. Spatulas must be wiped clean before and after inserting into reagent bottles.
20. Any accident and/or injury, however minor, to be reported to the instructor immediately.
21. All personal belongings should be placed in the bookcases while entering the laboratory.
22. The work area must be cleaned before leaving the laboratory.
23. Ensure that all gas, water, vacuum, and air valves are completely turned off.
24. For any assistance instructor must be contacted.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 2

 II. GENERAL SCHEME OF INORGANIC QUALITATIVE ANALYSIS

Preliminary Tests

Colour Red Cu2O, CrO3, Pb3O4

Pink Hydrated crystalline salt of Mn and

Co
Blue
Hydrated Cu(II) salts, anhydrous
Co salt
Green Cr2O3, Cr(OH)3, Fe(II) salts, Ni-
salts, CrCl3.6H2O, CuCl2.2H2O,
CuCO3, K2MnO4
Yellow
CdS, SnS2, PbI2, K4[Fe(CN)6],
chromates, Fe(III) chlorides and
nitrates, PbO
Black
PbS, CuS, CuO, FeS, MnO2,
Co3O4, CoS, NiS, Ni2O3etc
White / Colourless
Compound of Al, Zn, Ca, Ba, Sr,
Flesh Mg, K and ammonium salts

Fe or Mn salts

Nature Crystalline Soluble salts of Na, K and

ammonium or salts of Cl-, Br-, I-,
SO42-, NO2-, NO3-
Amorphous Insoluble salts CO32-, S2-, O2-,
PO43- , BO33- may be present

Dry Tests for Basic Radicals: (Cations)

Test Observation Inference

Heating in a dry i) Change in colour:

test tube: Place a
small quantity of the a) White to yellow when hot, white when cold ZnO and many
given mixture in a Zn salts
b) White to yellow when hot, yellow when cold
dry test tube. Heat it PbO and some
cautiously and c) White to brown when hot, brown when cold

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 3

strongly and observe d) Red to black when hot, black when cold Pb salts
any changes
e) Coloured to black when hot, black when cold CdO and many
Cd salts

Fe2O3
f) White to white when hot, white when cold
Cu, Mn, Ni, Co
salts may be
ii) Crackling sound present

Al, Ba, Ca, Sr,

Mg, Na, K,
iii) Substance fuses NH4+ salts

iv) Water vapour condenses on cooler part of

test tube
Crystalline salts
like KBr,
NH4Cl,
v) Formation of white sublimate
Pb(NO3)2,
vi) A gas or vapour is evolved Ba(NO3)2

Colourless and odourless gas Alkali salts,

nitrates of
Colourless gas with pungent odour turning alkaline earth
moist turmeric paper red metals
Brown fumes or red-brown vapour Hygroscopic
salts or salts
Violet vapour turns starch paper black
containing
water of
crystallization

NH4+ salts,
mainly halides

Nitrate or
chromate salts

Ammonium
salts

Nitrates or
nitrites or

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 4

 bromides of
heavy metals

Iodide salts

Charcoal Block i) White infusible residue was obtained (take Ca2+, Sr2+,
Reduction (Heating cobalt nitrate test) Ba2+, Al3+,
in a charcoal Mg2+, Zn2+
cavity): Make a Cobalt nitrate test: Moisten above residue salts may be
small fresh cavity with 2 drops cobalt nitrate solution and heat in present
(with the help of oxidizing flame (blue) with blow pipe-
tongs) on a piece of Residue turns
charcoal block, take
a little substance in Deep blue
the groove, moisten
Bright green
with a drop of water
Al salts
and heat in a Pale pink or rosy mass
reducing flame Zn salts
(yellow flame) using Grey mass
blow pipe Mg salts
ii) Coloured residue was obtained (take sodium
carbonate test) Ca, Sr, Ba salts

Sodium carbonate test: Add 10 mg Na2CO3 to Transition

the above coloured residue, moisten with one metal salts like
drop water and heat in yellow reducing flame Cu2+, Fe2+,
using blow pipe- Fe3+, Mn2+
present
Red flakes

Yellow incrustation when hot and white when

cold

Coloured residue left a)Cu salts

White infusible residue b)Zn salts

c) Cu, Fe, Cr,

Ni, Co, Mn
salts

d) Ca, Sr, Ba,

Mg, Al, Zn
salts

Action of NaOH: Strong smell of ammonia turning moist NH4+

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 5

Take a small amount turmeric paper red (ammonium)
of the given mixture salts may be
in a dry test tube and present
add few drops of
NaOH solution. Heat
and hold moist
turmeric paper at the
mouth of the test
tube without
touching inner wall

Dry tests for Acidic Radicals: (Anions)

Experiment Observation Inference

Action of dil HCl: Take a small amount of a) Brisk effervescence of a) CO32-

the given mixture (around 10 mg) in a dry CO2 gas (without heating) (carbonate)
test tube. Add 3-4 drops of dil HCl and may be present
observe carefully b) The colourless gas
turns freshly prepared b) CO32- may
Then heat gently lime water milky be present

[(b) Lime Water Test: Cover the test tube c) Evolution of colourless
with another blank test tube, hold it in 45ο H2S gas smell like rotten
angle and then heat gently. As the vapours eggs and turns lead acetate
enter into the blank test tube, cover the paper black c) S2- may be
mouth of the blank test tube with the
thumb. Slightly open the test tube, add d) Brown fumes acidic to present
fresh lime water and shake it vigorously] litmus

d) NO2- may
be present

Action of MnO2 + conc H2SO4: Take a a) Greenish-yellow gas a) Cl-

small amount of the given mixture in a dry evolves which turns blue (chloride) may
test tube heated with small amount of litmus red and bleaches it be present
MnO2 and 1 ml conc. H2SO4.
b) Reddish-brown vapours
i) Observe [observation (a) to (d)] the color evolve which turn starch-
of the vapors if any. iodide paper blue and b) Br-
starch paper yellow (bromide) may
be present
c) Violet vapours of

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 6

 iodine evolve which turns
starch paper blue
c) I- (iodide)
d) No coloured fumes may be present

ii) Dense white fumes of d) Halides

ii) Also bring a rod dip in NH4OH solution NH4Cl absent
to the mouth of the test tube
Cl- (chloride)
may be present

Nitrate Test: Brown fumes which NO3- (nitrate)

become more intense on may be present
Take a small quantity of the given mixture adding Cu-filings
in a dry test tube is heated with conc.
H2SO4 and to it add Cu-filings or small
paper-ball of filter paper

Preparation of Solution:

I] Water soluble mixture: - Take about 50 mg of mixture, add water, shake well. If it
dissolves completely, mixture is completely water soluble. Use this as an original solution
and use it in the detection of basic radicals and also for wet tests of acidic radicals.

II] Partly water soluble mixture:- Take mixture, adds little water, shake well and filter
through filter paper. Take a drop of clear filtrate on a glass slide and one drop of water to its
side and evaporate gently. If appreciable residue left, mixture is partly soluble in water. Use
the above filtrate as Water solution (W.S) for the detection of Group I and Group VI and wet
tests of acid radicals

Preparation of Acid Solution: For partly soluble or completely insoluble mixture add dilute
HCl to 100 mg mixture, boil to get a clear solution. This is called as Acid Solution (A.S.) and
should be used for the analysis of Gr. II to Gr. V.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 7

 E) Wet Tests for Basic Radicals: Systematic analysis to separate metals (cations) into
Groups.

