DIVISION OF NATURAL SCIENCES AND MATHEMATICS

UNIVERSITY OF THE PHILIPPINES VISAYAS

TACLOBAN COLLEGE
Magsaysay Boulevard, Tacloban City

LABORATORY MANUAL
in

CHEMISTRY 31.1
Elementary Organic Chemistry
Laboratory
(Source: Lab Manual on Chem 40.1, IC, CAS, UPLB)

1
 Laboratory Rules

1. Come to the laboratory prepared by studying the exercises

beforehand. Students will be questioned on their understanding of
the exercise any time during the performance. Know the hazards and
precautions of the chemicals indicated in the procedure. Some of the
steps may be altered as the need arises, but the principles involved
will be the same.

2. Students should come to the laboratory class in proper laboratory

attire; wearing of sleeveless tops, shorts, slippers, and open-toes
sandals are strictly not allowed. Students must wear protective
eyewear (safety goggles) and laboratory gowns during experiments.

3. Materials such as markers, masking tape, liquid detergent, rags and

tissue paper shall be provided by the students.

4. Experimental work is to be done on a group basis. Distribute the

laboratory work within the group but make sure everybody has a
chance to observe the experimental results.

5. Observe proper laboratory conduct. Minimize talking and avoid

disturbing other groups during experiments. Smoking, eating and
drinking inside the laboratory are absolutely prohibited. Honesty is
expected from everybody.

6. Handling of chemicals.

a. Pipets and droppers should not be placed inside bottles containing

stock solutions. Excess reagents should not be returned to the
main supply unless so directed; they could be given to the other
groups.
b. Beakers, test tubes and vials containing reagents, solutions, etc.
should be properly labeled.
c. If you must smell a substance, hold the container at a distance
and, with a cupped hand, waft the fumes toward your nose. Avoid
excessive inhalation.
d. If chemicals are spilled on your skin or on your clothing, flush them
off with water immediately. Inform your laboratory instructor.
e. Precautions must be observed to minimize fire hazards. Many
organic solvents are volatile and flammable.

2
7. Cleanliness

a. In the laboratory, reagents and special equipment are usually

located in the fume hood. Reagent bottles must not be brought
outside the hood. Keep all reagent bottles covered after use and
make sure they are returned to their proper locations.
b. Wipe spilled chemicals right away. After using any equipment such
as the balance, melting point determination apparatus, hot plate,
etc., make sure it is clean and ready for the next user.
c. Throw away solid trash – discarded filter paper, broken glass,
match sticks, etc. – into the waste containers provided and not
into the sink or bench drain.
d. Throw liquid waste – organic solvent waste, discarded reaction
mixtures, etc. – into the designated waste bottles provided and
not into the sink or bench drain unless instructed to do so.
Carefully follow waste management procedures written on the
laboratory manual.
e. Clean used glassware thoroughly before storage. All glasswares
must be finally rinsed with distilled water.

8. Report any accident, however minor, to the laboratory instructor.

3
 Exercise #1
Solubility as a Physical Property

Objectives

1. To observe and describe the dissolution process.

2. T establish that solubility is a physical property which is dependent
on some factors like the nature of the solute and solvent, pH and
temperature.
3. To perform and observe the salting-out process.

Introduction

A solution is a clear homogenous mixture of two or more

substances; the term is most commonly applied to those in liquid form, for
which the test for homogeneity is a uniform appearance and total lack of
cloudliness. The solvent is the main liquid component of a solution, while
the other present substances are said to be dissolved and are collectively
referred to as the solute. Solubility then, is the comparative ability of the
substances to dissolve in a given solvent and is usually measured as grams
solute per 100 mL solvent at a specific temperature.

Polarity and molecular (or ionic) size are the main factors that govern
solubility. When one substance dissolves in another, particles of the solute
(either molecules or ions) must be distributed throughout the solvent and
in a sense, the solute particles in the mixture occupy positions that are
normally taken by solvent molecules. Thus, ease with which a solute
particle may replace a solvent particle depends on the relative forces of
attraction of solvent particles for each other, solute particles for each other,
and the strength of solute-solvent interactions.

When an ionic substance dissolves in water, the ions that are adjacent
to one another in the solid become separated and are surrounded by water
molecules. In the immediate vicinity of the positive ion, the surrounding
water molecules are oriented so that the negative ends of their dipole points
in the direction of the positive charge. The water molecules surrounding a
negative ion have their positive ends directed at the ion. An ion, enclosed
with this “cage” of water molecules is said to be hydrated and generally,
when a solute particle becomes surrounded by molecules of solvent we say
that it is solvated. In a sense, the solvent insulates the ions from each
other.

4
 On the other hand, molecular compounds dissolve in water only if
they contain sufficient portion of hydrophilic or “water loving” bonds –
polar bonds involving oxygen or nitrogen atoms bearing a partial negative
charge whereby hydrogen bonding to water molecules is possible.
Otherwise, any attraction between solute and solvent molecules is too weak
to disrupt the hydrogen bonding that holds the water molecules together,
the mixing is not possible.

The solvent power of organic liquids is very different from that of

water because of the greater molecular size and the presence of a number
of nonpolar bonds in their molecules. Compared to water, organic solvents
are capable of dissolving a much wider range of organic compounds. “Like
dissolves like” is a general rule which emphasizes the fact that the solvent
power of a given organic liquid is greatest for substances of polarity similar
to its own.

In this experiment, the exact solubility values of the different solutes

will not be measured, but it is important to note whether the indicated
amounts of solute and solvent mix to give a solution or not.

