Extraction: A Separation and Isolation Technique

Before performing this lab, please review the safety rules that you agreed to follow in the safety
training portion of the course.

Your instructor will indicate any additional safety features of this lab activity and indicate
disposal issues.

Introduction:

In synthesis, the desired product must be separated from the by-products, excess reactants,
impurities, and other substances that may be present in the reaction mixture. Similarly,
substances in nature are always mixed with other substances. Extraction is the most common
technique used to separate a desired organic product from a reaction mixture or to isolate an
organic substance from its natural source.

Objectives:
1. Separation of a mixture containing an acidic and a neutral compounds by extraction.
2. Purification of the solid component by crystallization.
3. Identify those components from its IR spectrum and its melting point.

General Principles
Extraction usually involves shaking a solution that contains the desired substance with an
immiscible solvent in which the desired substance is more soluble than it is in the starting
solution. Upon standing, the solvents form two layers that can be separated. Extraction may
have to be repeated several times to effect complete separation.

Most commonly, one of the solvents is organic and the other is aqueous. Inorganic compounds
can usually be separated from organic compounds in this way: The former dissolve in the
aqueous phase and the latter in the organic solvent. In such cases, a single extraction may suffice
to effect a satisfactory separation. However, many organic compounds (particularly oxygen-or
nitrogen-containing compounds, such as aldehydes, alcohols, esters, and amines, which can form
hydrogen bonds) are partially soluble in water. They distribute themselves between the aqueous
phase (w, for water) and the organic solvent (0) in proportion to their relative solubilities (S) in
the two solvents.

The ratio of the concentrations (Expressed as mass of solute per unit volume of solvent (g/100,
for example).) of a substance in the two solvents (CO/Cw) at equilibrium is called its distribution
coefficient, KD:

SO CO
KD
SW CW

For example, suppose the solubility of compound A is 0.60 g/100 mL in ether and 0.12 g/100 mL
in water. KD is then 0.60/0.12 =5.
2009 Dr. Robert R. Klepper
To illustrate how the distribution coefficient KD can be used, let us calculate the amount A that is
removed from a solution containing 60 mg of A in 60 mL of water by extracting with 100 mL of
ether. If we let x be the number of milligrams of A extracted into the ether layer, then 60 -x
represents the milligrams of A remaining in the water. The equation for KD x is, therefore,

x
CO 100
KD 5
CW 60 x
60

x=53.5 mg

Solving for x, we find that 53.5 mg of A will be extracted by the ether and, consequently, that 6.5
mg of A (60 -x) will remain in the water.

It is easy to show that if we had extracted A twice with 50 mL of ether instead of once with 100
mL of ether, we would have removed 48.4 + 9.4 mg = 57 .8 mg of A from the water. In general,
performing several extractions using smaller volumes of solvent is more efficient than
performing a single extraction with a larger volume of solvent.

When equal volumes of the two solvents are used, one can solve for the amount of material
extracted by simple inspection because in this case, KD is simply the ratio of the mass of the
solute in each solvent. For example, if KD = 5, then 5/6 of the solute will be in the organic phase
and 1/6 in the aqueous phase when the volumes of the two solvents are equal.

Practical Considerations

An extraction solvent must readily dissolve the substance to be extracted, yet it must be only
sparingly soluble in the solvent from which the desired substance is to be extracted. Also, it
should extract only the desired substance or as small an amount as possible of any other
substance present; it should not react chemically with the solute in an undesirable way, and it
should be easily separated from the desired solute after extraction. This last requirement can be
met if the solvent is low-boiling and easily removed by distillation.

Common organic solvents that fulfill these requirements include many hydrocarbons and their
chloro derivatives, such as benzene, petroleum ether (a mixture of low-boiling alkanes),
dichloromethane, chloroform, and carbon tetrachloride. If benzene or chlorinated hydrocarbons
are used, however, it is important to avoid breathing their vapors because these compounds are
toxic and some are carcinogenic. They can be used safely if we carry out operations
in an efficient hood and take care to avoid getting them on the skin . Diethyl ether is another
common extraction solvent, but here, too, care is necessary. Diethyl ether (usually referred to
simply as ether) is highly flammable and, upon standing in air, its solutions may develop
dangerous concentrations of explosive peroxides. Furthermore, ether is slightly water-soluble
(about 7 g/100 mL). Nevertheless, because most organic compounds are highly soluble in it and
because of its low boiling point (35C), ether is frequently used despite its drawbacks.
2009 Dr. Robert R. Klepper
Sometimes we can use desirable, easily reversed chemical reactions such as acid-base reactions
to effect separations by extraction. For example, dilute sodium hydroxide (an inorganic base)
converts organic acids to their sodium salts:

RCO2H + Na+ OH- RCO2- Na+ + H2O

Although a particular acid may not be soluble in water, its more polar sodium salt usually is.
When a mixture of a neutral compound and an acidic, water-insoluble compound in an organic
solvent is shaken with dilute aqueous sodium hydroxide, the acid is converted to its sodium salt,
which dissolves in the aqueous layer, and the neutral compound remains in the organic layer.
After the layers are separated, the acid is recovered by acidifying the aqueous layer with a strong
acid.

RCO2- Na+ + HCl RCO2H + Na+ Cl-

Thus acids can easily be separated from neutral (or basic) contaminants by extraction with
aqueous alkali.

Phenols also form salts when treated with an inorganic base:

ArOH + Na+ OH- ArO- Na+ + H2O

However, most phenols are less acidic than the carboxylic acids. Aqueous sodium bicarbonate is
usually alkaline enough to convert carboxylic acids to their salts, but not alkaline enough to
convert phenols to their salts. Thus carboxylic acids and phenols can sometimes be separated by
extraction first with aqueous sodium bicarbonate (to remove carboxylic acids) and then with
aqueous sodium hydroxide (to remove phenols).

Dilute aqueous acid can be used to extract basic compounds, particularly amines, from neutral or
acidic substances by converting them to water-soluble alkyl ammonium salts:

RNH2 + H+ Cl- RNH3+ Cl-

After separating the organic and aqueous layers, we can recover the amine from the aqueous
layer by making the solution alkaline with a strong base.

RNH3+ Cl- + Na+ OH- RNH2 + H2O + Na+ Cl-

In this experiment we will apply these ideas in order to separate a three-component mixture of
organic compounds by extraction.

Use of the Separatory Funnel

Extractions are usually performed with a separatory funnel. Improperly handled, this moderately
expensive piece of glassware is easily broken. Always follow proper handling technique.

2009 Dr. Robert R. Klepper

Lubricate the stopcock of the funnel with a thin layer of grease so that the stopcock turns easily,
but be sure that the bore does not become clogged. (If the separatory funnel has a Teflon plug,
lubrication is unnecessary.) Support the funnel in an iron ring padded with plastic or rubber
tubing (Figure 1) . Close the stopcock, and add the liquids to be separated. Insert the stopper
(which has been lightly greased if it is not Teflon) and invert the funnel , being sure to hold the
stopper in with one hand and the stopcock in with the other (Figure 2). Then, with the barrel
pointed up and away from you and from anyone else in the vicinity, slowly open the stopcock.
This will relieve any pressure that may have built up inside the funnel. Pressure buildup is quite
common when volatile solvents such as ether are used because the vapor pressure of the solvent
adds to the atmospheric pressure already present. This situation is further aggravated when the
funnel is warmed by the heat of your hands or when a gas is generated during the extraction, as
happens when an ether solution of an acid is extracted with sodium bicarbonate.

After the pressure is released, close the stopcock, shake the funnel gently two or three times , and
again invert the funnel and release the pressure by opening the stopcock. Repeat this process
until the pressure buildup is slight. Then shake the contents vigorously to complete the
extraction.

Replace the funnel in the iron ring and remove the stopper immediately. Allow the funnel to
stand until the layers separate cleanly. Then slowly draw off the lower layer through the stopcock
into a flask or beaker of appropriate size. As the boundary between the two layers approaches the
stopcock, slow the flow. Close the stopcock just as the upper layer enters the stopcock bore. If
the upper layer is to be transferred to another vessel, pour it out through the top of the funnel.
Do not run the upper layer through the stopcock.

The relative positions of the aqueous and organic layers in the separatory funnel depend on their
densities . The more dense solvent forms the lower layer. Hydrocarbons and ether are less dense
than water, whereas chlorinated hydrocarbons (chloroform, dichloromethane, and carbon
tetrachloride) are more dense than water. If you have any doubt about which is the organic
layer and which the aqueous layer, withdraw a few drops of the lower layer and determine
whether or not they dissolve in water.

Sometimes, especially with alkaline solutions, it is difficult to obtain a sharp separation of layers
because an emulsion has formed . Gentle swirling of the funnel in a near-upright position, gentle
stirring with a glass rod, addition of salt to the aqueous layer, or addition of certain defoaming
agents may overcome this difficulty.