In the original solution (water solution) add few drops of dilute HCl.

Residue/ Centrifugate / Filtrate(if residue obtained) otherwise Acid Solution→ warm → pass H 2S. If ppt comes
centrifuge.
ppt
Residue/ppt Centrifugate / Filtrate
(white)
(Coloured) (If Group II present) boil well to expel H2S completely (tested with lead acetate
Metals of Group II paper) or take A.S. (Original solution) if Gr. II is absent
Group I present
present 2 drops on tile + K3[Fe(CN)6] → Blue ppt → Fe2+ present
Black ppt:
Cu2+, Pb2+, If Fe2+ present add 3-4 drops of Conc. HNO3 in the clear filtrate and boil to oxidize
Hg2+ Fe2+ to Fe3+ (solution will turn yellow)
Analyze
ppt Brown ppt: To the above solution add solid NH4C1 and 1:1 Ammonia (NH4OH) until the
according Bi3+ solution is alkaline (add a piece of red litmus to check alkalinity) → if ppt comes
to Table I centrifuge.
Yellow ppt:
Sn2+, As3+, Residue/ppt Centrifugate / Filtrate
Cd2+
Group IIIA To the above filtrate add solid NH4C1 and 1:1 Ammonia (NH4OH)
Orange ppt: present and pass H2S → if ppt comes centrifuge.
Sb3+
White ppt: Residue/ppt Centrifugate / Filtrate
Al3+
Group IIIB Transfer to a porcelain dish, boil well to expel H2S.
Analyze ppt by Brown ppt: present Add solid NH4C1, 1:1 Ammonia (NH4OH) and
Group Fe3+ then add, with stirring, (NH4)2CO3 (ammonium
Black ppt: carbonate) solution in slight excess. Filter
separation
Blue-Green Ni2+, Co2+
Table IIA and
ppt: Cr3+ White Centrifugate / Filtrate
Table IIB White ppt: Residue/ppt
Zn2+ To the filtrate add solid NH4C1,
Group IV 1:1 Ammonia (NH4OH) and then
Analyze ppt Pink ppt: present add Na2HPO4 (sodium
by Group Mn2+ hypophosphate also called as
separation Ca2+, Ba2+, sodium phosphate) solution.
Table IIIA Sr2+ Filter
Analyze ppt
White If no ppt till
by Group
Residue/ppt Group V and
separation Analyse ppt
only one cation
by Group Group V
Table IIIB is detected then
separation (Mg2+) present Group VI is
Table IV
present
Analyse ppt by
Group
separation
Table V

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 8

 DETECTION OF BASIC RADICALS:

Test Observation Inference

OS + dil HCl I
OS + dil HCl + H2S II
OS+ K3[Fe(CN)6] Fe2+
OS + NH4Cl(s)+ NH4OH till alkaline (Red IIIA
litmus-Blue)
OS + NH4Cl(s)+ NH4OH till alkaline (Red IIIB
litmus-Blue) + H2S
OS + NH4Cl(s)+ NH4OH till alkaline (Red IV
litmus-Blue)+NH4CO3
OS + NH4Cl(s)+ NH4OH till alkaline (Red V
litmus-Blue)+ Na2HPO4
VI
Separation table for Group I Cations Precipitate may contain PbCl2, AgCl, and Hg2Cl2.

Transfer the ppt of Group I metal halides in a test tube and add 5 drops of water, warm and
centrifuge immediately.

Residue Centrifugate

Add 5 drops 1:1 NH4OH, warm and centrifuge May contain PbCl2

Residue Centrifugate 2-drops of

Centrifugate + KI
Add aqua-regia Acidify with dil HNO3→ white ppt→ solution → yellow
Ag+ present ppt of PbI2 soluble
(3:1 conc HCl:conc
HNO3). Boil and add Above filtrate + 2-drops KI → yellow when hot reappears
SnCl2→ black ppt ppt when cold

Hg+ present and confirmed Ag+ present and confirmed Pb2+ present and
confirmed

Separation table for Group II Cations

The ppt. may consist of the sulphides of the Group IIA metals(HgS, PbS, Bi2S3, CuS, CdS)
and those of Group IIB (As2S3, Sb2S3, SnS).

Transfer the ppt of Group II metal sulphides in a porcelain dish and add 5-drops yellow
ammonium sulphide (NH4)2Sxand 2-drops NaOH. Warm and centrifuge.

Residue Centrifugate

Contains sulphides of Gr IIA; HgS, PbS, Bi2S3, CuS Contains thiosalts of IIB metals
and CdS. Use Table IIA for separation As, Sn and Sb. Use Table IIB for
separation

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 9

Separation table for Group IIA: Add 2-3 drops of conc. HNO3 to the above ppt, boil and
centrifuge.

Residue Centrifugate

Dissolve in aqua- Add few drops of dilute H2SO4, centrifuge

regia (3:1 conc
Residue Centrifugate
HCl:conc HNO3).
Boil and dilute with Contains PbSO4. Add 1:1 Ammonia till alkaline. Centrifuge.
water Boil with
ammonium Residue Centrifugate
Above solution + acetate, add acetic
White ppt dissolve If Blue contain Cu2+. Add dilute acetic acid
SnCl2→ white ppt acid and K2CrO4
in dilute HCl and (till acidic) + K4[Fe(CN)6]→ chocolate
turns grey → yellow ppt
add excess H2O brown ppt
Hg2+ present and Pb2+ present and
→ white ppt Cu2+ present and confirmed
confirmed confirmed
Bi3+ present and To the remainder of the filtrate, add KCN
confirmed solution dropwise until

colour discharged. Pass H2S → Yellow ppt

Cd2+ present and confirmed

Separation table for Group IIB: Transfer ppt in a porcelain dish and add 4-drops conc HCl,
warm and centrifuge.

Residue Centrifugate

If yellow contains As2S3. May contain Sb3+ or Sn2+. Neutralize with dil NH4OH till alkaline. Add solid
Boil with conc. HNO3 and oxalic acid, heat, pass H2S and centrifuge.
dilute with water. Divide
into two parts: Residue Centrifugate

a) One part + H2S → Orange coloured contains Sb2S3. Boil well to expel H2S, add Zn dust and dil.
yellow ppt Dissolve in few drops of conc. HCl, Sn is re-precipitated. Dissolve again in
HCl and divide into two parts: few drops of conc. HCl. Divide into two
b) Another part + few parts:
drops of ammonium a) One part dilute with
molybdate (NH4)2MoO4 water→white turbidity a) One part + few drops Hg solution → white
ppt of Hg2Cl2
→ yellow ppt b) Another part on tile +
Rhodamine B→Red to blue spot b) Another part + few drops dilute I2 solution
As3+ present and → Colour of I2
confirmed Sb3+ present and confirmed
disappeared

Sn2+ present and confirmed

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 10

Separation table for Group IIIA: The ppt contains Fe(OH)3, Al(OH)3 or Cr(OH)3. Transfer
the ppt in a porcelain dish and add 10-drops NaOH and 5-drops H2O2. Boil and centrifuge.

