East West University

LiquidLiquid Extraction
PHRM 309

Tareq Hasan 7/25/2011

Table of Contents
Liquid Liquid Extraction ........................................2 Introduction ..........................................................2 Definition ..........................................................2 Properties of Solvents .......................................2 Properties of Solute ..........................................2
Organic Solute ......................................................3 Inorganic Solutes ..................................................3 Avoiding High pH Range ................................ 12 Removal of Surfactant by Using Adsorbents . 13 Processes of Breaking Emulsion ......................... 13 Mechanical Means ........................................ 13 Centrifugation ............................................... 13 Addition of Mono or Di valent Cations .... 13 Addition of Ethanol or Higher Alcohol .......... 13 Silicone Defoaming Agent .......................... 14 Sudden Cooling or Thermal Shock ................ 14 Alteration of Solvent Ratio ............................ 14 Thin Bed of Adsorbent ............................... 14

Principles of Liquid Liquid Extraction .................3 Different Portions of Liquid Liquid Extraction 3
Feed Phase ...........................................................3 Extractant and Extract ..........................................3 Raffinate ...............................................................4

Drying Agents ..................................................14

Definition ........................................................... 14 Classification of Drying Agents........................... 14 Magnesium Sulfate (Mg2SO4) ........................ 14 Advantages of Magnesium Sulfate ........... 15 Disadvantages of Magnesium Sulfate ...... 15 Sodium Sulfate (Na2SO4) ............................... 15 Advantages of Sodium Sulfate ................. 15 Disadvantages of Sodium Sulfate ............. 15

Theory of Liquid Liquid Extraction or Nernst Distribution Law ................................................4

Statement and Expression ....................................4 Limitation .............................................................4

Effectiveness of Liquid Liquid Extraction .......5 Criterias of Solvent Selection ............................6

Miscibility .............................................................6 Density .................................................................6 Density Difference ................................................7 Solubility ...............................................................8

Liquid Liquid Washing ........................................ 15 Definition ........................................................15 Type of Impurities removed in Liquid Liquid Washing ..........................................................15 Steps of Liquid Liquid Washing ....................15
Reaction Work-up in Liquid Liquid Washing ... 16 Separation of Target Solute from the Reaction medium .............................................................. 16 Removal of Impurities ........................................ 16

Factors influencing Liquid Liquid Extraction ..8

Effect of Temperature ..........................................8 Effect of pH...........................................................9

Emulsion Problem encountered in Liquid Liquid Extraction ...............................................9

Factors Influencing Emulsion Stability ................10 Finely Divided Powders .................................10 Surfactants ....................................................11 Ionic Species ..................................................11 Prevention of Emulsion Formation.....................12 Moderate Shaking .........................................12 High Density Difference .................................12 Removal of Finely Divided Solids ...................12

Problem of Sparingly Soluble Solvents ...........17

Solution .............................................................. 18

Difference between Liquid Liquid Extraction and Liquid Liquid Washing ...........................18

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Liquid Liquid Extraction

In tro d u cti o n

has boiling point 500C, 61.20C and 76.80C respectively) , and Since, after extraction the solvent 2 is

Definition
Liquid Liquid Extraction is a method of separation where an aqueous solution is usually brought into contact with another immiscible organic solvent, so as to affect a transfer of one or more solute from the aqueous solution into the organic solvent. This is a fast, easy and convenient method of Separation. It is usually performed by moderately shaking the 2 liquids inside a separating funnel for a few minute and can be used for either large volume or trace level.

evaporated to collect the targeted solute, there will be a less chance for the targeted solute to be degraded at high temperature. 3. Solvent 2 must be chosen accordingly. So that, The Target Compound must have higher affinity towards Solvent 2, so that, during extraction maximum amount of solute is transferred to Solvent 2. 4. Density of both of the Solvents must be known. So, that in the separating funnel it is possible to know which solvent is at the lower phase and which one is at the

Properties of Solvents
Solvents are selected based on the

upper (Usually, the Solvent with higher density is at the lower phase). 5. Density Difference between Solvent 1 and Solvent 2 should be high, so that there will be a less chance of emulsion formation and even if emulsion forms, it will easy to breakdown. 6. The Kp value / Partition Coefficient of immiscible solvent pair must be higher, since it is important for the

following criteria 1. Both Solvents must be immiscible to each other. 2. Solvent 1 can be aqueous / H2O and Solvent 2 can be Organic in nature. It is because Most Targeted Compounds (or Solutes in Solvent 1) are soluble in Organic Solvents (If the solutes are organic). Most organic Solvents has lower boiling point (E.g. Methanol,

effectiveness of extraction.

