Liquid -Liquid Extraction

Presented by –Gitanjali Sahu

M Pharm pharmaceutical analysis
Anurag university
Definitions
• The separation of constituents (solutes) of a liquid solution by contact
with another insoluble liquid. Solutes are separated based on their
different solubilities in different liquid. The separation process of the
components of a liquid mixture by treatment with a solvent in which
one or more desired components is soluble.
Requirements
• There are two requirements for liquid – liquid extraction to be feasible:
• component (s) to be removed from the feed must preferentially distribute in the
solvent.
• The feed and solvent phases must be substantially immiscible. 
Characteristics of solvent
• The two liquid phases must be either immiscible, or partially miscible.
• usually isothermal and isobaric
• can be done at low temperature (good for thermally fragile solutes, such as large organic
molecules or biomolecules)
• can be very difficult to achieve good contact between poorly miscible liquids (low stage
efficiency)
• extracting solvent is usually recycled, often by distillation (expensive and energy-intensive)
• can be single stage (mixer-settler) or multistage (cascade ). 
Steps involved in liquid-liquid extractions
• In the simplest case, three components are involved:
• transition component A
• solvent B
• carrier liquid C
• The transition component A is combined with the carrier liquid C as the initial mixture (feed). If
the initial mixture and the solvent B are mixed together, the transition component A is transferred
into the solvent B.
• The requirement for this is that the solubility of the transition component A in the solvent B is
higher than in the carrier liquid C. In turn, the carrier liquid C should be almost insoluble in the
solvent B
Cont…
This means that the actual separation process results in two phases after settling:
• extract phase (mainly A and B, with residue of C)
• raffinate phase (mainly C, with residue of A and B)
To obtain the purest possible transition component, the extraction is normally followed by a
separating stage that takes the form of rectification, in which the solvent is separated from the
transition component. The solvent can be recirculated and is then available for the extraction again.
Principle of LLE
• The two phases are put into a device called a separatory funnel, and compounds in the system
will distribute between the two phases. There are two terms used for describing this
distribution, one of which is called the distribution coefficient (DC), the other of which is called
the partition coefficient (DM).
•  The distribution coefficient is the ratio of the concentration of solute in the organic phase over
the concentration of solute in the aqueous phase (the V-terms are the volume of the phases).
This is essentially an equilibration process whereby we start with the solute in the aqueous
phase and allow it to distribute into the organic phase. 
• soluteaqueous= soluteorganic
• DC=[solute]org molorg/Vorg mol Vaq
org*

• ________ = ________ = ________

• [solute]aq molaq/Vorg molaq*Vorg

• The distribution coefficient represents the equilibrium constant for this process .
Partition coefficient
• The partition coefficient is the ratio of the moles of solute in the two phases, and is a
more effective means of measuring whether you have achieved the desired goal. The
larger the value of DM, the more of the solute we have extracted or partitioned into the
organic phase.
•
Dm=molorg
• _________

• molaq
• molorg*molaq
• DM= _____________ =DC(Vaq/Vorg)
• molaq*molorg
Henderson hasselbalch equation
• The formula for the Henderson–Hasselbalch equation is: pH=pKa+log([A−]/[HA]) 
• where pH is the concentration of [H+]
• pKa is the acid dissociation constant, and [A–] and [HA] are concentrations of the conjugate base
and starting acid. 
• The balanced equation for an acid dissociation is: 
• HA⇌H++A− 
• The acid dissociation constant is: 
• Ka=[H+][A−]/[HA] 
• After taking the log of the entire equation and rearranging it, the result is: 
• log(Ka)=log[H+]+log([A−]/[HA]) 
Cont….
• log(Ka)=log[H+]+log([A−]/[HA]) 
• This equation can be rewritten as: 
• −pKa=−pH+log([A−]/[HA]) 
• −pKa=−pH+log([A−]/[HA]) 
• Distributing the negative sign gives the final version of the Henderson-Hasselbalch equation: 
• pH=pKa+log([A−]/[HA]) 
• pH=pKa+log([A−]/[HA])  
• where pKa= dissociation constant of acid 
Cont….

• Kb is the dissociation constant for base

Drug absorption in relation to pH and pka
Ph distribution in human body
Application
• Major applications exist in the biochemical or pharmaceutical industry, where
emphasis is on the separation of antibiotics and protein recovery.
• In the inorganic chemical industry, they are used to recover high – boiling
components such as phosphoric acid, boric acid and sodium hydroxide from
aqueous solution. 
Back extractions
• First extracted at low pH in organic phase
• Polar interferences that is neutral, protonated bases are left behind in
aqueous phase.
• Fresh portion of high pH aqueous buffer is added to organic phase.
• Back extractions takes place .
• Ionized organic acid is transferred back on to aqueous phase leaving
non- polar interference in aqueous phase.
Cont….
• Thus, two step back extractions allows the removal of both basic and
neutral interfernces.
• Whereas one step extractions can eliminate one or the other of these
interferences but not both.
Liquid liquid extractor
Case study-sample preparation for liquid-liquid extraction
for simultaneous determination of antineoplastic drugs in
plasma

• Aliquots of 1 ml of blank plasma spiked with the drugs

• Extraction with 5ml of Ethylacetate

• Shaking of tubes for 15 mins at 50 rpm

Cont….

• Centrifugation for 10min at 4000rpm .

• The upper organic layer transferred to conical tubes and separated.

• Aqueous layer reextracted with 5ml of Ethylacetate.

Cont….
• Organic phase were combined and evaporated at 40◦c.

• The residues were reconstituted in 200µl mobile phase and 50µl

were injected in to the HPLC system.
•Thank you