CHM 171 : Isolation and Purification of Organic Compounds

Isolation of Organic Compounds

The Isolation involve separation of mixtures of compounds to give the pure components is of
great practical importance in chemistry. Many synthetic reactions give mixtures of products
and it is necessary for you to have a reasonably clear idea of how mixtures of compounds can
be separated. Almost all compounds of biochemical interest occur naturally as components
of very complex mixtures from which they can be separated only with considerable difficulty.

The process of Isolation include Extraction, Separation o and purification of organic

compounds which be achieved by differences in physical properties, such as differences in
boiling point, or by chemical means, where differences in physical properties are enhanced
by chemical reactions. All techniques for isolation and purification of products use phase
equilibria:

• isolate the desired product into one phase that is in equilibrium with another phase

• The second phase ideally contains all other components of the mixture

• Equilibrium constants for distribution of materials between two phases are rarely either
infinite or zero, so partial separation of the components usually occurs

• phase equilibria used for purification may involve gas-liquid, gas-solid, liquid-liquid, or
liquid-solid interfaces

Extraction procedures make use of phase equilibria at liquid-liquid and liquid-solid

interfaces

Extraction

Extraction is the first step to separate the desired organic compounds from the raw materials.
Extraction methods include solvent extraction, distillation method, pressing and sublimation
according to the extraction principle

"Extraction" refers to transference of compound(s) from a solid or liquid into a different solvent
or phase.

When a tea bag is added to hot water, the compounds responsible for the flavor and color of
tea are extracted from the grounds into the water (Figure1a). Decaffeinated coffee is made by
using solvents or supercritical carbon dioxide to extract the caffeine out of coffee beans.
Bakers use the extract of vanilla, almond, orange, lemon, and peppermint in their dishes,
essences that have been extracted from plant materials using alcohol (Figure 1b).

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Figure 1: Examples of extraction: a) Tea, b) Baking extracts, c) Plant pigments extracted
into water droplets after sprinklers hit a fallen leaf on the sidewalk.

In the chemistry lab, it is most common to use liquid-liquid extraction, a process that occurs
in a separatory funnel (Figure 2). A solution containing dissolved components is placed in the
funnel and an immiscible solvent is added, resulting in two layers that are shaken together. It
is most common for one layer to be aqueous and the other an organic solvent.
Components are "extracted" when they move from one layer to the other. The shape of the
separatory funnel allows for efficient drainage and separation of the two layers.

Figure 2: Schematic of extraction.

Compounds move from one liquid to another depending on their relative solubility in each
liquid.

A quick guide to solubility is the "like dissolves like" principle, meaning that nonpolar
compounds should be readily extracted into nonpolar solvents (and vice versa).

The compounds responsible for the taste and color of tea (Figure 1a) must be polar if they are
readily extracted into hot water. When allowed to equilibrate between two liquids in a

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separatory funnel, the majority of a compound often ends up in the layer that it is more
soluble.

What Is Solvent Extraction?

Solvent extraction is a process in which compounds are separated based on their relative
solubility. This treatment method involves using a solvent - a fluid that has the ability to
dissolve another substance. Solvent extraction does not destroy a compound. It instead
separates it, a process that can provide several valuable benefits depending on the industry in
which the process is used.

Solvent extraction is the act of removing something or separating it. This must be done
through force and this process occurs over the course of two different immiscible phases.
Immiscible liquids (liquids that do not dissolve in one another) form layers when put
together. This is because each liquid differs in polarity, or orientations. The order of the
phases, whether a particular liquid is on top or on the bottom, is determined by its density
(figure 2).

For example, if you use petroleum ether and water during the extraction process, water has a
higher density than ether. Therefore, water will be the bottom phase.

Your method for extraction may vary depending on your equipment, but in its most simplistic
form, extractions often use some kind of separatory funnel ( Fugure 2). The liquid mixture
that is to be separated is first added to the funnel. When the two extraction solvents are then
added to the funnel, they should separate into two phases. After forcibly shaking the funnel
and allowing it to stand for several minutes, the two layers can be collected. In the end, the
target molecule for extraction will be located in one of the solvents.