Properties of Solute
Solutes / Target Compounds can be Organic Solute 2|Page

Chloroform, Carbon Tetrachloride

 Inorganic Solute

as Impurities or as Pharmaceutical Ingredients. And before their extraction, it is absolutely necessary to form ion association complexes or metal chelates (by using organic ligands), so that they may be extracted by an appropriate organic solvent. For example, Cu (II) can be extracted by acetyl acetone by forming Cu (II) acetyl acetone complex.
P rin cipl es o f Liqu id Li quid Ext r action

Organic Solute
Most Organic Solutes are soluble in Organic solvents, which can be

extracted with any organic solvents (E.g. Extraction Caffeine by

Chloroform). However, for both pharmaceutical and therapeutical purpose, some organic solutes are introduced as salts in order to increase their solubility in Water, (E.g. Promethazine hydrochloride, a hydrochloric salt of Promethazine). So, before performing liquid liquid extraction of such organic solutes, they are converted into their original organic forms by means of a chemical reaction, so that they can be extracted by any organic solvents (E.g. Promethazine Hydrochloride Promethazine is converted they into are

Different Portions of Liquid Liquid Extraction Feed Phase

Feed Phase is the initial solution containing the target compound / Solute and Solvent 1. In the feed phase, the solvent 2 added to perform the extraction. So,

before

extracted with organic solvents like n hexane or Chloroform). However, it is to say that upon chemical reaction the concentration of original compound does not changes.

Extractant and Extract

Extractant is the Solvent 2 that is added to the Feed Phase for extraction. After extraction, the Extractant is

Inorganic Solutes
Inorganic solutes are frequently

called Extract which contains the target compound.

encountered in aqueous solvents either

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 The Extract is evaporated to collect and measure the amount of the solute. So, Before extraction, And, after extraction,

And if the extraction process is repeated, the Raffinate becomes the feed phase to which the Solvent 2 is added again. So,

And, if the extraction process is

Raffinate
Raffinate is generally, termed for the Feed Phase after extraction.

repeated, then

Theory of Liquid Liquid Extraction or Nernst Distribution Law Statement and Expression
Liquid Liquid extraction follows the theory of Nernst Distribution Law / Partition Co efficient Law which states that At a Constant / Fixed temperature, the ratio of concentration of solute in both immiscible phases is found to be a constant. The Law can be expressed as

Here, Kp = Partition Co efficient

Limitation
The Nernst distribution Law comes with 2 limitations 1. This law is only applicable for very dilute solutions. 2. The law does not hold good when the target compound or solute is

freely soluble / distributed in both solvents. That means, for this law to be applicable, the solutes must be affined to either of the Solvent especially to the Solvent 2.

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Effectiveness of Liquid Liquid Extraction

Based on the Appropriate Partition

Coefficient of an immiscible solvent pair, it is possible to calculate the Effectiveness of Liquid Liquid Extraction. Let us assume that An Initial Feed Phase Solution

[Because, Concentration = Amount of Substance or Solute / Volume]

_______(1)

contains a Solute (Target Compound) of X mole in an Aqueous Solvent, Aaq with a volume of V2. A second Organic Solvent, Borg with a Volume of V1 is added to the Feed Phase for extraction of the solute. After 1
st

( ( (

) )

extraction, the amount of

solute remaining in Aqueous Solvent, Aaq is Y mole. So, after extraction, amount of solute transferred to the Organic Solvent , Borg is (X Y) mole. Now according to Nernst Distribution Law,

Since, X mole is the total amount of solute in Solvent Aaq as Feed Phase and Y mole is the remaining amount of solute in solvent Aaq as Raffinate then in Equation 1, value of Y / X mole will be a fraction or less than