Residue Centrifugate: Divide into two parts:

Contains Fe(OH)3Dissolve If colourless contains NaAlO2-

the residue in minimum
amount hot dilute HCl and Filtrate acidify with dilute HCl and add NH4OH till
divide into two parts: alkaline. Heat to boiling → white gelatinous ppt

a) One part + few drops Filtrate + one drop of Alizarin + Expose to ammonia (or
ammonium add one drop of NH4OH).
thiocyanate→blood red Al3+ present and confirmed
colouration
b) If centrifugate is yellow Cr3+ may be present
b) Another part + few drops
(potassium ferrocyanide) Filtrate acidify with dilute CH3COOH (acetic acid)
K4[Fe(CN)6] and

→ Deep blue ppt add few drops of lead acetate solution→ yellow
ppt of
Fe3+ present and confirmed
PbCrO4

Cr3+ present and confirmed

Space left blank

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 11

Separation table for Group IIIB: The ppt may contain sulphides of Ni, Co, Mn or Zn.
Transfer ppt of Gr. IIIB into a test tube and add 10 drops of dilute HCl, shake well, allow it to
stand for a minute, centrifuge.

Residue: Black residue indicates Centrifugate: May contain MnCl2 or ZnCl2.

NiS or CoS. Above residue
dissolve in aqua-regia (3:1 conc Boil this solution to expel H2S. Add dropwise excess
HCl: conc HNO3). Boil, cool it of dilute NaOH. Centrifuge.
and add 6 drops of water. Then Residue: Centrifugate:
add solid NH4C1 and 1:1
Ammonia (NH4OH) till alkaline White and slow turned to May contain
(tested with litmus paper) and brown due to atmospheric Na2ZnO2. Acidify
divide this solution into two parts: oxidation. Dissolve this ppt in (tested with litmus)
few drops of conc. H2SO4 and this solution with
To one part add K3[Fe(CN)6]→ add excess PbO2. Boil well acetic acid and
red colour or red-brown ppt and dilute with water → divide into two
Co2+ present and confirmed purple colouration of parts:One part + 4-
supernatant on standing. drops of
Another part + excess of K3[Fe(CN)6] + 4-
dimethyl glyoxime (DMG)→ Mn2+ present and confirmed. drops diphenylamine
reddish pink or rosy red ppt in acetic acid
→green ppt
Ni2+ present and confirmed.
Another part, pass
H2S → white ppt

Zn2+ present and

confirmed

Separation table for Group IV:

The ppt may contain carbonates of Ba, Sr or Ca.

Transfer white ppt of Gr. IV into a test tube and dissolve in 8-10 drops of acetic acid, boil to
expel CO2. Clear solution is known as acetate solution. Divide this solution into three parts:

Test Observation Inference

Acetate solution + K2CrO4 (Potassium Yellow ppt Ba2+present and confirmed

chromate)

Acetate solution + (NH4)2SO4 White ppt Sr2+ present and confirmed

(Ammonium sulphate)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 12

Acetate solution + (NH4)2C2O4 White ppt Ca2+ present and confirmed
(Ammonium oxalate)

Separation table for Group V:

The ppt contains Mg2+.Transfer the white ppt of Gr. V in a test tube and dissolve it in few
drops of dilute H2SO4.

Test Observation Inference

Two drops of above solution + 1 ml Reddish brown ppt Mg2+ present and confirmed
Hypoiodite reagent (freshly prepared) or colour of
Mg(OI)2

[Hypoiodite reagent: 5-drops NaOH

+5-drops KI solution + Iodine
solution dropwise till the solution
becomes just yellow]

Separation table for Group VI: (Use water solution only for NH4+, Na+, K+)

Test Observation Inference

Two drops of water solution + few drops of Evolution of NH3 gas NH4+present
NaOH and boil which turns moist
turmeric paper red
C.T. for NH4+

Two drops of Nessler’s reagent + 2 drops of

water solution [Nessler’s reagent: 2-drops Brown ppt or NH4+ present
HgCl2 + dropwise KI solution till scarlet red colouration and confirmed
ppt first formed then dissolve and add equal
amount NaOH]

Test for Na+ and K+:

Two drops of water solution + two drops of Yellow ppt K+ present

sodium cobaltinitrite, Na3[Co(NO2)6]

C.T. for K+

Two drops of water solution + few drops of

picric acid → rub the inner side of the test tube
with a glass rod Yellow ppt K+ present and
confirmed

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 13

Detection of Acidic Radicals (Anions)(Use water solution only)

Test Observation Inference

2-Drops of water solution a) Curdy white ppt soluble in Cl- (chloride) present
+ AgNO3 NH4OH but insoluble in dilute
HNO3

b) Pale yellow ppt soluble in Br- (bromide) present

ammonia

c) Yellow ppt insoluble in dilute I- (iodide) present

HNO3 and also in NH4OH

d) White ppt soluble in dilute

HNO3with effervescence of CO2 CO32- (carbonate) present

e) No ppt Cl-, Br-, I-,CO32- absent

2-Drops of water solution White ppt insoluble in dilute SO42- (sulphate) present
+ Ba(NO3)2 HNO3

Water solution + dilute Brown coloured solution NO2- (nitrite) present

acetic acid + FeSO4
solution

Water solution + conc. Brown ring at the junction of the NO3- (nitrate) present
H2SO4 (cool thoroughly) + two layers
FeSO4 solution dropwise
from the side of test tube

Confirmatory tests for Anions

Test Observation Inference

For Chloride (Cl-): Water solution CCl4 + Colourless CCl4 layer at Cl- present and
chlorine water → shake well the bottom confirmed

For Bromide (Br-): Water solution CCl4 + Brown colour ofCCl4 Br- present and
chlorine water → shake well layer at the bottom confirmed

For Iodide (I-): Water solution + CCl4 + Pink-violet colour to the I- (iodide) present
chlorine water → shake well CCl4 layer at the bottom and confirmed

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 14

For Sulphate (SO42-): 2-drops of water White ppt of PbSO4 SO42- (sulphate)
solution + two drops of lead-acetate soluble in warm
solution ammonium acetate present and
confirmed

For Sulphide (S2-): 2-drops of sodium Violet colour

carbonate extract + two drops of NaOH S2- (sulphide)
solution and then add sodium nitroprusside present and
solution confirmed

For Nitrite (NO2-): Water solution (or Red colour NO2-(nitrite) present
mixture) + 1 ml dilute acetic acid + pinch and confirmed
of thiourea. Shake well and add few drops
of aqueous FeCl3 solution.
Brown ring at the
-)
For Nitrate (NO3 - Brown Ring Test: junction of the two NO3- (nitrate)
Water solution (or Na2CO3 extract) + conc. layers formed due to present and
H2SO4 (cool under water), filter if FeSO4NO complex confirmed
necessary + few drops of saturated solution
of FeSO4 from the side of the test tube
Effervescence of CO2 CO32- (carbonate)
For Carbonate (CO32-) –10 mg mixture +
turns lime water milky present and
2-drops dilute HCl and then heat
confirmed

Result Table:

Basic Radicals (Cations) Acidic Radicals (Anions)

1. 1.

