CHM 2210L

SEPARATION OF AN ACID-NEUTRAL MIXTURE

THE PROBLEM TO BE INVESTIGATED:

The acidic and neutral components of a mixture will be separated and purified by classical methods
involving extraction and recrystallization procedures. The components will be identified by melting point
determinations.

BACKGROUND INFORMATION:

It is generally quite easy to carry out the conversion of one organic compound to another. The problems arise
when one attempts the isolation and purification of the desired material. Techniques used include
chromatography, fractional distillation, fractional crystallization, and extraction procedures. This last
procedure is extremely general when attempting to isolate components with differing pKa values. The overall
procedure is so commonly used that it is frequently described in chemical journals as "...working up a
reaction in the usual manner" rather than presenting a detailed description of the process. For example, let
us consider the reaction of benzoyl chloride with aniline to form N-phenylbenzamide.1

After allowing the reaction to stand for a period of time in some solvent (perhaps with an acid or base

O
H2N
Cl
+ + HCl
N
O H

catalyst), the reaction is quenched by addition of ice/water. Ideally, the product should crystallize from the
mixture. We can then isolate our product by suction filtration, recrystallize to constant melting point, and we
are through. Unfortunately, life is not always this kind to us. Either nothing crystallizes, or products "oil"
from the solution. We could throw the reaction mixture away and start over, hoping that this time we will be
blessed with good results, or we could "work it up in the usual manner."

We begin by realizing that everything in the reaction mixture should dissolve in either an organic solvent
(diethyl ether, methyl t-butyl ether, chloroform, toluene, etc.) or in water. Note that water is not miscible
with the organic solvents cited. In a separatory funnel, two layers would form and we could discard the
aqueous layer which carries with it any inorganic materials or traces of organic acids or bases. Considering
the reaction above, we would now have the organic materials - unreacted aniline, the amide, and benzoic
acid resulting from hydrolysis of benzoyl chloride -dissolved in the organic layer.

1
In CHM 2210L, a similar experiment using acetic anhydride and aniline was performed.

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 O O

Cl H2O
OH
+ HCl

To separate the basic amine and the acidic carboxylic acid from the neutral amide, we take advantage of
acid-base reactions and the water solubility of the resulting salts at room temperature. Addition of aqueous
acid (10% HCl) to the organic layer will convert the residual aniline to water-soluble anilinium chloride.
Discarding this acidic aqueous layer discards the unwanted aniline. Addition of aqueous base (10% NaOH)
converts the benzoic acid to water-soluble sodium benzoate.

+ -
NH2 HCl NH3 Cl

NaOH

O O
NaOH - +
OH O Na
HCl + H2 O

Again, separation of the layers leaves only the desired amide dissolved in the ether in the separatory funnel.
A wash with a saturated sodium chloride solution removes most of the residual water-soluble acid or base.
Drying the ether layer over anhydrous magnesium sulfate or sodium sulfate completes the preliminaries. The
drying agent is removed by decanting or filtering. The organic solvent is removed by evaporation (or
distillation). At this time, recrystallization usually provides a pure sample.

The process just described could be easily modified to allow isolation of the acidic material and/or/instead of
the neutral. If the acidic aqueous layer were now made basic (add NaOH), aniline would be regenerated from
anilinium hydrochloride. The aniline would be insoluble in the aqueous media and could be separated from
it; for example, by extraction with ether. If the basic aqueous layer containing sodium benzoate were now
acidified (HCl), benzoic acid (insoluble in water) would precipitate and could be collected by suction
filtration. Therefore, this "working-up in the usual manner" is a general technique for separating materials
with different pKa's. As a rule, carboxylic acids will generally react with even the weak base, sodium
bicarbonate (NaHCO3). Phenols, as well as carboxylic acids, dissolve in the stronger base, sodium
hydroxide. Amines are the only compounds basic enough to dissolve in hydrochloric acid. All other organic
compounds are essentially neutral. See figure below for a graphical representation of a flow diagram for
separation of a mixture of a neutral compound, a phenol, an amine, and a carboxylic acid.