Here, COrganic = Concentration of Solute in Organic Solvent, Borg And, CAqueous = Concentration of Solute in Aqueous Solvent, Aaq And, Kp = Partition Co efficient

1 and it is the amount of solute remaining in the Raffinate as the unextracted form. Since, value of Kp is a constant for a particular solvent pair and Volume of solvents V1 and V2 are also known then it is possible to find out value of Y / X even before performing the extraction. Thus, it is

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possible to find out the Effectiveness of the Solvent Pair used in the Liquid Liquid Extraction. If the extraction process is repeated a second time by using the same solvents in the same volume, then equation 1 can be

_____ (2)
Figure 1: A Water Alcohol System A One Phase

And, for n number of extractions the equation can be _____ (3)

System (Not suitable for Liquid Liquid Extraction)

Criterias of Solvent Selection Miscibility

Solvents which are miscible to each other in all proportions (E.g. Water Alcohol mixture) have no possibility to be separated into 2 phases; rather they form a one phase system. And, these types of Solvent pairs cannot be used in Liquid Liquid Extraction Process. Solvent pairs which are immiscible to each other and form a 2 phase system (E.g. Water Benzene System or Water Chloroform System) must be used in Liquid Liquid Extraction, so that, the solute effectively transfers to the organic phase. known. So that, in the separating funnel it is possible to identify which solvent is at the lower phase and which one is at the upper. Usually, the Solvent with higher density is at the lower phase. E.g. Density of water is 0.9982 gm / ml and of n hexane is 0.6548 gm / ml. 6|Page
Figure 2: A Water Benzene System A Two Phase System (A Suitable Solvent Pair for Liquid Liquid Extraction)

Density
Density of both of the Solvents must be

 So, in the separating funnel, Water will form the lower phase and n hexane will be at the Upper phase.

gm / ml and 0.6548 gm / ml respectively. In case of Water benzene system there is a very low density

difference (0.1217 gm / ml); so the system is very susceptible to

emulsion formation even upon moderate shaking. But, in case of Water n-hexane system, there is a high density difference (0.3434 gm / ml); so there is less chance of emulsion formation.
Figure 3: A Water - n - hexane System (Water is at the lower Phase due to higher density and n hexane is at the upper phase due to lower density)

Density Difference
Density difference between the

solvents must be high. It is because In Liquid liquid Extraction for pharmaceutical purpose, a frequent number of substances are encountered which are very susceptible to emulsion formation. There will be a less chance of emulsion formation upon shaking and even if emulsion E.g. Density of Water, Benzene and n hexane are 0.9982 gm / ml, 0.8765 forms, it will easy to

breakdown.
Figure 4: Emulsion formation in water - benzene system due to low density difference and moderate shaking

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Solubility
Though solvents used in Liquid Liquid Extraction are immiscible to each other, there is chance that the solvents are soluble / distributable in each other to a slight percentage. This is called Partial Solubility or Percentage Solubility. E.g. 1. In a CH2Cl2 Water system, 1.6% CH2Cl2 is soluble in water and, 0.24% Water is soluble in CH2Cl2. 2. In a Carbon tetrachloride Water system, 0.03% of Carbon

So, solvents must be both immiscible and insoluble to each other at any volume, otherwise the following

problems will occur 1. Volume Change may occur which will give error in the result. 2. Aqueous Contamination in the Organic Extract may increase its boiling point close to 1000C, so during evaporation of the extract (to collect and measure the solute), the solute can be degraded due to high temperature, thus resulting in content loss. In case of Law solubility, is Partition as

tetrachloride is soluble to Water and 0.03% of Water is soluble in Carbon tetrachloride.

Coefficient following

expressed

Here, SOrganic = Solubility of Solute in Organic Solvent And, SAqueous = Solubility of Solute in Aqueous Solvents

Factors influencing Liquid Liquid Extraction Effect of Temperature

Solubility of a compound changes
Figure 5: Distribution of Water and CH 2 Cl 2 in each other.

according to the changes in the temperature.

