Solid organic compounds prepared in the laboratory or extracted from natural sources,

such as leaves, are often impure. One of the steps usually involved in the purification of such
solid compounds is crystallization. It is the slow formation of a crystalline solid. In general, the
crude compound is first dissolved in a minimal amount of hot solvent. At this stage, insoluble
impurities can be separated from the hot solution by gravity filtration. If colored impurities are
present, the solution is treated with decolorizing charcoal and then filtered. The hot, saturated
solution is then allowed to cool slowly so that the desired compound crystallizes at a moderate
rate. When the crystals are fully formed, they are isolated from the mother liquor (the solution) by
filtration.
The success of this process is based on the difference in the solubilities of the compound
and the impurities, and on the fact that most compounds are more soluble in a hot solvent than in
a cold one.
Crystallization is different from precipitation since the latter is a rapid formation of solid
material. If the hot, saturated solution is cooled too quickly, the compound may precipitate
instead of crystallizing. The precipitate may still contain impurities. However, if the solution is
allowed to slowly cool down, impurities are likely to be excluded. The molecules in the crystal
structure are in equilibrium with the molecules still in solution. Molecules unsuitable for the
crystal lattice tend to return to the solution. Thus, crystallization is a more selective process. Since
impurities are often found in low concentrations, they tend to remain in solution as cooling
continues.
To obtain really pure compounds, recrystallization of the harvested crystals is often
employed. But this is with the disadvantage of some loss of the desired compound, which remains
in the solution.

Solvents for Crystallization

The ideal solvent for the crystallization of a particular compound is one that:
1) does not react with the compound;
2) boils at a temperature below the compound’s melting point;
3) dissolves a moderately large amount of compound when hot;
4) dissolves only a small amount of compound when cool;
5) is moderately volatile so that crystals can be readily dried;
6) is nontoxic, nonflammable, and inexpensive.

Of course, as with all ideal things, such an ideal solvent does not exist. However, the
main consideration for choosing a solvent is this: that the compound should be moderately
soluble in it. Crystallization brings us back to the rule of thumb: like dissolves like. Polar
compounds are soluble in polar solvents. So how do we make sure that our target compound is
just moderately soluble in a given solvent? Through experiment. Fortunately for us, information
about good solvents or solvent systems for the crystallization of certain compounds can be found
in literature.

Mixed Solvent Technique

Sometimes, it is hard to find a solvent that bests fits the solubility criteria for a particular
compound. In such case, a mixed solvent or solvent pair may be used. Table 1 lists the common

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solvent mixtures. Initially, the crude compound is dissolved in a minimum amount of the boiling
solvent in which it is soluble. Following this, a second hot solvent, which is miscible with the
first and in which the solute is relatively insoluble, is added dropwise to the boiling mixture until
the mixture becomes cloudy. Cloudiness indicates precipitation. When extensive cloudiness
occurs, more of the first solvent should be added to clear the cloudy solution. At this point, the
solution is saturated, and as it is cooled, crystals should separate. It is important not to add an
excess of the second solvent or to cool the solution too rapidly. Doing so will only cause the
solute to oil out or separate as a viscous liquid. If this happens, the solution should be reheated
and added with more of the first solvent to dissolve the oil.

Table 1 Common solvent mixtures.

Methanol-Water Ether-Acetone
Ethanol-Water Ether-petroleum Ether
Acetic acid-Water Benzene-Ligroin
Methylene Chloride-
Acetone-Water
Methanol
Ether-Methanol Dioxane-Water

Safety Using Solvents

Most organic solvents are either flammable or toxic, or even both. And since
crystallization involves heating these solvents, care must be taken. For example, if a solvent is
very flammable or is low-boiling, the safest heat source would be a hot plate. Heating must also
be done under a hood. Bunsen burners or any direct flame may only be used in solvents that are
not too flammable.
Toxic solvents must not be sniffed, and should be handled under a hood.
Boiling chips should be used when heating solvents to reduce the risk of uneven boiling,
which may result in bumping or splattering of hot liquid. These chips release bubbles when hot,
ensuring even heating. Fresh boiling chips must be used at all times.

Different Crystallization Techniques

Ways to crystallize a compound include the following:
1. Heating the solution then cooling it down gradually;
2. Dissolving the product then letting the solvent evaporate at room temperature;
3. Dissolving sample then cooling it in the refrigerator/cooling refrigerator (a special
refrigerator that cools down very slowly automatically);
4. Allowing a “non-solvent” to diffuse into the solution of interest thereby changing the
solubility of the solute in the new solvent system (refer to Figure 1); and

Figure 1. Non-solvent diffusing into the solution

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 5. Simply placing the system in a cold room since some materials are best crystallized
at low temperatures.

