CHEMISTRY HONOURS

[Choice Based Credit System]

(Semester Programme)
SEMESTER-I
C1P1: Organic Chemistry Lab

1. Separation of components of a binary solid mixture

Basic Concept: Synthesis of organic compounds often involves reactions that result in the
formation of a mixture of products. Again, when some parts of a tree are extracted in an attempt
to isolate a targeted compound, invariably a mixture of compounds appears. Isolation of the
desired compound, therefore, needs exclusion of the unwanted companions. This constitutes
the principle of separation of a mixture of compounds into its components. Wide variety of
techniques including solvent extraction, fractional crystallisation and steam distillation,
chromatography etc. is employed for this purpose. Increase in the number of components in
the mixture requires technique with enhanced sophistication and clinical accuracy for
separation. In the following section separation of a mixture of only two components utilising
the difference in solubility of components in a suitable solvent is discussed.
If one component dissolves in a particular solvent and the other does not, the mixture may
be shaken with appropriate solvent to dissolve the soluble component. Filtration separates the
insoluble component from the soluble one. Then the soluble component has to be regenerated
from its solution. Acidification, basification, evaporation etc. may be needed for this
regeneration purpose.
General Procedure of Separation:
1. Test the solubility of the supplied Binary Mixture by adding a pinch of the sample in
(~ 2 mL) each of the following two solvents and shake well to find the solvent in which
a part of the mixture is soluble.
Solvents: (i) 10% NaOH solution, (ii) 6 (N) HCI.
2. Add (~ 25 mL) of the appropriate solvent (identified in step 1) to the rest of the mixture
taken in a 100 mL beaker and stir well with a glass rod for 15 - 20 minutes.
3. Filter under suction and collect the filtrate in a 250 mL beaker.
 4. Wash the residue obtained in step 3 with the same solvent used in step 2. Then wash
the residue with water, dry and collect as the ‘Insoluble Part’
5. Precipitate the soluble component from the filtrate obtained in step 3 either by
acidifying with conc. HCl (if 10% NaOH solution is used for dissolution) or basifying
with 20% NaOH solution (if 6(N) HCl solution is used for dissolution) as the case may
be. Filter off the precipitate, wash with water, dry and collect the solid as the soluble
parts.
Reporting of Separation of Components of Unknown Binary Solid Mixture:

Chemical Apparatus Chemical Apparatus

required required required required
6(N) HCl Test tube 20% NaOH Glass rod
Conc. HCl 100 ml beaker Distilled water Suction filtering
10% NaOH Measuring apparatus with
cylinder small Buchner
funnel

TYPE I [Mixture of Acidic (PART A) and Neutral (PART B) Components]: 0.2 g of the
given binary mixture (preferably in powdered form, if crystalline sample is given, it should be
ground thoroughly with the pestle to make it powder) is vigorously shaken separately with 2-
3 mL of each of 6(N) HCl and 10% NaOH solution at room temperature for 5 minutes.

6(N) HCl 10% NaOH

Insoluble Partly Soluble
Melting point of part A=
Melting point of part B=

TYPE II [Mixture of Basic (PART A) and Neutral (PART B) Components] : 0.2 g of the
given binary mixture (preferably in powdered form, if crystalline sample is given, it should be
ground thoroughly with the pestle to make it powder) is vigorously shaken separately with 2-
3 mL of each of 6(N) HCl and 10% NaOH solution at room temperature for 5 minutes.
10% NaOH 6(N) HCl
Insoluble Partly Soluble
Melting point of part A=
Melting point of part B=

2. Boiling point determination: Here are varieties of methods by which a sample's

boiling point can be determined, including distillation, reflux, and by using a Thiele tube. The
most straightforward method uses a Thiele tube.

Distillation Method

There are simpler methods than a distillation to measure a compound's boiling point, and it is
recommended to explore other options (e.g. Thiele tube) if this is the only goal. However, if
materials are limited, or if purification is planned anyhow, a distillation can be used to
determine a compound's boiling point (Figure 1).

A simple distillation should suffice for most situations, and at least 5 mL of sample should be
used in the distilling flask along with a few boiling stones or stir bar. As the bulk of the material
distills, the highest temperature is noted on the thermometer corresponds to the boiling point.
A major source of error with this method is recording too low a temperature, before hot vapors
fully immerse the thermometer bulb. Be sure to monitor the thermometer periodically,
especially when the distillation is active. Record the barometric pressure along with the boiling
point.

Figure 1

Reflux Method

A reflux setup can also be used to determine a compound's boiling point. Reflux is when a
liquid is actively boiling and condensing, with the condensed liquid returning to the original
flask. It is analogous to a distillation setup, with the main difference being the vertical
placement of the condenser.

If materials are available, the best reflux setup for this application is shown in Figure 2 and
uses a microscale condenser and digital thermometer. The setup uses ~5 mL of liquid, and a
few boiling stones or stir bar. The condenser is attached to the round bottomed flask, with the
lower water hose connected to the water spigot and the upper water hose draining to the sink.
It is important to check that the joint connecting the flask and condenser is securely fastened.
The liquid is brought to a boil on a sand bath, and the thermometer is placed low into the
apparatus such that an inch separation exists between the boiling liquid surface and the bottom
of the thermometer. In this position, the thermometer can accurately measure the hot vapors
and the temperature will stabilize at the compound's boiling point. Record the barometric
pressure along with the boiling point.

Although it might seem prudent to plunge the thermometer directly into the boiling liquid, it is
possible the liquid may be superheated, or hotter than its boiling point. After determining the
boiling point, the flask should be raised from the sand bath to cool, and condenser kept running
until the flask is only warm to the touch. At this point the setup can be dismantled.

If a microscale condenser is not available, an alternative reflux method can also be used as
shown in Figure 2. Roughly 5mL of sample is placed in a medium test tube (18 x 150 mm )
with thermometer clamped inside so it does not touch the sides of the glass. The apparatus is
carefully heated on a sand bath such that reflux happens controllably and vapors do not escape
from the tube. The temperature during reflux will eventually stabilize (this takes some time),
and the highest temperature noted corresponds to the compound's boiling point. The boiling
points measured with this method may have significant error if the boiling point is very low or
high ( <70oC or >150oC ) as low boiling compounds boil away too easily and high boiling
compounds tend to cool too easily.

Figure 2

Thiele Tube Method

Thiele Tube Theory

The Thiele tube method is one of the simplest methods to determine a compound's boiling
point, and has the advantage of using small amounts of material (less than 0.5 mL of sample).
The sample is placed in a small tube along with an inverted capillary tube. The setup is attached
to a thermometer and heated inside a Thiele tube to slightly higher than the compound's boiling
point (which is evidenced by a continuous stream of bubbles emerging from the capillary tube).
The tube is then allowed to cool, and the moment liquid is drawn into the capillary tube, the
temperature is the compound's boiling point.

This method utilizes the definition of boiling point: the temperature where the compound's
vapor pressure equals the applied (atmospheric) pressure. The inverted capillary tube acts as a
reservoir to trap the compound's vapors. As the apparatus is heated, the air initially trapped in
the capillary tube expands and causes bubbles to emerge from the tube (Figure 3). With further
heating, the compound's vapors eventually displace all of the trapped air, which is why heat is
applied until there is a continuous stream of bubbles.

When the apparatus is cooled, eventually the pressure inside the capillary tube (due solely to
the compound's vapors) will match the atmospheric pressure, at which point the bubbles will
slow and liquid will be drawn into the tube. The temperature where this begins is the
compound's boiling point.

Thiele Tube Procedure

1. Obtain a Thiele tube and clamp it to a ring stand in the fume hood. The tube is normally
filled with clear mineral oil, but it may have darkened from oxidation or spilled
compounds. If the oil is quite dark, it should be replaced. The oil should be filled to at
least 1cm higher than the top triangular arm and if too low the oil will not circulate as
needed (Figure 3a).

2. Insert a thermometer into a one-holed rubber stopper with a slit down one side. Attach
a small glass vial ("Durham tube", or 66 x 50 mm culture tube) to the thermometer with
a small rubber band. The bottom of the vial should be flush with the bottom of the
thermometer.

3. Fill the vial about half-full with sample.

4. Insert a capillary tube into the sample (the same type that is used for melting points),
open end down and sealed end up.

5. Place the rubber stopper and thermometer assembly into the Thiele tube, adjusting the
height so that the sample is midway (if possible) inside the tube. The rubber band should
be higher than the top of the mineral oil, keeping in mind that the oil may expand
somewhat during heating. The thermometer should not touch the sides of the glass, and
if it does it should be clamped in such a way that it no longer touches.
6. Heat the oil gently on the side arm of the Thiele tube with a micro-burner if available,
or Bunsen burner using a back and forth motion. As the oil warms and becomes less
dense, it will rise and travel up the triangular portion of the tube. The cooler, denser oil
will sink, thereby creating a current. This method is an excellent way to indirectly and
slowly heat the sample.
7. Although bubbles should not be seen in the Thiele tube as it warms, they commonly are
seen if the tube had been used previously for boiling point determinations. In this
method, the rubber band occasionally breaks causing the sample to fall into the oil and
contaminate it. If the oil is not subsequently changed, the sample may boil when heated
in the tube. It is okay to continue heating a Thiele tube if bubbles are seen.
8. Studies of this method have determined that it is best to heat the oil gently and in a
continual manner, as stopping and starting have caused the results to suffer.
9. Continue heating until a vigorous stream of bubbles emerges from the tip of the
capillary tube (Figure 3d), such that individual bubbles can barely be distinguished. The
purpose of this step is to expunge the air originally present in the capillary tube and
replace it with the sample's vapor. Do not heat so vigorously that the entire sample boils
away. When bubbles are vigorously emerging from the capillary tube, the vapor
 pressure inside the tube is greater than the atmospheric pressure (the oil is at a higher
temperature than the boiling point).
10. Turn off the burner and allow the apparatus to cool. The bubbles will slow and
eventually stop. At some point the vapor pressure inside the capillary tube will equal
the atmospheric pressure and liquid will be drawn into the tube (Figure 3e). The boiling
point should be recorded as the temperature when liquid just begins to enter the
capillary tube.

Figure 3

3.Identification of a Pure Organic Compound

The sample supplied may be any one of the following types:

1. Pure solid

2. Pure liquid

The following preliminary tests will be performed with the given sample;
 Organic sample

Solids
Liquids
Miscibility in cold water

Miscible Formic acid, Acetic Immiscible

acid, Phenol Warmed with HCl
Blue litmus test

Acid Nutral Immiscible

Miscible
ic
Formic acid, Acetic acid, Phenol
Methyl, Ethyl and Benzyl alcohol
Phenol
Diazo-coupling test
NaHCO3

Effervescence No Effervescence Brilliant red dye Does not respond

Formic acid, Acetic acid, Phenol K4Fe(CN)6 solution is added to the solution in HCl

HgCl2

White crystalline ppt No ppt

White or grey ppt No. ppt
Formic acid Acetic acid

Methyl, Ethyl and Benzyl alcohol, Viscous

Light acetone, acetaldehyde
Glycerol
Iodoform test

+ve Ethyl alcohol, acetone, acetaldehyde -ve

Iodoform test with NH4OH Methyl and Benzyl alcohol,
Ignition test
+ve Acetone, acetaldehyde Ethyl alcohol, -ve
Fehling’s test
Blue non- Yellow
sooty sooty
Reductio No reduction
flame flame
n
Acetaldehy
Acetone Methyl Benzyl alcohol,
de
Preparation of neutral solution: Solid sodium carbonate is added pinch by pinch to
aqueous solution of suspension of organic sample taken in a beaker with continuous stirring
with a glass rod till the effervescence ceases and a little sodium carbonate remains in excess.
Then dilute nitric acid is added to it till the solution becomes distinctly acidic and finally
ammonium hydroxide solution is added till the smell of ammonia persists after thorough
mixing. Excess of ammonia is then boiled off completely and the solution is concentrated if
necessary. The one-third portion of O.S is then used for special and confirmatory tests to
identify a particular compound.