2. 2.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 15

 INORGANIC QUALITATIVE ANALYSIS
A) Preliminary Tests
Test Observation Inference
Colour
Nature

B) Dry Tests for Basic Radicals:

Test Observation Inference
Heating in a dry test tube

Heating in a charcoal cavity:

Cobalt nitrate test:

OR
Sodium carbonate test:

Action of NaOH: NH4+

C) Dry tests for Acidic Radicals:
Test Observation Inference
2-
Action of dil. HCl: a) CO3
c) S2-
d) NO2-
Action of MnO2 + Conc. H2SO4: a) Cl-
b) Br-
c) I-
Nitrate Test: NO3-

D) Preparation of Solution:

Test Observation Inference

Mixture+ Water
Glass slide test: Partly water soluble
Water Solution (Mixture+ Water+ Filter) used for Group I, Group VI, Wet Tests of Anions
Acid Solution (Mixture+ dil. HCl) used for Group II to Group V

E) DETECTION OF BASIC RADICALS:

Test Observation Inference

OS + dil HCl I

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 16

OS + dil HCl + H2S II

OS+ K3[Fe(CN)6] Fe2+

OS + NH4Cl(s)+ NH4OH till alkaline IIIA

(Red litmus-Blue)

OS + NH4Cl(s)+ NH4OH till alkaline IIIB

(Red litmus-Blue) + H2S

OS + NH4Cl(s)+ NH4OH till alkaline IV

(Red litmus-Blue)+NH4CO3

OS + NH4Cl(s)+ NH4OH till alkaline V

(Red litmus-Blue)+ Na2HPO4

VI

GROUP ANALYSIS:
Test Observation Inference
Separation Table of

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 17

Separation Table of

F) Detection of Acidic Radicals or Wet tests for Anions

Test Observation Inference
WS+ AgNO3 Cl , Br , I-
- -

WS+ Ba(NO3)2 SO42-

WS+ dilute acetic acid + FeSO4 NO2-

Water solution + conc. H2SO4 (cool ) + NO3-

FeSO4
G) Confirmatory tests for Anions
Test Observation Inference
CT for__________

CT for___________

Result Table:
Basic Radicals Acidic Radicals
+ -
1. NH4 (Ammonium) 1. Cl (Chloride)
2. Ba2+ (Barium) 2. CO32- (Carbonate)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 18

 IV: Inorganic Qualitative Analysis of Known Binary mixture (MgCO3 + NH4Br)

E) Preliminary Tests

Colour White / Colourless Compound of Al, Zn, Ca, Ba, Sr, Mg, K and ammonium
salts
Nature Mixture of crystalline Soluble salts of Na, K and ammonium or salts of Cl-, Br-, I-,
and amorphous SO42-, NO2-, NO3-
or
Insoluble salts of CO32-, S2-, O2-, PO43-, BO33- may be present

F) Dry Tests for Basic Radicals: (Cations)

Experiment Observation Inference

Heating in a dry test tube: Place a small i) Change in colour: Al, Ba, Ca, Sr, Mg, Na,
quantity of the given mixture in a dry test White to white when hot, K, NH4+ salts
tube. Heat it cautiously and strongly and white when cold
observe any changes
ii) White sublimate NH4+ salts, mainly
formed halides

iii) Decrepitation or
Crackling sound Crystalline salts like
KBr, NH4Cl, Pb(NO3)2,
Ba(NO3)2
iv) Water vapour
condenses on cooler part Hygroscopic salts or salts
of test tube containing water of
crystallization

Charcoal Block Reduction (Heating in a White infusible residue Ca2+, Sr2+, Ba2+, Al3+,
charcoal cavity): Make a small fresh was obtained (Therefore Mg2+, Zn2+ salts may be
cavity (with the help of tongs) on a piece cobalt nitrate test was present
of charcoal block, take a little substance in performed)
the groove, moisten with a drop of water
and heat in a reducing flame (yellow
flame) using blow pipe
Cobalt nitrate test: Moisten above Residue turned pink Mg salts may be present
residue with 2 drops cobalt nitrate
solution and heat in oxidizing flame (blue)
with blow pipe

Action of NaOH: Strong smell of ammonia NH4+ (ammonium) salts

Take a small amount of the given mixture turning moist turmeric may be present
in a dry test tube and add few drops of paper red was observed
NaOH solution. Heat and hold moist
turmeric paper at the mouth of the test
tube without touching inner wall

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 19

 G) Dry tests for Acidic Radicals: (Anions)

Experiment Observation Inference

Action of dil HCl: Take small amount of Brisk effervescence of CO32- (carbonate) may
the given mixture in a dry test tube. Add 3- CO2 gas (without heating) be present
4 drops of dil HCl and observe
[Cover the test tube with another blank test The colourless gas turned
tube, hold it in 45ο angle and then heat freshly prepared lime
gently. As the vapours enter into the blank water milky
test tube, cover the mouth of the blank test
tube with your thumb. Slightly open the
test tube, add fresh lime water and shake it
vigorously]

Action of MnO2 + conc H2SO4: Small Reddish-brown vapours Br- (bromide) may be
amount of the given mixture in a dry test evolved that turned moist present
tube heated with small amount of MnO2 starch-iodide paper blue
and 1 ml conc. H2SO4. [Observe the and moist starch paper
colour of vapours and further test with yellow
suitable test papers]

Nitrate Test: No intense brown fumes NO3- (nitrate) may be

Small amount of the given mixture in a dry obtained absent
test tube is heated with conc H2SO4 and to
it add Cu-filings /small piece (rolled) of
filter paper

D] Preparation of Solution: Appreciable amount of Mixture is partly

Take small amount of the mixture, add residue left soluble in water
little water, shake well and filter through
filter paper. Take a drop of clear filtrate on
a glass slide and one drop of water to its
side and evaporate gently
Use the above filtrate as Water solution
(W.S.) for the detection of Group I and
Group VI and wet tests for acid radicals

Preparation of Acid Solution: Add dilute

HCl to small amount of fresh mixture; boil
to get a clear (transparent) solution.
This is called Acid Solution (A.S.) and use
it for the analysis of Group. II to Group V.

