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SEPARATION METHODS
General Aspects of
Separation Methods
Structure
1.1
Introduction
Objectives
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
1.10
1.11
Summary
Terminal Questions
Answers
1.1
INTRODUCTION
Classical Methods
Objectives
After studying this Unit, you should be able to
1.2
The separations play a key role in the various branches of science and technology but
they themselves form a unified branch of science. The subject of separation science
essentially deals with the physical and chemical phenomena involved in achieving the
separations. The outcome of separations is very much dependent upon the
physicochemical principles resulting into separations. It also involves the
development, application and reproducibility of various separation processes. The
separations have assumed such a paramount importance that a common man has
become familiar with the meaning of word separation. However, with its requirement
and usage, the definition of separation has become a little complex. In the simplest
terms, separation is defined as an operation in which a mixture is divided into at least
two components having different compositions. But this particular definition has a
limitation as it does not cover chiral separations in which molecules of same
composition and chemical structure are involved. The molecules differ only in their
stereochemistry. Therefore, a broader definition of separation will be as under:
Separation is a process by which a mixture is divided in at least two
components with different compositions or two types of molecules with the same
composition but different stereochemistry.
After having learnt the definition of separation in true chemical sense, you should be
clear about the different objectives for achieving separations:
i)
ii)
10
Under (ii), for obtaining the pure materials from complex mixtures, the constituent
with the desired purity may be obtained by applying a single process or using a
number of separation techniques. In some cases, in order to attain the required level of
purity, the separation steps of the same process may have to be repeated.
General Aspects of
Separation Methods
The mixtures to be separated vary largely in terms of their complexity. They may
contain constituents which differ in their molecular weights, solubility in a solvent,
volatility or other properties. The sizes of species may range from atomic dimensions
through organic molecules and macromolecules to molecular aggregates.
A large number of separation methods are available that utilize selected
characteristics as means of separation. Each of these methods can be further
subdivided into different techniques using unique characteristics. In certain cases, the
properties of the constituents may be so different that very simple techniques of
separation can be applied. A very simple example, in this regard, is the recovery of
common salt from sea water. However, in other cases, the properties of constituents
may be so similar that the separation becomes a tedious job. A very well known
example of a difficult separation is the separation of Zn (IV) and Hf (IV). The other
example in this category is the separation of optical isomers.
Another important parameter which is critical in choosing the separation is the
amount of mixture available. In some cases, the amount may be a few molecules.
However, in industrial processes it may run in tonnes.
SAQ 1
What are the main applications of separations?
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SAQ 2
Define separation in the real chemical sense.
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1.3
We have learnt that the separation science deals with a variety of problems at hand.
Now, it may be necessary to illustrate the utility of separations by citing example from
daily life to various branches of science and technology. The number of even the
important examples is so large that they cannot be recounted here. However, by citing
a few, you will be able to appreciate the significance of separations.
11
Classical Methods
The art of separations is not new to the present day community of scientists. Even our
ancestors were familiar with the various separation methods and were using them for
their daily needs. A very typical example is the distillation of alcohol for drinking and
other purposes. Isolation of various dyes for coloring purposes from natural materials
is a good testimony of their skill for achieving separations. They were also quite
proficient in isolating metals for their use from ores by applying different separation
procedures. A number of drugs used to be isolated from plants and herbs. The
treatment of waters by solid adsorbents is as old as the civilization. There are records
available that in the time of Aristotle, sand filters were used for the purification of sea
water and impure drinking water. Moses used tree branches to make bitter water
sweet.
Separation processes play a key role in our daily life. We remove undesirable gases
and particles from the air we breathe. The municipal drinking water undergoes several
purification steps. It is well known that the identification and removal of contaminants
from food are largely possible due to separation processes. One of best examples of
use of separations in industry is the availability of a variety of products from crude
petroleum. The nuclear age did really take off due to improvements in the methods of
separation of U238 and U235. The requirement of high purity materials in industry,
particularly for semiconductor, is met due to advancements in separation processes.
It has been possible to understand the different biochemical processes taking place in
our body due to advancements in separation processes. The separation processes have
given a unique gift in the form of artificial kidney. The success in the studies on
human genome and proteomics owes a great deal to advancements in separation
sciences. The synthesis of different medicines is possible due to efficiency of
separation processes. The identification of explosives has been possible due to a key
role of separations.
In a nutshell, there is hardly any walk of life where separations do not play their
dominant role.
