CHEM 361 SEPARATION METHODS

Introduction

Separation is a technique used in isolating and / or purifying a chemical constituent (component)

or a group of chemically related substances. During separation operations, either the interfering
substances are removed or the substance of interest is isolated.

For a successful separation, the substances to be separated must differ in a chemical or physical
property due to which the separation can occur.

Some of the common separation methods are

 Distillation  Extraction
 Precipitation  Centrifugation and cyclones
 Absorption  Crystallization
 Adsorption  Decantation
 Ion exchange  Sieving
 Precipitation  Winnowing
 Chromatography
 Electrophoresis
 Dialysis
 Electro-deposition

Recovery Coefficient

The recovery coefficient indicates how much a component is isolated from a sample (mixture).
The recovery coefficient of a component A designated (RA) from a mixture is expressed as:

QA
RA =
(QA)o

Where QA = quantity of component A recovered (obtained)

(QA)o = initial quantity of component A in the mixture

Separation Coefficient

The separation coefficient expresses the level of separation between two components in a
mixture. The separation coefficient SB/A between two components, A and B in a mixture is
expressed as:

RB QB / QA
SB/A = 
RA (QB )o /(QA)o

Where QA and QB = quantities of A and B that are removed

(QA)o and (QB)o = initial quantities of A and B present in the sample.
Relative Error in Separations

The relative error resulting from the incomplete recovery of a component A from an impurity
component B in a separation process is given by

KB(QB)o
Er = (RA – 1) + RB
KA(QA)o

Where KA and KB are proportionality constants relating Q A and QB respectively

with the measured analytical parameter

Minimizing Relative Error in Separations

1. If RA ≈ 1 and RB ≈ 0 then the two components must be separated as completely as

possible
KB (QB )o
2. The ratios and should be small absolute numbers
KA (QA)o

QUESTION

To 3 ml of serum containing 200 µg ml-1 of an impurity B was added 200 µl of a solution

containing 1000 µg ml-1 of a drug. The mixture was shaken thoroughly. This mixture was then
taken through a separation process and 10 ml of a solution C was obtained. Chromatographic
analysis of solution C gave 19.50 µg ml-1 of A and 3.05 µg ml-1 of B. Calculate the recovery
and the separation coefficient of the separation method.

SOLUTION

(QA)o = (200 µl x 10-3 ml) x 1000 µg ml-1 = 200 µg

(QB)o = 3 ml x 200 µg ml-1 = 600 µg
QA = 10 ml x 19.50 µg ml-1 = 195 µg
QB = 10 ml x 3.05 µg ml-1 = 30.5 µg

QA 195
RA = = = 0.975
(QA)o 200

RB QB / QA 30.5 / 195
SB/A =  = = 0.0521
RA (QB )o /(QA)o 600 / 200

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 2

SOLVENT EXTRACTION

Solvent extraction is a very common laboratory separation procedure used when isolating or
purifying a product. It is extremely useful for very rapid and “clean” separations of both organic
and inorganic substances. It involves the distribution of a solute (e.g., product) between two
immiscible liquid phases. For example, in the extraction of a polar organic substance from an
organic solvent using an aqueous solution, the inorganic ions and polar organic compounds are
extracted into the aqueous phase whereas the non-polar organic molecules go into the organic
phase. However, in most cases a solute is extracted from an aqueous solution into an immiscible
organic solvent. After the mixture is shaken for about a minute, the phases are allowed to
separate and the bottom layer (the heavier solvent) is drawn off in a completion of the
separation. A separatory funnel is used for this process. The density of the two phases must
differ appreciably so that they can separate easily after mixing.

The organic solvent used for extraction must meet the following criteria:

1. Should readily dissolve substance to be extracted.

2. Should not react with the substance to be extracted.
3. Should not react with or be miscible with water(the usual second solvent)
4. Should have a low boiling point so it can be easily evaporated
5. Should be less viscous

Distribution Coefficient or Partition Coefficient

A substance A will distribute itself between two phases (after shaking and allowing the phases
to separate), and the ratio of the amounts or concentrations of the substance in the two phases
will be a constant. Distribution coefficient KD of substance A partitioned between two phases,
1 and 2, at equilibrium is given as:

S1
KD =
S2

Where S1 represents solvent 1 (e.g., an organic solvent) and S2 represents solvent 2 (e.g., water).
KD applies only if the solute is present in the same form in the two phases. It also depends on
temperature. If the distribution coefficient is large, then the substance will tend towards
quantitative distribution in solvent 1.

Deviation from the Distribution Law

It should be noted that during the distribution between the phases, many substances are partially
ionized in the aqueous layer as weak acids causing certain factors as pH to affect the extraction.
Thus by changing such parameters as pH, extraction agent, concentration of reagents, etc, the
extraction process could be made selective for the removal of a particular substance.

The distribution coefficient, KD, is used to describe distribution equilibrium of a system in

which the solute is present in one and the same form in both solvents. If, however, the solute is
present in more than one form in one or both solvents (multiple equilibra) apparent deviations
from the distribution law occur because quantitative separation may not be achieved. In order
to eliminate these deviations, distribution ratio, D, is used instead of distribution coefficient.

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 3

Distribution Ratio or Partition Ratio

If there is dissociation, dimerisation or complexation during the partitioning process, then it is

more meaningful to describe a different term, the distribution ratio, D, instead of partition
coefficient. It is the ratio of the concentrations of all the species of the solute in each phase.

C1
D=
C2
Where C1 and C2 = total concentrations of substance A in phases 1 and 2.