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 Solubility of a Solute increases with increasing temperature. In case of Liquid Liquid Extraction, this is crucial because, differences in

pH is important in Liquid Liquid Extraction because, most drugs to be extracted are either weak acids or weak bases which donate or accept protons respectively. Since, the solvent for extraction is organic, the solute / Target Compound must be in their extractable form (A neutral or unionized form of the solute regardless whether the solute is a weak base or a weak acid). So, pH of the Feed Phase must be adjusted by using a suitable buffer, so that the solute remains in their neutral / unionized form.

temperature at any sequence of the extraction process may change the solubility of solute in the both aqueous and organic phases. And if the solubility of the solute decreases in Organic Phase but

increases in Aqueous Phase due to change in temperature change then, it will be disastrous, because for the extraction process to be effective, the solute must have higher solubility towards Organic Solvent. So, temperature must be kept fixed during the extraction process.

Emulsion Problem encountered in Liquid Liquid Extraction

Emulsion can be defined as a dispersed

Effect of pH
pH is a measurement of concentration of proton present in a solution. It determines whether the solution is basic, acidic or neutral. The higher the pH value of a solution the fewer protons are present in it and the less acidic or more basic it is, and vice versa. pH value of neutral solution is 7. pH is expressed as the negative logarithm of Concentration of Proton.

system where one liquid phase is dispersed as droplets into another immiscible liquid phase and the dispersion is stabilized by using an appropriate emulsifying agent. In Liquid Liquid Extraction, the phases are dispersed in each other through moderate shaking so that they can come in each others contact for solute transfer. But, they must not form emulsion and after shaking they must form separate layers again. It is to remember that a good emulsion may destabilize into 9|Page

1. Coalescence 2. Flocculation

3. Creaming 4. Breaking

Figure 6: Instabilities in an Emulsion

Factors Influencing Emulsion Stability

Emulsion formation is a problem when dealing with the extraction of drugs from biological or pharmaceutical

Finely Divided Powders have a tendency to coat the droplets formed in an emulsion which ultimately prevent them from

forming coalescence. They are used as emulsifying agents and as binders in pharmaceutical liquid and solid dosage forms respectively. Such examples are Albumin Gelatin Natural Gums (Such as Acacia)

formulations which tend to emulsify even upon moderate shaking due to presence of emulsion stabilizers in the formulation. These Emulsion Stabilizers are 1. Finely Divided Powders 2. Surfactant 3. Ionic Species Finely Divided Powders

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Figure 7: Coating of Finely Divided Powders on the Droplets which prevents Emulsion Breaking

Figure 8: Surfactants Molecules Surrounding the Droplets

Surfactants Surfactants are used to increase H2O solubility of a compound which can either be solid / liquid. Presence of such surfactants is very frequent formulations in pharmaceutical which prevent

Ionic Species Ionic Species get absorbed at the interface of the droplets resulting in a net charge on the droplets. Because all droplets essentially bear the similar charge, they will repel one another thereby preventing coalescence.

coalescence by surrounding the droplets formed. Examples of such surfactants are Cetosteryl alcohol Glyceryl Monooleate

Figure 9: Anionic Species Absorbed at the Interface of Droplets builds up a net charge on the Droplets and repelling each other preventing Coalescence

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Prevention of Emulsion Formation

Moderate Shaking The solvents must be shaken moderately, so that the phases are dispersed in each other for solute transfer but then again forms separate layers. It is because vigorous shaking can form emulsion. High Density Difference Density Difference between the solvents chosen must be high so that they do not form emulsion upon dispersion. E.g. Density of Water, Benzene and n hexane are 0.9982 gm / ml, 0.8765 gm / ml and 0.6548 gm / ml respectively. In case of Water benzene system there is a very low density difference (0.1217 gm / ml); so the system is very susceptible to emulsion

there is less chance of emulsion formation.