It is well known that a large number of separation methods fall under the category of
chromatography and that is why the separation methods have become synonymous
with chromatography. Thus, before proposing a classification of separation methods,
it may be important to give here a brief description of evolution of chromatography.
SAQ 3
Give two examples of separations beneficial for our environment.
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1.4
EVOLUTION OF CHROMATOGRAPHY
12
been presented. Following this unit, some of the units discuss important
chromatographic methods in detail. In order to keep the text of this particular unit
properly jointed, a brief idea about the evolution of chromatography is being
presented here.
The subject of chromatography had a very modest beginning. The principles and the
applications were reported in 1906 by a Russian botanist, Mikhail Tswett. He
described the resolution of chlorophylls and other plant pigments in a plant extract. A
petroleum ether solution of chlorophyll was passed through a calcium carbonate
column. By passing the solvent through the column, the pigments were resolved into
various zones. This separation became practical if after the pigment solution, the pure
solvent was allowed to pass through the column. Such a preparation was termed as the
chromatogram and the corresponding method, the chromatographic method. The
term chromatography is derived from the Greek words chroma and graphy meaning
colour writing. The discovery of chromatography was made with the separation of
coloured compounds but its potential for colourless compounds was realized. Tswett
himself anticipated the potential of this technique for a wide variety of compounds.
This technique is now termed as liquid-solid adsorption chromatography.
General Aspects of
Separation Methods
Mikhail Tswelt
(1872-1919)
R. L. M. Synge
(1914-1994)
SAQ 4
Define chromatography.
A. J. P. Martin
(1910-2002)
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13
Classical Methods
SAQ 5
Cite the main processes which are responsible for separations by chromatography.
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1.5
The subject of separation science has grown very large in its dimensions. A variety of
methods have come up on the forefront to meet the different needs. A number of well
known techniques have undergone modifications. The growth in the number of
separation methods can be attributed to the following factors:
The separation methods are generally named after the forces or phenomena that give
rise to separation. In this respect, we can cite the examples of precipitation,
crystallization, extraction, adsorption and ion exchange. At times, the name is used to
reflect upon a distinct form of operation. Here, you can mention techniques like
filtration, distillation, and chromatography. Chromatography, for example, can
employ any number of forces such as adsorption, partition, ion exchange and size
exclusion.
The above discussion makes it clear that one of the classifications of the separation
methods may be based on the property which results into separations. The other
approach that can be adopted for classification is based on the physicochemical
phenomena responsible for separation. To simplify, we can further divide these
phenomena in two major categories: equilibrium processes and rate processes.
Thus, we can propose two types of classifications for separation methods.
It may be pertinent to point out that the details of the methods will not be discussed
here because either some of the methods are well known or a few of them are being
discussed in details in other units of this course.
SAQ 6
What are the main reasons for the growth in the number of separation mehods?
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14
General Aspects of
Separation Methods
SAQ 7
What are the two main criteria employed for classifying the different separation
methods?
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1.6
The main properties which result into segregation of components of the mixture are:
Volatility,
Solubility,
Partition,
Ion exchange,
Surface activity,
Electromigration.
1.6.1 Volatility
The methods based on volatility mainly include vaporization and distillation in its
different forms. Sublimation is a special case of distillation where a solid is directly
vaporized without passing through a liquid state. Vaporization is simple where solvent
is removed by using heat or air currents such that the material concentrates to a solid.
In separation by distillation, all the components of interest in the mixture are volatile.
Sublimation is an exception to this because of the physical state of the component.
Distillation depends on the distribution of constituents between the liquid mixture and
the vapor in equilibrium with the mixture. The more volatile component is
concentrated in the vapor while the less volatile components remain in the liquid
phase in greater concentration.
There are various forms of distillation such as fractional distillation, flash distillation,
vacuum distillation, steam distillation and azeotropic distillation.
Fractional distillation involves the return of condensate to the distillation unit under
conditions such that this condensate is continuously and counter currently in contact
with the vapors. By this method, you can achieve greater enrichment of vapors of
more volatile component than that obtained by simple distillation.
In flash distillation, there is an instantaneous and continuous vaporization of a
definite component of the mixture such that the total vapor produced is in equilibrium
with the remaining liquid.
Vacuum distillation is actually distillation under reduced pressure. It is used to
separate high boiling mixtures or liquids that decompose below their normal boiling
points. Low pressure reduces the boiling point.
15
Classical Methods
SAQ 8
What type of distillation is used to separate components of a mixture if they
decompose below their normal boiling point?
