3.

Some Analytical Techniques

3.1 Introduction
There has been a systematic development The insoluble part remaining on the filter paper
in the techniques used for analysis of chemical is called residue and the liquid collected in the
substances. In this chapter we are going to beaker is called filtrate.
look into basic analytical techniques, namely, Filtration under suction : When filtration is
purification and separation techniques. carried out using a vacuum pump it is called
Chemical substances occur in nature in impure filtration under suction. It is a faster and more
stage. Also, when synthesized in the laboratory efficient technique than simple filtration.
they are obtained in crude and impure form. The assembly for filtration under suction
Before investigating their composition and consists of a thick wall conical flask with a
properties it is essential to obtain them in side arm. The flask is connected to a safety
the pure form. Methods of purification and bottle by rubber tube through the side arm. The
separation of compounds depend on the safety bottle is used to prevent sucking of the
difference in their physical properties. filtrate into suction pump. A special porcelain
3.2 Purification of solids funnel called Buchner funnel is fitted on the
A solid substance may contain two types of conical flask with the help of a rubber cork as
impurities, those (i) which are soluble in the shown in Fig. 3.2.
same solvent as the main substance and (ii) The Buchner funnel has a porous circular
which are not soluble in the same solvent as bottom. A circular filter paper of correct size
the substance. The second type of impurity is placed on the circular porous bottom of the
can be separated easily using a suitable Buchner funnel and the funnel is placed on the
solvent to dissolve the main compound when flask. It is moistened with a few drops of water
the impurities remain undissolved and can be or solvent. Suction is created by starting the
separated by a simple process called filtration. pump and filtration is carried out. Crystals are
This process is similar to separating tea leaves collected on the filter paper and filtrate in the
from a decoction of tea, or sand from mixture flask.
of sand and water. Filtration is carried out with
the help of a filter paper cone placed in a funnel
as shown in the Fig. 3.1. A circular piece of
filter paper is folded to form a cone and fitted
in the funnel. The funnel is fixed on a stand
and a beaker kept below it. The paper is made
moist, the solution to be filtered is poured on
the filter paper.

Mixture of Filter
water and paper Residue
sand

Filter
funnel
Filtrate

Fig. 3.1 : Separation of mixture of water and

Fig. 3.2 : Setup for filtration under suction
sand by simple filtration
27
3.2.1 Crystallization : When a crude solid is Choice of the solvent : The solvent to be
made of mainly one substance and has some used for crystallization must have following
impurities, it is purified by the process of properties :
crystallization. It is done in four steps: 1. The compound to be crystallized should
(i) Prepartion of saturated solution : A be least or sparingly soluble in the solvent
saturated solution is a solution which cannot at room temperature but highly soluble at
dissolve addtional quantity of solute. A high temperature.
saturated solution of the crude solid is prepared 2. Solvent should not react chemically with
by boiling it in a small but sufficient quantity the compound to be purified.
of a suitable solvent. On doing so the main 3. Solvent should be volatile so that it can be
solute forms an almost saturated solution, removed easily.
but the solution is not saturated with respect Water, ethyl alcohol, methyl alcohol, acetone,
to the soluble impurties, as they are in small ether or their combinations are generally used
proportion. as solvent for crystallization. The choice is
(ii) Hot filtration : The above solution is done by trial and error method.
quickly filtered while hot. Filtration under Activity : Crystallization of common salt from
suction allows rapaid filtration. Undissolved impure sample.
impurities get removed in this process as Many times the common salt obtained from the
residue. market may contain some siliceous matter and
(iii) Cooling of the filtrate : The hot filtrate other impurities. These can be removed and
is allowed to cool. Solubility of a substance larger crystals of pure NaCl can be obtained.
decreases with lowering of temperature. As a Apparatus : Beaker, glass rod, funnel, filter
result, the filtrate becomes supersaturated with paper and stand, evaporating dish, etc.
respect to the main dissolved solute. The excess Materials : Impure market salt or a mixture of
quantity of the dissolved solute comes out of the salt and fine sand, etc.
solution in the form of crystals. The dissolved Procedure : Arrange apparatus as shown in
impurities, however, do not supersaturate Fig. 3.3. Take some water in a beaker. Add salt
the solution, as their quantity is small. These to the beaker and stir it with a glass rod. Add
more salt and stir till no more salt dissolves.
continue to stay in the solution in dissolved
Heat the solution. Filter the hot solution,
state even on cooling. The separated crystals
insoluble impurities will remain on the filter
are, therefore, free from soluble impurities as
paper. Collect the filtrate in an evaporating
well.
dish and allow to cool. Crystals of pure salt
(iv) Filtration : The crytals of the pure
NaCl will separate leaving soluble impurities
substance are separated by filtration. The
in the mother liquor. Filter the solution and
filtrate obtained is called mother liquor. The
collect the crystals on the filter paper and dry
crystals so formed are free from soluble as well
them.
as insoluble impurities.
Filter paper