Detection of Basic Radicals:

W.S. + dil HCl No precipitate obtained Group I (Ag+, Hg22+)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 20

 absent
A.S. + pass H2S No precipitate obtained Group II (Hg2+, Pb2+,
Cu2+, Bi3+, Sn2+, Sn4+,
As3+, Cd2+, Sb3+)
absent

2 drops A.S. on a tile + 2 drops No blue colour obtained

K3[Fe(CN)6] Fe2+ absent

To the acid solution add solid NH4C1 and No precipitate obtained

1:1 Ammonia (NH4OH) until the solution Group IIIA (Fe3+, Al3+,
is alkaline (add a piece of red litmus into Cr3+) absent
the solution till red litmus turns blue
completely)
Part of the above solution + pass H2S No precipitate obtained Group IIIB (Ni2+, Co2+,
Mn2+, Zn2+) absent
In the other part of the alkaline solution No precipitate obtained Group IV (Ca2+, Ba2+,
add (NH4)2CO3, centrifuge Sr2+) absent
Original A.S. + NH4Cl + NH4OH till White precipitate Group V (Mg2+) is
alkaline(add a piece of red litmus to check obtained present
alkalinity) + Na2HPO4

Since only one group was detected (Group V), therefore Group VI must be present

Analysis of Group V:
Transfer precipitate of Group V in test tube + 2 drops of dilute H2SO4 and warm slightly. Use the
clear solution for confirmatory test of Mg2+ ion
Two drops of solution + 4 drops of Reddish brown colour/ Mg2+ present and
freshly prepared Hypoiodite solution precipitate obtained confirmed

[Hypoiodite reagent: 5-drops NaOH +5-

drops KI solution + Iodine solution
dropwise till the solution becomes just
yellow]

Separation table for Group VI: (Use water solution only for NH4+, Na+, K+)

Test Observation Inference

+
Two drops of water solution + few drops of Evolution of NH3 gas NH4 present
NaOH and boil which turns moist
turmeric paper red
Confirmatory Test for NH4+
Two drops of freshly prepared Nessler’s Brown precipitate or NH4+ present and
reagent + 2 drops of water solution colouration obtained confirmed
[Nessler’s reagent:2-drops HgCl2 +
dropwise KI solution till scarlet red
precipitate first formed then dissolve and
add equal amount NaOH]

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 21

 G) Detection of Acidic Radicals (Anions) (Use W.S only)

Test Observation Inference

2-Drops of water solution + AgNO3 Pale yellow precipitate Br- (bromide) present
soluble in ammonia

2-Drops of water solution + Ba(NO3)2 No white precipitate SO42- (sulphate) absent

obtained
Water solution + dilute acetic acid + FeSO4 No brown coloured NO2- (nitrite) absent
solution solution

Water solution + conc. H2SO4 (cool No brown ring seen NO3- (nitrate) absent
thoroughly) + FeSO4 solution dropwise
from the side of test tube

H) Confirmatory tests for Anions

Test Observation Inference

For Bromide (Br-):
W.S. + CCl4 + Brown colour of CCl4 Br- present and
chlorine water → shake well layer at the bottom confirmed

For Carbonate (CO32-) –10 mg mixture + Effervescence of CO2 CO32- (carbonate)

2-drops dilute HCl and then heat turns lime water milky present and
confirmed

Result Table:
Basic Radicals (Cations) Acidic Radicals (Anions)
1. Mg2+ (Magnesium) 1. Br – (Bromide)
2. NH4+ (Ammonium) 2. CO32- (Carbonate)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 22

 V: GENERAL SCHEME OF ORGANIC QUALITATIVE ANALYSIS

AIM:To determine the type and perform functional group test for the given organic
compound.

(1) NATURE: Solid or Liquid

Test Observation Inference

Volatility Test for Liquid

Take small amount of substance in Bubbles seen a)Substance is volatile

small test tube and add few
porcelain piece and heat in water No bubbles seen b)Substance is non-volatile
bath.

Conclusion:The given organic substance is_____________(Volatile/ non-volatile)

(2)TYPE DETERMINATION

Test Observation Inference

Solubility Test

Take small amount of substance in a Clear solution not obtained Substance is insoluble in
test tube and add 1ml water. Shake water
well

(A)Type determination for water soluble (miscible) substances

Test Observation Inference

(a)Litmus Test Litmus paper turns red Acid or Phenol is present

Dissolve few crystals of the Base is present

substance in water and add 2 drops
of this solution on wet litmus paper. Red Litmus paper turn blue

No change on either litmus Neutral substance is present

paper
(b) Distinguishing Test for a) Acid is present
Acid/Phenol Effervescence of CO2
b) Phenol may present
Take small amount of substance and No effervescence of CO2
add 10 drops 10% NaHCO3

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 23

(c) Phenol test

Take small amount of substance and Blue/green/violetcolouration Phenol is present

add 2 drops FeCl3(i.e.Fe2Cl6)
obtained

(B) Type determination for water insoluble (immiscible) substances

Test Observation Inference

Take small amount of substance and

add 1ml 10% NaHCO3..Shake well.
Substance dissolves with Acid is present
effervescence of CO2 and

re-precipitated by conc. HCl

*(HCl should be added till

solution becomes acidic)

Take small amount of substance and

add 1 ml 2 N NaOH..Shake well.
Substance dissolves and Phenol is present

re-precipitated with conc.HCl

Take small amount of substance and Substance dissolves and Base is present.
add 1 ml 1:1 HCl and shake
well.(1:1 HCl= equal amount of re-precipitated with 2NNaOH
water and conc. HCl) *(NaOH should be added till
solution becomes alkaine)

*If all the above test are negative then the substance is neutral

Conclusion: - The type of given organic substance is_____________(Acid

/Phenol/Base/Neutral)

(3) DETECTION OF SATURATION /UNSATURATION

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 24

Test Observation Inference

(a)Br2 in CCl4 test a) Brown colour disappears a) Unsaturated substance

present
10g substance+2drops of Br2in CCl4
solution. Shake well
b) No decolourization
b) Saturated substance present

(b)Baeyer’s (KMnO4) Test

10mg substance + 4 drops of dilute a) Unsaturated substance

KMnO4+ shake well. a) Pink colour of KMnO4 present or easily oxidisable
disappears *functional group present
(dil KMnO4=1 drop diluted to Half
test tube)
b)No decolourization b)Saturated substance present

*Aldehyde /Ketones undergo oxidation reaction with KMnO4 to give acids.Therefore shows
decolourization.

4) DETECTION OF AROMATIC / ALIPHATIC NATURE

Test Observation Inferences

Place little substance on Copper a) Sooty flame seen a) Aromatic substance present
gauge and heat it on flame

b) Non sooty flame seen b)Aliphatic substance present

Conclusion: Given organic compound is ________________________________

i) Saturated /Unsaturated ii) Aromatic / Aliphatic

5) PHYSICAL CONSTANT OF THE SUBSTANCE………………….oC

6) ELEMENTS PRESENT:

Conclusion: Given organic substance contains C, H, (O) and ______elements

7) FUNCTIONAL GROUP TEST

(A) TYPE ACID

(i) Functional group test for acids with C, H, (O) elements
Test Observation Inference

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 25

(a)NaHCO3 Test:

Take small amount of Effervescence of CO2 Carboxylic group (-COOH)

substance and add 1ml 10% obtained and substance group obtained
NaHCO3. Shake well. dissolves and then
reprecipitated by conc. HCl

(b)FeCl3 test:

Take small amount of Blue / violet /green colour Phenolic –OH (Ar-OH)group
substance and add 1mlethyl obtained is present
alcohol .Shake well and add
2-3 drops of neutral FeCl3
(Fe2Cl6)

(ii) Functional group test for acids with C, H, (O) N elements

Test Observation Inference

Perform above test a,b along with test c given below for nitro acids

(c)Neutral reduction test:

Take small amount of Black or gray precipitate Nitro group (-NO2 group) is
substance and add 1ml ethyl obtained present
alcohol +6 drops of CaCl2
+pinch of Zn dust .Boil Well
and Filter it hot into Tollen’s
reagent through filter paper.