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1.6.2 Solubility
When we refer solubility as a means of separation it essentially implies precipitating
the constituent of interest from the solution. When the solubility limit of the solute in
the solvent exceeds, the material appears as a precipitate. If this process is carried out
in an appropriately selected solvent and in a controlled manner, it can lead to
crystallization. Precipitation can be brought about in a number of ways out of which
some important ones are discussed below.
Solvent precipitation is achieved by adding another miscible solvent to the solution
such that the solubility of the component of interest is reduced and the material
appears as a precipitate in the mixed solvent. The precipitation brought about by a
chemical reaction is well known. Right from the beginning in a chemistry laboratory,
you have been precipitating different ions by adding appropriate reagents. A typical
example is precipitating Ba2+ by adding SO42-. There are many selective reagents
known which can precipitate one ion in the presence of other ions. A detailed
discussion about this is given in Units 14 and 15 of the Course on Basic Analytical
Chemistry. However, some organic compounds can be precipitated by suitable
adjustment of pH. It is possible to precipitate weakly basic organic compound from an
aqueous solution by making the solution more basic. Similarly, organic acids can be
precipitated by making the solution more acidic by strong acids.
1.6.3 Partition
The methods based on partition require two phases and there has to be a redistribution
of components between these two phases. Some important types of methods fall under
this category and they are as follows:
Liquid-liquid extraction
Gas-liquid chromatography
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General Aspects of
Separation Methods
SAQ 9
Name the two types of liquid-liquid partition chromatography.
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Classical Methods
SAQ 10
Name the type of ion exchanger which shows both cation and anion exchange
properties.
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potential of this technique for the purification and isolation of biological molecules is
unlimited. Specific adsorbents can be designed for the purification of enzymes,
antibiotics, nucleic acids and proteins.
General Aspects of
Separation Methods
Molecular sieves
Gel filtration
Microporous membranes,
Homogenous membranes,
Thin membranes.
There are numerous applications of these membranes. Some of these are listed below;
Ultrafiltration,
Reverse osmosis,
Dialysis, and
Electrodialysis.
Unit 11 of Block 5 of this course is assigned for the detailed discussion on membrane
separations.
Gel filtration has taken the form of a regular chromatography. This is also known as
gel permeation or size exclusion chromatography. The column packing materials are
polymer beads and silica based particles containing a network of uniform pores into
which the solute and solvent molecules can diffuse. In a column operation, the solute
molecules which are small enough to enter the pores of gel, are retarded. While the
molecules large enough not to enter the pores, will spend all their time in the mobile
phase and move rapidly through the column. This is a useful technique for the
separation of high molecular weight natural products from low molecular weight
species and salts. It is also used for the rapid determination of molecular weight. The
separation by gel permeation can be carried out at high pressure with an HPLC
instrument. The details of this branch of chromatography are given in Unit 10.
SAQ 11
What are the main requisites of a semi-permeable membrane to be used for various
applications?
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19
Classical Methods
1.6.7 Electromigration
The simplest example of separation by migration under electric field is controlled
potential electrodeposition. This method is quite effective for the separation of certain
metal ions in the presence of other metal ions. But in the normal classification of
separation methods, it is not given its due recognition. However, the techniques like
electrophoresis and electrochromatography in which an electric field is utilized to
produce or affect the relative motion of charged species in solution, have gained more
prominence. The two frontline techniques included under this head are
electrophoresis and electrochromatography.
In electrophoresis, only the electric field causes the motion while in
electrochromatography, the movement is caused by the resultant of an electric and a
gravitational (or other non-electrical) forces. Electrophoretic separations are
performed in two different ways. One is called slab electrophoresis and the other
capillary electrophoresis.
In slab electrophoresis, a filter paper, porous glass or a gel cast in the form of a bed
can be used. Samples are introduced as spots or band and a dc potential is applied
across the slab for a fixed period. The separated species are visualized by staining in a
way similar to TLC.
In capillary electrophoresis, a buffer filled capillary tube is used. The tube extends
between two buffer reservoirs that also holds the platinum electrodes. The sample is
introduced at one end and a dc potential is applied. The separated analytes are
observed by a detector at the other end.
The simplest form of electrochromatography is by fusion of electrophoresis and
paper chromatography. The sheet of paper is usually suspended vertically with the
solvent (buffer) descending from the top. The electric field is applied horizontally
along the sides of paper. The capillary electrochromatography is a more sophisticated
version of the paper electrochromatography. It is a hybrid of capillary electrophoresis
and HPLC and offers some of the advantages of the two techniques. The different
aspects of separation by electromigration are discussed in Unit 12.