Liquid Compounds

1. Formic acid

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Strong pungent odour
4. Miscibility : Miscible with water in all proportions
5. Litmus : Blue litmus paper turns red
6. Action of heat : Completely volatilises without
leaving any residue
7. Action of H2SO4 : Evolution of carbon monoxide
in the cold and there is brisk
evolution of carbon monoxide
on warming which burns with a
blue flame
8. Ignition test : Blue non-sooty flame

Special Tests

Experiment Observation
 1. Silver mirror test: Silver mirror 1. White ppt. of silver formate -
test is performed with silver nitrate. blackens on warming due to
This is added to O.S or neutral reduction to metallic silver.
solution of O.S. N.B: The reduction is prevented in
presence or ammonia.
(difference from acetic acid)

2. FeCl3 test: A few drops of 2. A red colour develops.

freshly prepared feric chloride
solution is added to neutral solution
of O.S. The resulting solution is
divided into two parts.

(a) Dilute HCI is added to another (a) The red colour is discharged and
part. yellow colour of FeCl3 appears.

(b) One part is boiled. (b) A brown ppt. of basic ferric formate.

3. Denige’s reagent: Denige's

reagent is added to a neutral 3. A white sand-like ppt.
solution or O.S. and boiled.

4. Mercuric chloride test: A few

drops of neutral solution of O.S. is 4. White ppt. of mercurous formate.
added to a little HgCl2 solution and
warmed.

Then excess of neutral solution is Grey ppt. of metalic mercury.

added and warmed. (difference from acetic acid)

2. Acetic acid

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Characteristics smell of vineger
4. Miscibility : Miscible With water
5. Litmus : Blue litmus paper turns red
6. Action of heat : Completely volatilises without
 leaving any residue
7. Ignition test : Blue non-sooty flame

Special Tests

Experiment Observation
1. Denige’s reagent: Denige's 1. No ppt. from cold or hot solution.
reagent is added to cool neutral
solution of O.S. and then boiled.
Very dilute solution of KMnO4 is No change of permanganate color.
added to the resulting solution.

2. FeCl3 test: A few drops of 2. A red colour develops.

freshly prepared feric chloride
solution is added to neutral solution
of O.S. The resulting solution is
divided into two parts.

(a) Dilute HCI is added to another (a) The red colour is discharged and
part. yellow colour of FeCl3 appears.

(b) One part is boiled. (b) A brown ppt. of basic ferric formate.

3. AgNO3 test: AgNO3 solution is 3. A white ppt. which does not turn
added to concentrated neutral black on boiling.
solution of O.S. and boiled. (difference from acetic acid)

4. Mercuric chloride test: HgCl2 4. No ppt.

solution is added to solution of O.S. (Distinction from formic acid)
[N.B. : A white ppt. is obtained with
ammonium acetate but no grey or
black ppt. comes down.]
5. Cacodyl oxide test (C.T): The 5. A characteristic obnoxious
neutral solution of O.S. is heated to smell of cacodyl oxide.
dryness and than thc residue is [N.B: The vapour should not be
mixed with equal parts of arsenious inhaled directly as it is highly
oxide in a test tube and then heated. poisonous. The test tube must be
Thumb is held at the mouth of the discards as the test is over.]
test tube and the thumb is inhaled. (Adv.: Better do not perform this
test)
 3. Methyl Alcohol

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Pungent but faint alcoholic
4. Miscibility : Miscible With water
5. Litmus : Neutral
6. Action of heat : Volatilises
7. Ignition test : Blue non-sooty flame

Special Tests

Experiment Observation
1. Oxidation test : A Cu-spiral is made 1.
repeatedly red hot and introduced into 2
ml of O.S. kept in a test tube dipped in a
beaker containing cold Water. The
solution is then divided into four pan ts.
(a) Schiffis test : One part is added to (a) Pink color which deepens slowly.
Schiff s.
(b) Tollen’s test: Another part is added (b) Bright mirror of silver on the side of
to Tollen's reagent and heated in a water- the test tube.
bath.
(c) Resorcinol test: A mixture of 0.5 ml (c) A reddish -violet ring appears at the
of oxidised liquid and a drop of 0.5% aq. junction of two liquid layers. A white
Solution of resorcinol is added carefully ppt. changing to reddish-violet appear in
down the side of the test tube held in an the aq. Layer above the ring after a little
inclined position containing 2ml of conc. while.
H2SO4.

2. Denige’s reagent: To about 5ml of aq. 2. A violet color develops deepening on

Solution of o.s. in a porcelain basin standing.
placed on ice-water, 2-3 ml of 2.5% soln.
of KMnO4 soln. is added followed by
few drops of conc. H2SO4 when a brown
color developes. The brown color is
destroyed by addition of saturated soln.
of oxalic acid. A freshly prepared soln.of
schiffs reagent is added dropwise to it
with stirring.

3. Oil of wintergreen test: 0.5 ml of o.s. 3. Characteristics smell of methyl

and 3 drops of conc. H2SO4 are added to salicylate.
0.5 g of salicylic acid and heated for one
minute and finally the reaction mixture is
poured into 50 ml of water taken in a
beaker. 4. The test is negative i.e., no yellowish
4. Iodoform test in NaOH is performed ppt. of iodoform
(vide test 3a for ethanol) (distinction from ethyl alcohol and
acetone)

4. Ethanol
CH3CH2OH
Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Pungent but faint alcoholic
4. Miscibility : Miscible with water
5. Litmus : Neutral
6. Action of heat : Volatilises
7. Ignition test : Blue non-sooty flame

Special Tests

Experiment Observation
1. Oxidation test : A Cu-spiral is made 1.
repeatedly red hot and introduced into 2
ml of O.S. kept in a test tube dipped in a
beaker containing cold Water. The
solution is then divided into four parts.
(a) Schiff’s test : One part is added to (a) Pink color which deepens slowly.
Schiff s.
(b) Tollen’s test: Another part is added (b) A silver mirror is disposited on the
to Tollen's reagent and heated in a water- walls of the test tube.
bath.
(c) To another part equal volume of very (c) Wine-red color develops.
dilute solution of sodium nitroprusside
solution is added followed by a few drops
of NaOH solution.
(d) To another part a very dilute solution (d) A deep red color develops.
of sodium nitroprusside solution is added
followed by a few drops of piperidine.

2. Ethyl acetate formation: About 0.2

ml of O.S. and an equal volume of glacial
acetic acid and a little conc. H2SO4 ( or 2. Pleasant fruity smell.
0.2 g of fused sodium acetate) are taken
in a test tube and warmed in a water-bath
for about 5 min. the mixture is then
poured into water taken in a beaker to
which a little Na2CO3 has been added.

3. Iodoform test: (a) To 2 ml of aq. soln.

of o.s. an equal volume of a conc.
solution of iodine in potassium iodide is 3. (a) Yellow ppt. of iodoform and its
characteristic smell.
added and then dil. NaOH solution is
added dropwise with stirring until violet (distinction from methanol)
color of iodine disappears. Then the
mixture is warmed and cooled under tap
with shaking. (b) No ppt. of iodoform.
(b) The iodoform test is repeated by (distinction from acetone)
adding NH4OH instead of NaOH.

5. Chloroform
CHCl3

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Sweet
4. Miscibility : Immiscible with water
and much heavier than water
5. Litmus : Neutral
6. Action of heat : Volatilises
7. Ignition test : Yellow sooty flame

Special Tests

Experiment Observation
1. Hydrolysis test: O.S. is treated with 1. Curdy white ppt. soluble in NH4OH
aq. KOH or NaOH solution and boiled. but reappears on addition of conc. HNO3.
The mixture is acidified with conc.
HNO3 and then AgNO3 is added. 2. A yellowish red ppt.
2. Fehlings test: O.S and Fehlings
solution (I + II equal vol.) is warmed with
constant shaking. 3. A brilliant reddish colouration is
3. Resorcinol test: A little powdered developed in aq. layer.
resorcinol and few drops of O.S. is taken
in a basin, about 1 ml of conc. solution of
NaOH is added to it, then the mixture is
warmed gently. 4. Intolerable obnoxious smell of
carbylamine.
(Not recommended to perform)
4. Carbylamine test: Few drops of O.S.
aniline and alc. KOH are warmed in a diy
test tube and the ensuing gas is smelt by
placing the thumb at the mouth of the test
tube and then holding tfie thumb near the
nostrils.

6. Nitrobenzene

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Pale yellow
3. Odour : Characteristic smell of bitter
almonds
4. Miscibility : Immiscible with water
5. Litmus : Neutral
6. Action of heat : Volatilises without leaving any
residue
7. Ignition test : Yellow sooty flame

Special Tests

Experiment Observation
1. Caustic soda test : About 0.2 ml of 1. The color darkens.
O.S. is heated with conc. solution of
caustic soda.
2. Reduction and diazocoupling test:
0.5 ml is reduced by tin or zinc and dil.
HCl for 5 minutes. The decanted 2. Brilliant scarlet-red dye.
solution is cooled and very dilute cold
solution of NaNO2 is added to it. The
diazotised soln. is added to cold alkaline
soln. of. -naphthol. 3. Grey or black ppt..
3. Muliken-Barker test: An aq.
ethanolic solution of few drops of given
sample is boiled with a pinch of zinc dust
 and little solid NH4Cl. The mixture is
cooled and filtered into Tollen’s reagent.

7. Acetone
CH3COCH3

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Pleasant ethereal odour
4. Miscibility : Miscible with water
5. Litmus : Neutral
6. Action of heat : Completely Volatilises without
leaving any residue
7. Ignition test : Blue non-sooty flame

Special Tests

Experiment Observation
1. Brady’s test : A small amount of O.S. is 1. Yellow ppt. on mixing.
added to 2-4-dinitrophenylhydrazine solution
2. Denige’s test: An equal volume of
Denige’s reagent is added to an aqueous 2. Heavy white ppt. of double compound of
solution of O.S and the test tube is then kept acetone and basic mercuric sulphate.
in a boiling water bath for few minutes.

3. Legal’s test: A few drops of a very dil. 3. A ruby red (reddish-purple) color
solution of sodium nitroprusside is added to develops which disappears on warming but
2-3 ml of aqueous solution of O.S. followed reappears on cooling.
by a few drops of dil. NaOH solution.

4. Iodoform test: (a) iodoform test is

performed with I2 in KI solution and NaOH 4. (a) Yellow ppt. of CHI3 having
characteristics sweeet smell.
solution. (b) the iodoform test is repeated by
adding NH4OH solution instead of NaOH (b) Yellow ppt. of CHI3 having
solution. characteristics sweet smell.
(Distinction from ethanol)

8. Aniline

Physical characteristics and preliminary test :

 1. State : Liquid
2. Colour : Colorless when freshly distilled
but turns brown on exposure to
light and air
3. Odour : Characteristic aromatic smell
4. Miscibility : Immiscible with water but soluble
in dil. HCI
5. Litmus : Neutral
6. Action of heat : Completely volatilises without
leaving any residue
7. Ignition test : Yellow sooty flame

Special Tests

Experiment Observation
1. Bleaching powder test: A few drops of 1. Purple-violet colouration.
bleaching powder solution is added to a dilute
solution of O S. in HCl.
2. Potassium dichromate test: A drop of 2. Intense blue colour develops.
O.S. is added to 5 to 6 drops of conc. H2SO4
taken in a spot plate and the mixture is stirred
well with glass rod. Then a drop of K2Cr2O7
solution is added to it. 3. Brilliant scarlet-red dye.
3. Diazocoupling test: 5-6 drops of O.S are
dissolved in dil. HCl in a test tube and cooled
in ice-water. Then 3-4 drops of very dilute ice
cold solution of sodium nitrite are added to it.
Then the solution is added dropwise to 2 mL
of ice-cold alkaline solution of -naphthol;
finally solution should remain mild alkaline

9. N,N-Dimethylaniline

Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless when freshly distilled
 but turns brown on exposure to
light and air
3. Odour : Characteristic bad smell
4. Miscibility : Immiscible with water but
soluble in dil.HCI

5. Litmus : Neutral
6. Action of heat : Completely volatilises without
leaving any residue
7. Ignition test : Yellow sooty flame

Special Tests

Experiment Observation
1. Bleaching powder test: A few drops of 1. Purple-violet colouration.
bleaching powder solution is added to a dilute
solution of O S. in HCl.
2. Potassium dichromate test: A drop of 2. Intense blue colour develops.
O.S. is added to 5 to 6 drops of conc. H2SO4
taken in a spot plate and the mixture is stirred
well with glass rod. Then a drop of K2Cr2O7
solution is added to it. 3. A colour appears at this stage.
3. Nitrous acid test: 5-6 drops of O.S are (No red dye after addition to alkaline 2-
dissolved in dil. HCl in a test tube and cooled naphthol solution)
in ice-water. Then 3-4 drops of very dilute ice (distinction from aniline)
cold solution of sodium nitrite are added to it.
4. Malachite green test: See test of 4. An intense green colouration
benzaldehyde below. Use benzaldehyde and
this sample.