Tollen’s reagent: 5drops AgNO3 +2 drops NaOH(gray precipitate obtained)+ NH4OH till
precipitate dissolves

(B) TYPE PHENOL

(i) Functional group test for phenols with C, H, (O) elements
Test Observation Inference

(b)FeCl3 test:

Take small amount of Blue / violet /green colour Phenolic –OH (Ar-OH)group
substance and add 1mlethyl obtained is present
alcohol .Shake well and add
2-3 drops of neutral
FeCl3(Fe2Cl6)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 26

(ii) Functional group test for phenols with C, H,(O) N elements

Test Observation Inference

Perform above test (a), (b) along with test (c) given below for nitro acids

(c)Neutral reduction test:

Take small amount of Black or gray precipitate Nitro group (-NO2 group) is
substance and add 1ml ethyl obtained present
alcohol + 6 drops of CaCl2 +
pinch of Zn dust .Boil Well
and Filter it hot into Tollen’s
reagent through filter paper.

Tollen’s reagent: 5drops AgNO3 +2 drops NaOH + NH4OH till ppt. dissolves

(C) TYPE BASES

(i) Functional group test for primary/ secondary/ tertiary amines with C, H, (O) Nelements
Test Observation Inference

(a) Take small amount of a) If yellow solid separates a) Secondary amine (Ar2NH)
substance and add 10 drops present
of conc.HCl +boil and cool
in ice water + 3 drops of b) Tertiary amine (Ar3N)
b) If red colour obtained and present
NaNO2 solution on adding NaOH green solid
is obtained

If above observations (i) and c)Orange dyestuff obtained c)Primary amine(ArNH2)

(ii) are absent then add above present
solution dropwise into 10
drops of ice cold solution of
β-naphthol in NaOH (taken
into another test tube)

If given substance is yellow in colour then…..

Perform above test (a) along with test (b) given below for nitroamines

(c)Neutral reduction test:

Take small amount of Black or gray precipitate Nitro group (-NO2 group) is

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 27

substance and add 1ml ethyl obtained present
alcohol + 6 drops of CaCl2 +
pinch of Zn dust .Boil Well
and Filter it hot into Tollen’s
reagent through filter paper.

Tollen’s reagent: 5drops AgNO3 +2 drops NaOH + NH4OH till ppt. dissolves

(D) TYPE NEUTRAL

(i) Functional group test for neutral substances with C, H, (O) elements
Test Observation Inference

FOR ALDEHYDES: Liquids

(a)Caro’s Test

Take 4 drops of schiff’s Pink colour slowly developed Aromatic aldehyde(Ar-CHO)

reagent* in a test tube, add 4 in the organic layer(at the is present
drops of the substance. Shake bottom)
well and keep it for sometime

*Schiff’s reagent used should be colourless

FOR KETONES: Liquids /solids

(b)Nitroprusside test

10mg substance + 4drops of Red colour solution obtained Ketone (R-CO-R’) is present
sodium nitroprusside +4drops
of NaOH.Shake well

FOR ESTERS: Water insoluble Liquids

(c)Phenolphthalein test

5drops of substance + 1ml Pink colour slowly Ester group(R-COOR’) is

water +2 drops of disappears present
phenolphthalein indicator+
very dilute NaOH(0.1 N) (Ester on hydrolysis gives an
drop by drop till faint pink acid. phenolphthalein
colour persists then heat the indicator is colourless in
solution acidic medium)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 28

(ii) Functional group test for neutral substances with C, H, (O), N elements

Test Observation Inference

FOR AMIDES:Water soluble solids

(a)10mg substance + 10 Evolution of ammonia gas Amide group (R-CONH2)is

drops of NaOH solution and turns moist turmeric paper present
boil well red

FOR ANILIDES

Take small amount of Orange dyestuff obtained Anilide group (Ar-NHCOR)

substance and add 10 drops is present
of conc.HCl + boil and cool
in ice water + 3 drops of
NaNO2 solution. Add above
solution dropwise into 10
drops of ice cold solution of
β-naphthol in NaOH(taken
into another test tube)

FOR NITRO-HYDROCARBONS: Water insoluble yellow solid/liquids

(c)Neutral reduction
test:Take small amount of
substance and add 1ml ethyl Black or gray precipitate Nitro group (-NO2 group) is
alcohol + 6 drops of CaCl2 + obtained present
pinch of Zn dust .Boil Well
and Filter it hot into Tollen’s
reagent through filter paper.

Tollen’s reagent: 5drops AgNO3 +2 drops NaOH + NH4OH till ppt. dissolves

(ii) Functional group test for neutral substances with C, H, (O) N and S elements

Test Observation Inference

Take small amount of Thioamide group

substance in a test tube and
fuse it. Add 1 ml water and 3 Blood red colouration
drops of FeCl3 solution.
is present

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 29

(iii) Functional group test for neutral substances with C,H,(O) X(halogen) elements

Test Observation Inference

FOR HALOHYDROCARBONS : Water insoluble LIQUIDS

(a) Take small amount of a) White precipitate settles at a) Side chain/Aliphatic halide
substance and add 5 drops the bottom (R-X) is present
ofNaOH,boil well and
cool,acidify with
dil.HNO3+2drops of AgNO3 b)No white precipitate b) Aromatic halide (Ar-X) is
solution Shake well formed present

(ii) Functional group test for neutral substances with C, H elements

Test Observation Inference

FOR HYDROCARBONS : No Functional group

If all above tests for neutrals - Hydrocarbon(-CH) is present

are absent

(8) RESULT TABLE

Nature Type Aliphatic/ Saturated/ M.P/B.P Name of the General

functional formula
(solid/Liquid) aromatic Unsaturated oC group

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 30

 ORGANIC QUALITATIVE ANALYSIS
(1) NATURE: Solid or Liquid

Test Observation Inference

Volatility Test for Liquids

Conclusion: (Volatile/ Non-volatile)

(2) TYPE DETERMINATION

Test Observation Inference

Solubility Test Soluble/ Insoluble
Miscible/Immiscible
(A)Type determination for water soluble (miscible) substances

Test Observation Inference

(a) Litmus Test

(b) Test for

Acid/Phenol

(c) Phenol test

(B) Type determination for water insoluble (immiscible) substances

Test Observation Inference

Substance + 10% NaHCO3. Acid
re-precipitated by conc. HCl
Substance + 1 ml 2 N NaOH Phenol
re-precipitated with conc. HCl..
Substance + 1 ml 1:1 HCl Base
re-precipitated with 2N NaOH
*If all the above test are negative then the substance is neutral
Conclusion: - The type of given organic substance is_____________ (Acid
/Phenol/Base/Neutral)

(3) TEST FOR SATURATION/ UNSATURATION

Test Observation Inference

(a)Br2 in CCl4

(b)Baeyer’s (KMnO4)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 31

4) TEST OF AROMATIC / ALIPHATIC NATURE

Test Observation Inferences

Substance on Copper Gauze+
heat

Conclusion: Given organic compound is ________

i) Saturated /Unsaturated ii) Aromatic / Aliphatic

5) PHYSICAL CONSTANT _________oC

6) ELEMENTS PRESENT:

7) FUNCTIONAL GROUP TEST:

Test Observation Inference

RESULT TABLE

Nature Type Saturated/ Aliphatic/ Physical Functional General

Unsaturated Aromatic Constant group formula

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 32

VII: DETERMINATION OF PHYSICAL CONSTANT OF ORGANIC COMPOUND

MELTING POINT

To Determine Melting Point of the given organic compound

Apparatus: Thieles tube, Thermometer 360oC, Thread, glass capillary tube.

