Liquid-liquid Extraction

PHARM 309

Introduction:
Liquid-liquid extraction is a versatile and
dependable separation technique wherein an
aqueous solution is usually brought into
contact with another organic solvent,
exclusively immiscible with the former, so as
to affect a legitimate and actual transfer of
either one or more solutes into the latter.
The separation technique is superior to others
due to ease of use, faster extraction times,
decreased volumes of solvent, and their
superior ability to concentrate the analytes.

Invariably
such
separations
may
be
performed by shaking the two liquids in a
separatory funnel (separating funnel) for a
few minutes; and may be extended either to
large quantities of pharmaceutical substances
or trace levels.
Liquid-liquid
extractions
are
usually
accomplished with a separatory funnel. The
two liquids are placed in the separatory
funnel and shaken (moderately) to increase
the surface area between the phases. When
the extraction is complete, the liquids are
allowed to separate, with the denser phase
settling to the bottom of the separatory

The
solvent
is
chosen so that the
solute
in
the
solution has more
affinity toward the
added solvent.

Fig: Separatory

In the case of pharmaceutical chemicals that

are mostly organic solutes, the liquid-liquid
extraction system may very often make use
of two immiscible organic solvents (e.g.,
alcohol and ether) instead of the aqueousorganic type of extraction.
On the contrary, the inorganic solutes
normally encountered are invariably in
aqueous solutions; therefore, it has become
absolutely necessary to produce such neutral
substances out of them, for instance ionassociation complexes and metal-chelates
(using organic-ligands) that may be extracted
into an appropriate organic solvent.

Liquid-liquid
extraction
principles:
Feed phase contains a component, i, which is
to be extracted. Addition of a second phase
(solvent phase) which is immiscible with feed
phase but component i is soluble in both
phases. Some of component i (solute) is
transferred from the feed phase to the solvent
phase.
After extraction the feed and solvent phases
are called the raffinate (R) and extract (E)
phases respectively.

Normally one of the two phases is an organic

phase while the other is an aqueous phase.
Under equilibrium conditions the distribution
of solute i over the two phases is determined
by the distribution law.
After the extraction the two phases can be
separated because of their immiscibility.
Component i is then separated from the
extract phase by a technique such as
distillation and the solvent is regenerated.
Further extractions may be carried out to
remove more component i.
Liquid liquid extraction can also be used to
remove a component from an organic phase
by adding an aqueous phase.

Theory:
The Nernst Distribution Law or the Partition
Law states that at constant temperature, a
solute
distributes
itself
between
two
immiscible solvents only in a particular ratio.

Kp= Co/Caq
Kp is the distribution constant or the partition
coefficient or partition ratio (Solvent-to-Feed
Ratio)
Co is the concentration of the analyte in the
organic phase (Solvent)
Caq is the concentration of the analyte in the

Solute
Acetone
Acetone
Acetone

Feed solvent
Water
Water
Water

Extraction solvent
Kp (wt% basis)
1-Pentanol
1.14 (at 300 C)
2-Octanol
0.66 (at 300 C)
Chloroform
1.83 (at 250 C)

Extraction is an equilibrium process, and

therefore a finite amount of solute might be in
both phases, necessitating other processing
steps or manipulation of the chemical
equilibria.
The Partition Law offers the following two
limitations, namely:
- It is solely applicable to very dilute solutions.
- It does not hold good when the distributing
substances
encounters
association
or

In liquid-liquid extractions the following

two aspects are very crucial and
important, namely:
(a) Error due to the volume change
(b) Effectiveness of an extraction
(a) Error due to the volume change
In a situation wherein two immiscible solvents
are employed in an extraction, the volumes of
the two individual phases after attainment of
equilibrium may be appreciably different in
comparison to the initial volumes of the
solvents used.