10. Benzaldehyde
C6H5CHO
Physical characteristics and preliminary test :

1. State : Liquid
2. Colour : Colorless
3. Odour : Characteristic smell of bitter
almonds
4. Miscibility : Immiscible with water
5. Litmus : Neutral
6. Action of heat : Volatilises
7. Ignition test : Yellow sooty flame
 Special Tests

Experiment Observation
1. 2,4-D.N.P test: A few drops of 2,4-D.N.P 1. Reddish yellow ppt. forms immediately
solution is added to 1 mL alcoholic solution simply on mixing.
given sample.
2. Tollen’s test: An alcoholic solution of the 2. Black ppt.
given sample is added to 2 mL of Tollen’s
reagent, warmed gently. 3. A intense green coloration.
3. Malachite green test: 0.5 ml of O.S is
heated with 1 ml of dimethyl aniline and a
small bit of anhydrous ZnCl2 in a dry test tube
for one minute. The leucobase produced is
oxidised with lead dioxide in a solution of
acetic acid and excess conc. HCl is added.

Solid compounds

1. Oxalic acid [(COOH)2]

Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystaline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Blue litmus turns red
6. Action of heat : Volatilises completely without charring when heated slowly and
a sublimation is formed but decomposed with evolution of
gases when heated strongly
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. Lime water turns milky.
heated with conc. H2SO4 and the evolved
gas is passed into lime water.
2. Soda-lime test: A small amount of 2. The gas burns at the mouth of the test
O.S. is heated in a hard glass test tube tube.
with sodalime.
3. Calcium chloride test: Calcium 3. A white ppt. forms immediately.
chloride solution is added to neutral
solution of O.S. The ppt is divided into
two parts
(i) Dil. acetic acid is added to one (i) The ppt does not dissolve.
portion. (Distinction from tartaric and succinic
acids)
(ii) The ppt. dissolves
(ii) Dil. HCl or dil. HNO3 is added to 4. A white crystalline ppt.
another portion.
4. Silver nitrate test : AgNO3 solution
is added to neutral solution of O.S. The (i) The ppt. dissolves
ppt. is divided into three parts : (ii) The ppt. dissolves
(il Dil. NaOH is added to one portion. (ii) (iii) Metallic silver is formed
Dil. HNO3 is added to another portion.
(iii) The third portion is warmed, dried
and heated strongly in a dry test tube 5. White ppt. in the cold which does not
5. Denige's test (C.T.): Denige's reagent dissolve
is added to neutral solution or aqueous
solution of O.S. 6. The pink colour of permanganate
6. Potassium permanganate test: A disappears.
little of dil. H2SO4 is added to a solution of
O.S or neutral soln. of O.S. The solution is
warmed and dil. KMnO4 solution is
added drop by drop with shaking.

2. Citric acid
Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystaline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Blue litmus turns red
6. Action of heat : Charring with evolution of acid-
smelling vapour
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. Slow charring.
heated with conc. H2SO4 .
2. Calcium chloride test: An equal 2. No ppt. appears even on shaking and
volume of calcium chloride solution is scratching but white crystalline ppt.
added to neutral solution of O.S. The appears after boiling for several
mixture is shaken vigorously inner side minutes.
of the test tube is scratched with glass rod
and finally boiled. 3. A curdy white ppt

(i) The ppt. dissolves

3. Silver nitrate test : AgNO3 solution (ii) The ppt. dissolves
is added to neutral solution of O.S. The (iii) No metallic silver is formed
ppt. is divided into three parts :
(i) Dil. NH4OH is added to one portion.
4. No ppt. in the cold but the color of
(ii) Dil. HNO3 is added to another permanganate discharges with
portion. appearance of heavy white turbidity on
(iii) The third portion is warmed, dried boiling.
and heated strongly in a dry test tube
4. Denige's test (C.T.): Denige's reagent
is added to neutral solution or aqueous
solution of O.S. and then a drop of 2%
KMnO4 solution is added.

3. Succinic acid
Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystaline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Blue litmus turns red
6. Action of heat : Melts and then boils giving off extremely irritating vapours.
A sublimate is forme d at the cooler part of the test tube
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. H2SO4 test: Few crystals of O.S. is 1. O.S dissolves without charring. Slight
warmed conc. H2SO4 and then heated chairring occurs on strong heating and
strongly. the solution turn brown with evolution of
SO2.
2. Calcium chloride test: An equal 2. A white ppt. soluble in acetic acld.
volume of Calcium chloride solution is (Distiction from oxalic macid)
added to neutral solution of O.S. The
mixture is shaken vigorously inner side
of the test tube is scratched with glass rod
and finally boiled. 3. Buff-colored ppt. soluble in dil. HCl.
3. Silver nitrate test : AgNO3 solution 4. No ppt. in the cold but the color of
is added to neutral solution of O.S. permanganate presists.
4. Denige's test (C.T.): Denige's reagent
is added to neutral solution or aqueous
solution of O.S. and then a drop of 2% 5. A beautiful reddish green
KMnO4 solution is added. fluorescence, intesifies to a beautiful
5. Fluorescein test: A small quantity of green fluorescence on addition of
the sample ix mixed with resorcinol and alkali..
 2 / 3 drops of c. sulfuric acid and heated.
Then poured into large volume of water.

4. Resorcinol
Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystaline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : feebly acidic
6. Action of heat : Melts and volatiles
7. Ignition test : Yellow sooty flame

Experiment Observation
1. H2SO4 test: Few crystals of O.S. is warmed 1. O.S dissolve without charring.
with conc. H2SO4.
2. Ferric chloride test: A few drops of FeCl3 2. Bluish-violet color.
solution is added to aqueous solution of O.S.
3. Bromine writer test: Bromine writer is 3. White ppt.
added to aq. solution of O.S. and shaken. 4. Black ppt of silver mirror.
4. Tollen's test: Aqueous solution of O.S is added
to Tollen’s reagent and warmed. 5. An intense reddish-green
5. Fluorescein test: A small quantity of O.S. is fluorescein, intesifies to a beautiful
mixed with succinic acid / phathalic acid and 2 green fluorescence on addition of
drops conc. Sulfuric acid and heated. Then poured alkali.
into a large volume of water. 6. A brilliant reddish colouration is
6. O.S. is mixed with 3 - 4 drops CHCl3 and developed in aq. layer.
gently heated on a basin with NaOH solution.

5. Urea
Physical characteristics and preliminary test :
1. State : Solid
2. Colour : Colorless
3. Texture : Crystaline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Neutral
6. Action of heat : Melts and gives off characteristic smell of NH3
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. Nitrous acid test: Dil. HCl and dil. NaNO2 1. Effervescence with evolution of
solution is added to aq. Solution of O.s in a test gas.
tube and shaken.
 2. A white crystalline ppt.
2. Nitric acid test: 2 mL of conc. HNO3 is added
to aqueous concentrated solution of O.S. 3. A white crystalline ppt.
3. Oxalic acid test: A concentrated solution of
oxalic acid is added to conc. solution of O.S.
4. A pink or violet colour develops.
4. Biuret test: About 0 5 g of O.S. is heated
gently# to melt. The heating is continued till the
molten mass solidifies at once when test tube is [#strong and rapid heaing leads to formation
of cyanuric acid and then the test will fail]
taken out of flame. If there is no solidification the
process is repeated. The residue is cooled and
dissolved in I ml of NaOH solution by warming
and shaking. The solution is cooled again and a
drop or two of a very dilute copper sulphate
solution is added to it.

6. Benzoic acid
Physical characteristics and preliminary test :

1. State : Solid
2. Colour : White
3. Texture : Plate-shaped or needle shaped
Crystal
3. Odour : Odourless
4. Solubility : Insoluble in cold water but
Soluble in hot water
5. Litmus : Blue litmus turns red
6. Action of heat : Melts and sublimates
7. Ignition test : Yellow sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. Dissolves without charring.
warmed with conc. H2SO4.
2. Soda-lime test: A small amount of 2. Characteristics smell of benzene.
O.S. is heated in a hard glass test tube
with sodalime. 3. White ppt. dissolves on boiling but
3. Denige’s test: Denige’s reagent is appears on cooling.
added to neutral solution of O.S. 4. (a) a buff-colored ppt.
4. Ferric chloride test: (a) A drop or two (Distinction from salicylic acid)
of freshly prepared solution of Fecl3 is
(b) buff-colored ppt dissolves with
added to neutral solution of O.S.
appearance of white ppt.
(b) Dilute HCl is added to it.

5. Esterification test: About 0.1 g of O.S 5. Characteristics fruity smell of ester.

and 2 mL of dehydrated alcohol are taken
in a clean test tube and shaken well. Then
a few drops of conc. H2SO4 is added and
the test tube is waremd in a hot water
bath. The test tube is cooled and content
of the test tube is poured into 50 mL of
dilute sodium cxarbonate solution taken
in a beaker.

7. Salicylic acid

Physical characteristics and preliminary tests:

1. State : Solid
2. Colour : White
3. Texture : needle shaped Crystal
3. Odour : Odourless
4. Solubility : Sparingly soluble in cold water
but readily Soluble in hot water
5. Litmus : Blue litmus turns red
6. Action of heat : Melts and sublimes
7. Ignition test : Yellow sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. The solid dissolved, charring occurs
warmed with conc. H2SO4. after some time, solution darkens and gas
evolve.

2. Soda-lime test: A small amount of 2. Characteristics smell of phenol.

O.S. is heated in a hard glass test tube
with sodalime. 3. White ppt. dissolves on boiling but
3. Denige’s test: Denige’s reagent is appears on cooling.
added to neutral solution of O.S. 4. An intense violet color
4. Ferric chloride test: A drop or two of (Distinction from benzoic acid)
freshly prepared solution of Fecl3 is
(a) The color discharges
added to neutral solution of O.S.
(a) Dilute HCl is added to it. (b) The color persists
(b) acetic acid is added to another part. 5. A characteristics pungent fragnant
5. Oil of wintergreen test: About 0.1 g odor which intensifies on pouring the
of O.S and 1 mL of methanol and few mixture to dil. Sodium carbonate
drops of conc. H2SO4 are warmed in solution.
aclean dry test tube.

8. Tartaric acid
 Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystalline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Blue litmus turns red
6. Action of heat : Chars and gives off smell
of burnt sugar
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. charrs immediately.
warmed with conc. H2SO4.
2. Calcium chloride test: Excess of 2. A white crystalline ppt. soluble in hot
calcium chloride solution is added to dilute acetic acid.
neutral solution of O.S. shaken and
inside of the test tube is scratched with a
glass rod. 4. (a) A white crystalline ppt. (b)
4. Silver nitrate test : (a) AgNO3 shining silver mirror forms at the
solution is added to neutral solution of inner of the test tube.
O.S. (b) Addition of AgNO3 solution is (Distinction from oxalic, citric,
continued till the precipitation is succinic acids)
complete. Then dil. NH4OH is added
dropwise with constant shaking till the
ppt. almost and not completely dissolved
as revealed by slight turbidity. 5. Permanganate color discharges
5. Denige's test (C.T.): Denige's reagent immediately without turbidity.
is added to neutral solution or aqueous
solution of O.S.

9. Glucose
Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystalline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Neutral
6. Action of heat : Charrs and gives off smell
of burnt sugar
7. Ignition test : Blue non-sooty flame
 Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. No charring in cold but darkens on
warmed with conc. H2SO4. heating immediately.
2. Sodium hydroxide test: 2 ml of 3%
solution of NaOH is added to conc. 2. The solution turns first yellow and
solution of O.S. and the mixture is then raddish brown.
heated. The mixture is then acidified with
dil. HNO3.
3. Tollen’s test: 5 ml of aqueous solution 3. Black or grey precipitate or shining
of O.S. is added to equal volume of silver mirror.
Tollen’s reagent taken in a clean test tube
and then the test tube is placed in a
beaker of boiling water bath for a few
minutes. 4. Brick red precipitate.
4. Fehling's test: The sample is added to
a mixture of Fehling A and Felling B and
5. The white ppt. turns salmon-pink in
heated for 1 minute.
color.
5. Lead acetate test: Lead acetate
solution is added to solution of o.s and
the mixture is boiled for few seconds.
Then dil. NH4OH solution is added
dropwise till just sufficient to produce a
permanat white ppt. then the mixture is
boiled again.