Solvent extraction is the most widely used method. The extraction of organic compounds
progresses through the following stages:

1. the solvent penetrates into the solid matrix;

2. the solute dissolves in the solvents;
3. the solute is diffused out of the solid matrix;
4. The extracted solutes are collected.

Any factor enhancing the diffusivity and solubility in the above steps will facilitate the
extraction such factors include

i. The properties of the extraction solvent,

ii. the particle size of the raw materials,
iii. the solvent-to-solid ratio.
iv. the extraction temperature and
v. the extraction duration which affect the extraction efficiency

Solvent extraction is used across multiple industries, including during the processing of
perfumes and vegetable oil. It’s also commonly used in petrochemical refining. Industries.
Solvent extraction is used to separate hazardous contaminants from sludge and sediments as

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well. This can be especially beneficial for hazardous waste generators since solvent
extraction ultimately reduces the amount of hazardous waste that must be treated.

What Is a Solvent?

A solvent is a molecule that can dissolve other molecules, known as solutes. A solvent can
be solid, liquid or gas. A solvent’s molecules pull apart the solute’s molecules, and eventually
the solute’s molecules become evenly distributed throughout the solvent. This homogeneous
mixture cannot be separated physically. That means heat or another chemical process must be
applied to the solution in order to separate the solvent from the solute. Popular solvents
include water and organic compounds like benzene, tetrachloroethylene and turpentine.

Extraction is the first step to separate the desired natural products from the raw materials.
Extraction methods include solvent extraction, distillation method, pressing and sublimation
according to the extraction principle.

Principle of Extraction

Generally, the finer the particle size is the better result the extraction achieves. The extraction
efficiency will be enhanced by the small particle size due to the enhanced penetration of
solvents and diffusion of solutes. Too fine particle size, however, will cost the excessive
absorption of solute in solid and difficulty in subsequent filtration.

High temperatures increase the solubility and diffusion. Temperatures that too high, however,
may cause solvents to be lost, leading to extracts of undesirable impurities and the
decomposition of thermo labile components. The extraction efficiency increases with the
increase in extraction duration in a certain time range. Increasing time will not affect the
extraction after the equilibrium of the solute is reached inside and outside the solid material.

The greater the solvent-to-solid ratio is, the higher the extraction yield is; however, a solvent-
to-solid ratio that is too high will cause excessive extraction solvent and requires a long time
for concentration.

Method of Extraction

The conventional extraction methods, include:

Maceration, percolation, Decoction and reflux extraction, usually use organic solvents and
require a large volume of solvents and long extraction time.

Maceration
This is a very simple extraction method with the disadvantage of long extraction time and
low extraction efficiency. It could be used for the extraction of thermolabile components.

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Percolation
Percolation is more efficient than maceration because it is a continuous process in which the
saturated solvent is constantly being replaced by fresh solvent.

Decoction
The extract from decoction contains a large amount of water-soluble impurities. Decoction
cannot be used for the extraction of thermolabile or volatile components.

Reflux extraction
Reflux extraction is more efficient than percolation or maceration and requires less extraction
time and solvent. It cannot be used for the extraction of thermolabile natural products.

The modern Extraction methods modern or greener extraction methods such as include :

super critical fluid extraction (SFC), pressurized liquid extraction (PLE) and microwave
assisted extraction (MAE), have also been applied in natural products extraction, and they
offer some advantages such as lower organic solvent consumption, shorter extraction time
and higher selectivity. Some extraction methods,

Soxhlet extraction
The Soxhlet extraction method integrates the advantages of the reflux extraction and
percolation, which utilizes the principle of reflux and siphoning to continuously extract the
herb with fresh solvent. The Soxhlet extraction is an automatic continuous extraction method
with high extraction efficiency that requires less time and solvent consumption than
maceration or percolation. The high temperature and long extraction time in the Soxhlet
extraction will increase the possibilities of thermal degradation.

Pressurized liquid extraction (PLE)

Pressurized liquid extraction (PLE) has also been described as accelerated solvent extraction,
enhanced solvent extraction, pressurized fluid extraction, accelerated fluid extraction, PLE
applies high pressure in extraction. High pressure keeps solvents in a liquid state above their
boiling point resulting in a high solubility and high diffusion rate of lipid solutes in the
solvent, and a high penetration of the solvent in the matrix. PLE dramatically decreased the
consumption of extraction time and solvent and had better repeatability compared to other
methods.

