Extraction

Taken from various sources

• Why separation and isolation are
important?
 Steps of separation of natural
products
General
isolation
strategy of
natural
products…
• Crude aqueous
extracts of certain
plants (and animals)
provided pigments,
such as indigo and
alizarin.
• Other examples of natural
products:

• ephedrine
from Ephedra sinica
(for respiratory
ailments) tetrahydrocann
abinol (marijuana)
geraniol (rose oil)

• cinnamaldehyde
(cinnamon) diallyl
disulfide (garlic)
• morphine
(narcotic analgesic)
1817
• strychnine
(poison) 1818
• cocaine
(narcotic stimulant)
1859
• nicotine
(toxic) 1828
Liquid-liquid extraction is a useful method to separate components
(compounds) of a mixture
Let's see an example.

Suppose that you have a mixture of sugar in vegetable oil (it tastes
sweet!) and you want to separate the sugar from the oil. 

You
observe that the sugar particles are too tiny to filter and you
suspect that the sugar is partially dissolved in the vegetable oil.

What will you do?

 How about shaking the mixture
with water
Will it separate the sugar from the
oil? Sugar is much more soluble in
water than in vegetable oil, and,
as you know, water is immiscible
(=not soluble) with oil.

Did you see the result?The water

phase is the bottom layer andthe oil
phase is the top layer, because
water is denser than oil.

*You have not shaken the mixture

yet, so sugar is still in the oil phase.
By shaking the layers (phases) well, you
increase the contact area between the
two phases.The sugar will move to the
phase in which it is most soluble: the
water layer

Now the water phase tastes

sweet,
because the sugar is moved to
the water phase upon shaking.**You
extracted sugar from the oil with
water.**In this example,water was
the extraction solvent ;the original
oil-sugar mixture was the solution to
be extracted; and sugar was the
compound extracted from one phase
to another. Separating the two layers
accomplishes the separation of the
sugar from the vegetable oil
Did you get it? .....the concept of liquid-liquid extraction?

Liquid-liquid extraction is based on the transfer of a solute

substance from one liquid phase into another liquid phase according
to the solubility.Extraction becomes a very useful tool if you choose
a suitable extraction solvent.
You can use extraction to separate a
substance selectively from a mixture, or to remove unwanted
impurities from a solution.In the practical use, usually one phase is a
water or water-based (aqueous) solution and the other an organic
solvent which is immiscible with water.

The success of this method depends upon the difference in solubility

of a compound in various solvents. For a given compound, solubility
differences between solvents is quantified as the "distribution
coefficient"
Partition Coefficient Kp (Distribution Coefficient Kd)

When a compound is shaken in a separatory funnel with two immiscible

solvents, the compound will distribute itself between the two solvents.

Normally one solvent is water

and the other solvent is a
water-immiscible organic
solvent.

Most organic compounds are

more soluble in organic solvents,
while some organic compounds
are more soluble in water.
Here is the universal rule:

At a certain temperature, the ratio of concentrations of a solute

in each solvent is always constant.ﾊAnd this ratio is called the
distribution coefficient, K.

(when solvent1 and solvent2 are immiscible liquids

For example,Suppose the

compound has a distribution
coefficient K = 2 between
solvent1 and solvent2

By convention the organic

solvent is (1) and waater is
(2)
 (1) If there are 30 particles
of compound , these are
distributed between equal
volumes of solvent1 and solvent2..

(2) If there are 300

particles of compound , the
same distribution ratio is
observed in solvents 1 and 2

(3) When you double the

volume of solvent2 (i.e., 200
mL of solvent2 and 100 mL of
solvent1),
the 300 particles of
compound distribute as shown

If you use a larger amount of extraction solvent, more solute is

extracted
What happens if you extract twice with 100 mL of solvent2 ?
In this case, the amount of extraction solvent is the same volume as was
used in Figure 3, but the total volume is divided into two portions and
you extract with each.