10. Cane sugar

Physical characteristics and preliminary test :

1. State : Solid
2. Colour : Colorless
3. Texture : Crystalline
3. Odour : Odourless
4. Solubility : Soluble in cold water
5. Litmus : Neutral
6. Action of heat : Charrs and gives off smell
of burnt sugar
7. Ignition test : Blue non-sooty flame

Experiment Observation
1. H2SO4 test: A small amount of O.S. is 1. Immediate charring even in cold.
warmed with conc. H2SO4.
2. Sodium hydroxide test: 2 ml of 3% 2. No chharing but sometimes a light
solution of NaOH is added to conc. yellow color develops.
Solution of O.S. and the mixture is
heated. The mixture is then acidified with
dil. HNO3.
 3. Fehling's test: (a) The sample is 3. (a )No ppt.
added to a mixture of Fehling A and
Felling B and heated for 1 minute. (b) Red ppt of Cu2O.
(b) A little of aq. Solution of o.s is boiled
with dil. HCl or dil. H2SO4 for few
minutes. The solution is then made
alkaline with NaOH solution. The
solution is added to 2 ml of Fehling 4. The white ppt. does not change color.
solution and warmed.
4. Lead acetate test: Lead acetate
solution is added to solution of o.s and
the mixture is boiled for few seconds.
Then dil. NH4OH solution is added
dropwise till just sufficient to produce a
permanat white ppt. then the mixture is
boiled again.

C2P2: Physical Chemistry Lab

Experiment 1: Determination of pH of unknown solution (buffer), by color

matching method:

Theory:
PH of a solution is defined as the negative logarithm of its H+ ion activity i.e,

PH = -logaH+

In case of solution of low ionic strength

PH = -logc H+

The solution having applicable capacity to resist the change in its PH due to addition of acid
or base are called buffer solutions. Usually they consist of a mixture of a weak acid or base
and its salt.

In case of a mixture of a weak acid and its salt [provided the solution is not very acidic
(PH>4)]. PH of the solution can be expressed as
[Salt]
PH= pKa+ log [acid]
And I case of a mixture of weak base and its salt [provided the solution is not very basic
(PH<10)].
[salt]
PH = pKb+log[base]

Both the equations are known as Henderson equation for weak acid and weak base
respectively and will be valid only within the PH range 4 to10.

As indicator is a substance which can indicate, generally by colour change, the specific
physic-chemical condition of a chemical system.

The acid-base indicator or neutralization indicators are substance which exhibit different
colours according to the hydrogen ion concentration of their environment. It is therefore
possible to obtain a idea of the PH of a given solution by adding a little of a suitable indicator
to the same. Colour changes are believed to be due to some structural changes. In solution the
acidic form (InA) and the basic (InB) of an indicator will be in equilibrium.

InA ⇌ H+ + InB

The equilibrium constant called indicator constant (KIn) corresponding to this equilibrium is

KIn = aInaH+/aInA= [H+][InB]/[ InA], in solution of low ionic strength

[InB]
Or, PH= pKIn + log [ InA]

The colour of the indicator as perceived by the eye is determined by the ratio of the
concentrations of the acid and alkaline forms i.e, by [InB]/[ InA]

The value of [InB]/[ InA] determines the actual shade of colour as a mixture of two colours (
predominant colours in acidic and basic medium). Thus by changing the PH of the solution
one can developed by an indicator. If a solution of known PH exhibits a particular shade of
colour, with a definite amount of indicator, then another solution (of unknown PH) which
exhibits the same shade of colour, with the same amount of indicator, must also posses the
same PH.

Apparatus required:
1. 100 mL volumetric flask-3
2. 500 mL bottle-2
3. Burette-1
4. 10 mL pipette-1
5. 250 mL conical flask-1
6. Hard glass test tubes of equal diameter-10
Procedure:
1) Prepare 100 mL standard 0.5(N) oxalic acid solution in a volumetric flask by accurate
weighing.
2) Prepare 250 mL ̴ 0.5(N) NaOH solution.
3) Standardize the prepare NaOH solution against the 0.5(N) oxalic acid solution, taking
10 mL of the acid as aliquot and using phenolphthalein as indicator.
4) Determine the strength of the alkali: Prepare 100 mL of exact 0.4(N) NaOH solution
by proper dilution.
5) Prepare 250 mL̴ 0.5(N) CH3COOH solution and Standardize it against prepared ̴
0.5(N) NaOH solution.
6) Determine the strength of CH3COOH solution. Prepare 100 mL of exact 0.4(N)
CH3COOH solution by proper dilution.
7) Take 10 hard glass test tubes by approximately equal diameter, lebel them from 1 to 9
and prepare the following buffer solutions by proper mixing of exact 0.4(N) NaOH
and 0.4(N) CH3COOH.

Test tube Volume of Volume of Volume of Total PH

No. CH3COOH(mL) NaOH(mL) H2O(mL) volume(mL)

8) In the remaining test tube pipette out exactly 10 mL of the unknown buffer. To each of
these test tubes add 5 drops of the indicator. Mix thoroughly to develop uniform colour in
each test tube. Identify the pH of the buffer solution by colour matching.

Experimental Result:
1) Room temperature
2) Preparation of 100 mL 0.5(N) oxalic acid:
Required weight (3.15 gm) of oxalic acid was taken in a 100 mL volumetric flask and
volume was made up to the mark by deionized water.
3) Preparation of 250 mL approximately 0.5(N) NaOH solution:
5 gm NaOH solid was taken in a glass bottle and ̴ 250 mL deionized water was added
(using measuring cylinder) to make 250 mL̴ 0.5(N) NaOH solution.
4) Standardization of the prepared NaOH solution:
 Volume of Burette reading Volume of Average volume
0.5(N) Oxalic NaOH(mL) of NaOH(mL)
acid (mL) Initial(mL) Final(mL)

5) Preparation of 100 mL exact 0.4(N) NaOH solution:

100×0.4
Required volume of prepared NaOH solution = strength of NaOH solution mL
…… mL prepared NaOH solution was taken (using burette) in a 100 mL volumetric
flask and rest of the volume was made by adding deionized water up to the mark.
6) Preparation of 250 mL̴ 0.5(N) acetic acid:
7.4 mL of the acetic acid was taken in a glass bottle and ̴ 250 mL deionized water was
added (using measuring cylinder) to make 250 mL ̴ 0.5(N) CH3COOH solution.

7) Standardization of the prepared CH3COOH solution:

Vol. of ̴ 0.5(N) Burette reading Vol. of Average vol. of
acetic acid standardization NaOH (mL)
Initial(mL) Final(mL)
(mL) NaOH solution
used (mL)

Strength of CH3COOH solution:

8) Preparation of 100 mL exact 0.4(N) CH3COOH solution:

𝟏𝟎𝟎×𝟎.𝟒
Required volume of prepared CH3COOH solution= 𝑺𝒕𝒓𝒆𝒏𝒈𝒕𝒉 𝒐𝒇 𝐂𝐇𝟑𝐂𝐎𝐎𝐇 𝐬𝐨𝐥𝐮𝐭𝐢𝐨𝐧 mL.
…..mL prepared CH3COOH solution was taken (using burette) in a 100 mL
volumetric flask and rest of the volume was made by adding deionized water up to the
mark.
9) Preparation of buffer solution:
Test Volume of Volume of Volume of Total PH
Tube No. CH3COOH(mL) NaOH(mL) H2O (mL) volume
(mL)
1 0.5 0.5 4.5 10.0 3.72
2 0.5 1.0 4.0 10.0 4.05
3 0.5 1.5 3.5 10.0 4.27
4 0.5 2.0 3.0 10.0 4.45
5 0.5 2.5 2.5 10.0 4.63
6 0.5 3.0 2.0 10.0 4.80
7 0.5 3.5 1.5 10.0 4.99
8 0.5 4.0 1.0 10.0 5.23
9 0.5 4.5 0.5 10.0 5.57
 Conclusion:
The colour of the test tube containing unknown buffer matched with
the colour of the test tube no …… having pH …… Therefore pH of the unknown
buffer solution is…..

Experiment 2: Determination of heat of neutralization of a strong acid by a

strong base:

Theory:
The molar heat of neutralization of an acid can be defined as the heat change
associated with the complete neutralization of its dilute aqueous solution
+
containing one mole of H ions by a dilute aqueous solution of a base containing one mole
OH- ions.

Let us consider a neutralization reaction of strong acid (HA) by a strong base (BOH) in
aqueous solution. Since both are completely dissociated in aqueous solution we can represent
the neutralization reaction as

H+ (aq)+ A- (aq) + B+ (aq) → A- (aq) + B+ (aq) +H2O

Or, H+ (aq)+ OH- (aq) → H2O

Thus the neutralization of strong acid with strong base can be considered as the
combination or reaction of H+ ions with OH- ions. In other words molar heat of neutralization
is the formation of 1 mole of water from one mole each H+ and OH- ions. Therefore molar
heat of neutralization is constant irrespective of the strong acid or the strong base used.

Heat measurements are performed by carrying out the reaction in a special container called a
calorimeter. The heat given off by the neutralization reaction is absorbed by the reaction
solution and the calorimeter. Both the solution and calorimeter increases in temperature due
to the absorbed heat and this increase can be measured with a thermometer.

Total Heat of neutralization (-ΔHneu)

 = Heat gained by salt solution + heat gained calorimeter

= [Mass of salt solution × Sp. Heat capacity of salt solution× Δt of salt

solution]+[Calorimeter constant × Δt of calorimeter]

Molar heat of neutralization

𝑇𝑜𝑡𝑎𝑙 ℎ𝑒𝑎𝑡 𝑜𝑓 𝑛𝑒𝑢𝑡𝑟𝑎𝑙𝑖𝑧𝑎𝑡𝑖𝑜𝑛(−𝚫𝐇𝐧𝐞𝐮)

= 𝑁𝑜.𝑜𝑓 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑎𝑐𝑖𝑑𝑠 𝑛𝑒𝑢𝑡𝑟𝑎𝑙𝑖𝑧𝑒𝑑

To evalute the calorimeter constant ( also known as its heat capacity) in J/⁰C, adds a known
mass of hot water to a known mass of cold water which is in the calorimeter. Heat is lost by
the hot water and is absorbed by the cold water and the calorimeter.

Heat lost by Hot water = Heat gained by cold water + Heat gained by calorimeter

Or, Mass of hot water × Sp. Heat capacity of water × Δt of Hot water = [Mass of cold
water × Sp. Heat capacity of water × Δt of cold water]+[Mass of calorimeter × Sp. Heat
capacity of calorimeter × Δt of calorimeter].

where, [ Mass of calorimeter × Sp. Heat capacity of calorimeter]=Calorimeter constant

Δt is found by plotting temperature versus time for the system in the calorimeter and
extrapolating the results to find Δt the instant of mixing.

Apparatus required:
1) Glass calorimeter,
2) 100 mL conical
3) 100 mL beaker,
4) 25 mL pipette
5) Stop watch
6) 500 mL glass bottle.

Chemicals required: Oxalic acid , HCl, NaOH and Phenolphthalein

Procedure:
a) Determination of calorimetric constant:
1) Pipette out 25/50 mL of distilled water into a clean and dry calorimeter with stirrer.
This is the cold water. Suspend a digital thermometer into water.
 2) Pipette out 25/50 mL of distilled water into a 100 mL conical flask. Heat this water to
20-30⁰C above the temperature of the cold water. Remove the heat source and place a
digital thermometer within the flask.
3) Be sure to note the starting time when measuring temperatures. Starting with the hot
water, simultaneously measure the hot and cold water (in the calorimeter ) and record
the temperature of the mixture at 1 minutes intervals for 8-10 minutes.
4) Clean and dry the beaker and calorimeter for the neutralization reaction.
5) Plot graphs of temperature vs time (in minutes as abscissa) for hot water, cold water
and mixture. Extrapolate the three straight lines beyond the data points. Indicate the
temperature of hot
water and cold
water and that of
the mixture at the
time of mixing in
the graph as shown
in Figure 3.1.Evalute
the calorimeter
constant.

b) Acid-Base neutralization:

1) Prepare ̴ 2(N) HCl and ̴ 2(N) NaOH solutions. Standardize the prepared NaOH solution by
taking 25 mL supplied oxalic acid solution [ strength 1(N)] using phenolphthalein indicator.
Determine the strength of NaOH is molarity.