Supercritical fluid extraction (SFE)

Supercritical fluid extraction (SFE) uses supercritical fluid (SF) as the extraction solvent. SF
has similar solubility to liquid and similar diffusivity to gas, and can dissolve a wide variety
of natural products. Their solvating properties dramatically changed near their critical points
due to small pressure and temperature changes. Supercritical carbon dioxide (S-CO 2) is
widely used in SFE because of its attractive merits such as low critical temperature (31 °C),
selectivity, inertness, low cost, non-toxicity, and capability to extract thermally labile
compounds. The low polarity of S-CO 2 makes it ideal for the extraction of non-polar natural

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products such as lipid and volatile oil. A modifier may be added to S-CO 2 to enhance its
solvating properties significantly.

Ultrasound assisted extraction (UAE)

Ultrasonic-assisted extraction (UAE), also called ultrasonic extraction or sonication, uses
ultrasonic wave energy in the extraction. Ultrasound in the solvent producing cavitation
accelerates the dissolution and diffusion of the solute as well as the heat transfer, which
improves the extraction efficiency. The other advantage of UAE includes low solvent and
energy consumption, and the reduction of extraction temperature and time. UAE is applicable
for the extraction of thermo labile and unstable compounds. UAE is commonly employed in
the extraction of many types of natural products

Microwave assisted extraction (MAE)

Microwaves generate heat by interacting with polar compounds such as water and some
organic components in the plant matrix following the ionic conduction and dipole rotation
mechanisms. The transfers of heat and mass are in the same direction in MAE, which
generates a synergistic effect to accelerate extraction and improve extraction yield. The
application of MAE provides many advantages, such as increasing the extract yield,
decreasing the thermal degradation and selective heating of vegetal material. MAE is also
regraded as a green technology because it reduces the usage of organic solvent. There are two
types of MAE methods: solvent-free extraction (usually for volatile compounds) and solvent
extraction (usually for non-volatile compounds)

Pulsed electric field (PEF) extraction

Pulsed electric field extraction significantly increases the extraction yield and decreased the
extraction time because it can increase mass transfer during extraction by destroying
membrane structures. The effectiveness of PEF treatment depends on several parameters
including field strength, specific energy input, pulse number and treatment temperature. PEF
extraction is a non-thermal method and minimizes the degradation of the thermolabile
compounds.

 In that physical region the high temperature enables high solubility and high diffusion
rate of lipid solutes in the solvent, while the high pressure, in keeping the solvent
below its boiling point, enables a high penetration of the solvent in the sample.
 PFE permits a high extraction efficiency with a low solvent volume (15-40 ml) and a
short extraction time (15-20 min).
 That procedure is also known as “Accelerated Solvent Extraction” (ASE).

Enzyme assisted extraction (EAE)

The structure of the cell membrane and cell wall, micelles formed by macromolecules such
polysaccharides and protein, and the coagulation and denaturation of proteins at high
temperatures during extraction are the main barriers to the extraction of natural products. The
extraction efficiency will be enhanced by EAE due to the hydrolytic action of the enzymes on
the components of the cell wall and membrane and the macromolecules inside the cell which

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facilitate the release of the natural product. Cellulose, α-amylase and pectinase are generally
employed in EAE.

Hydro distillation and steam distillation

Hydro distillation (HD) and steam distillation (SD) are commonly used methods for the
extraction of volatile oil. Some natural compounds encounter decomposition in HD and
SD.The chemical composition essential oil from plants are usually affected by distillation
methods.

Separation methods

The components in the extract from above methods are complex and contain a variety of
natural products that require further separation and purification to obtain the active fraction or
pure natural products. The separation depends on the physical or chemical difference of the
individual natural product. Chromatography, especially column chromatography, is the main
method used to obtain pure natural products from a complex mixture.

Separation based on adsorption properties

Adsorption column chromatography is widely used for the separation of natural products,
especially in the initial separation stage, due to its simplicity, high capacity and low cost of
adsorbents such as silica gel and macroporous resins. The separation is based on the
differences between the adsorption affinities of the natural products for the surface of the
adsorbents. The selection of adsorbents (stationary phase) as well as the mobile phase is
crucial to achieve good separation of natural products, maximize the recovery of target
compounds and avoid the irreversible adsorption of target compounds onto the adsorbents.