Figure 10: Formation of Emulsion in water benzene system due to low density difference

Removal of Finely Divided Solids Finely divided Solids tend to

stabilize the emulsion Formation by coating the droplets. So, before extraction these solids must be removed from feed phase by filtration. Avoiding High pH Range Since, Emulsification occurs more frequently and easily in Extreme pH range (i.e. below 4 Extremely Acidic and above 9 Extremely Basic); these ranges should be avoided and kept between 4 to 9.

formation even upon moderate shaking. But, in case of H2O n-hexane system, there is a high density difference (0.3434 gm / ml); so

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Removal of Surfactant by Using Adsorbents To remove Surfactants, a thin bed of adsorbents (E.g. Alumina, Silica Gel) can be used through which the feed phase can be passed through. However, use of such adsorbents must be avoided which can adsorb the solute.

Centrifugation Centrifugation is a process of separation by the application of Centrifugal Force based on the density difference between the phases. So, when the density difference between the phases / solvents is considerably high, centrifugation can be applied to break the emulsion.

Processes of Breaking Emulsion

Mechanical Means In case of simple emulsion,

Addition of Mono or Di valent Cations Saturated Solution of Mono valent or Di valent Salts (E.g. NaCl or CaCl2) can be added to break down the emulsion by increasing the ionic strength of Water / Aqueous Phase / Feed Phase. Upon increasing the ionic strength of Feed Phase, solubility of organic solute or Complex / Chelate of inorganic solute in aqueous phase / Feed Phase decreases and they will return to their own more soluble organic phase. Addition of Ethanol or Higher Alcohol Addition of Ethanol or higher alcohol increases coalescence in

coalescence can be achieved by mechanically creating turbulence on the surface of the droplets by stirring with a glass rod.

Figure 11: Emulsion Breaking by Mechanical Means

emulsion

by

increasing

the

interfacial tension between phases. 13 | P a g e

Silicone Defoaming Agent Addition of Silicone Defoaming Agent helps in breaking the

Drying Agents are compounds that are used to absorb H2O from the extract, since presence of H2O in the extract can cause Volume Change giving error in the

emulsion. Sudden Cooling or Thermal Shock Sudden temperature drop or

result. Water in the Organic Extract may increase its boiling point close to 1000C, so during evaporation of the extract (to collect and measure the solute), the solute can be degraded due to high temperature, thus resulting in content loss.

freezing (i.e. giving a thermal shock) of emulsion mostly enhances the interfacial tension between the two immiscible phases thereby causing coalescence. Alteration of Solvent Ratio Coalescence of an emulsion can be achieved by altering the ratio of solvents prevailing in the dispersed phase. It can be done by slowly adding solvents. Thin Bed of Adsorbent A thin bed of adsorbents (E.g. Alumina, Silica Gel) can be used through which the feed phase can be passed through to break the emulsion.

Upon addition of drying agents and following stirring, they will react with water present in the extract to form a clump and settle to bottom. When there will be no water in the extract, drying agents will float on the surface of the extract, since there is no water to react with.

Classification of Drying Agents

Drying Agents used in Liquid Liquid Extraction can be of 2 types 1. Magnesium Sulfate (Mg2SO4) 2. Sodium Sulfate (Na2SO4) Magnesium Sulfate (Mg2SO4) Mg2SO4 can be used as Drying Agent to remove Water from the extract, in Liquid Liquid Extraction 14 | P a g e

Drying Agents Definition

 If water is present in the extract, then, Mg2SO4 will form Magnesium Sulfate Water complex

Disadvantages of Sodium Sulfate Sodium Sulfate is a granular compound, so it has low surface area. As a result, it absorbs water slowly from the extract. Liquid Liquid Washing

(Mg2SO4.2H2O). Advantages of Magnesium Sulfate Magnesium Sulfate is granule type material, so it has large surface area. As a result it provides quick absorption of H2O from the extract. Disadvantages of Magnesium Sulfate Magnesium sulfate is acidic in nature, so it easily reacts with basic target solutes. Sodium Sulfate (Na2SO4) Na2SO4 can be used as Drying Agent to remove Water from the extract, in Liquid Liquid Extraction If water is present in the extract, then, Na2SO4 will form a clump of Sodium Sulfate Water complex (Na2SO4.2H2O) which will settle at the bottom. Advantages of Sodium Sulfate Sodium Sulfate is neutral

Definition
Liquid Liquid Washing is a process of separation of impurities from the extract or reaction medium containing the target solute and also the impurities. The process can be used to remove impurities from a reaction medium.