As seen previously, with 200 mL

of solvent2 you extracted 240
particles of compound . One
extraction with 200 mL gave a
TOTAL of 240 particles
You still have 100 mL of solvent1,
containing 100 particles. Now you
add a second 100 mL volume of
fresh solvent2. According to the
distribution coefficient K=2, you
can extract 67 more particles
from the remaining solution
An additional 67 particles are
extracted with the second portion
of extraction solvent
(solvent2).The total number of
particles extracted from the first
(200 particles) and second (67
particles) volumes of extraction
solvent is 267.This is a greater
number of particles than the
single extraction (240 particles)
using one 200 mL portion of
solvent2!

It is more efficient to carry out

two extractions with 1/2 volume
of extraction solvent than one
large volume!
If you extract twice with 1/2 the volume, the extraction is more
efficient than if you extract once with a full volume. Likewise,
extraction three times with 1/3 the volume is even more efficient….
four times with 1/4 the volume is more efficient….five times with 1/5
the volume is more efficient…ad infinitum

The greater the number of small extractions, the greater the

quantity of solute removed. However for maximum efficiency the
rule of thumb is to extract three times with 1/3 volume
Chemically active (acid-base) extraction

Can you change the solubility property of a compound? How?

Most organic compounds are more

soluble in organic solvents than in
water,usually by the distribution
coefficient K > 4
However, specific classes of
organic compounds can be
reversibly altered chemically to
become more water-soluble.

This is a powerful technique and allows you to separate organic

compounds from a mixture -- if they belong to different solubility
classes
What type of organic compounds can be made water-soluble?
Compounds belonging to the following solubility classes can be
converted to their water-soluble salt form

(1) Organic acids include carboxylic acids (strong organic acids)

and phenols (weak organic acids).

(2) Organic bases includes amines

How can organic acids or bases be converted to a water-solubleform?
1. Organic Acids can be converted to their salt form when treated with an
aqueous solution of inorganic base (e.g., NaOH (sodium hydroxide) and
NaHCO3 (sodium bicarbonate)).Salts are ionic, and in general, ions are
soluble in water but not soluble in water-immiscible organic
solvents.Remember: water is a very polar solvent thus salts (i.e., ionic
species) are well dissolved in it.

A. Carboxylic Acids are

converted to the salt form
with 5% NaOH aqueous
solution. NaOH is a strong
inorganic base.

Carboxylic acids are strong

organic acids (pKa = 3 to 4),
so they can also be ionized
with weak inorganic bases
(e.g., NaHCO3 (sodium
bicarbonate)) aqueous
solution.
Let's try a sample problem.
Here is a mixture of naphthalene and benzoic acid, dissolved in
dichloromethane.

You want to separate

these two compounds.
What will you do?

You may use an aqueous solution of

either 5% NaOH or sat. NaHCO3, to
extract benzoic acid as a salt form
B. Phenols are considered to be weak organic acids. Phenol, the parent
compound, is partially water-soluble (1 g will dissolve in 15 mL of water),
whereas substituted phenols are not.Sodium bicarbonate (NaHCO3) aqueous
solution, a weak inorganic base, will not deprotonate phenols to make it
ionic, because it is not strong enough.However, treatment with NaOH, a
strong inorganic base, can change phenol to its ionic (salt) form.
 Let's try a another sample problem.
Here is a mixture of benzoic acid and p-methoxyphenol, dissolved in
dichloromethane.

You want to separate

these two compounds.
What will you do?

You cannot use 5% NaOH to separate these two

compounds. NaOH will react with both benzoic acid and p-
methoxyphenol, thus both compounds will be extracted into the
aqueous layer.
 Let's try this problem again.
Here is another mixture of benzoic acid and p-methoxyphenol, dissolved in
dichloromethane.