Fig. 3.3 : Process of crystallization

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 Impure copper sulphate can be purified 3.3.1 Simple distillation :
by crystallization using water as solvent. Liquids which boil without decomposition
Similarly, Benzoic acid can also be purified by at atmospheric pressure are purified by the
crystallization using water as solvent. process of simple distillation. In this process
3.2.2 Fractional crystallization : the liquid is first converted into its vapour by
Two or more substances in a mixture boiling and then the vapour is condensed back
can be separated by fractional crystallisation into liquid by cooling, and the pure liquid
process. Fractional crystallisation is a process trickles down in the reciever.
wherein two or more soluble substances The apparatus used for simple distillation
having widely different solubilities in the same is shown in Fig. 3.4. It consists of round bottom
solvent at room temperature are separated by flask fitted with a cork having a thermometer.
crystallization. The flask has a side arm through which it is
Mixture of two solutes A and B are connected to a condenser. The condenser has
dissolved in a suitable hot solvent to prepare a jacket with two outlets through which water
a saturated solution. The saturated solution is circulated. The liquid to be distilled is taken
is filtered to remove dust particles and then in the round bottom flask fixed by clamp. The
allowed to cool. As the solution cools, the flask is placed in a water bath or oil bath or
solute which is less soluble crystallizes out first. sometimes wire gauze is kept on a stand as
The crystals are filtered, washed with solvent shown in Fig 3.4.
and dried. The mother liquor is concentrated The condenser is connected to receiver to
by evaporating the solvent. The second solute collect the purified liquid. Care is taken that
crystallizes from the mother liquor. These the bulb of the thermometer is just below the
crystals are filtered to obtain the separated and side arm of the round bottom flask.
purified second component. The flask is heated. As the boiling point
3.3 : Distillation : Distillation is an important of the liquid is reached it starts boiling and
method used to separate. (i) Volatile liquids the vapors rise to the neck of the flask and
from non-volatile impurities (ii) Liquids having pass through the side arm into the cooler
sufficient difference in their boiling point. parts of the condenser, which is kept cool
by circulating water through its jacket. The
vapours condense and the liquid is collected
in the receiver.
Activity : To separate the components of a
liquid mixture containing acetone (b. p. 560C)
and water (b. p. 100 0C)
Apparatus : Distillation flask with condenser,
two receivers, thermometer, etc.
Chemicals : mixture of acetone and water.
Principle : Acetone and water are two
miscible liquids having a wide difference
in their boiling points. Acetone boils at 56 0
C while boiling point of water is 100 0C.
When the mixture of acetone and water is
heated and temperature of the mixture reaches
56 0C acetone would distil off. When all
acetone distils out and when the temperature
rises to 100 0C water would to distil out.
Fig. 3.4 : Simple distillation
29
Procedure : Take the mixture of water and Suppose we have a mixture of two liquid
acetone in the distillation flask and arrange (A) and (B) having boiling points 363 K and
the apparatus. Heat the flask on a water bath 373 K respectively. A is more volatile and B is
carefully. At 56 0C acetone will distil out, less volatile. As the mixture is heated, vapors
collect it in reciever number 1. After all acetone of (A) along with a little of (B) rise up and
distilled, change the receiver. Discard a few come in contact with the large surface of the
ml of the liquid. As the temperature reaches fractionating column. Vapors of (B) condense
100 0C water will begin to distil. Collect this in rapidly into the distillation flask. While
reciever number 2. passing through the fractionating column
3.3.2 Fractional Distillation : there is an exchange between the ascending
If in a mixture the difference in boiling vapors and descending liquid. The vapors of B
points of two liquids is not appreciable, they are scrubbed off by the descending liquid, this
cannot be separated from each other using the makes the vapors richer in (A). This process is
simple distillation assembly. repeated each time the vapors and liquid come
To separate such liquids, the process in contact with the surface in the fractionating
called fractional distillation is employed in column. Rising vapors become richer in (A)
which a special assembly is used (see Fig. and escape through the fractionating column
2.5(a)). In this assembly the distillation flask and reach the condenser while the liquid in the
is fitted with a fractionating column (Fig. 3.5 distillation flask is richer in B. The separated
(b)). Hence, the vapours first pass through components are further purified by repeating
the fractionating column. Vapours of more the process. Mixtures of acetone (b.p. 329 K)
volatile liquid with lower boiling point rise up and methyl alcohol (b.p. 337.7 K); acetone
more than the vapours of liquid having higher and benzene (353 K) can be separated by
boiling point. fractional distillation.