Steps in Crystallization

Dissolving the Compound

The solid compound will be dissolved in a minimum amount of hot solvent in an
Erlenmeyer flask. A flask is preferred over a beaker since it minimizes the possibility of
splattering liquids. It can also be corked and stored.
Solid lumps must be pulverized first before dissolving it in the solvent. To make
sure that only a minimum amount of solvent is used, small portions of the solvent are
added until the compound completely dissolves. Usually, excess solvent can be reduced
by boiling it off. There will be no need for gravity filtration if the resulting solution looks
clean and clear. Colored impurities can be eliminated by adding decolorizing charcoal.

Filtering Insoluble Impurities

This portion involves gravity filtration (refer to section in Filtration). In the case
of filtering solid impurities, it must be ensured that premature crystallization or
precipitation does not occur in the process. Thus, the glassware immediately in contact
with the hot solution must also be maintained at a high temperature.
The glassware (an Erlenmeyer flask and a funnel) must be preheated in an oven,
and used while very hot at the time when the solution to be filtered is ready. The
receiving flask will also serve as the site for crystallization later on. Keep the solution
being filtered hot. While waiting for some of the solution poured on the funnel to drain
into the receiving flask, heat the remainder of the unfiltered solution to keep it hot.
If premature crystallization occurs in the funnel, the crystals can be washed down
with fresh, hot solvent. To make the washings easier, it would be better to add only small
portions of the crystallization solution instead of filling the funnel to the brim. After all of
the solution is filtered, the empty flask can be washed with a small amount fresh, hot
solvent to dissolve remaining traces of crystals that may form in the flask. This wash is
then poured into the filter paper.
If crystals immediately form in the receiving flask, it can be reheated to
redissolve the crystals. If the crystals remain undissolved, additional hot solvent can be
added to dissolve these formations. At this point, excess solvent may then be boiled off.

Crystallizing the Compound

The crystallization flask must be covered with a watch glass during cooling down
to prevent dust contamination and solvent evaporation. The flask must be set aside in a
place where it cannot be disturbed. Bumping the flask may induce precipitation instead of
crystallization. Placing the hot flask in an ice-bath also produces precipitates rather than
crystals. Thus, it is important to take note that crystallization is a slow cooling process
rather than a fast and rapid one. If there is an abrupt change in temperature, the impurities
may be trapped inside the crystal lattice making the isolated crystals smaller and impure,
as shown in Figure 2.

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 Figure 2. Diagrams showing the fate of a solution that is cooled down quickly; the
triangle represents the impurities trapped inside the crystals formed by the hexagon.

On the other hand, if the solution is cooled slowly, as shown in figure 3, the
impurities trapped inside the crystal lattice of hexagons may be displaced by particles
which fit better in the crystal lattice, which are the hexagons as well. This results to a
large and pure crystallized product. It is important that the solution be cooled gradually
but not too slowly that impurities adhere in the crystal matrix again.

Figure 3. Diagrams showing the fate of a solution that is cooled down slowly

If several days are required for crystallization, the solution must first be cooled to
room temperature before corking the flask and storing it. Rubber stoppers are not to be
used when organic solvents are involved to prevent the former from being dissolved.
During crystallization, several problems may be encountered:
Case 1: No crystallization occurs.
It may be that the solution was supersaturated. Scratching the
sides of the flask will produce microcrystals of glass which may serve as
templates for crystallization. A seed crystal may also be used as a
template. Your instructor may have a few crystals previously prepared.
Seed crystals can also be obtained from the tip of your stirring rod as the

40
 solvent evaporates. Note that this stirring rod was previously immersed
in the solution where crystals are to grow.
It may also be that too much of the solvent was used, especially
if scratching or adding a seed crystal did not work. In this case, boiling
off excess solvent may be employed.
Case 2: Oiling out
Sometimes, when the melting point of the compound is lower
than the boiling point of the solvent, the compound separates as an oily
substance instead of crystallizing. This may also be the case if impurities
present in the compound depress its melting point. Reheating the mixture
to dissolve the oil is encouraged. Additional hot solvent may be added,
boiling off excess when necessary.
If oiling out still occurs, let this oil solidify. The solidified oil
may then be separated and redissolved in fresh, hot solvent. By this time,
much of the impurities may have been reduced to allow crystallization to
proceed more smoothly.

Isolating the Crystals

At this point, the crystallization solution would have already cooled to room
temperature, and stayed at that temperature for quite some time. Placing the flask in an
ice-bath may also increase crystal yield. The crystals may be separated from the mother
liquor through suction filtration (refer to section on Filtration).
This part is optional. The mother liquor collected from suction filtration may still
contain some dissolved compound and provide another batch of crystals, aptly called the
second crop. The crystals may be obtained by boiling off excess solvent, and allowing
the solution to cool down as before. Each successive crop, though, will be less pure than
the previous one. Thus, only one to two crops are usually taken. All harvested crystals
can be purified further by recrystallization in fresh solvent.