NaOH was too strong a base,

thus it does not differentiate
the strong and weak organic
acids. Use of weak inorganic
base such as NaHCO3 will
differentiate between the
compounds

Strong organic acids such as benzoic acid would be

deprotonated and ionized, while weak organic acids such
as phenols would NOT be deprotonated
2. Organic Bases (amines) can be converted to their salt form when treated
with an aqueous solution of an inorganic acid such as HCl (hydrochloric acid).

Recall that salts are ionic and generally soluble in water but not soluble in
water-immiscible organic solvents.
Let's try a third sample problem.
Here is a mixture of benzoic acid and p-chloroaniline, dissolved in
dichloromethane.

You want to separate

these two compounds.
What will you do?

You may use an aqueous solution of

either 5% HCl, to extract the amine as
a salt form and benzoic acid has
remained in the organic layer
You can separate four different classes of compounds from a mixture based on
differing solubility properties. The four classes are:1.Amines (organic
base)2.Carboxylic acids (strong acid)3.Phenols (weak acid)4.Neutral
compounds.
After the separation of the mixture of four components, we will have four
solutions: each solution contains one component.

The first three compounds are chemically altered, existing in their salt form
dissolved in aqueous solution. The fourth compound is not chemically altered,
but it is dissolved in an organic solvent.We now want to recover each compound
in its original state (i.e., in the non-ionic form) to complete the experiment. We
call this step isolation or recovery.

Let's see, one by one, how to recover each compound obtained from the
separation process
Isolation (Recovery) of amines
An amine is a basic compound. It is protonated in the presence of excess HCl
forming a salt that is soluble in aqueous solution. This is how you separated the
amine from the original mixture containing it.

An amine is soluble in acidic aqueous

solution because it forms a salt, an
ionic form.

However, if you change the pH

of the solution to basic the
amine can no longer stay
dissolved because it is no longer
ionic! This process is called
basification.

Basification is done by carefully

adding concentrated NaOH
solution to the solution containing
the amine salt until it becomes
basic.
･In the basification step, you use
concentrated NaOH solution to
minimize the volume of the final
solution. Recall that a dilute
solution of HCl was used to
extract the amine as its water-
soluble salt (see the picture on
the right side).


･Basification must be done

carefully, portion by portion, with
swirling each time because the
acid-base neutralization reaction
is exothermic.


･Check the pH of the solution to

ensure that it is basic. (~pH 10)
 Isolation (Recovery) of Acids
There are two different groups of organic acids: carboxylic acids (strong acids) and
phenols (weak acids).In the separation procedure, acids were extracted using (weak or
strong) basic aqueous solutions
Both acids can be returned to the original form in the same
manner! Organic acids are currently dissolved in a basic aqueous
solution, because the acid forms a salt, an ionic form. When you
make the aqueous solution acidic, the organic acids no longer
remain dissolved because they are no longer ionic and usually
precipitate out of solution. This process is called acidification.

Acidification is done by carefully adding

concentrated HCl solution until the mixture
becomes acidic,

When the weak base, NaHCO3, was the

extracting solution, CO2 gas will evolve
during acidification.
･The recovery of organic acids
requires acidification with
concentrated HCl solution. Recall
that in the extraction step for
the separation of an organic acid
either dilute NaHCO3 or NaOH
was used. Concentrated HCl will
now help minimize the volume of
the final aqueous solution


･Acidification must be done

carefully, portion by portion, with
swirling each time because the
acid-base neutralization reaction
is exothermic.