Fig. 3.5 (a) :Fractional distillation Fig 3.5 (b) : Fractionating Columns

30
 This process is used in petroleum industry If the solute is less soluble in organic phase
to separate different fractions of crude oil. then a technique called continuous extraction
3.3.3 Distillation under Reduced Pressure : is used where the same amount of organic
Liquids having very high boiling point or solvent is used repeatedly for extraction. This
those which decompose on heating are purified technique involves continuous distillation of
by carrying out distillation under reduced the solvent within the same assembly. Thus use
pressure. In this method the liquid is made to of large quantity of organic solvent is avoided.
boil at a temperature which is below its normal
boiling point by reducing the pressure on Internet my friend
the surface of the liquid. Pressure is reduced
using a water pump or vacuum pump. In soap Get more information about continuous
industry glycerol is separated from soap by extraction/soxhlet extraction from
using this technique. YouTube.Royal Scociety of chemistry
3.4 Solvent Extraction : When an organic Soxhlet extraction.
substance is present in an aqueous solution, it
can be extracted from that solution by shaking it 3.5 Chromatographic techniques :
with an organic solvent in which the substance Chromatography is a technique used to
is more soluble. The organic liquid should be separate components of a mixture, and also
immiscible with water and be able to form two purify compounds. The name of the technique
layers. In this process the solute distributes comes from the Greek word Chroma meaning
itself between two immiscible liquids. From the Colour.
aqueous phase the solute gets extracted in the In 1903, Tswett discovered this technique
organic phase. Extraction of compound takes for separating the coloured components
place based on the difference in solubility of found in plants. The principle of separation
compound in two liquids (See Fig. 3.6). On of substances in this technique is similar to
shaking for a few times with small volumes of solvent extraction i. e. distribution of the
organic phase, most of the solute gets extracted solutes in two phases. In chromatography we
into the organic phase. The organic solvent is, use two phases for separation. (a) Stationary
then, removed by distillation and the solute is phase and (b) Mobile phase. This technique
collected. is based on the difference in rates at which
The solvent extraction process is important components in the mixture move through the
as it helps clean separations in a short time span. stationary phase under the influence of the
mobile phase. First the mixture of components
is loaded at one end of the stationary phase
and then the mobile phase, which is a pure
solvent or a mixture of solvents, is allowed to
move over the stationary phase. Depending on
the relative affinity of the components toward
the stationary phase and mobile phase they
remain on the surface of the stationary phase
or move along with the mobile phase, and
gradually get separated.
The stationary phase can be a solid or
a liquid. Depending on the stationary
phase, chromatography is classified into
Adsorption Chromatography and Partition
Fig. 3.6 : Solvent Extraction
Chromatography.
31
3.5.1 Adsorption Chromatography : This The solutions of these components are collected
type of Chromatography is based on the separately. On evaporating the solvent the
principle of Differential Adsorption. Different solutes can be recovered.
solutes are adsorbed to different extent on the ii. Thin Layer Chromatography : A thin
stationary phase. Adsorption Chromatography layer (0.2 mm thick)of adsorbent silica gel or
is of the following two types. alumina spread over a glass plate acts as the
i. Column Chromatography : This type stationary phase. The plate is called the TLC
involves the separation of components over plate or chromplate. The mixture of solutes
a column of stationary phase. The stationary is applied on the Chromplate as a small spot
phase material can be Alumina, Silica gel. about 2 cm from one end of the plate as shown
A slurry of the stationary phase material is in Fig. 3.8.
filled in a long glass tube provided with a The plate is then placed in a closed jar
stopcock at the bottom and a glass wool plug containing the mobile phase such that the
at the lower end. The mixture to be separated spot is well above the mobile phase. As the
is dissolved in a small amount of appropriate mobile phase rises up the components of
solvent and is then loaded on top of the the mixture move along with it. They move
adsorbent column. A suitable mobile phase upto different distances depending upon their
which could be a single solvent or a mixture degree of adsorption and thus get separated.
of solvents is then poured over the adsorbent If the components are colored they appear as
column. The mixture along with the mobile separated colored spots on the plate. If the
phase slowly moves down the column. The components are not colored but have property
solutes get adsorbed on the stationary phase of fluorescence they can be visualised under
and depending on the degree to which they are UV light, or the plate can be kept in a chamber
adsorbed, the solutes get separated from each containing a few iodine crystals. The Iodine
other. The most strongly adsorbed component vapors are adsorbed by the components and
is retained on the column and others move
down the column to various distances forming
bands as seen in Fig. 3.7. The component
which is less strongly adsorbed is desorbed
first and leaves the column first, while the
strongly adsorbed component is eluted later.
Fig. 3.8 (a)Stages in Thin Layer
Chromatography