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 aqueous layer Basify Isolate Pure Amine
Mixture + - RNH2
Extract with aqueous RNH3 Cl Aqueous Layer
Neutral Compound;
Phenol; Amine;
Carboxylic Acid dissolved 10% HCl Organic Layer
in an organic solvent Neutral Compound;
Phenol;
Carboxylic Acid

Extract with
Isolate Pure Phenol aqueous 10%
ArOH NaHCO3

Acidify Aqueous
Isoloate Pure
Layer aqueous layer Acidify
Carboxylic Acid
RCO2-Na+ Aqueous Layer
aqueous layer RCO2H
ArO-Na+ Organic Layer
Extract with aqueous
Neutral Compound;
10% NaOH Phenol
Organic Layer
Pure Neutral
Compound

Figure 1. Graphical representation of an extraction scheme based on acid-base chemistry.

THE NATURE OF THIS INVESTIGATION:

In this experiment, you will obtain 4.0 g of a mixture containing 2.0 g of acidic material and 2.0 g of neutral
material. Each 2.0 g portion will contain 1.8 g of a major component which is to be purified and identified,
and 0.2 g of impurity which is to be removed and rejected. The mixtures will be composed as follows:
Acidic Mixtures
o o
Major Component MP ( C) Impurity MP ( C)

Cinnamic Acid 134 Glutaric Acid 95-98

o-Chlorobenzoic Acid 140 Glutaric Acid 95-98

m-Nitrobenzoic acid 140 Glutaric Acid 95-98

2-Chloro-5-nitrobenzoic acid 165 Glutaric Acid 95-98

p-t-butylbenzoic acid 162-165 Glutaric Acid 95-98

Neutral Mixtures
o o
Major Component MP ( C) Impurity MP ( C)

Methyl m-nitrobenzoate 78 Napthalene 80

p-Dibromobenzene 89 p-Dichlorobenzene 53
m-Dinitrobenzene 90 2,4-Dinitrotoluene 70
Acenaphthene 94 Napthalene 80
Acetanilide 114 Benzamide 128

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In each case the impurity selected is very similar in chemical type to the major component while having a
lower melting point. Accordingly, this impurity will have a greater solubility in any solvent than will the
major component. Crystallization will necessarily change the composition in the desired direction.

You will first separate the acidic mixture from the neutral mixture by extraction from ether solution with a
base, followed by isolation of these mixtures. From your two mixtures, you will then crystallize to constant
melting points (see Helpful Hints) your purified compounds. Of those compounds listed in the tables,
decide which compounds are eligible, and use mixture melting points with those compounds for final
identification. (Be sure you realize that your mixture may contain any one of the four acidic major
components along with any of the four neutral ones.

PROCEDURE
Part I. Separation by Neutralization and Extraction

Obtain a 4.0 g sample of your acid-neutral mixture, note the number and then dissolve it in a 125 mL
Erlenmeyer flask with a total of 20 mL (or more if needed) of ether (methyl t-butyl ether will be provided).
After the entire sample has dissolved, pour the solution directly into your separatory funnel. Use an
additional 5 mL of ether to rinse the bottle and funnel, pouring the rinse solution into the separatory funnel.

Caution: Low boiling ethers are the most easily ignited of the common solvents. No flames
or hot plates should be in the immediate vicinity. Also, all aqueous HCl and NaOH solutions
should be treated as dangerous.

A. The Acidic Fraction:

Add to the ethereal solution 20 mL of 10% sodium hydroxide solution, and shake the funnel gently for 2
minutes, remembering to release the pressure in the funnel frequently. (Caution: The high vapor pressure
of ethers will increase the pressure inside the stoppered separatory funnel when shaken!!) If the
aqueous solution is colorless or nearly so, use pH paper to find if the aqueous layer is still sufficiently basic.
Draw off the aqueous layer and save it in a 50 mL Erlenmeyer flask. If the solution is not basic, add an
additional 10 ml of the 10% sodium hydroxide solution to the separatory funnel, shake as before, and save
both portions of aqueous solution in a 50 mL Erlenmeyer flask.

To the ethereal solution in the funnel, add 5 mL of water and shake briefly. Separate and add this aqueous
washing liquid to the previously collected aqueous material. Then transfer the organic layer to a dry 50 mL
Erlenmeyer flask.