Separation based on partition coefficient

Partition chromatography (PC) follows the liquid–liquid extraction principle based on the
relative solubility in two different immiscible liquids. In the early stage, one liquid phase was
coated to a solid matrix (silica gel, carbon, cellulose, etc.) as the stationary phase and another
liquid phase was employed as the mobile phase. The disadvantage of an easily removed
stationary phase and unrepeatable results has led to this kind of PC being rarely used today.
The bonded-phase, in which the liquid stationary phase is chemically bound to the inert
support, which is used as the stationary phase overcomes those drawbacks. Commercially
available alkyl such as C8 and C18, aryl, cyano and amino substituted silanes are often used
as bonded phases, which are widely used to separate a variety of natural products, especially
in the final purification step.

Separation based on the molecular size

The separation of natural products by membrane filtration (MF) or gel filtration
chromatography (GFC) is based on their molecular sizes.

Membrane filtration (MF)

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In MF, the semipermeable membrane allows smaller molecules to pass through and retains
the larger molecules. MF of natural products could be characterized as microfiltration,
ultrafiltration, and nanofiltration based on the pore size of the membrane applied.

Membrane filtration has been a powerful tool for the concentration, clarification and removal
of impurities in the lab, as well as in the food and pharmaceutical industries

Gel filtration chromatography (GFC)

Gel filtration chromatography is also known as gel permeation chromatography or size
exclusion chromatography. The small molecules have a longer retention time in GFC than
large molecules.

Sephadex is formed by cross-linking dextran, and the G-types of Sephadex were used for the
separation of hydrophilic compounds such as peptides oligosaccharides and polysaccharides

Separation based on ionic strength

Ion-exchange chromatography (IEC) separates molecules based on the differences in their net
surface charge. Some natural products, such as alkaloids and organic acids possessing a
functional group capable of ionization, might be separated by IEC. The charged molecules
could be caught and released by ion-exchange resin by changing the ionic strength of the
mobile phase (e.g., changing pH or salt concentration). Cation ion-exchange resins were used
for the separation of alkaloids, while the anion ion-exchange resins were used for the
separation of natural organic acids and phenols.

Other modern separation techniques

Molecular distillation (MD)
Molecular distillation separates the molecular by distillation under vacuum at a temperature
far below its boiling point. It is a suitable distillation method for separating thermo sensitive
and high-molecular-weight compounds.

Preparative gas chromatography (Prep-GC)

Gas chromatography (GC) with high separation efficiency and fast separation and analysis
makes it potentially the ideal preparative method for the separation of volatile compounds.
The injection port, column, split device and trap device of GC equipment must be modified
for preparative separation due to a lack of commercial Prep-GC.

Supercritical fluid chromatography (SFC)

SFC uses supercritical fluid as the mobile phase. SFC integrates the advantages of both GC
and liquid chromatography (LC) as the supercritical fluids possess properties of high
dissolving capability, high diffusivity and low viscosity, which allows rapid and efficient
separation. Thus, SFC can use a longer column and smaller particles of the stationary phase
than HPLC, which provides greater numbers of theoretical plates and better separation. SFC
can be used for the separation of non-volatile or thermally labile compounds to which GC is
not applicable.

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Molecular imprinted technology
Molecular imprinted technology has been an attractive separation method in the last decade
due to its unique features, which include high selectivity, low cost and easy preparation.
Many complementary cavities with the memory of size, shape, and functional groups of the
template molecules are generated when the template molecules are removed from the
molecular imprinted polymer (MIP). Thus, the template molecule and its analogs will have
the specific recognition and selective adsorption for the MIP. MIPs have been widely used in
the separation of natural products or as solid-phase extraction sorbents for sample preparation
of herbal materials to enrich the minor compounds.

Simulated moving bed chromatography

Simulated moving bed (SMB) chromatography uses multiple columns with stationary phases
(bed). The counter current movement of the bed is simulated through rotary valves, which
periodically switch the inlet (feed and eluent) and outlet (extract and caffeinate). The SMB
process is a continuous separation method and a powerful tool for the large-scale separation
of natural products with the advantage of lower solvent consumption over a shorter period of
time.