2) Standardize the prepared acid solution against standard NaOH solution, taking 25 mL of
the acid as aliquot and using phenolphthalein as indicator. Calculate the strength of acid in
molarity.

3) Pipette out 25/50 mL of NaOH solution into a 100 mL beaker.

4) Pipette out 25/50 mL of assigned acid solution into the clean dry calorimeter. Suspend
digital thermometers in both the acid and the base solution. Start the timer.
5) Note the temperature of both acid and base at 1 minute interval (total 5 readings for each
acid and base). [Since both the acid and base have been at room temperature for several
hours, we can safely assume that both are at he same temperature.(Check the thermometer)].

6) Quickly add the base to the acid in the calorimeter, stir gently and note the time. Record
the solution temperature every minute for 8-10 minutes.

7) Plot the graphs of temperature vs.time (in minutes as abscissa) for acid, base and the
mixture. Extrapolate the straight lines beyond the data points. Indicate the temperature of the
acid, base and the mixture in the graph as shown in Figure 3.2. Evalute the heat of
neutralization.

8) Be sure to neutralization all of the acidic hydrogens of the acid ( strength of base should be
slightly higher than that of acid as same volume of acid and base are mixed to react).
Detremine the moles of water formed in the system from the molarity and volumes of acid
and base used and calculate molar heat of neutralization.

Experimental result:
1) Determination of strength of NaOH using 1N oxalic acid:

Vol. of 1(N) Burette reading Vol. of NaOH Average vol. of

oxalic acid (mL) Initial(mL) Final(mL) used (mL) NaOH(mL)
25
25

Strength of NaOH solution:

2) Determination of strength of HCl:

Vol. of HCl (mL) Burette Vol. of NaOH used Average vol. of

reading (mL) NaOH(mL)
 Initial Final
(mL) (mL)
25
25

Strength of HCl solution:

3) Determination of calorimeter constant:

Time (min) Hot water temp(⁰C) Cold water(⁰C) Mixed water

temp(⁰C)
0
0.5
1.0
1.5
2.0
2.5 Mixing of Hot and Cold water

Time(min) Hot water temp Cold water temp Mixed water temp
(⁰C) temp (⁰C) (⁰C)
3.0
4.0
5.0
6.0
7.0
8.0

4) Determination of Heat of neutralization:

Time ( min) Acid solution Base solution Salt solution

temperature (⁰C) temperature (⁰C) temperature (⁰C)
0
1.0
2.0
3.0
4.0
5.0 Mixing of Acid and Base
6.0
7.0
8.0
9.0
10.0

Calculation:
 1) Calorimeter constant: Heat lost by Hot water = Heat gained by cold water + Heat
gained by calorimeter
 Mass of hot water × Sp. Heat capacity of water × Δt of Hot water = [Mass of cold
water × Sp. Heat capacity of water × Δt of cold water]+[Mass of calorimeter ×Sp.
Heat capacity of calorimeter × Δt of calorimeter]
 [Mass of calorimeter × Sp. Heat capacity of calorimeter]= Calorimeter constant =
… J/⁰C.
2) Heat of neutralization:
a) If acid and base have different temperature: Heat gained by acid solution +
Heat gained by base solution + Heat gained by Calorimeter.
 Total Heat of neutralization (-ΔHneut) = [Mass of acid solution × Sp. Heat
capacity of acid solution × Sp. Heat capacity of calorimeter ]
b) If acid and base have same temperature: Heat of neutralization = Heat gained
by salt solution + heat gained by calorimeter.

 Total Heat of neutralization (-ΔHneut) = [Mass of salt solution × Sp. Heat
capacity of salt solution × Δt of salt solution]+[Calorimeter constant × Δt of
Calorimeter]

Calculation of moles of acid used in neutralization reaction :

Strength of HCl (say) = M

So, 1000 mL HCl solution contains M mole HCl

M×(25 or 50)
25/50 mL HCl solution contains mole HCl.
1000

M×(25 or 50)
Heat of neutralization for mole HCl= X joule (say)
1000

Therefore, Heat of neutralization for 1 mole HCl,

= X/ M × (25 or 50) KJ mol-1

Experiment 3: Study of kinetics of acid-catalyzed hydrolysis of methyl
acetate:

Theory:
The acid catalysed hydrolysis reaction of ester can be represented as

R1COOR2+ H2O+H+ ⇄ R1COOH +R2OH

The rate equation is represented as

𝑑[R1 COOR2 ]
- = k[R1COOR2][ H+][ H2O]
𝑑𝑡
= k1[R1COOR2], where k1=k[ H+][ H2O] = constant, (in large excess of
water,[H2O] remains constant and H+ being the catalyst [ H+] is constant)

Therefore the reaction becomes first order w.r.t ester. Integration of this equation with the
boundary conditions, when t= 0, [R1COOR2] =C0 and at time t, [R1COOR2] =C, gives

K1=(2.303/t)log (C0/C)

The progress of the catalysed reaction may be studied by withdrawing measured volume of
aliquot from the reaction mixture at different intervals of time and titrating with standard
alkali solution using phenolphthalein indicator. The volume of alkali required at any instant is
equivalent to be the sum of weak acid (produced as a result of hydrolysis) and the acid used
as catalyst (a constant quantity).

If V0, Vn and Vꝏ be the volumes of alkali required for the same volume of aliquot at the
beginning, at time t= tn and at the end of the reaction (at infinite time) respectively then,

C0 ∝ (Vꝏ- V0) and C∝ (Vꝏ- Vn)

 2.303 (Vꝏ− V0)
Then, k1= 𝑙𝑜𝑔 (Vꝏ− Vn)
𝑡𝑛

To avoid the measurement of V0 the equation may be represented in the form

2.303 (Vꝏ− V1)
Δtn= tn-t1= 𝑙𝑜𝑔 (Vꝏ− Vn)
𝑘1

Where Vn and V1 are the volumes of alkali required at times tn and t1 respectively.

(Vꝏ− V1)
Plot of 𝑙𝑜𝑔 (Vꝏ− Vn) vs Δtn will give a straight line passing through the origin and the slope
k1 may be determined by

K1=slope× 2.303

Apparatus required:
1) 100 mL dry conical flask-1
2) 500 mL bottle for NaOH.
3) 2 mL pipette; 5 mL pipette-1 each
4) 250 mL conical flask-5
5) Sufficient ice cold water; water bath
6) Stop watch

Chemicals required: Methyl acetate, NaOH, and Phenolphthalein

Procedure:
1) Prepare 250 mL of approximately 0.1(N) NaOH solution.
2) Prepare 100 mL 1(N) HCl solution
3) Pipette out 50 mL of prepared HCl solution (catalyst solution) in a 100 mL dry
conical flask, add 5 mL of ester using a pipette. Start the stop watch at the time of half
discharge. Mix the solution thoroughly by swirling motion.
4) At 5-7 minutes intervals take 2 mL aliquot and add to 50 mL ice cold water taken in a
250 mL conical flask. Note the time of half discharge. Titrate rapidly against the
prepared ̴ 0.1(N) NaOH solution taken in a burette, using phenolphthalein as
indicator. Take at least 6 readings.
5) The remaining solution is heated at about 60⁰C in a water bath ̴ for 40 minutes with an
air condenser fitted in the mouth of conical flask. The solution is allowed to cool to
room temperature. Pipette out 2mL of it in 50 mL water taken in a 250 mL conical
flask and titrate with ̴ 0.1(N) NaOH solution; using phenolphthalein as indicator. The
titre value correspond to Vꝏ.
 (Vꝏ− V1)
6) Plot a graph of 𝑙𝑜𝑔 (Vꝏ− Vn) vs Δtn and draw the best fit straight line passing through
the origin.
7) Calculate the value of ‘k1’ from the slop of the graph.

Experimental result:
1) Room temperature
2) Preparation of 250 mL ̴ 0.1(N) NaOH solution: Dissolve 1 g NaOH in 250 mL
deionized water.
3) Preparation of 100 mL 1(N) HCl solution: Dissolve ̴ 9mL conc. HCl in 91 mL
deionized water
4) Recording of data for ester hydrolysis:

Time(t) Time(tn)in Δtn= tn-t1 Volume of (Vꝏ-V1) (Vꝏ- (Vꝏ− V1)

𝑙𝑜𝑔 (Vꝏ− Vn)
sec NaOH(Vn mL Vn)mL
mL)

5) Determination of Vꝏ:
Vꝏ = Volume of NaOH required to standardize 2 mL reaction mixture after
heating at about 60⁰C in a water bath.

6) Plotting of graph:
(Vꝏ − V1)
𝑙𝑜𝑔
(Vꝏ − Vn)
Δtn(Sec)

Calculation: Observed rate constant, k1=slope×2.303

Conclusion: Threfore observed rate constant for the acid catalyzed hydrolysis of
methyl acetate at …..⁰C is…..sec-1.
Experiment 4: Study of kinetics of decomposition of H2O2

Theory:
The reaction between H2O2 and KI in dilute H2SO4 medium is represented as

H2O2+2KI+ H2SO4= I2+2H2O+ K2SO4

Suggested reaction mechanism is

H2O2+I-= H2O+OI- ….(slow)

OI- +2H++I-= H2O+I2 …….(fast)

The first step being the slowest step is the rate determining step. The differential rate
becomes
𝑑[𝐻2𝑂2 ]
- = k[H2O2][I-], where k is the rate constant and other terms have their usual
𝑑𝑡
significance.

Therefore kinetically the reaction is of second order (overall). But if the concentration of
iodide ion is kept constant [by adding Na2S2O3 continuously whereby the S2O3- ions react
with the liberated iodine (I2) and regenerate I- according to the equation, I2+2 S2O32- = S4O62-
+2I-] the reaction becomes kinetically first order w.r.t.[ H2O2] only. Under this condition the
rate equation may be expressed as
𝑑[𝐻2𝑂2 ]
- = k1[H2O2], where k1=k[I-]
𝑑𝑡

If the initial (at time t=0) concentration of H2O2 is a and at time t, [H2O2] = (a-x), (where, x=
amount of H2O2 reacted), integration of the above equation gives
2.303 a
K1= log a−x
t

If both ‘a’ and (a-x) are represented in terms of their Na2S2O3 equivalent , we have

V0 ∝ a (where V0 is the titre value of thiosulphate for the total iodine liberated by a fixed
volume of H2O2 solution of concentration ‘a’ after complete decomposition).

And Vt ∝ x (where Vt is the titre value of the same thiosulphate solution for the iodine
liberated by the same H2O2 solution undergoing reaction at time t)
 The rate equation may then be express as
2.303
K1= log V0/V0-Vt
t

Or, log V0/V0-Vt=k1t/2.303

A plot of V0/V0-Vt vs. t will give a straight line passing through the origin and from the slope
k1 may be determined, k1= slope× 2.303.

Apparatus required:
1) 500 mL conical flask-2
2) 500 mL glass bottle for thiosulphate-1
3) Burette-1
4) 250 mL volumetric flask for KI-1
5) 10 mL pipette
6) Watch glass
7) Stop watch

Chemicals required: H2O2 , Sodium thiosulphate,KI,12N H2SO4,1% Ammonium

molybdate solution, Starch

Procedure:

1) Prepare 100 mL ‘2 volume’ H2O2 solution. in a glass bottle.

2) Prepare 250 mL of 0.1(N) sodium thiosulphate solution in glass bottle. Fill the burette
with sodium thiosulphate solution.
3) Take 10 mL 12 (N) H2SO4 in a 500 mL conical flask. Add 10 mL of prepared H2O2
solution followed by addition of 50 mL of water. Then add approximately 2 gm soli KI.
Add 2 mL 1% ammonium molybdate solution. Cover the conical flask with a watch
glass and keep it in dark for 1 minute. Add about 50 mL of water and titrate the
liberated iodine with sodium thiosulphate solution using starch as indicator. Record the
volume of thiosulphate (V0).
4) Take 250 mL of 0.4% KI solution (dissolve 1 gm KI in 250 mL deionized water) in a
500 mL conical flask. Add 15 mL 12(N) sulphuric acid and 5 L of freshly prepared
starch solution. Add 10 mL of prepared H2O2 solutions and at the time of half discharge
start stop watch.
Run immediately sodium thiosulphate solution from the burette from the burette into
the mixture in excess to discharge the blue colour. Wait for appearance of blue colour
and record the time (t) of reappearance of colour, volume of thiosulphate used (Vt).
Again run thiosulphate into the mixture in excess to discharge the blue colour, record
time (t) of reappearance of colour, and total volume of thiosulphate used (Vt).Take at
least 6 readings.
 5) Plot log [V0/( V0-Vt)] vs. t in millimeter graph paper and draw the best fit straight line
passing through the origin. Calculate ‘k1’ value from the slope of the graph.