By convention, when one of the solvents is water, then the aqueous concentration is made the
denominator whilst the organic solvent concentration is made the numerator. D depends on
experimental conditions and many separation methods are based on controlling its value by
varying the conditions.

Trial

A chemist dissolves 155 mg of iodine (I2) in 1L of distilled water. This solution is equilibrated
at 20oC with 50 ml of carbon tetrachloride (CCl4 ). Titration of the organic solvent requires
22.50 ml of 0.050M sodium thiosulphate (Na2S2O3) solution. Calculate the distribution
coefficient KD of iodine between the two solvents (CCl4/H2O).

[2S2O32- + I2 → S4O62- + 2I-]

Distribution of a Weak Acid between Organic Solvent and Water

Consider, for example, the extraction of benzoic acid from an aqueous solution (a) into an
organic phase (o). Benzoic acid (HBz) is a weak acid in water with a particular ionization
constant Ka.

The ionization of Benzoic acid in the aqueous phase is represented as

HBz = H+ + Bz-

and the ionization constant is given by

[ H  ]a[ Bz  ]a
Ka  [1]
[ HBz ]a

The distribution coefficient KD is given by

[ HBz ]o
KD  [2]
[ HBz ]a

The distribution ratio is given by

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 4

 [ HBz ]o
D [3]
[ HBz ]a  [ Bz  ]a

From the equilibra involved, a relationship between the distribution ratio (D) and the
distribution coefficient (KD) can be derived as follows:

[ H  ]a[ Bz  ]a
From the ionization constant of the acid Ka 
[ HBz ]a

The concentration of the benzoic ion in the aqueous phase is given as

Ka[ HBz ]a
[ Bz  ]a  [4]
[ H  ]a

[ HBz ]o
From the distribution coefficient KD of the benzoic acid KD 
[ HBz ]a

The concentration of benzoic acid in the aqueous phase is given as [ HBz ]o  KD[ HBz ]a [5]

Substitution of equations [4] and [5] into equation [3] gives

KD[ HBz ]a
D [6]
[ HBz ]a  Ka[ HBz ]a /[ H  ]a

KD
D [7]
1  Ka /[ H  ]a

This equation predicts that when [H+]a >> Ka, D is nearly equal to KD
If KD is large, the benzoic acid will be extracted into the organic layer. That is D is maximum
under these conditions.

If on the other hand, [H+] << Ka, then D reduces to KD[H+]a/Ka, which will be small, and the
benzoic acid will remain in the aqueous layer.

In summary, in alkaline solution, the benzoic acid is ionized and cannot be extracted, while in
acid solution it is largely undissociated.

Thus, by regulating the pH, the course of the extraction is shifted to the desired direction.

One of the attractive features of solvent extraction is that, as indicated by equation [7] the
extraction efficiency will be independent of the original concentration of the solute.

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 5

Question

At pH of 5, an acid has a dissociating constant of 1.0 X 10-4 and distribution coefficient of 100.
Find the distribution ratio of this acid.

Efficiency of Extraction

This expresses the fraction of the analyte extracted in one extraction process. Extraction
efficiency increases with increasing number of extractions. For a given volume of extractant,
successive extractions with smaller individual volumes of extractant are more efficient than a
single extraction with the whole volume of the extractant.

If Wo grams of substance A is initially present in V1 ml of solvent 1(the raffinate) and A is

extracted successively with equal volumes of V2 of solvent 2(the extractant). The quantity Wn
of the substance A that remains in the layer after n of such extraction is given by:

 V1  n
Wn =   Wo
 DV 2  V 1 
Fraction of A remaining in solvent after n extractions, ƒ n, is given as:
Wn  V 1 n  1 n
ƒn = =   =  
W o  DV 2  V 1   D(V 2 / V 1)  1 

the percentage of the substance that is extracted in a single extraction is given by:
%E = 100D
D  (V 1 / V 2)

Question

The distribution ratio of aspirin between toluene and water is 4.70. if 100ml of an aqueous
solution of aspirin is extracted with
a) One 120ml and
b) Four 30ml portions of toluene,
Calculate the percentage of aspirin extracted in each case.

Solution

 V1 n
(a) Wn =   Wo
 DV 2  V 1 

 100 1
=   Wo
 (4.70 X 120)  100 

=0.1506Wo

% aspirin is given by;

W 0 W1
%E =   100
 Wo 

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 6

  1  0.1506 
= 100
 1 

= 84.9%

Or using
100 D
%E =
D  (V 1 / V 2)

100 X 4.70
=
4.70  (100 / 120)

= 84.9%

(b) For four extractions, n = 4

 V1 n
Wn =   Wo
 DV 2  V 1 

 100 4
=   Wo
 (4.70 X 30)  100 

= 0.0296Wo

W 0 W1
%E =   100
 Wo 

 1  0.0296 
=  100
 1 

= 97.0%

Question 2

If the distribution ratio of drug A is 9.0, what is the minimum number of 5.00ml aliquots of
ether that must be used in order to extract 99.9% of drug A from 5.00ml of an aqueous
solution that contains 0.0400g of the drug?

Solution

Wn  V1 n
ƒn = =  
Wo  DV 2  V 1 

CHEM 361 SEPARATION METHODS R. B. Voegborlo Page 7

  100  99.9   5.00 n
=  =  
 100   (9.0 X 5.00)  5.00 

1 1
=  n
1000  10 
n=3
Question

The capacity factor of a given solute in a water-chloroform system is 10. calculate the
percentage of the solute extracted from 50ml of water by 100ml of chloroform where:
a) the chloroform was used all at once
b) the 100ml chloroform is divided into five 20ml portions which are used one after the
other.

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