Procedure:

1. Seal one end of the capillary tube by heating on the flame.

2. Place the substance in the capillary tube.

3. Tie the capillary with a thread to the thermometer.

4. Place this set up in Thieles tube and heat the Thieles tube using a burner

gently in clockwise manner.

5. Stop heating when half of the substance is melted.

6. Allow the substance to melt completely and record this temperature as

its melting point.

7.

Result Table

Melting point of the given substance: ……….. oC

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 33

BOILING POINT:

Aim: To Determine Boiling Point of the given organic compound

Apparatus: Thieles tube, Thermometer 360oC, Boiling point capillary, Sodium fusion tube
Thread.

Procedure:

1. Seal one end of the capillary tube by heating on the flame.

2. Take 4 -5 drops of the liquid organic compound in the sodium fusion tube.

3. Place the capillary with sealed end on the top and open end inside the sodium fusion
tube with liquid.

4. Now, tie this sodium fusion tube with a thread to the thermometer.

5. Place this set up in Thieles tube in line with the upper side arm and heat the Thieles
tube using a burner gently.

6. Stop heating when there is continuous stream of bubbles inside the sodium fusion
tube.

7. When the bubbles disappear and the liquid rises in the capillary, record this
temperature as the boiling point of the given liquid.

Result:

Boiling point of the given substance: ……….. oC

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 34

VIII: ORGANIC QUALITATIVE ANALYSIS OF KNOWN COMPOUND

AIM: To determine the type and perform functional group tests for the given known
organic compound (Benzoic acid)

(1) TYPE DETERMINATION

Test Observation Inference

(a)Solubility Test
Take small amount of substance in Clear solution not obtained Substance is insoluble in
a test tube and add 1ml water. water
Shake well

Type determination for water insoluble substances

Test Observation Inference

Take small quantity of substance in Substance dissolved with Acid is present

a test tube and add10 drops 10% effervescence of CO2 and
NaHCO3.Shake well. reprecipitated with conc. HCl

Conclusion: - The type of given organic substance is Acid

(2) DETECTION OF SATURATION /UNSATURATION

Test Observation Inference

(a)Br2 in CCl4 test Saturated substance is

Take small quantity of substance in No decolourization present
a test tube and add2drops of Br2
Water.Shake well

(b)Baeyer’s (KMnO4) Test No decolourization Saturated substance/

Take small quantity of substance in Not easily oxidisable is
a test tube and4 drops of dilute present
KMnO4+ shake well and compare
the colour with original dilute
KMnO4 as Reference
(dil KMnO4=1 drop KMnO4diluted
with water to Half test tube)

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 35

3) DETECTION OF AROMATIC / ALIPHATIC NATURE:

Test Observation Inferences

Heat a little substance on Sooty Flame seen Aromatic Compound is
Copper Gauge Present

Conclusion: Given organic compound is Aromatic

4)FUNCTIONAL GROUP TEST FOR ACIDS WITH C, H (O) ELEMENTS

Test Observation Inference

(a)NaHCO3 Test:
Take small quantity of Effervescence of CO2 -COOH group is present
substance in a test tube and obtained and substance
add dropwise NaHCO3 dissolved and then
reprecipitated by conc. HCl

(b)FeCl3 test:
Take small quantity of Blue / violet /green colour not Phenolic –OH (Ar-OH)group
substance in a test tubeand obtained is absent
add 1ml of ethyl alcohol.
Shake well and add 2-3 drops
of neutral FeCl3 (Fe2Cl6)

RESULT TABLE

Nature Type Aliphatic/ Saturated/ M.P/B.P Name of the General

(solid/Liq) Aromatic Unsaturated oC functional formula
group
Solid Acid Aromatic Saturation 122 carboxylic Ar-COOH

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 36

IX: PURIFICATION BY Crystallisation:

Crystallisation is one of the easiest & common method of purification of organic compounds.
It is the method in which impure solid organic compound is converted in to the pure
crystalline form using suitable solvent.

Recrystallisation is also known as fractional crystallisation, is a procedure for purifying an an

impure compound in a solvent. The method of purification is based on the principle that the
solubility of the solid increases with increased temperature & vice versa.

Important thing in crystallisation is selection of a solvent. Solvent must be inert with the
compound to be crystallised. Compound should be insoluble in the solvent at the room
temperature & soluble at the high temperature. Water is the best solvent but we may require
alcohol –water or alcohol depending on the solubility of the compound. Another very
important part is saturated solution of the compound is to be prepared in that particular
solvent (saturated means no more solid dissolves in the solvent). And the clear solution
obtained after crystallisation should not be cooled in ice or under cold water to get the
crystals. Solution should come to room temperature naturally. In this method insoluble
impurities & soluble less concentrated impurities are removed.

Important steps: Selection of a solvent. Preparing saturated solution by heating on a wire

gauge or small blue flame or heating in a water bath (For inflammable organic solvents like
alcohol) electrically heated water bath is more preferable. The container may be small or big
test tube or small beaker or a conical flask depending on the size of the compound. Filtering
this boiling solution through cotton plug, filter paper cone or fluted filter paper cone. Filtering
the crystals obtained. Drying the crystals on a watch glass or on a clean paper boat.

Diagram:

1. Preparing saturated solution in a test tube

2: Fluted filter paper: Advantage of fluted filter paper is it increases surface area and filtration
is fast.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 37

 Figure: Filtration unit for solution

By crystallisation we get pure crystalline solid with improved qualities like uniform crystal
size, color , appearance, physical constants. Impurities reduces physical constant like melting
point. But by crystallisation this is improved. It always advisable to carry out experiments &
reactions on pure sample to obtain good results.

Fig: Filtration unit for crystals

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 38

 3.B) Purification of the given organic compound by recrystallization. (Acetanilide)

Principle: Recrystallisation is a technique in which solid impure crystals which have to be

crystallised are dissolved in a suitable solvent to form its saturated solution which on filtering
& cooling recrystallises to give solid crystals improved in qualities like appearance &
physical constants.

Chemicals: Given organic compound 250 mg, suitable solvent.

Apparatus: Clean dry test tubes, glass rod, cotton, filter paper, watch glass, porcelain chip
hot plate or water bath, tripod stand, test tube holder, wire gauge, burner, m.p apparatus etc.

Procedure:

1)Clamp the thiel’s Tube

2)Take M.p of the compound using thiel’s tube & thermometer 360 oC

3)Prepare saturated solution of this powder in water by heating it on a burner small blue
flame (about 2-3 ml water is sufficient).Put a small porcelain chip in the solution while
heating.

4)Filter it in a clean test tube.

5)Allow the filtrate to cool it to room temperature.

6)Allow complete recrystallisation. Wait for some time.

7)Filter the crystals obtained at filter pump.

8)Dry the crystal under IR lamp or in air at room temperature.

9)Take the weight of the crystals W2.

10) Take m.p of the compound after crystallisation.