Experimental result:
1) Room temperature
2) Preparation of 250 mL ̴ 0.1(N) sodium thiosulphate solution: Dissolve ̴ 6.2 gm
Na2S2O3 in 50 mL deionized water.
3) Determination of V0 :
V0 = titre value of thiosulphate for the total iodine liberated by 10 mL of’’2 volume’
H2O2 solution after complete decomposition.

4) Titre value of thiosulphate at different times:

Time(t) Time (t) in sec Volume of (V0-Vt) mL log V0/V0-Vt

S2O32- used (Vt
mL)

5) Plotting of graph:

log V0/V0-Vt
t(sec)

Calculation: Observed rate constant, k1=slope× 2.303

Conclusion: Therefore observed rate constant for the decomposition of H2O2 at …..
⁰C is …….Sec-1.

Experiment 5: Determination of heat of solution of oxalic acid from

solubility measurement
Theory:

The dissociation of a solid into a liquid is usually accompanied by heat change (heat is either
evolved or absorbed). For equilibrium between a solid non-electrolyte and its saturated
solution (pure solvent+ pure solute ⇌ Saturated solution) the thermodynamics equilibrium
constant (k) is given by

K=a2=S, solubility of the non electrolyte (assuming the saturated solution to be

ideal).

Since the solubility of a solid non electrolyte in its saturated solution at a particular
temperature, is a special case of the equilibrium constant we can determine the heat of
solution using the van’t Hoff equation, i.e,

……..(1)

Integrating equation (1) we get,

logS = - 2.303RT + constant

where R is gas constant, S is the solubility in moles per 1000g of solvent at temperature T,
and is the average heat of solution per mole over the temperature range used at the
saturation concentration.

Thus, by obtaining the solubility at different temperatures ,and by applying the van’t
Hoff equation,it is possible to determine the heat of solution ( ) from the slope of logS vs
1/T plot.In this experiment oxalic acid dihydrate is assumed to be non-electrolyte (being
weak acid) and its dissotiation is considered to be ideal.

Here, obtained differs from the heat of solution at infinite dilution,which is the heat of
solution usually given in literature,by an amunt equivalent to the heat of dilution from
saturation to infinite dilution.

Apparatus required:
1) 500 mL glass bottle and 100 mL volumetric flask
2) 10 mL pipette-1
3) 2 mL pipette-1
4) Burette-1
5) 250 mL stoppered bottle-1
6) 100 mL conical flask-1
7) Temperature controlled water bath
 Chemicals required: NaOH, Oxalic acid, Phenolphthalein

Procedure:
1) Prepare 100 mL 0.1(N) oxalic acid and 500 mL ̴ 0.2(N) NaOH solution. Determine the
strength of NaOH solution using standard Oxalic acid solution 0.1(N) and
phenolphthalein indicator.[Pipette out 10 mL 0.1(N) oxalic acid in a 100 mL conical
flask. Titrate the solution with prepared NaOH solution using 2 drops of
phenolphthalein until the solution becomes pink].
2) Prepare 50 mL saturated solution of oxalic acid in a 100 mL conical flask (or in a 250
mL stopper bottle) at room temperature. Pipette out 2 mL of the solution into 100 mL
conical flask using a tip (stuffed with cotton) attached to the pipette end( so as to
prevent any solid particle from entering into the pipette).Add 2 drops of
phenolphthalein and titrate using standard NaOH solution.
3) The saturated solution is brought to equilibrium at 3-4 different (higher) temperatures
(at a gap of 3⁰C) using temperature controlled hot water bath and at a lower
temperature using ice cooled bath.
4) At each temperature pipette out 2 mL of the solution into 100 mL conical flask using a
tip (stuffed with cotton) attached to the pipette end (so as to prevent any solid particle
from entering into the pipette). Add 2 drops of phenolphthalein and titrate using
standard NaOH solution.
5) From the titre value calculate solubility of oxalic acid in normality at different
temperatures. Transform the solubility into moles of oxalic acid /1000 gm of water for
each temperatures using the following equation:

Solubility (S)= moles/100 gm of water at temperature T

6) Plot log(S) vs 1/T and obtain ΔH (J/mol) from the slope.

Experimental data:
Determination of strength of NaOH using 0.1(N) oxalic acid:

Volume of 0.1(N) Burette reading Volume of NaOH Average volume of

oxalic acid (mL) Initial(mL) Final(mL) used(mL) NaOH(mL)

Strength of NaOH solution:

Determination of heat of solution of oxalic acid:

Sl.No. Temperature(⁰C) Burette reading log(S)

 Volume Initial(mL) Final(mL) Solubility (S) 1/T(K-
1
of NaOH moles/1000gm )
consumed

Calculation:

A straight line is obtained on plotting log(S) vs 1/T. From the slope = /2.303R, we can
calculate the average heat of solution per mole ( ) over the temperature range used.

GE1: Lab

1. Estimation of Fe(II) Ions by Titrating it with K2Cr2O7 Solution using Internal Indicator
Principle:
In acid medium Fe(II) in a solution may be estimated by direct titration with a standard solution
of K2Cr2O7 in presence of either H3PO4 or ammonium bifluoride (NH4HF2) using barium
diphenylaminesulphonate (BDS) as indicator. Under this condition K2Cr2O7 quantitatively
oxidizes Fe2+ to Fe3+:

Chemicals required:
a) standard N/20 K2Cr2O7 solution (to be prepared by accurate weighing)
b) N/20 Mohr’s salt solution
c) 4 N H2SO4
d) Syrupy H3PO4 or NH4HF2
e) saturated aqueous solution of BDS indicator

Procedure:
Pipette out an aliquot of 25 ml from the supplied Mohr’s salt solution in a 250 ml conical flask.
Add 25 ml of 4 N H2SO4, 3 ml syrupy H3PO4 (or 1-2 gm of NH4HF2) and 3-4 drops of BDS
indicator. Titrate with standard N/20 K2Cr2O7 solution until the colour of the solution just
changes from green to violet. Calculate the amount of iron in gm per litre present in the supplied
solution.
.

2. Estimation of Cu (II) ions iodometrically using Na2S2O3.

Principle:
Cu(II) is Iodometrically estimated by treating with an excess of KI solution and titrating the
liberated iodine with a sodium thiosulphate (Na2S2O3.5H2O) solution which is standardized
against a standard K2Cr2O7 solution.

Chemicals required:
a) Standard N/20 K2Cr2O7 solution (to be prepared by accurate weighing)
b) N/20 Na2S2O3.5H2O solution
c) 15% KI solution
d) 4 N H2SO4
e) Freshly prepared 1% starch solution

Procedure:
1. Standardization of the supplied Na2S2O3 solution:
Pipette out an aliquot of 25 ml from the supplied N/20 K2Cr2O7 solution in a 500 ml conical
flask. Add 25 ml of 4 N H2SO4 and 10 ml 15% KI solution. Cover the flask with a watch glass
and keep it in the dark for 5 minutes. Take it out and dilute the mixture with 150 ml distilled
water by properly rinsing the watch glass and the inner side of the conical flask. Titrate rapidly
the liberated iodine by the supplied thiosulphate solution until the color of the solution turns
straw-yellow. Add 2 ml of freshly prepared 1% starch solution and shake to obtain a
bluecoloured solution. Continue the titration until the blue colour is just discharged and a clear
light-green solution appears. Repeat the titration twice.
2. Estimation of Cu(II):
Pipette out 25 ml of the supplied solution in a 500 ml conical flask, dilute to 100 ml with
distilled water. Add 10 ml 15% KI solution and titrate the liberated iodine with standard
thiosulphate solution till the solution assumes a straw-yellow colour. Add 2 ml of starch
indicator. The solution turns intense blue. Continue the titration till the milky-white precipitate
of CuI appears. Record the titre value and calculate the amount of Cu(II) in the supplied
solution.
3. Estimation of oxalic acid by titrating it with KMnO4.
Principle:
In dilute H2SO4 medium MnO4- quantitatively oxidizes C2O42- to CO2 and it is reduced to
Mn2+:

It is an example of autocatalytic reaction in which Mn2+, a product of the reaction, acts as the
catalyst.
KMnO4 solution must be standardized against standard oxalic acid solution in 4 N H2SO4
medium at 70-80 0C. Purple coloured KMnO4 acts as a self-indicator. Its strength may be
calculated using the relation:

Chemicals required:
a) Standard N/20 oxalic acid, H2C2O4.2H2O [F.W. = 126.05 gm]; strength = (W/0.7879) N/20
where W is the weight of H2C2O4.2H2O per 250 ml of the solution
b) N/20 KMnO4 solution
c) 4 N H2SO4
d) N/20 oxalic acid solution
Procedure:
1. Standardization of KMnO4 solution:
Pipette out an aliquot of 25 ml of N/20 standard oxalic acid in a 250 ml conical flask. Add 25
ml of 4 N H2SO4 and heat to about 70-80 0C. Titrate the hot solution with supplied KMnO4
solution until the solution turns light pink colour that is stable for 30 seconds. Repeat the
titration to have a concordant reading. Calculate the strength of KMnO4 solution.
2. Estimation of oxalic acid solution:
Pipette out an aliquot of 25 ml of N/20 oxalic acid in a 250 ml conical flask. Add 25 ml of 4 N
H2SO4 and heat to about 70-80 0C. Titrate the hot solution with standard KMnO4 solution
until the solution turns light pink colour that is stable for 30 seconds. Repeat the titration to
have a concordant reading. Calculate the amount of the supplied oxalic acid in gram per litre.
4. Estimation of water of crystallization in Mohr’s salt by titrating with KMnO4.
Principle:
In dilute H2SO4 medium MnO4- quantitatively oxidizes Fe2+ to Fe3+ and itself is reduced to
Mn2+:

Mohr’s salt (Fe2+) solution may be estimated by titrating it in 4 N H2SO4 medium with a
standard solution of KMnO4 at room temperature in presence of H3PO4. Theoretical molecular
weight of Mohr’s salt [FeSO4.(NH4)2SO4.nH2O] = (284 + 18n); where n = number of
molecules of water of crystallization in Mohr’s salt. Since equivalent weight of Mohr’s salt is
equal to its molecular weight, so (284 + 18n) = E; where E = equivalent weight of Mohr’s salt.
By accurate weighing of Mohr’s salt and expressing the strength of its aqueous solution in gm
per litre, the number of water molecules present as solvent of crystallization can easily be
calculated.

