Cebu Doctors’ University

College of Arts and Sciences

Physical Sciences Department

Name: _________________________________ Date: __________________

Program, Year and Section: ________________ Group No.: _____________

Experiment 2 – Separation and Purification Techniques of Organic Compounds

Intended Learning Outcomes:

At the end of this experiment, you will be able to:
1. Preform the following separation and purification methods:
i. Recrystallization
ii. Sublimation
iii. Liquid-liquid extraction
2. Discuss the principles behind the different separation and purification
techniques.

Apparatus and Equipment:

Bunsen burner, test tubes, beakers, Erlenmeyer flask, stirring rod, funnel, water bath, test
tube holder, tripod, wire gauze, filter paper, evaporating dish, top loading balance

Materials:
Benzoic acid, impure benzoic acid, distilled water, hexane, acetone, naphthalene, ether,
10% NaOH solution, 3M HCl solution

I. RECRYSTALLIZATION

Recrystallization involves the dissolution of the solid to be purified in an appropriate solvent at an

elevated temperature and subsequent reformation of the crystals upon cooling. During this cooling
process, impurities may stay in solution.

The following diagram shows the steps conducted during recrystallization:

Explanation of the different steps:

1. Solvent selection. Listed below are the guidelines in selecting recrystallization solvent:
a. The desired compound should be insoluble in cold solvent but soluble in hot solvent.
b. Impurities should either be insoluble in the solvent or remains in the solution in the
cold solvent.
c. Solvent should not chemically react with the solid being purified.
2. Dissolution. Dissolution of the impure solid is done by heating the mixture once the solid
is mixed with the appropriate solvent. A beaker should not be used during heating because
it has a wide opening. Erlenmeyer flask is used for this to avoid splashing.
3. Decolorization. Activated charcoal is added to the mixture for decolorization when it
becomes colored after dissolution. Activated charcoal is added to the hot solution, not

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 boiling. When you add activated charcoal in boiling solution, it will produce frothing; thus,
you will lose your crystals.
4. Hot filtration. Hot filtration is important especially when using activated charcoal in the
procedure because activated charcoal is so fine that it cannot be removed by simple
decantation. This is performed by heating the filtration set up in a hot plate to get the funnel
hot. The recrystallization mixture is then poured through the filter paper while it’s hot.
5. Formation of Crystals. The hot solution of solute is allowed to cool slowly to room
temperature, and crystallization should occur. During cooling and crystallization, the
solution should be protected from airborne contaminants by covering the opening with a
piece of filter paper, an inverted beaker, or by loosely plugging it with a clean cork. Rapid
cooling by immersing the flask in water or an ice-water bath tends to lead to the formation
of very small crystals that may adsorb impurities from solution. Generally, the solution
should not be disturbed as it cools, since this also leads to production of small crystals.
The formation of crystals larger than about 2 mm should also be avoided because some
of the solution may become occluded or trapped within the crystals. The drying of such
crystals is more difficult, and impurities may be left in them. Should overly large crystals
begin to form, brief, gentle agitation of the solution normally induces production of smaller
crystals.
6. Filtration and Collection of Crystals. The crystals can be separated by simple gravity
filtration. After filtration, crystals are washed with cold solvent to ensure that no filtrate is
occluded in the crystals.

Procedure:
In this experiment, you will be purifying an impure benzoic acid using recrystallization.

Selecting Recrystallization Solvent

1. Add a small amount of pure benzoic acid in 3 separate test tubes.
2. Add 5 mL of water in the first test tube. Label accordingly.
3. Do the same for the second and third test tubes but use acetone and hexane instead of
water, respectively.
4. Shake the test tubes vigorously. Observe and record at what solvent does the acetanilide
dissolve.
5. Place the test tubes in a boiling water bath for 2 minutes. Observe if acetanilide has
dissolved in the solvents after heating.

Table 1. Solvent Selection

Solvent Before Heating After Heating

Water

Acetone

Hexane

Based on the results of the test above, what solvent shall be used in recrystallizing benzoic acid?

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Recrystallization of Impure Benzoic Acid
1. Place 1 g of impure benzoic acid in an Erlenmeyer flask.
2. Add 10 mL of water into the flask. Heat the mixture to a gentle boil.
3. Continue adding water in 0.5-mL portions until no more solid appears to dissolve in the
boiling solution.
4. If the resulting solution is colored, do a decolorization step:
a. Cool the solution slightly and add a microspatula-tip full of carbon.
b. Reheat to boiling for a few minutes.
5. If there are insoluble impurities or decolorizing carbon in the solution, perform a hot
filtration using the setup in figure below.