Multi-dimensional chromatographic separation

The components in the extract subjected to separation were complex, and generally, no pure
compound will be separated in one column chromatography. Multi-dimensional separation
based on the solid phase extraction and coupling of multiple columns with different
stationary phases greatly improves the separation efficiency. With more commercial multiple
dimensional separation equipment entering the market, the separation of natural products is
becoming more rapid, efficient and automated.

Extracting Natural Compounds

Fruit and plant leaves are primarily composed of cellulose and water, but also contain
"essential oils." a greasy mixture of compounds that capture the "essence" of the plant
material's smell and taste. Orange oil is roughly 95%95% limonene (Figure 4.3b), and due to
its nonpolar structure, can be extracted from its rind into an organic solvent like hexanes or
dichloromethane (Figure 4.3a). The oil can then be concentrated and used to flavor or scent
foods, cleaning supplies, and candles.

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Figure 4.3: a) Orange rind extracted into dichloromethane, b) GC spectrum of orange oil.

In the chemistry lab, essential oils are often extracted from their source using solvents, and
analyzed using gas chromatography or spectroscopy.

Transferring Compounds From Layers

Another method for extracting essential oils from fragrant plant materials is through steam
distillation (Figure 4.4b). This process often results in the lovely smelling compounds
suspended in the aqueous distillate (Figure 4.4c). In order to concentrate the oil, the aqueous
suspension is often extracted with a low-boiling organic solvent (Figure 4.4d), which can
then be easily removed from the oil.

Figure 4.4: a) Whole cloves, b) Steam distillation of cloves, c) Milky distillate composed of
oil and water, d) Using extraction to separate the oil from the water.

Selective Removal of Components

When conducting an experiment that synthesizes a chemical product, a reaction is often

complete whenever stirring or heating is ceased. And yet, there are always more steps in the
procedure! What commonly happens directly afterwards is to "work-up" the reaction in some
way. A work-up refers to methods aimed at isolating the product from the reaction mixture,
and often begins by using a separatory funnel and extractions.

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 Reaction to make isopentyl acetate.

For example, imagine that acetic acid and isopentanol have been heated in the presence of an
acid catalyst for one hour (Figure 4.5) in order to make isopentyl acetate, an ester that smells
of bananas (see reaction scheme in Figure 4.6). After the one-hour time period, there is
unfortunately not just the banana-smelling ester in the flask. The flask will also contain
byproducts (the water in this case), leftover starting materials if the reaction is incomplete, as
well as any catalysts used (H2SO4H2SO4 in this case). In this example, there could
be five compounds in the reaction flask after heating is ceased (Figure 4.7)!

Figure 4.6: Reaction scheme to produce isopentyl acetate.

When "working up" this reaction, the resulting mixture is often poured into a separatory
funnel along with some water and organic solvent. This produces two layers in the separatory
funnel: an aqueous layer and an organic layer.

After shaking this heterogeneous mixture, the compounds distribute themselves based on
their solubility. Compounds that have high water solubility favor the aqueous layer while less
polar compounds favor the organic layer. In this example, the acid catalyst and residual
carboxylic acid or alcohol would likely be drawn into the water layer. The ester would have a
greater affinity for the organic layer than the aqueous layer, causing it to be isolated from the
other components in the reaction mixture (Figure 4.7).

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Figure 4.7: Extraction using water and an organic solvent to isolate isopentyl acetate from
the reaction mixture.

In this example, it is possible that small amounts of alcohol are also drawn into the organic
layer, but they could likely be removed with a water "wash." In a wash, the desired
compound (e.g. the isopentyl acetate), remains in its current layer of the separatory funnel (in
this example the organic layer), and unwanted compounds are removed, or "washed away"
into another layer (e.g. the aqueous layer). A wash is different than an extraction, because in
an extraction the desired compound moves from its current location (i.e. moves from an
aqueous layer to an organic layer), while in washing the desired compound stays in its current
layer

Purification of Organic Compounds

The purification of organic compounds is necessary, though complex, step after its extraction
from a natural source or synthesis in the laboratory. The method of purification of the
organic compound depends mainly on the nature of the compound and the impurities present.
One easy method to check the purity of an organic compound is to either melt or boil it as
organic compounds tend to have sharp melting and boiling points.
Purification means the removal of unwanted impurities present in an organic compound. The
general methods of purification are:

 Sublimation

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  Crystallisation
 Distillation
 Differential Extraction
 Chromatography

Methods of Purification

Sublimation
Some solids can directly pass to the vapour state without going through the liquid phase. The
purification technique which exploits this property is called sublimation. It is helpful in
separating sublimable compounds from non-sublimable ones.