Therefore, a number of procedures have been

adopted to avoid error due to the volume
change incurred thereby, namely:
(i) Measure the volume of the phase
employed for the analysis and incorporate
this volume in the calculations
(ii) Separate the phase quantitatively and
subsequently dilute to a known volume
(iii) Separate the phase quantitatively and
make use of the entire volume in the
remaining steps of the ongoing analysis
(iv) Carry a marker substance through the
extraction to automatically compensate for
volume changes (used in chromatographic

(b) Effectiveness of an extraction

Based on the appropriate partition coefficient
of an immiscible solvent pair it is possible to
calculate the effectiveness of an extraction.
Let us assume that x moles of solute present
initially in a volume V2 of solvent b. Now, this
particular sample undergoes extraction with a
volume V1 of solvent a and subsequently y
moles of compound are left in V2 at
equilibrium.
Substituting
these
values
coefficient equation we have:

in

partition

Kp = (

x-y
V1

after simplifying and

rearranging-

V2

y V2
x
)
Kp = (
V1 V1 y
=

=
or

or
or

Kp

Kp
y
x

x V2 V2
y V1 V 1
V2
V1
V1
V2
V1
V2
=

-1)

y
=

x
y

+1 =
( Kp

-1
x
y
V1
V2

+1 )

-1

where,
f = fraction not
extracted

From the above equation it is quite evident

that the fraction extracted is absolutely
independent
of
the
initial
solute
concentration.
Hence, the fraction left unextracted after n
extraction may be given by the following
expression-

fn = ( Kp

V1
V2

+1 )

-n

Solvent
Miscibility:
criteria:

selection

Solvents defined as miscible if the two

components can be mixed together in all
proportions without forming two separate
phases.
Solvents miscible with water in all proportions
include
acetone,
acetonitrile,
dimethyl
acetamide, N,N-dimethylformamide, dimethyl
sulfoxide, 1,4-dioxane, ethyl alcohol, glyme,
isopropyl
alcohol,
methanol,
2methoxyethanol,
N-methylpyrrolidone,
npropyl alcohol, pyridine, tetrahydrofuran, and

Density:
Another consideration when selecting an
extraction solvent is its density. Solvents that
are more dense than water will form the lower
layer of the pair when mixed together, while
solvents that are less dense than water will
form the upper layer or float on water.
For example, ethyl ether has a density of
0.7133 g/mL at 20oC and would constitute the
upper phase when combined with water,
which has a density of 0.9982 g/mL at that
temperature.
On the other hand, the density of chloroform
is 1.4892 g/mL at 20oC. Therefore, water

Ethyl ether (0.7133

g/mL)
Water (0.9982 g/mL)

Water (0.9982 g/mL)

Chloroform (1.4892 g/mL)

- Should have high density difference

Water (0.9982 g/mL) and Hexane (0.6548 g/mL) They are
less prone to emulsion problems.
Water (0.9982 g/mL) and Benzene (0.8765 g/mL) Should
not be used in the extraction process.

Solubility:
Although immiscible solvents may form two
visibly distinct phases when mixed together,
they are often somewhat soluble in each
other and will, in fact, become mutually
saturated when mixed with each other.
For example, 1.6% of the dichloromethane
(solvent) is soluble in water. Conversely,
water is 0.24% soluble in dichloromethane.
- Should be insoluble to each other.

When the phases are separated for recovery

of the extracted analyte, the organic solvent
layer will contain water.
Similarly, after extraction the depleted
aqueous phase will be saturated with organic
solvent and may pose a disposal problem.

Factors
influence
extraction:
Effect of temperature

solvent

The effect of temperature on the partition

coefficient may be estimated conveniently
from its effect on the solubilities of the
substance in the two respective solvents.
Considerations: solvents used should be
immiscible and the concentrations of solutes
are fairly S
low
in both the
Solubility
ofphases.
solute in solvent a
1
KP =
=
S2 Solubility of solute in solvent b
Temperature should be constant throughout
the process.

Effect of pH on extraction
Generally, it has been found that the organic
acids and bases in solution may exist as
equilibrium mixtures of their respective
neutral as well as ionic forms.
Thus, these neutral and ionic forms may not
have the same identical partition coefficients
in a second solvent; therefore, the quantity of
a substance being extracted solely depends
upon the position of the acid-base equilibrium
and ultimately upon the pH of the solution.

In conclusion, it may be observed that the

pH for an extraction system must be selected
in such a fashion so that the maximum
quantity of the analyte is present in the
extractable form, that obviously suggests that
the analyte should always be in the form of
either a free base or a free acid.

Emulsion problem encountered

in extraction:
Emulsion:
Emulsion may be defined as- a dispersed
system containing at least two immiscible
liquid phases and the system is stabilized by
emulsifying agents.
The effective and meaningful extraction of a
solute is rendered almost impossible when
there is an emulsion formation during an
extraction process. Emulsion formation makes
the separation of the two phases difficult.