Result Table,
i. Solvent used for the crystallisation =
o
ii. M.P before crystallisation = C
o
iii. M.P after crystallisation = C

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 39

 X: PURIFICATION BY SUBLIMATION

Aim: 1. To Purify the given organic compound by sublimation method.

2. To report the Melting Point of the given sample before and after sublimation.

Principle:

Sublimation is a process in which the solid substance on heating directly passes into vapour
state without first passing into the liquid state. The vapours, on cooling of the funnel give
pure cystals.

Apparatus: Evaporating dish, Glass funnel, Sand bath, Tripod stand, Filter paper, Cotton,
Pencil

Chemicals: Given Organic compound (solid)

Diagram:

Procedure:

1. Take the given substance in a dry evaporating dish and keep on a sand bath supported by a
tripod stand.

2. Cover the dish with a filter paper and perforate the filter paper with pencil.

3. Place an inverted funnel over the perforated filter paper as shown in the figure.

4. Place a small piece of cotton at the mouth of the funnel stem.

5. Heat the evaporating dish on low blue flame.

6. The vapours on cooling will be collected as pure crystals on the inner side of side of the
funnel.

7. The crystals are also collected on the filter paper.

8. Scrap the crystals from the funnel and filter paper with the help of glass rod.

9. Find out the melting point of the substance before and after crystallization.

Precautions:

 Trim the edges of the perforated filter paper before heating.

 Heat the evaporating dish containing the substance with low oxidizing flame/ blue
flame
Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 40
Result table

1 Method of Purification of compound = Sublimation

2 M elting point of the substance before purification =____oC
3 Melting Point of the substance after purification =____oC

XI: Thin Layer Chromatography (TLC)

Title: Separation of a mixture of o-and p-nitroaniline by thin layer chromatography (TLC).

Aim: To find the Rf values for o-and p-nitroaniline by thin layer chromatography (TLC).

TLC is a type of planar chromatography.

 It is routinely used by researchers in the field of phyto-chemicals, biochemistry and so

forth, to identify the components in a compound mixture, like alkaloids,
phospholipids, and amino acids.
 It is a semi quantitative method consisting of analysis for purity.
 High performance thin layer chromatography (HPTLC) is the more sophisticated or
more precise quantitative version.

Principle:
Similar to other chromatographic methods, thin layer chromatography is also based on the
principle of separation.

1. The separation depends on the relative affinity of compounds towards stationary and
the mobile phase. (Adsorption and partition)
2. The compounds under the influence of the mobile phase (driven by capillary action)
travel over the surface of the stationary phase. During this movement, the compounds
with higher affinity to stationary phase travel slowly (generally polar compound/s)
while the others travel faster (generally non-polar compound/s). Thus, separation of
components in the mixture is achieved.
3. Once separation occurs, the individual components are visualized as spots at a
respective level of travel on the plate. Their nature or characters are identified by
means of suitable detection techniques.
4. There are various techniques based on this principle like Preparative TLC,
Fluorescent TLC, Glass plate TLC.

Advantages:
 It is a simple process with a short development time.
 It helps with the visualization of separated compound spots easily.
 The method helps to identify the individual compounds.
 It helps in isolating of most of the compounds.
 The separation process is faster and the selectivity for compounds is higher (even
small differences in chemistry is enough for clear separation).
 The purity standards of the given sample can be assessed easily.
 It is a cheaper chromatographic technique.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 41

Applications

1. To check the purity of given samples.

2. Identification of compounds like acids, alcohols, phenols.
3. To evaluate the reaction process by assessment of intermediates, reaction course and
so forth.
4. To purify samples.
5. To keep a check on the performance of other separation processes.
6. To check the authenticity of compound.

System Components

TLC system components consists of

1. TLC plates. The TLC plates are prepared by using simple glass plates immersed into
the jar containing TLC silica, binder and the volatile solvent like chloroform (Silica
Gel Slurry).
2. TLC chamber. This is used for the development of TLC plate. The chamber
maintains a uniform environment inside for proper development of spots. It also
prevents the evaporation of solvents, and keeps the process dust free.
3. Mobile phase. This comprises of a solvent or solvent mixture The mobile phase used
should be particulate-free and of the highest purity for proper development of TLC
spots. The solvents recommended are chemically inert with the sample, a stationary
phase.
4. A filter paper. This is moistened in the mobile phase, to be placed inside the
chamber. This helps develop a uniform rise in a mobile phase over the length of the
stationary phase.

Procedure:
The stationary phase is applied onto the plate uniformly and then allowed to dry and stabilize.
These days, however, ready-made plates are preferred e.g. Merck F-256 TLC plates.

1. With a pencil, a thin mark is made at the side of the plate to apply the sample spots.
2. Then, samples solutions are applied on TLC baseline in equal distances. Generally,
for comparison of two compounds say compound A and compound B. The samples
spots applied on the plates are Compound A (at left side), Compound A and B (at the
center) and Compound B (at the right side). However, the Rf values are always
recorded for individual compounds A and B.
3. The mobile phase is poured into the TLC chamber to a leveled few millimeters above
the chamber bottom. A moistened filter paper in mobile phase is placed on the inner
wall of the chamber to maintain equal humidity (and also thereby avoids edge effect
this way).
4. Now, the plate prepared with sample spotting is placed in TLC chamber so that the
side of the plate with the sample line is facing the mobile phase. Then the chamber is
closed with a lid.
5. The plate is then immersed, such that the sample spots are well above the level of
mobile phase for development.

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 42

 6. Allow sufficient time for the development of spots. Then remove the plates and with a
pencil, a thin mark is made at the side of the plate to mark solvent front and allow
them to dry. The sample spots can now be seen in a suitable UV light chamber, or any
other methods as recommended for the said sample e.g. Iodine complex.

Being a semi quantitative technique, TLC is used more for rapid qualitative measurements
than for quantitative purposes. But due its rapidity of results, easy handling and inexpensive
procedure, it finds its application as one of the most widely used chromatography techniques.
Rf values recorded are always less than 1 and cannot be treated as intrinsic property of the
compound as it is depending on various factors.

Solvent
front

Base line

Before developing TLC

o-nitroaniline
mixture of o-nitroaniline and p-nitroaniline
p-nitroaniline

Glass lid

Glass jar

Solvent

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 43

 Developing TLC

Solvent
front

z
x

y
Base line

After developing TLC

Note: The Rf Values are only recorded from individual spot and not from mixture

Calculation: Rf: Retention factor

𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑡ℎ𝑒 𝑠𝑜𝑙𝑢𝑡𝑒 𝑜𝑟 𝑠𝑢𝑏𝑠𝑡𝑎𝑛𝑐𝑒 𝑖𝑛 𝑐𝑚

Rf = 𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑡ℎ𝑒 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑖𝑛 𝑐𝑚

𝑥 𝑦
Rf of o-nitroaniline = 𝑧 Rf of p-nitroaniline = 𝑧
Result Table:
1. Rf of o-nitroaniline
2. Rf of p-nitroaniline
3. Solvent or solvent composition used for the TLC

Reference:

1. http://hubpages.com/education/tlc-thin-layer-chromatography-Principle-Procedure

Lab Manual F.Y. B. Sc. CHEMISTRY, SEM I 2019-20 Page 44