Methods of Purification – Sublimation

The substance is heated in a china dish above which an inverted funnel is kept to collect the
sublimable compounds. The funnel is kept cool so as to hasten the process. Vapours of the
substance solidify on the funnel.

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Crystallisation
The principle here is that the compound and the impurities have different solubilities in a
solvent. A solvent is chosen where the compound to be purified is sparingly soluble, that is, it
is sparingly soluble at lower temperature and soluble at a higher temperature. The solution is
heated to get a saturated solution, and on cooling, the crystals of the compounds are removed
via filtration.
For example, crystals of benzoic acid can be crystallised with water. Benzoic acid is
sparingly soluble in cold water and soluble in hot water.
If the mixture contains impurities that have the same solubility as of the compound to be
purified, repeated crystallisation is performed.

What is Crystallization?
Crystallization is a technique used for the purification of substances. A separation technique
to separate solids from a solution.
Crystallization can be defined as the process through which the atoms/molecules of a
substance arrange themselves in a well-defined three-dimensional lattice and consequently,
minimize the overall energy of the system. When a substance is subjected to crystallization,
its atoms or molecules bind together through well-defined angles.
On adding a solid substance in a liquid and stirring it, the solid dissolves in the fluid. But
when added more and more solid to the liquid, a point comes after which no more solid
dissolves in the liquid. This point is called a saturation point and the fluid is called a
saturation solution.

Crystallization Process

1. The solution is heated in an open container

2. The solvent molecules start evaporating, leaving behind the solutes
3. When the solution cools, crystals of solute start accumulating on the surface of the
solution
4. Crystals are collected and dried as per the product requirement
5. The undissolved solids in the liquid are separated by the process of filtration
6. The size of crystals formed during this process depends on the cooling rate

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 7. Many tiny crystals are formed if the solution is cooled at a fast rate
8. Large crystals are formed at slow cooling rates

Separating Substances via Crystallization

Activity:
Here is an experiment to understand crystallization clearly:
Step 1: Take 50 ml water in a beaker
Step 2: Add sugar in it and stir it
Step 3: Now heat the solution
Step 4: Repeat the process continuously
Step 5: After some time there will be a point at which no more sugar can be dissolved in
water. This stage is the saturation point, and the solution is referred to as a saturated solution
Step 6: Now filter the sugar with the help of a filter paper
Step 7: Collect the filtrate in a glass bowl and cool it
Step 8: We will observe that some fine crystals are formed in the bowl

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Step 9: The process of filtration can separate these crystals. The liquid left after the removal
of crystals is known as mother liquor

Application of Crystallization:

 Purification of seawater
 Separation of alum crystals from impure samples
 In the pharmaceutical industry, crystallization is used as a separation and purification
process for the synthesis and isolation of co-crystals, pure active pharmaceutical
ingredients (API), controlled release pulmonary drug delivery, and separation of
chiral isomers.
This was just a brief layout of the process of crystallization. To know about crystallization,
other methods of purification of organic compounds and more, register with BYJU’S and
download our app.

Exercise

What is the definition of the term crystallization?

Crystallization can be defined as the solidification of a liquid substance into a highly
structured solid whose atoms or molecules are placed in a well-defined three-dimensional
crystal lattice. The smallest individual part of a crystal is called a unit cell. The crystal is
made up of millions of such unit cells.

What are the uses of crystallization?

Crystallization is primarily employed as a separation technique in order to obtain pure
crystals of a substance from an impure mixture. Another important application of
crystallization is its use to obtain pure salt from seawater. Crystallization can also be used to
obtain pure alum crystals from an impure alum. In such scenarios, crystallization is known to
be more effective than evaporation since it also removes the soluble impurities.

What are the two primary types of crystallization?