Procedure:

Separation of a Three-Component Mixture by Extraction

A mixture of m-nitroaniline (a base), benzoic acid (an acid), and naphthalene (a neutral
substance) will be separated into its components by extraction."

Caution Ether is highly flammable: No flames should be allowed in the laboratory when this
experiment is being performed. Also, wear disposable gloves to avoid skin contact with the
mixture being separated and with its components.

2009 Dr. Robert R. Klepper

Figure 1: Proper support for a separatory funnel. Note that the stem of the funnel touches the side
of the receiver so that the liquid will run down its walls and not splash. Grip the stopcock handle
with one hand and the barrel with the other, and push the stopcock in as you turn it.

Figure 2: Proper method for holding the separatory funnel during extraction.

Macroscale

Obtain from your instructor a 1.5-g sample of the three-component mixture and dissolve it in
Procedure 14 mL of ether. Pour the solution into a 60-or 125-rnL separatory funnel. To extract
the basic component (m-nitroaniline) from the mixture, add a solution of 1.5 mL of concentrated

2009 Dr. Robert R. Klepper

hydrochloric acid in 14 mL of water and shake thoroughly, using the technique described above.
Draw off the lower (aqueous) layer into a 125-rnL Erlenmeyer flask and repeat the extraction
two more times . Finally, extract with 5 mL of water to remove excess hydrochloric acid that
may be dissolved in the ether layer. Combine the three acid extracts with the water extract and
set them aside.

To separate the acidic component (benzoic acid) from the mixture, extract the remaining ether
solution three times with 14 mL of 10% aqueous sodium hydroxide and once with 5 mL of
water. Combine the alkaline and water layers and set them aside.

Pour out the remaining ether solution (which should contain only the neutral component,
naphthalene) through the top of the separatory funnel into a small Erlenmeyer flask . Add enough
anhydrous calcium chloride to cover the bottom of the small Erlenmeyer flask, and swirl the
mixture occasionally for 15 min. Then decant the ether into a small beaker of known mass. Rinse
the flask contents with a small amount (3 mL) of ether, and add the rinsing to the beaker. Then
place the beaker in the hood to allow the ether to evaporate.* Weigh the residue and determine
its melting point. Store the compound in a corked test tube if it is to be saved until a later lab
period .

*Evaporation of the solvent can be hastened by placing the flask in a warm (40-50C) sand bath
or on a warm hotplate and directing a gentle stream of air (or nitrogen) toward the upper inside
rim of the flask.

Neutralize the combined acidic extracts by adding 10% aqueous sodium hydroxide until the
solution is alkaline to litmus paper.* Extract the alkaline solution twice with 14-mL portions of
ether, Collecting and combining both ether extracts. Add anhydrous calcium chloride, and swirl
as described above. Then decant the ether solution into a small beaker of known mass, rinse as
described above, and place the beaker in the hood to allow the ether to evaporate. Weigh the
residue (m-nitroaniline, or the basic component) and determine its melting point. Store the
compound in a corked test tube if it is to be saved until a later lab period.

Neutralize the combined alkaline extracts by adding concentrated hydrochloric acid drop by drop
until the solution is acid to litmus paper. The solution may be kept cool with ice during
neutralization. Recover the precipitated benzoic acid (acidic component) by vacuum filtration
using a Buchner or Hirsch funnel, or by extraction with ether as described above. Allow the
product to air-dry, weigh it, and determine its melting point. Each component can be
recrystallized, if desired, to obtain a pure product with a sharp melting point.

Waste Disposal Neutralize the basic and acidic aqueous filtrates with dilute hydrochloric acid
and 10% aqueous sodium hydroxide, respectively. When the aqueous solutions are neutral to
litmus, you may discard them in the sink.

Microscale

The basic method for conducting this experiment on a microscale is the same as that for
macroscale. There are two major differences: (1) a test tube and cork or a conical centrifuge tube

2009 Dr. Robert R. Klepper

and screw-cap are used in place of the separatory funnel as the extraction vessel, and (2) a
Pasteur pipet is used to remove the bottom layer from the test tube or centrifuge tube .