6. Cover the opening of the flask with a piece of filter paper, an inverted beaker, or loose-
fitting cork to exclude airborne impurities from the solution and allow the filtrate to stand
undisturbed until it has cooled to room temperature and no more crystals form.
7. Complete the crystallization by placing the flask in an ice-water bath for at least 15
minutes.
8. Collect the crystals using gravity filtration and wash the residue with two small portions of
cold water. Spread the benzoic acid on a watchglass, protecting it from airborne
contaminants with a piece of filter paper, and air-dry it at room temperature.

Describe the crystal formed after recrystallization and compare it with the impure benzoic acid.
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II. SUBLIMATION

Like the vapor pressure of a liquid, the vapor pressure of a solid increases with temperature. If
the vapor pressure of a solid is greater than the ambient pressure at its melting point, then the
solid undergoes a direct-phase transition to the gas phase without first passing through the liquid
state. This process is called sublimation. To be purified by sublimation, a compound must have
a relatively high vapor pressure, and the impurities must have vapor pressures significantly lower
than that of the compound being purified.

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Sublimation is carried out by heating the impure material in a vessel that is covered with funnel
with an evaporating dish containing ice. This causes sublimation of the substance you want to
purify. The vapor of this substance will then be trapped in the filter paper and recrystallizes. The
crystal that is formed in the filter paper is already pure and is known as sublimate.

One advantage of sublimation is that no solvent is used, and therefore none needs to be removed
later. Sublimation also removes occluded material, such as molecules of solvent, from the
sublimed substance.

Sublimation is a faster method of purification than crystallization but not as selective. Similar vapor
pressures are often a factor in dealing with solids that sublime; consequently, little separation can
be achieved. For this reason, solids are far more often purified by crystallization. Sublimation is
most effective in removing a volatile substance from a nonvolatile compound, particularly a salt
or other inorganic material. Sublimation is also effective in removing highly volatile bicyclic or
other symmetrical molecules from less volatile reaction products.

Procedure:
1. Place 1 g of impure benzoic acid in a beaker.
2. Cover it with an evaporating dish with the convex side facing the interior of the beaker.
3. Place ice into the evaporating dish.
4. Heat the beaker in low heat.
5. Cool the beaker without removing the funnel.

Describe the crystal formed after sublimation and compare it with the impure benzoic acid.
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III. EXTRACTION

A common technique for isolating and purifying the product of a chemical reaction involves liquid-
liquid extraction, or simply extraction. This process involves transferring a solute from one sol-
vent into another, because of its greater solubility in the second. The two solvents must be
immiscible and form two distinct layers, and in general one layer is aqueous, and the other is an
organic solvent such as diethyl ether, hexane, or dichloromethane. The physical separation of the
two immiscible phases will be performed in separatory funnels.

Separatory funnels are available in many different shapes, ranging from pear- shaped to nearly
conical (See figure on the right); the more elongated the funnel, the longer the time required for
the two liquid phases to separate after the funnel is shaken. Although separatory funnels may be
equipped with either a glass or Teflon stopcock, the latter does not require lubrication and thus is
preferred because the solutions being separated do not get contaminated with stopcock grease.
Separatory funnels are most used during work-up procedures after completion of a chemical
reaction. For example, they are used for extracting the desired product from one immiscible liquid
phase into another and for “washing” organic layers to remove undesired substances such as
acids or bases from the desired organic compound.

There are a number of general guidelines for using separatory funnels that merit discussion:
1. Filling Separatory Funnels. The stopcock should be closed, and a clean beaker placed
under the funnel before any liquids are added to the funnel in case the stopcock leaks or

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 is not completely closed. A separatory funnel should never be more than three-quarters
full. The upper opening of the funnel is then stoppered with a ground-glass, plastic, or
rubber stopper; most separatory funnels are now fitted with a ground-glass or plastic
stopper.
2. Holding and Using Separatory Funnels. If the contents of the funnel are to be shaken,
it is held in a specific manner. If the user is right-handed, the stopper should be placed
against the base of the index finger of the left hand and the funnel grasped with the first
two fingers and the thumb. The thumb and the first two fingers of the right hand can then
be curled around the stopcock. Holding the funnel in this manner permits the stopper and
the stopcock to be held tightly in place during shaking. A left-handed person might find it
easier to use the opposite hand for each position. If you are lucky enough to be
ambidextrous, take your choice!
3. Shaking Separatory Funnels. A separatory funnel and its contents should be shaken to
mix the immiscible liquids as intimately as possible. The shaking process increases the
surface area of contact between the immiscible liquids so that the equilibrium distribution
of the solute between the two layers will be attained quickly; however, overly vigorous, or
lengthy shaking may produce emulsions.