･Check the pH of the solution to

ensure that it is acidic. (~pH 3)
 RCO2H / ArOH / RNH2 / RH General Acid/Base Extraction
dissolved in diethyl ether
Flow Diagram
extract with • separation of organic acids, bases, and neutrals
5% NaHCO3

Aqueous Phase #1 Organic Phase #1

RCO2Na dissolved ArOH / RNH2 / RH RCO2H = carboxylic acids
in aqueous base dissolved in Et2O ArOH = phenols
RNH2 = amines
acidify extract with RH = others (neutrals)
with HCl 5% NaOH

RCO2H Aqueous Phase #2 Organic Phase #2

ArONa dissolved RNH2 / RH
in aqueous base dissolved in Et2O

acidify extract with

with HCl 5% HCl

ArOH Aqueous Phase #3 Organic Phase #3

dissolved RNH3Cl RH dissolved in Et2O

make basic dry, filter &

with NaOH concentrate

RNH2 RH
 Acetamidophen Aspirin Caffeine

O OH
O CH3
H
N CH3 O CH3 H3C N
N
O O
HO O N N
CH3
ACE (phenol) ASP (carboxylic acid) CAF (amine)
[weak organic acid] [strong organic acid] [organic base]

extract extract extract

with with with
aqueous aqueous aqueous
NaOH NaHCO3 HCl

O O- Na+
O CH3
H
N CH3 O CH3 H3C +
N H Cl-
N
O O
Na+ -O O N N
CH3
phenoxide salt carboxylate salt ammonium chloride salt
polar, water soluble polar, water soluble polar, water soluble
 ACE / ASP / CAF
methylene chloride

gravity
filter

separates insoluble
solid #1 solid from solution
solution #1 {separatory funnel}

acetaminophen ASP / CAF in CH2Cl2

example extract with

10 mL NaHCO3

extraction solution #2
separates acids
from neutrals
solution #3

flow diagram lower layer in the

separatory funnel } (organic phase)
CAF dissolved
(aqueous phase)
ASP salt dissolved
(density of CH Cl
= 1.325 g/mL )
2 2
in CH2Cl2 in aqueous base

dry with acidify with

anh. MgSO4 2 mL 6N HCl
for separation of three components:
acetaminophen, aspirin & caffeine filter with glass filter ppt.
fritted funnel

solid #3
concentrate by
removing solvent aspirin

solid #2
 Separatory Funnel Extraction Procedure
Separatory funnels are designed to facilitate the mixing of immiscible liquids
 Separatory Funnel Extraction Procedure

1. Support the 2. Pour in liquid to 3. Add extraction 4. Add ground glass

separatory funnel in a be extracted solvent Stopper (well greased)
ring on a ringstand.
Make sure stopcock is
closed
 Separatory Funnel Extraction Procedure
Shake the separatory funnel.

Then, point the stem up and slowly open the

stopcock to release excess pressure. Close the
stopcock. Repeat this procedure until only a small
amount of pressure is released when it is vented
Pick up the separatory
funnel with the stopper in
palce and the stopcock
closed, and rock it once
gently.
 Separatory Funnel Extraction Procedure
Shake the separatory funnel vigorously.
Now, shake the funnel vigorously for a few seconds. Release
the pressure, then again shake vigorously. About 30 sec
total vigorous shaking is usually sufficient to allow solutes to
come to equilibrium between the two solvents.

Vent frequently to prevent pressure buildup,

which can cause the stopcock and perhaps
hazardous chemicals from blowing out. Take
special care when washing acidic solutions with
bicarbonate or carbonate since this produces a
large volume of CO2 gas
 Separatory Funnel Extraction Procedure
Separate the layers.

Let the funnel rest While waiting, remove the Carefully open the stopcock and
undisturbed until the layers stopper and place a beaker allow the lower layer to drain
are clearly separated or flask under the sep into the flask. Drain just to the
funnel. point that the upper liquid
barely reaches the stopcock
 Solvent Extraction
• Equilibrium constant for this partitioning is
K (partition coefficient)

[S]2
K=
[S]1
 Solvent Extraction
• Determination of solute concentration in
each phase
• Define some variables:
– V1 & V2 are volumes of solvents 1&2
– m = total # of moles of solute (S) present
– q = fraction of solute remaining in phase 1 at
equilibrium
 Solvent Extraction
• [S]1 = qm/V1
• [S]2 = (1-q)m/V2
 Solvent Extraction