Without rinsing the separatory funnel, add to it the combined aqueous material, as well as the rinse from the
flask (ca. 1-2 mL of water) along with an additional10 mL of ether. Shake the mixture, and then slowly run
only the aqueous layer into a 150 mL beaker containing 6 mL of concentrated hydrochloric acid (or more if it
was necessary to add a second portion of base) and 3 mL of water, stirring during the addition - use pH paper
to ensure the solution is acidic. Combine the ether layer with the bulk of the ethereal solution.

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Meanwhile cool the suspension of precipitated acid to 15-20oC, collect by Buchner funnel and wash with
cold water. Set the material aside for through air drying on a watch glass or piece of filter paper. Finally,
weigh the dry acid mixture.

B. The Neutral Fraction:

Decant the ethereal solution (or filter it, if necessary) into a 125 mL Erlenmeyer flask, using 1-2 mL of ether
for rinsing. Add a boiling chip and remove the ether by evaporation. Stop when 5-8 mL remains. Carefully
pour the solution into a tared 25 mL Erlenmeyer flask, using 2-3 mL of ether for rinsing.

Evaporate the remaining ether using a hot water bath (metal pan or beaker with small layer of water),
observing caution in not charring the solid. The resulting neutral mixture should crystallize on cooling (some
scratching may be necessary).

Part II. Selective Crystallization of a Compound from a Mixture

Transfer the desired mixture to a 25 mL, 50 mL, or 125 mL Erlenmeyer flask, selecting among these sizes
according to the amount of solvent you think may be needed. (Don't plan to have the liquid level above the
lower half of the conical part of the flask.) Erlenmeyer flasks are always to be used for crystallization
unless a good and specific reason exists for using a different container in a given instance. (Beakers are
unsatisfactory for crystallization or for any operation involving the boiling of organic liquids or solutions;
they are awkward to hold when hot and hard to swirl; loss by evaporation and the resulting fire hazard are
needlessly great; crust forms on the surface and wall; the material on the walls tending to "creep"; it is
impossible to tell when a solution is truly saturated.

A. Use of a Single Solvent. Add enough solvent to the flask to just cover the solids, and heat the
mixture to boiling, using an appropriate means of heating. If any lumps are left, crush them with the
end of a stirring rod that has been flattened (by heating and pressing onto an asbestos board). Add
more solvent in small portions until an approximately saturated solution is obtained. Don't hesitate to
shift (with rinsing) to a larger flask should this prove advisable. Let the solution stand undisturbed
except for scratching or possible seeding with a trace of the original mixture.

These directions assume that the solute melts at a temperature at least 15-20o above the boiling point
of the solvent selected. If this is not the case, saturate the solution at an appropriate lower
temperature.

If care has been taken in the earlier steps of the isolation, there should be present no insoluble matter.
If undissolved particles remain, ask your instructor for advice. This experiment intentionally aims to
avoid filtration of a hot solution and the use of decolorizing charcoal.

When the mixture has cooled nearly to room temperature, cool it in ice, and collect, wash, and dry
the product. Determine the melting point of the material, weigh it.

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B. Use of a solvent pair. If the use of a single solvent seems impractical, place the mixture in a 50 mL
Erlenmeyer flask, add about 5 mL of the overly effective solvent, and heat the mixture to boiling.
Now add the other member of the pair in small portions (use a disposable pipet) to the boiling
solution until a little solid (or oil) separates and will not redissolve on boiling. Clear the solution by
adding a few drops of the more potent solvent. Let the flask cool in the usual fashion, finally
collecting the product, as in the case above.

There are two hazards to avoid; the first is that already mentioned, the unfavorable melting
point/boiling point relationship and the second is peculiar to the use of solvent pairs. If a compound
is very soluble in one solvent and somewhat soluble in the other, and if too much of the first solvent
is used there is no amount of the second solvent which can give a saturated solution. A point is first
reached at which enough of the second solvent itself has been used to dissolve the compound without
assistance from the more potent solvent. This situation is recognizable by the failure of solid to
precipitate on the addition of a relatively large amount of the second solvent.

For this experiment, methanol or ethanol (neat or with added water) should serve as a good
recrystallizing solvent. The "solvent pair" is a mixture (of varying proportions) of alcohol-
water.