Type of Impurities removed in Liquid Liquid Washing

Impurities to be removed via Liquid Liquid Washing can be of 2 type Byproduct Catalyst For example, all of A and B reacts to form C and D which is catalysed by X. So, here the impurities are the byproduct D and catalyst X.

compound, so it can be used with any type of solute.

Steps of Liquid Liquid Washing

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 Steps involved in the Liquid Liquid Washing are 1. Reaction Work up 2. Separation of Target Solute from the Reaction medium by adding Organic Solvent 3. Removal of Impurities

So, after the reaction of all of Phenol and Acetic anhydride, the compounds that remain in the reaction medium are Phenol acetate Acetic Acid the Byproduct Triethylamine the Catalyst Here, Acetic Acid the Byproduct and Triethylamine the Catalyst are the impurities and Phenol acetate is the

Reaction Work-up in Liquid Liquid Washing

Reaction of Phenol and Acetic

product or Target Compound.

anhydride results in the formation of Phenol acetate and Acetic Acid which is catalysed by Triethylamine.

Separation of Target Solute from the Reaction medium

Before performing the separation, it must be considered that solubility of the target compound in organic solvent must be higher than the solubility of impurities. So, solvent should be selected in such a way. In case of this reaction, the target compound Phenol acetate is insoluble in water and soluble in Ether which is higher than that of both Acetic Acid and Triethylamine. So, Ether can be the organic solvent of choice to separate Target Solute from the Reaction medium.

Removal of Impurities
Figure 12: Reaction of Phenol and Acetic anhydride

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 Selection

of

Solvents

to

remove

Impurities of basic nature can be removed by adding dilute solution of Strong Acids in the organic extract. E.g. Triethylamine (A base) can be removed by adding 10% HCl aqueous solution in Ether extract of Phenol acetate. As a result, Triethylamine hydrochloride will form which is salt of Triethylamine and soluble in water.

impurities depends on the properties of impurities. Impurities of acidic nature can be removed by adding saturated aqueous solution of a Strong Base in the organic extract. E.g. Acetic Acid (A Weak Acid) can be removed by adding saturated aqueous solution of NaHCO3 in Ether extract of Phenol acetate. As a result, Sodium acetate will form which is salt of Acetic Acid and soluble in water from the saturated aqueous solution of NaHCO3.

Figure 14: Removal of Triethylamine

Problem of Sparingly Soluble Solvents

Use of sparingly soluble solvents must be avoided. It is because, severe content loss of target
Figure 13: Removal of Acetic Acid

compound occurs upon using sparingly soluble solvents especially when the organic 17 | P a g e

solvent is soluble in water (E.g. Solubility of Ether in water is 5.6 mg / 100ml). So, an amount of target compound that is soluble in organic solvent is lost every time, when the separation is performed, since an amount of organic solvent is sparingly soluble in water. Thus resulting in content loss of target compound.

Table

1:

Difference

between

Liquid

Liquid

Extraction and Liquid - Liquid Washing

Topic

Solution
The problem of content loss is solved by adding Brine solution / Saturated Solution of NaCl in water before performing separation of impurities which incorporates a high concentration of Na+ and Cl- ions in the water, thus increasing its ionic strength. As a result, the solubility of organic compound in aqueous phase will decrease. So, the organic compound will transfer to its own organic phase.

Liquid Liquid Extraction Target Extracted by Compound Organic Solvent Solvent Performed to Evaporation collect and measure the Target solute Solubility of Solubility of Target the target Solute compound in organic solvent must be higher than in aqueous solvent Selection of Depends on Solvents Miscibility, Solubility and Density of the solvent pair Contents in Only Target the extract Solute

Liquid Liquid Washing Remains in the Organic Solvent Not required because Impurities are removed Solubility of the target compound in organic solvent must be higher than the solubility of impurities Depends on the properties of impurities

Target Solute + Impurities (Byproduct + Catalyst)

Difference between Liquid Liquid Extraction and Liquid Liquid Washing

Difference between Liquid Liquid Extraction and Liquid Liquid Washing is as followed

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