[S]2 (1-q)m/V2 (1-q) /V2

K= = =
[S]1 qm/V1 q/V1
Rearrange:

V1 KV2
q= (1-q) =
KV2 + V1 KV2 + V1

fraction of S in: phase 1 phase 2

 Solvent Extraction
• If remove V2 and extract V1 with fresh
layer of V2, what fraction remains in V1?
– Initial moles = m
– after first extraction - qm
– after second extraction - q(qm)=q2m

V1 2
q(2) =
KV2 + V1
 Solvent Extraction
• Fraction in V1 after n extractions:

V1 n
q(n) =
KV2 + V1
 Solvent Extraction
• Example: Solute A has a partition
coefficient of 4.000 between hexane and
water. (K = [S]hexane/[S]water = 4) If 150.0
ml of 0.03000 M aqueous A is extracted
with hexane, what fraction of A remains if:
 Solvent Extraction
• a) one 600.0 ml aliquot of hexane is used?

150ml
q= = 0.05882 = 5.882%
4(600ml) + 150ml

# moles remaining
0.05882 (0.03M•0.150L) = 2.647x10-4 moles
 Solvent Extraction
• b) 6 successive 100.0 ml aliquots of
hexane are used?

150ml 6
q= = 0.0004115
4(100ml) + 150ml

# moles remaining
4.115 x 10-4 (0.03M•0.150L) = 1.852x10-6moles
 Solvent Extraction
• Although same volume of hexane is used,
it is more efficient to do several small
extractions than one big one!
– 1 600 ml extraction extracts 94.12%
– 6 100 ml extractions extract 99.96%
 Solvent Extraction (pH effects)
• with organic acids/bases:

Ka
HA H+ + A-
Kb
B + H2O BH+ + OH-
Generally, neutral species are more soluble
in an organic solvent and charged species
are more soluble in aqueous solution
 Solvent Extraction (pH effects)
• Partitioning of organic acids between two
phases:
very little here, ions
have poor solubility
organic HA H+ + A-

Ka
aqueous HA H+ + A-
 Solvent Extraction (pH effects)

• When the solute (acid/base) can exist in

different forms, D (distribution coefficient) is
used instead of K (partition coefficient)
 Solvent Extraction (pH effects)

total conc. in phase 2

D=
total conc. in phase 1
HA [HA]org
D=
K [HA]aq + [A-]aq
Ka
HA H+ + A-
 Solvent Extraction (pH effects)

• Substitute for [A-] in D eq. and rearrange

[H+][A-] Ka [HA]
Ka = [A-] =
[HA] [H+]
[HA]2
D=
Ka [ HA]1
[HA]1  
[H ]
 Solvent Extraction (pH effects)

[HA]2 [HA]2
D= 
Ka [ HA]1  K 
[HA]1   [HA]1  1   
a
[H ]  [ H ]

 Ka  [HA]2
D 1   = =K
 [ H ] [HA]1
Solvent Extraction (pH effects)

 Ka 
D 1   = K
 [ H ]


K K[ H ]
D=  
 Ka  [ H ]  Ka
 1  
 [ H ]
 Solvent Extraction (pH effects)
• pH effect on D for organic acids

[H+]=Ka
[H+]>>Ka
pH=pKa
K

mainly mainly
D HA A-

[H+]<<Ka

pH
 Solvent Extraction (pH effects)
• Example problem: Want to separate two organic
acids using a scheme based on pH. Acid 1 (pKa =
4), Acid 2 (pKa = 8)

K1 Acid 2 stays in
K2 organic phase,
acid 1 is extracted
D into aqueous phase

4 pH 8
 Solvent Extraction (pH effects)
• Analogous treatment for organic bases
(proton acceptors, not KOH)

[H+]=Ka
pH=pKa [H+]<<Ka
K
K Ka mainly mainly
D= D BH+ B
[H+] + Ka
[H+]>>Ka

pH
 Solvent Extraction (pH effects)
• In general:

acid base
K

pH