References:
Fessenden, R.J., Fessenden, J.S. Techniques and Experiments for Organic Chemistry. USA:
Merck & Co., 1983.
Most, Clark F. Jr. Experimental Organic Chemistry. USA: Wiley & Sons, 1988.
http://www.sinc.sunysb.edu/Class/orgolab/hotfiltration.PDF
http://orgchem.colorado.edu/hndbksupport/cryst/crystdiag.html

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 EXPERIMENT 1

On Benzoic Acid and Salicylic Acid

Benzoic acid and salicylic acid are two valuable organic compounds in the manufacturing
industry. Benzoic acid and its sodium salt are used as preservatives in food, fruit juices, and soft
drinks that are naturally in an acidic pH range. Salicylic acid, on the other hand, because of its
keratinolytic and fungicidal properties is applied topically in the treatment of dandruff,
ichthyosis, psoriasis, and fungal skin. There are a lot more uses of these two organic compounds.
Benzoic acid occurs naturally in many plants and animals. Due to the demand for high-
fructose corn syrup in carbonated beverages, sodium benzoate finds its major market in the soft
drink industry as a preservative. Undissociated benzoic acid is actually the more effective
antimicrobial agent for preservation purposes. However, sodium benzoate is used preferably due
to its higher solubility in water.
After a food containing benzoic acid or sodium benzoate is digested, the two are rapidly
absorbed from the gastrointestinal tract and metabolized in the liver by conjugation with the
amino acid glycine. Benzoic acid is first converted to benzoyl CoA which then conjugates with
glycine to hippuric acid. Hippuric acid is rapidly excreted through the urine. Owing to rapid
metabolism and excretion, an accumulation of the benzoates or their metabolites in the body is
not to be expected.

Figure 1. Formation of hippuric acid

Benzoic acid is also used together with salicylic acid in a formulation for destroying warts
and corns. Still, the major use of salicylic acid is in the preparation of its ester derivatives.
Several useful esters are formed by the reaction of salicylic acid with alcohols. The methyl ester,
methyl salicylate (oil of wintergreen), is formed with methanol. It is used in food flavorings and in
liniments. The phenyl ester, phenyl salicylate, or salol, is formed with phenol. It is used in
medicine as an antiseptic and antipyretic. This ester hydrolyzes, not in the acidic stomach, but in
the alkaline intestines, releasing free salicylic acid. The menthyl ester, menthyl salicylate, which
is used in suntan lotions, is formed with menthol.

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 Salicylic acid or its ester when taken orally, is metabolized in the liver in the same way as
benzoic acid. It conjugates with glycine to form salicyluric acid. When conjugated with glucuronic
acid, acyl and phenolic glucuronide are formed. A small amount of salicylic acid are also
hydroxylated to gentisic acid. These metabolites are also excreted through the urine as well.

Sources: http://www.bartleby.com/65/sa/salicyli.html
http://www.psoriasis.org/treatment/psoriasis/salicylic_acid.php
http://www.inchem.org/documents/cicads/cicads/cicad26.htm#SubSectionNumber:4.2.1
http://www.inchem.org/documents/pims/pharm/pim642.htm

Prelaboratory Questions:
1.) Benzoic acid has higher antimicrobial activity than its sodium salt. Why is sodium
benzoate preferred over benzoic acid as a preservative?
2.) Based on the reaction of glycine with benzoic acid, show the reaction of glycine with
salicylic acid to form salicyluric acid?
3.) Why should you not place the hot crystallization solution immediately in an ice bath?
4.) When the crystallization solution is allowed to slowly cool down, a watch glass or
beaker is used to cover the flask. This is to prevent solvent evaporation. Why must it
be prevented?
5.) Compute maximum recovery for the crystallization of 2.5 g of benzoic acid from 100
mL of water (assuming water is cooled down to 0°C). (Hint: know the solubility of
benzoic acid in water @ 0°C.)
6.) In a laboratory session, when is the best time to put the flask and funnel in the oven?
7.) How do you avoid premature crystallization when filtering off the insoluble
impurities from the hot solution? How do you keep the solution hot?
8.) How do you distinguish good crystals of benzoic or salicylic acid from bad ones?