1.7
Now we have seen that the different types of separation methods have been classified
under various categories based on property resulting into separation. Practically, all
these methods can be divided into various classes using equilibrium and rate
processes as criteria. The equilibrium processes are based on differences in the
properties of individual components. These processes are generally based on phase
equilibria and involve the distribution of substances between two phases. Rate
processes are based on the kinetic properties of the components. It is already clear that
a majority of important separation processes are chromatography based; therefore, it
may be reasonable to introduce a sub-classification of the method listed under this
head as chromatographic and non-chromatographic methods.
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Gas-liquid,
Gas-solid,
Liquid-liquid,
Liquid-solid.
a)
Gas-liquid
General Aspects of
Separation Methods
Some of the important methods in which gas and liquid are the two phases
involved are as follows:
1.
Non-chromatographic methods
i)
Distillation
ii)
Foam fractionation
When we say distillation, it means distillation in all its forms. The wellknown example of foam fractionation is concentration of ores by froth
floatation process.
2.
Chromatographic method
i)
b)
Gas-liquid chromatography
Gas-solid
When the two phases are gas and solid, a few examples of the methods are as
given below:
1.
2.
Non-chromatographic methods
i)
Sublimation
ii)
Adsorption
Chromatographic methods
i)
Gas-solid chromatography
ii)
Exclusion chromatography
In the case of sublimation, the solid is directly vaporised and the equilibrium
exists between the gas and the solid. Adsorption of gases by solids is a well
known method of separation and this forms the basis of gas-solid
chromatography. Exclusion of molecules based on shape and size of both gas
and liquid can take place.
c)
Liquid-liquid
In this case, two immiscible liquids come in contact with each other. Here, the
separation takes place because of favorable partition of one or more components
in one of the phases.
1.
Non-chromatographic method
i)
2.
d)
Liquid-liquid extraction
Chromatographic methods
i)
ii)
iii)
Liquid-solid
1.
Non-chromatographic methods
i)
Precipitation
21
Classical Methods
ii)
2.
Fractional crystallization
Chromatographic methods
i)
ii)
iii)
Affinity chromatography
iv)
a)
b)
i)
Ultrafiltration,
ii)
Reverse Osmosis,
iii)
Dialysis, and
iv)
Electrodyalysis.
Field separations
Electrical field is commonly used to affect separations and the different
methods which are based on the application of this type of field are
i)
Electrodeposition,
ii)
Electrophoresis,
iii)
iv)
Electrochromatography.
The examples for separations resulting due to application of gravitational and thermal
fields are ultracentrifugation and thermal diffusion, respectively.
After going through both types of classification, one can see that most of the known
techniques are included in one or the other category. But it may be mentioned that the
list of separation methods considered for classification is not exhaustive. Some of the
lesser known methods have not been taken into consideration. The other point which
has to be kept in mind is that in some of the methods, particularly chromatographic,
more than one mechanism is operative and the method may be classified under
different categories.
SAQ 12
Give one example each of a non-chromatographic separation process when the
following two phases are in equilibrium.
i)
22
Gas-liquid
ii)
Liquid-liquid
iii)
Liquid-solid
General Aspects of
Separation Methods
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SAQ 13
Name the separation methods which are performed under the influence of electrical
field.
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1.8
Now we have some idea about a majority of separation methods. The advantages,
applications and limitations of some of the important methods will be discussed in the
different units of this course. It may be a little premature to make a comparison of the
methods, particularly, when we do not have sufficient background of the different
separation methods. Moreover, the assessment of utility of a method is really a
situation based problem. To elaborate this point, one can cite that the selection of a
method will be determined by the physical state of mixture, its complexity, amount of
sample available, the speed required and the level of purity desired. Over and above
this, one important point that has to be kept in mind is that whether the method has to
be used for analysis or synthesis/recovery/purification. If a method has to be used on
an industrial scale, some different considerations also get prominence. In a situation
like this, it is a little difficult task to impart special importance to one or the other
criteria. However, having known about the utility of the separation methods, we can
talk about some of the general criteria for the selection of the separation methods.
They are
Selectivity,
Detectability,
1.8.1 Selectivity
The foremost and most important is the selectivity. The selectivity relates to the
capability of the technique to separate the desired component effectively from a
complex mixture particularly containing closely similar components. The term
effective separation is expressed in terms of resolution, separation factor,
decontamination factor and percentage purity. All these terms, in one form or the
other, will appear in different units of this course.