Chemicals required:
a) standard N/20 oxalic acid solution
b) N/20 KMnO4 solution
c) 4 N H2SO4
d) syrupy phosphoric acid
e) 19.6 gm per litre Mohr’s salt solution
Procedure:
1. Standardization of N/20 KMnO4 solution:
Pipette out an aliquot of 25 ml of N/20 standard oxalic acid in a 250 ml conical flask. Add 25
ml of 4 N H2SO4 and heat to about 70-80 0C. Titrate the hot solution with supplied KMnO4
solution until the solution turns light pink colour that is stable for 30 seconds. Repeat the
titration to have a concordant reading. Calculate the strength of KMnO4 solution.
2. Standardization of Mohr’s salt solution:
Pipette out an aliquot of 25 ml of Mohr’s salt solution in a 250 ml conical flask. Add 25 ml of
4 N H2SO4 and 3 ml of syrupy H3PO4. Titrate the solution with standard KMnO4 solution
until the solution turns light pink colour that is stable for 30 seconds. Repeat the titration to
have a concordant reading. Calculate the number of water molecules present as solvent of
crystallization in Mohr’s salt.
5. Estimation of sodium carbonate and sodium hydrogen carbonate present in a
mixture.
Method: Indicators change colour within well defined pH ranges: phenolphthalein changes
from colourless to red in the range 8.0 - 9.6; methyl orange changes from red to yellow in the
range 3.1 - 4.4. The pH just after the neutralization point depends upon the strength of the acid
/ base, ie., their pKa / pKb.
Hydrochloric acid is standardized against previously standardized sodium hydroxide using
phenolphthalein as indicator: HCl + NaOH = NaCl + H2O.
The decomposition of carbonate by acid takes place in two steps: (i) CO32– + H+ = HCO3–.
(ii) HCO3– +H+ = H2O + CO2. At the end of step (i), the pH will be near about 8 or 9. After
the complete neutralization, the next drop of HCl will bring down the pH to near about 3 or 4.
The Na2CO3 content of the given sample is determined by titration against standard
hydrochloric acid using methyl orange as indicator: Na2CO3 + 2HCl = 2NaCl + H2O + CO2.
The equivalent weight of Na2CO3 is half its molecular weight (M). This means M/2 g of
Na2CO3 is equivalent to 1000 ml of 1 N HCl. It also means that the molarity of a solution of
Na2CO3 is half its normality.
The total amount of carbonate and bicarbonate in the mixture is determined by titration against
standard hydrochloric acid using methyl orange as indicator: CO32– + HCO3 – + 3H+ = 2H2O
+ 2CO2. Another titration using phenolphthalein as indicator will give the concentration of
carbonate. CO3 2– + H+ = HCO3 –. If the above titre values are Vm and Vp respectively, then
carbonate is equivalent to 2Vp and bicarbonate is equivalent to Vm – 2Vp.
Note that in the second titration the pH change is relatively slow near the neutralization point,
therefore the colour change is not sharp and the titration has to be done slowly.
Procedure:
(i) Standardization of hydrochloric acid: Transfer 25 ml of sodium hydroxide solution to a 250
ml conical flask using a pipette. Dilute with a little water (about 10 ml) and add 1-2 drops of
phenolphthalein. Titrate against hydrochloric acid taken in a burette. Repeat till concordant
titre values are obtained. Calculate the normality of the acid using the given normality of
sodium hydroxide.
(ii) Estimation of Na2CO3 content of the given sample: A : Weigh accurately about 0.6 g of the
sample (w g) into a 100ml standard flask. Dissolve in distilled water and make upto the mark
with distilled water. Titrate 25 ml of this solution against standard hydrochloric acid (normality,
Na) using 1-2 drops of methyl orange as indicator. When the colour becomes very faint yellow,
continue the titration dropwise until the colour becomes orange or a faint pink. Repeat the
titrations till concordant values (Va) are obtained. Calculate the normality (Nb = VaNa/25) of
the
sodium carbonate solution and the percentage purity of the sample. [Amount of Na2CO3 in the
weighed sample, m = (100/1000) x (Nb/2) x M ; percentage purity = 100m/w ]. B : Repeat the
above experiment by accurately weighing about 0.15 g of the solid sample to a 250 ml titration
flask (ie., conical flask), dissolving it in water and diluting to about 40 ml and titrating as
before. Calculate the percentage purity. The amount of Na2CO3 in the weighed sample is, m =
(Na x Va /1000) x (M /2).
Estimation of Na2CO3 and NaHCO3 in a mixture: Accurately weigh about 2.0 g of the mixture
and prepare a solution in distilled water in a 250 ml standard flask. Slowly titrate 25 ml of this
solution against standard hydrochloric acid using phenolphthalein as indicator. Repeat to
concordance (Vp ml). Now titrate the same volume using methyl orange as indicator. Repeat
to concordance (Vm ml). Calculate the number of equivalents, in the titrated volume(ie., 25.0
ml), of Na2CO3 (Ecarb = Na x 2 x Vp/1000) and NaHCO3 [ Ebicarb = Na x (Vm – 2Vp)/1000
]. Hence calculate the respective amounts and percentages in the given sample.

Qualitative Analysis of Single Solid Organic Compound

Background

Organic chemists often must identify unknown compounds. In some cases, such as a reaction, you
may have a good idea of what the compound in question is. However in other cases, such as when
you isolate a compound from a natural source, you may have no idea what the compound might
be. In this experiment you will determine the identity of an unknown compound. First,you will
need to purify your compound, then you will need to identify its functional group (it will contain
only one), and finally you will need to make a derivative of the compound. You will confirm your
results with boiling or melting point, IR, and NMR.

Impurities in your compound will make it extremely difficult to identify. Thus, before you do
anything else, you will need to make sure your unknown compound is pure. Consider each of the
following purification techniques you have learned over the courseof the year.

1. Recrystallization: Works well for solid compounds. You will need to find an appropriate
recrystallization solvent. Consider a variety of solvents and mixed solvent systems.
2. Distillation: Works well for liquids that have a boiling point of <250 °C. (Note: Fractional
distillation may be required if you suspect impurities close to the boiling point of your
unknown.)

3. Column Chromatography: Works well for UV active compounds. You will need to use TLC to
identify a solvent system that will separate your unknown from any impurities.

After you have purified your unknown, verify that it is pure enough to proceed by measuring the
boiling or melting point. Note that while you will not know what the melting point or boiling point
of your unknown should be, the narrowness is an excellent indicator of whether or not your
product is pure. Also pay attention to the appearance of your unknown and see if it has changed
(hopefully for the better) during the course of the purification process.

Once your unknown is pure, you will need to identify its functional group. Your unknown will
have one major functional group (alcohol, ketone, aldehyde, amide, amine, carboxylic acid, or
ester). Additionally, your unknown compound may or may not contain an aromatic ring. To
determine the functional group, it is recommended that you start with solubility tests, and then
conduct functional group classification tests. IR spectroscopy may also be useful at this point.
Solubility can sometimes provide a surprisingly useful amount of information. First, you will test your
unknown’s solubility in water. Compounds with 4 carbons or less will easily dissolve in water, whereas
compounds with 8 carbons or more will be insoluble. Compounds containing 5-­­­7 carbons may or
may not dissolve (often they will display “partial” solubility). If your compound dissolves in water, you
will also want to check the pH of the solution. Amines will typically be basic, and carboxylic acids will
typically be acidic. Most other compounds will be neutral. Compounds that are insoluble in water
should then be subjected to a solubility test in 5% HCl. Typically, only amines will be soluble in HCl
because they form water-soluble hydrochloride salts when they react with HCl. Compounds that are
not soluble in HCl, should be subjected to testing in basic solutions (5% NaOH and 5% NaHCO3). Both
strong and weak acids (Carboxylic acids and phenols) will be deprotonated by NaOH to form
water­soluble alkoxides. Only strong acids like carboxylic acids will react with NaHCO 3. Compounds
that are not soluble in base should then be reacted with a very strong acid, sulfuric acid (note that in
the case of sulfuric acid, “solubility” is also indicated by any type of reaction such as heat, gas
generation, or a color change). Compounds that cannot become protonated by sulfuric acid at all (i.e.,
alkanes, alkyl halides, and aromatic carbons) will still remain insoluble. These solubility tests are
summarized in the flow charts below.

The results from the solubility tests can significantly help in determining which classification tests
should then be performed, or at least narrow down the list. By no means do you need to conduct all
classification tests. In fact, you should do your best to select only tests that will provide you with
additional information about your unknown and/or confirm results. Also, make sure that your
glassware is clean and dry so you do not get any false positive or false negative results. Keep in mind
that a negative result for a classification test provides useful information, so be sure to keep track of
negative results as well as positive results. Also, for each classification test that you

perform, be sure to run a blank, and one or more controls. These will help you to determine if a
reaction actually occurred. A blank includes everything but the unknown, and a control includes a
compound for which the outcome is known in place of the unknown. Controls can be positive (a
compound you know will react) or negative (a compound that you know will not react). The
classification tests are summarized in the table below.

Functional group Test Test no Notes

Elemental analysis Lassaigne test C-1 Test for nitrogen,

sulphur, halogens

Amine Basicity test C-2 Test for aromatic amine

Bleaching powder test C-3

Dye test C-4

Nitro Reduction test C-5 Test for aromatic nitro

group
Muliken and barker test C-6

Amido Nitrous acid test C-7 Test for amido group

Hydrolysis test C-8

Phenolic –OH Ferric chloride test C-9 Test for phenolic-OH

Back dye test C-10

Carboxylic acid Bicarbonate test C-11 Test for carboxylic acid

Esterification test C-12

Aldehyde Benedict test C-13 Test for aldehyde

Tollens test C-14

Ketone 2,4-Dinitrophenyl C -15 Test for ketone

hydrazine test
At this point, you should be able to use your boiling or melting point data combined with the results of
your functional group data to develop a hypothesis as to what your unknown might be or at least
narrow down the list to only a few candidates. Note that due to the accuracy (or lack thereof) of our
thermometers, your boiling or melting points may be up to 15 C lower than the literature values.
°

Once your functional group has been determined, you will prepare a derivative of your unknown. To
prepare a derivative, you will select a suitable reaction that converts your unknown into a different
functional group for which the boiling or melting point is known. This is particularly useful because
compounds that have similar boiling or melting points will often have derivatives that differ
significantly in terms of boiling or melting point. You should then be able to identify your unknown
using this information.

Finally, you can confirm the identity of your product using IR and NMR. Note that these measurements
can be taken at any time during the course of the lab after you purified your product. In fact, it is
recommended that you conduct them sooner rather than later as they may provide valuable
information as to the identity of your unknown (e.g., IR may reveal your functional group).

Lab Notebook Preparation A

Before coming to lab on the first day of this experiment, the following items must be in your lab
notebook:

1. Title of experiment

2. Date the experiment is to be performed

3. Outline of your plan for determining the identity of your unknown

4. Hazards of and appropriate precautions for the safe handling of unknown organic
compounds
5. References

Lab Notebook Preparation B

Before coming to lab on the day you plan to prepare a derivative, the following items must be in your
lab notebook:

1. Title of experiment

2. Date the experiment is to be performed

3. List of possible unknowns

 4. The chemical reaction(s) you are attempting (with skeletal structures…R groups are okay if
you do not know the identity of your unknown yet)
5. For each reaction you are attempting, include a table with information about your starting
materials. Include molecular weight, molar equivalents, and mmoles to be used. For solids
include grams. For liquids, include grams, density, and volume. For solutions, include the
concentration and volume. (Note: You will not be able to completely fill in the table if you do
not know the identity of your unknown yet. If that is the case, list whatever data you can.)

6. Any relevant physical properties (i.e., melting points or boilingpoints of possible unknowns

and their derivatives)

7. Hazards of and appropriate precautions for the specific reaction(s) you are conducting

8. References

Safety Notes

 Assume that all unknowns are flammable and harmful by inhalation, ingestion, and skin

absorption. Do not inhale their vapors and avoid contact with eyes, skin and clothing.

Directions

1. Purify your unknown using distillation, recrystallization, or column chromatography. It is

recommended that purify the entire unknown provided so that you have enough pure material for
all of the tests.

2. Measure the boiling or melting point of your unknown to confirm its purity.

3. Confirm with your instructor that the boiling or melting point you obtained for your unknown is
within 15 °C of the reported literature value before proceeding.
4. Test the solubility of your unknown in water. (If your unknown is a solid, crush it into a fine
powder.)

a. Add approximately 30 mg of your unknown to a test tube or smallvial.

b. Add 1 mL of water and shake vigorously for approximately 30 seconds. If the unknown
appears to be soluble, test the pH of the solution and then skip to step 9.

5. Test the solubility of your unknown in 5% HCl. (If your unknown is a solid, crush it intoa fine
powder.)
a. Add approximately 30 mg of your unknown to a test tube or small vial.
 b. Add 1 mL of 5% HCl and shake vigorously for approximately 30 seconds. If the unknown
appears to be soluble, skip to step 9.

6. Test the solubility of your unknown in 5% NaOH. (If your unknown is a solid, crush it into a fine
powder.)

a. Add approximately 30 mg of your unknown to a test tube or small vial.

b. Add 1 mL of 5% NaOH and shake vigorously for approximately 30 seconds. If the

unknown appears to be insoluble, skip to step8.
7. Test the solubility of your unknown in 5% NaHCO3. (If your unknown is a solid, crush it into a
fine powder.)

a. Add approximately 30 mg of your unknown to a test tube or small vial.

b. Add 1 mL of 5% NaHCO3 and shake vigorously for approximately 30 seconds.

8. Note whether your unknown is soluble or insoluble and then skip to step 9.

9. Test the solubility of your unknown in concentrated H2SO4. (If your unknown is a solid, crush it

into a fine powder.)

a. Add approximately 30 mg of your unknown to a test tube or small vial.

b. Add 1 mL of concentrated H2SO4 and shake vigorously for approximately 30 seconds.

c. Note whether your unknown is soluble or insoluble. (Any indication of a reaction such as
heat, gas generation, or a color change also indicates solubility.)