Equipment:
4.5 x 4.5 in. Filter papers
2.0 x 2.0 in. Filter papers
Melting-point apparatus
Boiling chips
Sample vial

Chemicals:
Benzoic acid
Salicylic acid

Procedure:

Dissolving the Compound

For this experiment, we will be purifying crude benzoic acid using recrystallization.
Weigh approximately 2.0 g of the solid into an Erlenmeyer flask. Record weight.

TIPS: Preheat the Erlenmeyer flask and stem funnel in an oven. Use when very hot and when solution for
gravity filtration is ready. Boil about 25 mL of distilled water and set aside. Meanwhile, flute a piece of porous
(fast speed) filter paper by folding it several times to create creases. Set this aside for use later.

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 Add 60 mL of distilled water and 2-3 boiling chips to the flask with the crude solid. Heat
the solution, while stirring, until all of the acid has dissolved. Observe the resulting solution. If
you’re lucky to end up with a clean and clear solution, you may do away with the gravity
filtration. But if you see undissolved particles or a colored solution, filter the solution through
gravity filtration. Do this quickly but carefully.

TIPS: Use a towel for handling hot glassware. Keep the solution hot by heating over a flame while
filtering.

If premature crystallization occurs on the filter paper, wash down the crystals with hot water.
Wash the flask with 10 mL of hot water (from the previously boiled water) and pour this into the
funnel to remove remaining traces of crystals that may form on the filter paper.
Boil off excess water from the filtered solution until the volume reaches about 50-60 mL.
Set the flask aside, cover it with a watch glass, and allow it to cool to room temperature in a place
where it will not be disturbed.

TIP: Do not disturb the solution as it cools down.

While waiting for your solutions to cool down, you can clean your glassware and other
equipment. You can also set up your suction filtration apparatus at this point. Prepare filter papers
and cut them to fit inside a Büchner funnel.

TIP: The size of the filter paper must just be enough to cover the holes of the funnel.

Moisten the paper to seal and prevent leakage. You may test the water aspirator and see how it
works. Check if the water aspirator creates a vacuum when turned on by putting your palm over
the Büchner funnel.
Constantly check on your crystallization solution for problems.
When the solution has reached room temperature, prepare an ice-water bath. Chill some
amount of distilled water in the ice-water bath (~25 mL).

Harvesting the Crystals

Allow the solution to stay at room temperature for about 10 minutes. Then place the flask
with the solution in the ice bath to allow further crystallization. Turn on the water aspirator and
check the vacuum created. Pour the solution into the Büchner funnel. Wash remaining traces of
crystals in the flask with cold distilled water. Pour the wash into the funnel.

TIP: Make sure that as you pour the wash into the funnel, the filter flask does not overflow with the
filtrate.

Press another filter paper onto the top of the crystals to squeeze out excess solvent.
Do not discard the filtrate (or mother liquor). If time allows you, do a second crop and
dry the crystals by suction filtration after removing the mother liquor.
If there is not much time, scoop out the filter cake and spread out the crystals on a large
filter paper. Separate a few crystals, about a spatula point, and air-dry in a different watch glass
(for melting point determination).
Weigh the crystals into a previously weighed vial. Record weight as weight of wet
crystals. Keep the vial inside your locker uncovered to further dry the crystals. Obtain the dry
weight of the crystals the next day. Compute percent recovery. Hand the vials containing the dry
crystals to your instructor.

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Postlaboratory Questions:
1.) If 1.75 g of benzoic acid is dissolved in 100 ml of water at 100ºC, how many grams
of crystals can crystallize at 25ºC? Compare the yield when the minimum volume of
water is used to crystallize the same amount of benzoic acid. The solubility of
benzoic acid at 25 ºC is 3.44 g/L while near 100 ºC is 56.31g/L.
2.) Why do you wash your crystals with cold solvent?
3.) Benzoic acid, which can be crystallized from water, is contaminated with sodium
chloride and sand. In which steps of the crystallization process are the salt and sand
removed? Make a flowchart for this experiment.
4.) The table below provides the solubilities of compounds A and B in ethanol.
Solubility (g/100 ml)
@ 0ºC @ the boiling point
A 0.1 7.0
B 0.9 1.3
You are purifying 10.0 g of compound A contaminated with 1.00 g of compound B.
a.) Calculate the minimum volume of ethanol that you will need to completely
dissolve the sample.
b.) Calculate the % Recovery of A upon crystallization, disregarding experimental
losses.
c.) Is A pure after crystallization?
d.) Is A pure after crystallization if A (10.0 g) is contaminated with 1.50 g of B?
What is the % of A in the crystallized product?

References:
Fessenden, R.J., Fessenden, J.S. Techniques and Experiments for Organic Chemistry. USA:
Merck & Co., 1983.
Most, Clark F. Jr. Experimental Organic Chemistry. USA: Wiley & Sons, 1988.

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