Actually, emulsion formation is a frequent and

serious problem when dealing with the
extraction of drugs from biological as well as
pharmaceutical formulations.

Emulsion formed
during extraction

Fig: Emulsion formation during liquid liquid extraction

Factor causes slow-coalescence

emulsion
The breaking of an emulsion (coalescence)
could be a slow process. There are a number of
factors which may be responsible for the slowcoalescence of an emulsion, namely:
(a) Finely divided powders of albumin,
gelatin and natural gums have a tendency to
coat the droplets formed in an emulsion which
ultimately prevent them from coalescing.
(b) Usually surfactants decrease the interfacial
tension (or surface tension) between the two
immiscible liquids which help in stabilizing an

(c) Ionic species may get absorbed at the

interface of two immiscible layers resulting in
the formation of a net charge on the droplets.
Because all droplets shall essentially bear the
similar charge, naturally they will repel one
another thereby preventing coalescence.
In fact, there are many natural and synthetic
substances that are profusely incorporated in
the formulation of drugs which are found to
stabilize emulsions either by coating the
droplets or by minimizing the interfacial
tension, namely:

(i) Coating the droplets: e.g., starch, acacia,

silica, gelatin, finely divided talc, and
(ii) Minimizing the interfacial tension: e.g.,
mono-and
di-glycerides;
stearates
and
sorbitan monoleate.

Prevention of emulsion formation:

It has been observed that once an emulsion is
formed it is difficult to break the emulsion.
Therefore, it is absolutely necessary to
concentrate to the following guidelines, as far
as possible, in order to avoid forming
emulsions in the course of an extraction
process:
(1) Very cautious & gentle agitation and
employing a sufficiently large liquid-liquid
interface
provides
a
reasonably
good
extraction.
When the two-liquid layers have a large
contact surface in an extraction process,

(2) The removal of any finely divided insoluble

material(s) in a liquid phase must be done by
filtration before carrying out the extraction
process.
(3) Always prefer and use such solvent pairs
that have a large density difference and a high
interfacial tension.
Water (0.9982 g/mL) and Hexane (0.6548
g/mL)
They are less prone to emulsion problems.
Water (0.9982 g/mL) and Benzene (0.8765

(4) When performing extraction from water

always ensure not to work at pH extremes and
particularly at high pH ranges (basic) to avoid
emulsification.
A major inconvenience of this method is the
extreme pH (either strongly acidic, <4, or
basic, >9)
(5) In cases of acute emulsion problems
substances like ion exchangers, alumina or
silica gel are used specifically to resolve the
problem by adsorption of the emulsifying
agents.

Process of breaking of an emulsion

(coalescence):
Following
are
the
various
techniques
invariably used to break an emulsion or to
achieve coalescence:
Mechanical means: Coalescence may be
achieved by mechanically creating turbulence
on the surfaces of the droplets either by
passing the emulsion through a bed of glasswool or simply by stirring with the help of a
glass-rod.

Centrifugation: In cases where the densities

of the two liquids are appreciably different
coalescence
may
be
achieved
by
centrifugation.
Addition of monovalent and divalent
ions: Relatively simple emulsions are broken
by adding monovalent salts like sodium
chloride; whereas charge-stabilized emulsions
are specifically sensitive to the divalent ions,
such as: CaCl2, MgCl2 etc.
Ethanol or higher alcohol: Addition of small
quantities of either ethanol or sometimes a

Silicone- defoaming agent: A few drops of

the silicone-defoaming agent sometimes help
in breaking of an emulsion.
Sudden cooling of emulsion (thermal
shock): Sudden temperature drop or freezing
(i.e., giving a thermal shock) of an emulsion
mostly enhances the interfacial tension
between the two immiscible phases thereby
causing coalescence.
Altering the ratio of solvents: Coalescence
of an emulsion may also be achieved either by
altering the ratio of the prevailing dispersed

Thin-bed of an adsorbent: Sometimes

simply passing of an emulsion through a thinbed of an adsorbent remarkably helps in
achieving coalescence.
The analyte must not be absorbed from either
solvent.

Liquid-liquid extraction (washing):