10. Conduct classification tests as needed. See directions for specific tests below.

11. Confirm the identity of your functional group with your instructor before proceeding.

12. Prepare one or more derivatives of your unknown. See directions for specific derivatives below.

13. Measure the melting point of any derivatives.

14. Confirm with your instructor that the melting point you obtained for your derivative is within 15
°C of the reported literature value.

Classification Tests
C-1 Elemental Analysis

This reaction tests for the presence of nitrogen, sulfur and halogens (Cl / Br / I).

Safety Notes: Sodium can cause serious burns and the sodium-lead alloy may react violently with some
substances. Wear gloves, avoid contact, and keep the sodium-lead alloy away from other chemicals.

Recommended Controls: butylamine, acetamide, bromobenzene

Procedure:

1. In the fume hood or under a snorkel, place 0.25 g of 10% sodium-lead alloy in a small, dry

test tube held vertically by a clamp.

2. Melt the alloy with a Bunsen burner flame and continue heating until the sodium vapor rises
about 1 cm up the tube.

3. Using a Pasteur pipet, add 2 drops of the unknown (or 10 mg of a solid) directly onto the
molten alloy so that it does not touch the sides of the tube.

4. Heat gently to start the reaction, remove the flame until the reaction subsides, then heat the
tube strongly for a minute or two, keeping the bottom a dull red color.
5. Let the tube cool to room temperature.

6. Dropwise add 1.5 mL of water and heat gently for a minute or so until the excess sodium has
decomposed and gas evolution ceases.

7. Filter the solution through a Pasteur pipet with a small plug of cotton, wash the cotton with 1
mL of water, and combine the wash water with the filtrate. (Use a rubber bulb to expel any
liquid that adheres to the cotton.) The filtrate should be colorless or just slightly yellow. If it is
darker, repeat the fusion with stronger heating or more of the alloy.

To test for nitrogen:

1. Put 5 drops of the sodium fusion solution into a small test tube.

2. While stirring, add enough solid sodium bicarbonate, to saturate it (a little excess solid
should be present).
 3. Add 1 drop of this solution to a test tube containing 10 drops of PNB reagent
(p­nitrobenzaldehyde in dimethyl sulfoxide) and note any color change.

To test for halogens:

1. Acidify 10 drops of the sodium fusion solution with dilute nitric acid.

2. Boil it gently under the hood for a few minutes.

3. Add a drop or two of 0.3 M aqueous silver nitrate, and note the color and volume of any
precipitate that forms. (If a voluminous precipitate forms, let the precipitate settle and then
remove the solvent using a pipet.)

4. Add 2 mL of 3 M aqueous ammonia to the solid, shake vigorously, and note your
observations.

5. To test further for bromine and iodine, acidify 1 mL of the original sodium fusion solution
with 1 M sulfuric acid, boil for a few minutes, and add 0.5 mL of dichloromethane and a then a
drop of freshly prepared chlorine water. Shake and look for a color in the dichloromethane
layer.
To test for sulfur:

1. Put 1ml of sodium-extract into a test tube.

2. Add 1 ml dil. NaOH solution followed by 2-3 drops of sodium nitroprusside (freshly prepared).

Interpretation: In the PNB test, a purple color indicates the presence of nitrogen (green indicates

sulfur). In the halogen tests, formation of a voluminous precipitate on addition of silver nitrate

indicates that a halogen is present, and the color of the precipitate (a silver halide) may suggest which

halogen: white for chlorine, pale yellow for bromine, and yellow for iodine. If only a faint turbidity is

produced, it may be caused by traces of impurities or by incomplete sodium fusion. If the precipitate

is silver chloride, it will dissolve in aqueous ammonia; silver bromide is only slightly soluble and silver

iodide is insoluble. In the chlorine water test, a red-brown color is produced by elemental bromine

and a violet color by elemental iodine. In the sulfur test, a violet or purple color indicates the presence

of sulfur.

C-2 Basicity Test

This test is useful if you have already determined that you have an amine. It is used to distinguish
alkyl amines from aromatic amines.
Recommended Controls: p-toluidine, dibutylamine

Procedure for water-soluble compounds:

1. Dissolve 4 drops of your unknown (0.10 g of a solid) in 3 mL of water.

2. Measure the pH of the solution using pH paper.

Procedure for water‑insoluble compounds:

1. Dissolve 4 drops of your unknown (0.10 g of a solid) in 3 mL of a pH 5.5 acetate-acetic acid

buffer.

2. Mix thoroughly.

Interpretation: Water-soluble alkyl amines give pH values above 11, whereas water-soluble aromatic
amines have pH values below 10. Water-insoluble alkyl amines should dissolve in the buffer, but
water­insoluble aromatic amines will not dissolve.

C-3 Bleaching powder test

Procedure:

1. Dissolve 0.05 g of your unknown in 5 mL of water.

2. Add 3-4 drops of bleaching powder solution.
3. Shake vigorously.

Observation: Transient purple color which soon turns brown or light purple color.

C-4 Dye test:

NH2 N2Cl

NaNO2

HCl

N N
O
N2Cl O
Procedure:

1. 0.1 g of organic sample is dissolved in 5ml of dil. HCl.

2. The mixture is cooled at 0-5 ºC in an ice-bath.
3. Then add 1ml of ice cold solution of dil. NaNO2.
4. The mixture is added to ice-cold alkaline solution of β-naphthol (excess).

Interpretation: Red or orange red dye (brown or reddish purple or violet dye indicates the presence of
two amino groups; soluble dye indicates the presence of SO3H or Ar-OH along with Ar-NH2 group).

C-5 Reduction followed by Dye test

NO2 NH2.HCl

Sn/HCl

Zn/HCl

1. A mixture of 0.1 g of organic sample, few pieces of granulated tin or zinc and 3ml of Conc. HCl is
warmed gently with occasional shaking till the reaction is complete.
2. The mixture is cooled.
3. Filtered, if required, diluted and diazo-coupling reaction is performed.

Observation: Brilliant red or scarlet dye obtained.

Interpretation: A group reducible to –NH2 group. Normally an aromatic nitro compound is indicated.

C-6 Mulliken and Barker Test

Procedure:

1. 0.1 g of organic sample is dissolved in 5 ml of 50% alcohol.

2. A little solid NH4Cl or 10% CaCl2 solution and a pinch of Zn-dust is added to it.
3. The mixture is boiled for a few minutes.
4. Then the mixture is cooled and allowed to stand for 5 minutes and then filtered.
5. With the filtrate following three tests are performed:
a) A portion of the solution is added to Tollen’s reagent and then warmed in a water bath.

b) Two drops of benzoyl chloride and 2 drops of conc. HCl are added to another portion of the filtrate
followed by 12 drops of FeCl3 solution.

c) The last portion of the filtrate is warmed with a little Fehling’s solution.

Observation: From the part (a), a silver mirror or black or grey precipitation is obtained. From part (b),
a wine-red color of ferric hydroxamate is present, from last part (c), a red precipitation is obtained.

Interpretation: A nitro group is reduced to –NHOH that oxidises Ag+ to Ag, and Cu++ to Cu+ (Cu2O)

C-7 Nitrous Acid Test

Procedure:

1. A little of the aqueous solution of organic sample is treated with a few drops of HNO2 (NaNO2
and HCl).

Interpretation: Effervescence due to evolution of N2 gas. Indicates presence of –NH2 (aliphatic) / -

CONH2

C-8 Hydrolysis Test

Procedure:

0.2 g of organic sample is heated with 2 ml of 50% NaOH solution.

Observation: Characteristics smell of NH3 which turns mercurous nitrate paper black or copper
sulphate paper deep blue.

Interpretation: Primary amide (-CONH2) is hydrolysed to organic acid and ammonia. The latter gives
the above observations.

C-9 Ferric Chloride [Must be freshly prepared]

This reaction tests for the presence of phenols.

Recommended Control: Phenol / resorcinol

Procedure:

1. Dissolve 1 drop of the unknown (40 mg of a solid) in 1 mL of water. If (you know based on the
results of your solubility tests that the unknown is insoluble in water, use 0.5 mL of water and
0.5 mL of methanol instead of 1 mL of water.)
2. Add two drops of 2.5% ferric chloride solution.

Interpretation: Formation of an intense red, green, blue, or purple color suggests a phenol or an easily
enolizable compound (such as an -diketone or -ketoester). Yhe colour specis is (ArO)6Fe3-

Some phenols do not react under these conditions.

C-10 Back Dye Test

This reaction tests for the presence of phenols.

Procedure:

1. A few drops of aniline dissolved in dil. HCl.

2. Few drops of cold dil. NaNO2 solution is added.
3. Then the clear solution is added to the cold solution of organic sample in NaOH.

Observation: A brilliant red dye is obtained.

Interpretation - Phenolic OH group present and confirmed; test is very good for resorcinol, naphthols
etc.

C-11 Bicarbonate Test

This reaction tests for the presence of carboxylic acid ( or any good acidic compounds e.g., picric acid,
2,4-dinitrophenol)
Procedure:

1. A small amount of organic sample is sprinkled over aqueous solution of sodium bicarbonate.

Interpretation: Effervescence due to the evolution of CO2.

C-12 Esterification Test

This reaction tests for the presence of carboxylic acid.

1. 0.5 g of organic sample is taken in a dry test tube.

2. To this, add 1 ml of dehydrated ethanol.
3. Then 2-3 drops of conc. H2SO4 is added and heated for 5 minutes in a water bath.
4. The mixture is then poured into a beaker containing large volume of Na2CO3 solution.

Interpretation: Characteristics sweet smell (fruity) of ester.

C-13 Benedict’s Test

This reaction tests for the presence of aldehydes. Note that most ketones and aromatic
aldehydes will not react.

Recommended Controls: butanal

Procedure:

67
 1. Add 2 drops of the unknown (80 mg if it is a solid) to 2 mL of water.

2. Add 2 mL of Benedict’s reagent.

3. Heat the mixture to a boil.

4. Observe if a precipitate forms, and note its color.

Interpretation: Benedict’s reagent contains copper(II) sulfate, sodium citrate, and sodium
carbonate. Aldehydes will react with the Cu2+ from the copper(II) sulfate to form copper(I) oxide
which appears as a yellow or orange precipitate (it may look a little green in the blue reaction
solution). Note that most ketones and aromatic aldehydes will not react.

C-14 Tollen’s Test

This reaction tests for the presence of aldehydes. [#a-Hydroxyketones e.g., benzoin responds to
this test]

Recommended Controls: benzaldehyde

Procedure:

1. Measure 2 mL of 0.3 M aqueous silver nitrate into a test tube and add 1 drop of 3 M
sodium hydroxide.

2. Add 2 M aqueous ammonia drop by drop, with shaking, until the precipitate of silver
oxide just dissolves (avoid an excess of ammonia).
3. Add 1 drop of the unknown (40 mg of a solid) to this solution, shake the mixture, and let
it stand for 10 minutes. (If a silver mirror is observed at this point, this is considered a
positive result.)

4. Heat the mixture in a 35 °C water bath for 5 minutes.

5. Immediately after the test has been completed, dissolve any solid residue in 1M nitric
acid and then dispose of the solution in the designated waste container.

Interpretation: Formation of a silver mirror on the inside of the test tube is a positive test for an
aldehyde. (Note that if the tube is not sufficiently clean, a black precipitate or a suspension of
metallic silver may form instead.)

68
C-15 2,4-Dinitrophenylhydrazine

This reaction tests for the presence of aldehydes and ketones.

Safety Notes: 2,4­Dinitrophenylhydrazine (DNPH) is harmful if absorbed through the skin. Wear
gloves and avoid contact.

Recommended Controls: cyclohexanone, benzaldehyde

Procedure:

1. Dissolve 1 drop of the unknown (40 mg of a solid) in 1 mL of 95% ethanol (use more
ethanol if necessary to completely dissolve the unknown).

2. Add this solution to 2 mL of the DNPH reagent.

3. Shake and let the mixture stand for 15 minutes or until a precipitate forms. (If a
precipitate is observed at this point, this is considered a positive result.)

4. Scratch the inside of the test tube and observe if a precipitate forms, and note its color.

Interpretation: Formation of a crystalline yellow or orange-red precipitate indicates an aldehyde

or ketone. The color of the precipitate may give a clue to the structure of the carbonyl compound
(unconjugated aliphatic aldehydes and ketones usually yield a yellow precipitate, while aromatic
and α,β-unsaturated aldehydes and ketones yield a orange-red precipitate).

69