The funnel must be vented every few seconds to avoid the buildup of pressure within the
funnel. Venting is accomplished by inverting the funnel with the stopcock pointing upward
and away from you and your neighbors and slowly opening it to release any pressure. If
the funnel is not carefully vented, liquid may be violently expelled, covering you and your
laboratory partners with the contents. Venting is particularly important when using volatile,
low-boiling solvents such as diethyl ether or methylene chloride; it is also necessary
whenever an acid is neutralized with either sodium carbonate or sodium bicarbonate,
because CO2 is produced. If the funnel is not vented frequently, the stopper may be blown
out accidentally; under extreme circumstances the funnel might blow up. Any sudden
release of pressure is likely to result in the contents being lost and spattered on you and
your coworkers—not a good way to make friends. The funnel may be vented simply by
holding it as described previously and opening the stopcock by twisting the fingers curled
around it without readjusting your grip on the funnel.

At the end of the period of shaking (1–2 min are usually sufficient if the shaking is
vigorous), the funnel is vented a final time. It is then supported on an iron ring, and the
layers are allowed to separate.

4. Layer identification. It is important to ascertain which of the two layers in a separatory

funnel is the aqueous layer and which is the organic. Because the layers should separate
so that the denser solvent is on the bottom, knowledge of the densities of the liquids being
separated provides an important clue as to the identity of each layer. This generalization
is not foolproof, however, because a high concentration of a solute in one layer may
reverse the relative densities of the two liquids. You must not confuse the identity of the
two layers in the funnel and then discard the layer containing your product. Both layers
should always be saved until there is no doubt about the identity of each and the desired
product has been isolated.

Because one of the layers is usually aqueous and the other is organic, there is a simple
and foolproof method to identify the two layers. Withdraw a few drops of the upper layer
with a pipet and add these drops to about 0.5 mL of water in a test tube. If the upper layer
is aqueous, these drops will be miscible with the water in the test tube and will dissolve,
but if the upper layer is organic, the droplets will not dissolve.

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Procedure:
In this experiment, you will be purifying benzoic acid with naphthalene impurity using extraction.

1. Obtain 2 g of a mixture of benzoic acid and naphthalene. Dissolve the mixture by swirling
it with 30 mL of diethyl ether in an Erlenmeyer flask. If any solids remain, add more diethyl
ether to effect complete dissolution. Transfer the solution to the separatory funnel.
2. Extract the solution with a 15-mL portion of 2.5 M (10%) aqueous sodium hydroxide. Be
sure to hold both the stopcock and the stopper of the funnel tightly and frequently vent the
funnel by opening its stopcock.
3. Identify the aqueous layer, transfer it to an Erlenmeyer flask labeled “Hydroxide Extract.”
4. Transfer the organic solution into a clean Erlenmeyer flask containing two spatula-tips full
of anhydrous sodium sulfate and labeled “Neutral Compound.” Let this solution stand for
about 15 min, occasionally swirling it to hasten the drying process. If the solution remains
cloudy, add additional portions of sodium sulfate to complete the drying process.

Separation of the Benzoic Acid

5. Cool the “Hydroxide Extract” in an ice-water bath. Carefully add 3M HCl until acid with
litmus paper.
6. Once acidified, cool further in the ice bath for 10-15 minutes to complete crystallization.
7. Collect the precipitate by gravity filtration. Wash the solid on the filter paper with a small
portion of cold water. Transfer the solid to a labeled watch glass, cover it with a piece of
filter paper, and allow the product to air-dry. This is your pure benzoic acid.

Separation of the Naphthalene

8. Separate the “Neutral Compound” from the drying agent by decantation into a 100-mL
beaker.
9. Remove the solvent by heating on a steam bath.
10. Allow the residue to cool to room temperature to solidify and scrape the contents of the
flask onto a piece of weighing paper to air-dry. This is your pure naphthalene.

Describe the crystals formed after extraction and compare them with the mixture benzoic acid
and napthalene.
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