23
Classical Methods
1.8.2 Detectability
The term detectability refers to the minimum amount that can be detected. It is also
termed as sensitivity. In the case of detectors, it can be calculated based on certain
assumptions such as detection at two or three times the noise level observed at the
baseline. Sensitivity can also be determined practically. Sensitivity can be improved
by properly optimizing the separation conditions using the same detector. Sensitivity
assumes paramount importance when the quantity of mixture to be analyzed is very
small or trace impurities for obtaining an ultrapure material are to be assayed.
1.8.3 Reproducibility
The method should be such that it gives reproducible results of separation. It is
usually expressed in terms of standard deviation or coefficient of variation based on
replicate measurements. Now at this point, it may be important to point out that the
separation conditions should not be such that a slight variation in the conditions may
alter the result significantly. A flexibility in the conditions of separation is always
preferred. This is particularly important if the process is to be scaled up on a
commercial scale.
24
General Aspects of
Separation Methods
SAQ 14
What is the usual way of expressing the reproducibility of a separation method?
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SAQ 15
In what particular type of separations, the speed of the separation becomes very
important?
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SAQ 16
What different parameters gain prominence when a separation method is to be scaled
up for plant production?
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1.9
SUMMARY
25
Classical Methods
Separations encompass different physicochemical principles and the subject has itself
matured to a unified science. Most of the well known methods of separations fall
under the category of chromatographic methods.
It is a little difficult task to categorize the variety of separation methods available
today. A simpler classification can be made on the basis of property which results into
separation. The other approach that can be adopted for classification is based on the
physicochemical phenomena responsible for separation. In other words, these
phenomena can be divided under two categories, the equilibrium and the rate
processes. In equilibrium processes, there have to be two competing phases. However,
in rate processes, the constituents move differently through a permeable barrier or
show different migration velocities under various fields, mainly electrical.
In both the classifications, it has been possible to classify most of the well known
separation methods in one or the other category. But some of the methods are such
that they can be put in more than one category.
As regards the criteria to be used for the assessment of utility of a separation method,
the whole problem is situation based. This point can be clarified by the fact that the
selection or the utility of a method will be determined by the physical state of the
mixture, its complexity, amount of sample available, the speed required and the level
of purity desired. It has to be kept in mind that whether the method is to be used for
analysis or synthesis/recovery. But this only does not necessarily determine the
importance of the different criteria cited in the text.
2.
Name the properties which are generally used for achieving separations.
3.
4.
5.
6.
Name the different criteria which are commonly used for the selection of
separation methods.
1.11 ANSWERS
Self Assessment Questions
1.
ii)
2.
3.
The two important examples beneficial for our environment are as follows:
i)
26
ii)
4.
5.
6.
7.
i)
Adsorption
ii)
Partition
iii)
Ion exchange
iv)
Size exclusion.
General Aspects of
Separation Methods
The different reasons for the growth in separation methods are as under.
i)
ii)
iii)
The two main criteria employed for classifying different separation methods are
as given below.
i)
ii)
8.
9.
Column chromatography
ii)
Planar chromatography
10.
The type of ion exchanger which shows both cation and anion exchange
properties is known as amphoteric exchanger.
11.
ii)
iii)
27
Classical Methods
12.
13.
Phases in Equilibrium
Examples of non-chromatographic
Separation Process
i)
Gas-liquid
distillation
ii)
Liquid-liquid
solvent extraction.
iii)
Liquid-solid
precipitation
Electrodeposition,
ii)
Electrophoresis,
iii)
iv)
Electrochromatography.
14.
15.
16.
ii)
iii)
iv)
Environmental friendliness.
2.
3.
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The different ways in which the separations help in chemical analysis are stated
below:
i)
ii)
iii)
The properties which are generally used for achieving separations are
i)
Volatility,
ii)
Solubility,
iii)
Partition,
iv)
Ion exchange,
v)
Surface activity,
vi)
vii)
Electromigration.
i)
Fractional distillation,
ii)
Flash distillation,
iii)
Vacuum distillation,
4.
iv)
v)
Azeotropic distillation.
General Aspects of
Separation Methods
ii)
iii)
Some weakly basic and acidic compounds are precipitated by altering the
pH of the solution.
5.
6.
Selectivity,
ii)
Detectability,
iii)
Reproducibility,
iv)
v)
vi)
Further Reading
1.
2.
Quantitative Analysis, By R.A. Day Jr. and A.L. Underwood, Prentice and Hall
of India.
3.
4.
5.
6.
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