FIG 3.7 Column Chromatography. Different

stages of separation. Fig. 3.8 (b) Developed chromatogram.
32
the spots appear brown. Amino acids are the colored components are visible as colored
visualised by spraying the plate with a solution spots and the colourless components are
of ninhydrin. This is known as spraying agent. observed under UV light or using a spraying
3.5.2 Partition Chromatography : In this agent.
type of chromatography the stationary phase Retention factor (Rf) : Migration of the solute
and mobile phase both are liquids. Separation relative to the solvent front gives an idea about
of components takes place by continuous the relative retention of the solutes on the
differential partitioning of the components stationary phase. This is termed as the Rf of
between the stationary and mobile phases. the solute.
For example, Paper Chromatography : In this Distance travelled by the solute from the base line
technique a special quality paper, Whatmann Rf = Distance travelled by the solvent from the base line
paper number 1, is used. The water trapped in
the fibres of the paper acts as the stationary
phase. The solution of mixture is spotted on
the strip of the Chromatography paper at the
about 2 cm from one end of the paper using a
glass capillary. The paper is then suspended in
a chamber containing the mobile phase taking
care that the spot does not dip in the mobile
phase (Fig. 3.9).
The mobile phase rises up by capillary
action and flows over the spot. Partitioning of Fig. 3.10 : Retention factor
the components takes place between stationary
phase (water) and the mobile phase.
Different solutes are retained differently Internet my friend
on the paper depending on their selective
partitioning between the two phases. This Column chromatography
developed paper strip is the chromatogram. https ://youtube/KqQITxvFDLB8
Similar to TLC (Thin layer chromatography)

Fig. 3.9 : Stages in Paper chromatography

33
 Exercises
1. Choose the correct option F. What will happen if the upper outlet of
A. Which of the following methods can be the condenser is connected to the tap
used to seperate two compounds with instead of the lower outlet?
different solubilities in the same solvent? G. Give names of two materials used as
a. Fractional crystallization stationary phase in chromatography.
b. Crystallization H. Which properties of solvents are useful
c. Distillation for solvent extraction?
d. Solvent extraction I. Why should spotting of mixture be done
B. Which of the following techniques is above the level of mobile phase ?
used for seperation of glycerol from soap J. Define : a. Stationary phase b. Saturated
in soap industry ? solution
a. Distillation under reduced pressure K. What is the difference between simple
b. Fractional distillation distillation and fractional distillation?
c. Filtration L. Define a. Solvent extraction b.
d. Crystallization Distillation.
C. Which technique is widely used in M. List the properties of solvents which
industry to seperate components of make them suitable for crystallization.
mixture and also to purify them ? N. Name the different types of
a. Steam distillation Chromatography and explain the
b. Chromatography principles underlying them.
c. Solvent extraction O. Why do we see bands separating in
d. Filtration column chromatography?
D. A mixture of acetone and benzene can be P. How do you visualize colourless
seperated by the following method : compounds after separation in TLC and
a. Simple distillation Paper Chromatography?
b. Fractional distillation
Q. Compare TLC and Paper
c. Distillation under reduced pressure
Chromatography techniques.
d. Sublimation
3. Label the diagram and explain the process
E. Colourless components on chromatogram
in your words.
can not be observed by the following :
a. Using UV light
b. Using iodine chamber
c. Using the spraying reagent
d. Using infrared light
2. Answer the following
A. Which of the following techniques is
used for purification of solid organic
compounds?
a. Crystallisation b. Distillation
B. What do you understand by the terms
a. residue b. filtrate.
C. Why is a condenser used in distillation
process?
D. Why is paper moistened before Activity :
filtration?
Use any one analytical technique in
E. What is the stationary phase in Paper
labroratory and discuss it in groups.
Chromatography?
34