Obtain from your instructor a 150-mg sample of a three-component mixture and dissolve it in 4
mL of ether in a 13 x 125 mm test tube (or similar size centrifuge tube). To extract the basic
component from the mixture, add 2 mL of 3 M hydrochloric acid, stopper the test tube with a
cork, and shake it vigorously sideways (not up and down) for several minutes, removing the cork
periodically to relieve pressure. Allow the layers to separate; then draw off the lower (aqueous)
layer with a Pasteur pipet and transfer it to a labeled 10-mL Erlenmeyer flask . (Using the pipet
bulb, expel air from the pipet as you lower the tip to the bottom of the test tube or centrifuge
tube. Then release the bulb slowly , drawing in the lower layer. Lift the pipet through the upper
layer without drawing in liquid from this layer.) Repeat this procedure one more time and add
the aqueous layer to the same 10-mL Erlenmeyer flask . Set this flask aside .

To separate the acidic component from the mixture, extract the remaining ether solution two
times with 2 mL of 3 M (12%) aqueous sodium hydroxide. Place both aqueous extracts in a
single labeled 10-mL Erlenmeyer flask and set this flask aside.

Extract the remaining ether solution (which should contain only the neutral component)
withtwo1-mLportions of water and discard the water extracts. Then add 4-6 pellets of anhydrous
calcium chloride (or 250 mg of anhydrous sodium sulfate) to the ether solution and swirl. Set it
aside to dry for 15-20 min, with occasional swirling.

Neutralize the combined acidic extracts (containing the basic component) by adding 6 M
aqueous sodium hydroxide dropwise until the solution is alkaline to litmus paper. Cool the flask
in an ice bath for 10--15 min. Collect the solid precipitate on a small Hirsch funnel, and wash it
on the funnel (vacuum on) with two l mL portions of distilled water. Dry and weigh the product
and determine its melting point.

Neutralize the combined basic extracts (containing the acidic component) by adding 6 M
hydrochloric acid dropwise until the solution is acidic to litmus paper. Cool the flask in an ice
bath for 10-15 min. Collect the solid precipitate on a Hirsch funnel, and wash it with two 1-mL
portions of distilled water. Dry and weigh the product and determine its melting point.

*The proper way to test with litmus paper is to dip a clean glass rod into the solution and then
touch it to the litmus g: paper. This method prevents sample contamination.

Using a Pasteur pipet, transfer the dried ether solution (make sure it is dry before continuing) to a
weighed 1O-mLErlenmeyer flask containing a boiling stone. (Avoid transferring the drying
agent.) Rinse the drying agent with an additional 1 mL of ether and transfer the rinse to the same
Erlenmeyer flask. Evaporate the ether, using a warm sand bath or hotplate (40-50C)in the hood.
Remove the last 0.5mL of solvent without heating. A gentle stream of nitrogen gas or air may be
used to hasten the evaporation process . Weigh the residue and determine its melting point.

Each component can be recrystallized (Craig tube), if desired, to obtain a pure product with a
sharp melting point.

2009 Dr. Robert R. Klepper

Neutralize the basic and acidic aqueou s filtrates with dilute hydrochloric acid and 10% aqueous
sodium hydroxide, respectively. When the aqueous solutions are neutral to litmus, you may
discard them in the sink.

Report:

Compound Mass recovered mp C found mp C (lit.)

m-Nitroaniline
Benzoic acid
Naphthalene

Questions
1. What are the advantages and the disadvantages of using ether as an extraction solvent?

2. What is the practical advantage of having the organic solvent be more dense than water when
one is extracting an organic compound from an aqueous solution in a separatory funnel?
2009 Dr. Robert R. Klepper
3. Why must the stopper at the top of the separatory funnel be removed before liquid can be
withdrawn through the stopcock?

4. Complete the following flow diagram for this extraction experiment, showing names and
structures of the materials present at each stage.

Ether solution of m-nitroaniline, benzoic acid, and naphthalene

|dilute HCI
|

| |
| |

lower layer upper layer

10% NaOH |10% NaOH

lower layer |upper layer

| |
| |

|concentrated
|HCl

5. Write four equations (showing structures) for the reactions used in this experiment to separate
and recover m-nitroaniline, benzoic acid, and naphthalene.

2009 Dr. Robert R. Klepper

6. Verify the conclusion (drawn in Sec. I) that two extractions of A with 50-mL portions of ether
yield 57.8 mg of A, whereas one extraction with 100 mL of ether yields only 53.5 mg of A.

7. The solubility of suberic acid is 0.14 g/l00 mL of water or 0.56 g/l00 mL of ether. Calculate
the volume of ether needed to remove 90% of the suberic acid from 100 mL of a saturated
aqueous solution in a single extraction.

2009 Dr. Robert R. Klepper