OVERVIEW
1. Dissolve solid in the minimum amount of alcohol using heat to help minimize the amount
2. Remove flask from heat and allow to cool to room temperature.
3. Chill solution in an ice-water bath and allow crystallization to occur. If crystallization occurs, skip to
step 7.
4. If no crystallization occurs, add water carefully until cloudiness persists. Use squirts, followed by
dropwise addition later.
5. Reheat until solution becomes clear.
6. Remove from heat, add seed crystals upon cooling in ice-water bath (if desired), and allow solution
to cool slowly.
7. Collect crystals.

Steps 1-3 describe a single solvent recrystallization, see Part A for details. Adding Steps 4-6 to the
procedure transforms the method to that of a solvent pair, i.e. methanol-water, as described in detail in Part
B.

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Part III. Mixture Melting Points.
Your second crystallization of each mixture should have given a product melting within a degree of the value
obtained after the first crystallization. (If such is not the case, consult your instructor as to the advisability of
doing a third crystallization). Consulting the melting point data for the four possible compounds in each
case, decide which ones should be tested by a mixture melting point determination.

Take very small and roughly equal amounts of your compound and an appropriate known substance, crush
and mix the combined sample very thoroughly, and then determine the melting point in the usual fashion.
Repeat with each of the substances that were possible choices. You should obtain broad and depressed
melting points in all but one case.

THE LABORATORY REPORT:

In this experiment there are three main considerations. In order of decreasing importance, these are: (i) the
correct identification of the substances, (ii) the purity of the isolated materials, and (iii) the amounts of
material recovered. Bottle your final samples, label them using the recommended identification system.
Include all of usual data except the percentage yield; substitute the word "Recovery" for "Yield".

Helpful Hints - Acid-Neutral Mixture

* The melting (or, boiling) point is always recorded as a range (initial and final).

* The choice of a single solvent should meet the following criteria: (i) dissolves solid when warm or at
its boiling point, but precipitates solid when cooled and (ii) bp generally lower than mp of solid.

* In the case of a solvent pair, the "overly effective" or "potent (strong)" solvent represents the chosen
solvent in which the solid to be crystallized is readily soluble at (or near) room temperature. The co-
solvent is chosen because it is miscible with the "potent" solvent. Crystallization or recrystallization
with a solvent pair necessitates dissolving the solid in a minimum amount of the potent solvent
followed by the addition of the "weaker solvent" to the point of cloudiness which indicates
precipitation. Heating to clarify the solution once again (more of the strong solvent may be
necessary), allowing the solution to cool slowly, and seeding the solution will normally result in
crystalline formation.

* Common solvent pairs used in our laboratories are: (i) alcohol (either methanol or ethanol) - water,
(ii) acetone - water, (iii) acetic acid - water, (iv) ether - methanol, and (v) ether - hexanes.

* Some of the common solvents, with properties of dielectric constant (ε), bp, and solubility in
water, are listed in the table below. With the exception of water, all are flammable and should be
treated with caution.

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 Summary Guideline for Acid-Neutral Report

The following should be recorded in your laboratory notebook

& turned in as part of lab report:

Unknown Number: _________________

Acid Component
Name: Structure

Gross weight:

Tare weight:

Net weight:

% recovery = 100 x =
1.8 g

mp: expected:

found:

Mixture:

Neutral Component
Name: Structure

Gross weight:

Tare weight:

Net weight::

% recovery = 100 x =
1.8 g

mp: expected:

found:

Mixture:

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 CHM 2211L

QUESTIONS

1. Provide and label all possible structures of the major and minor components of the acidic and neutral
mixtures.

2. Although acetic acid - water may be used as a "solvent pair", why are dilute solutions of an inorganic
acid/water (or, inorganic base/water) avoided in recrystallization attempts? [Hint: see McMurry:
sections 21.7, 21.8. Consider functional-group possibilities.]

3. Using a flow diagram, show the separation of a mixture containing p-bromoacetanilide,

N-cyclopentyl-N-methyl-N-propylamine (N means the group is attached to the nitrogen of the amine)
and m-nitrophenol. Specify the organic solvent and aqueous solutions to be used for the separation.
Provide the exact acid-base reaction that occurs during each step of the separation.

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