Crystallization processes can be broadly categorized into the following two types:

 Evaporative crystallization
 Cooling crystallization

List some examples of crystallization.

Some common examples of crystallization are listed below.

 The crystallization of water to form ice cubes and snow.

 The crystallization of honey when it is placed in a jar and exposed to suitable
conditions.
 The formation of stalagmites and stalactites (especially in caves).

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  The deposition of gemstone crystals.

What are the advantages of crystallization?

The key advantages of crystallization are listed below.

 A product of high purity can be obtained from one single step via the process of
crystallization.
 The dry products formed from crystallization can be directly packaged and stored.

Purification of Liquids

Distillation
The underlying principle behind distillation is that the mixture of liquids can be separated by
the difference in their boiling points. The boiling point is defined as that temperature at which
the vapour pressure of the liquid is equal to the atmospheric pressure. This method separates
volatile liquids from non-volatile liquids. The setup is given below.

Purification of Liquids – Distillation

The mixture is taken in the RB flask and boiled. The more volatile or the component with
lower boiling point evaporates faster and is collected in a separate container. A condenser is
used to hasten the process of condensation.

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For example, a mixture of chloroform and aniline can be separated by distillation. The boiling
point of chloroform is 60°C and that of aniline is 189°C. Therefore distillation can be used to
separate a mixture of chloroform and aniline.

Fractional Distillation
This method is employed when the difference between the boiling points of the liquids isn’t
much. Since the vapours of such liquids might condense together, a fractionating column is
fixed to the mouth of the RB.

Purification of Liquids – Fractional Distillation

Vacuum Distillation
Since the boiling point is dependent on the atmospheric pressure, the liquids will boil at a
temperature lesser than their boiling points if they were distilled in an atmosphere having
lower pressure. This is achieved by using a vacuum pump. Since the atmospheric pressure is
reduced, the liquids also boil faster and hence the whole process of distillation is made fast.

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Process of Vacuum Distillation

Steam Distillation
In this variant, steam is passed into the flask containing the liquids to be separated. The
principle here is that the liquids will boil faster because of aqueous tension (vapour pressure
of water) helps in equalising the atmospheric pressure.
Total pressure = Aqueous tension + vapour pressure of liquid components

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Purification of Liquids by Steam Distillation
In the absence of aqueous tension, the process of boiling would have been continued until it
equalises the atmospheric pressure. Now with the addition of steam, that process is expedited.

Differential Extraction
This method is used for immiscible liquids, that is, liquids that do not mix together. For
example, oil and water are immiscible.
The immiscible liquids are taken in a separating funnel and left undisturbed. After a while,
they separate out according to their specific gravities, with the heavier liquid at the bottom.
Then they are later collected.

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Substances can also be separated according to their preferential solubilities in the liquid. For
example, if phenol is to be extracted, it can be preferentially extracted using NaOH solution
as one of the liquids used.

Chromatography
Chromatography is an important separation technique used to separate constituent particles of
a mixture of substances, to purify the compounds and check the purity of organic compounds.
In this technique on a stationary phase (solid or a liquid) a mixture of substance is applied.
The mixture of gas or the pure solvent is allowed to move slowly on the stationary phase.
Due to which the components of the mixture start separating from one another.
Chromatography is of two types:

 Adsorption Chromatography
 Partition Chromatography

Adsorption Chromatography
It is based on the principle that the constituents are adsorbed on an adsorbent in varying
degrees. The adsorbents used are generally silica gel or alumina. When a mobile phase moves
over the fixed phase, different constituents of the mixture get adsorbed at various distances
over the fixed phase.
Adsorption Chromatography is further classified into:

 Column Chromatography
 Thin Layer Chromatography

Column Chromatography
Here, a mixture is separated over a column of either silica gel or alumina, packed in a glass
column. The constituent with the most affinity with the fixed phase is adsorbed at the top, and
so on. It is then retrieved by using an eluant. The solvent is then evaporated to get the
constituent.
Check: Column Chromatography

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Thin Layer Chromatography
Here a sheet of alumina is taken ( 0.2 mm thick) over which a small spot of the mixture is
placed and it is kept in a suitable solvent. The solvent rises due to capillary action and the
constituents also rise with the solvent depending on their differential adsorption, and thereby,
they are separated.

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