Procedure

CAUTION: Organic solvents are flammable and generally toxic. Stay away
from an open flame when working with them, and avoid inhaling their
vapors.

A. Preliminary Observations of the Dissolution Process

Drop a small crystal of potassium permanganate into about 50 mL
of water contained in a 250-mL Erlenmeyer flask that is placed on a
white background. Observe what happens.

B. Solubility in Organic Solvent at Room Temperature

Before doing this part, record the descriptions of the pure organic
compounds.

Place 2.0 mL of petroleum ether in a small, clean, dry test tube.

Add the organic compound (3 drops of a liquid or a spatula tip of solid),
and mix by thorough agitation. Examine the mixture and record its
description.

5
 Using the above procedure, test the solubility of each of the
following organic compounds.
Set A: acetic acid, benzoic acid, sodium benzoate
Set B: cyclohexanol, dextrose, cellulose (cotton fiber)

C. Solubility in Water at Room Temperature

Adapt the procedure given in the preceding section using water in
place of the organic solvent. Test the same set of compounds for their
solubility in water at room temperature. Save the test tube containing
benzoic acid-water mixture for part D).

D. Effect of Temperature on Solubility

Carefully without any spillage, pour the contents of the test tube
containing the benzoic acid-water mixture into a large test tube. Use 1.5
mL extra water to rinse out any solids adhering to the walls of the small
test tube into the larger one. The resulting mixture now contains a larger
portion of water. Is it enough to dissolve all the benzoic acid? If not,
heat the test tube over a small flame or in a water bath. Observe what
happens to the mixture. When all the solids have disappeared, remove
the test tube from the heat and allow it to cool slowly. Observe what
happens.

E. Effect of pH on Solubility
Using the procedure in part B, determine the solubility of the
following compounds in the following solvents: water, 10% HCl, 10%
NaOH, and 10% NaHCO3.
Set C: aniline, diethylamine, naphthalene
Set D: benzoic acid, phenol, toluene

F. Salting Out and its Effects

Add 10 drops of 1-butanol to 4 mL water in a large, clean test
tube, and mix thoroughly. If any butanol remains undissolved, add a few
drops of water and agitate the mixture until finally a true solution is
observed. Record its appearance. Now, add a spatula of sodium chloride
and agitate the contents of the test tube until the solids are completely
dissolved. Is there any difference between this mixture and the original
solution? If no difference is observed, add more sodium chloride and
shake as before. Allow the mixture to stand for a while and repeat the
examination. Record the appearance of the new mixture.

6
G. Separation of a Simple Ternary (1:1:1 by Weight) Mixture of
Benzoic Acid, Naphthalene and m-Nitroaniline
Dissolve 3.0 g of the mixture in 20 mL dichloromethane. Transfer
the mixture quantitatively into a separatory funnel using
dichloromethane for washing so that the total volume of
dichloromethane used is 30 mL. Extract the dichloromethane mixture
with 20 mL of 10% HCl twice. Perform water test to determine which
layer is organic or aqueous. Separate the layers and reserve the organic
layer. Neutralize the combined aqueous layers with 20% NaOH while
using an ice bath for cooling the mixture. Collect the solids obtained and
wash with cold water.

Extract the organic layer with 20 mL of 10% NaHCO 3 twice.

Combine the aqueous layer and neutralize with concentrated HCl while
using an ice bath for cooling the mixture. Collect the crystals and wash
with cold water.

Evaporate dichloromethane from the remaining organic extract

using a steam bath (Do this under the fumehood!). Determine the
mass of the three components of the mixture and obtain the percentage
recovery. Transfer the recovered solids into their designated containers
(Consult your instructor!).

7
 Exercise #1
Solubility as a Physical Property

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 1.1 Preliminary observations on the dissolution process.

Reagents/Actions Taken Observations
KMnO4 crystals

KMnO4 crystals + 50 mL water

Mixture after swirling

Table 1.2 Description of organic compounds.

Compounds Description
Petroleum ether

Acetic acid

Benzoic acid

Sodium benzoate

Cyclohexanol

Dextrose

Cellulose (cotton fiber)

8
Table 1.3 Solubilities at room temperature.
Sample In petroleum ether In water
Mixture Solution Mixture Solution
description (+/-) description (+/-)
Acetic acid

Benzoic acid

Sodium
benzoate
Cyclohexanol

Dextrose

Cellulose

(+) soluble; (-) insoluble; (+/-) slightly soluble

Table 1.4 Effect of pH on solubility.

Sample In water In 10% HCl In 10% NaOH In 10% NaHCO3
Description +/- Description +/- Description +/- Description +/-

Aniline

Diethylamine

Naphthalene

Toluene

Benzoic acid

Phenol

Table 1.5 Salting-out and its effects on solubility.

10 drops 1-butanol + 1-butanol-water solution treated with sodium
4 mL H2O chloride
Freshly shaken After standing

9
Table 1.6 Effect of temperature on solubility: benzoic acid + water.
Action Observations
Freshly shaken

Hot mixture

Cooled after heating

Table 1.7 Separation and purification of a ternary mixture.

Action Observations
Ternary mixture

Addition of dichloromethane

Addition of 20% HCl

- Organic layer

- Aqueous layer

- Aqueous layer + 20% NaOH

- Recovered crystals

Addition of 20% NaHCO3

- Organic layer

- Aqueous layer

- Aqueous layer + 20% NaOH

- Recovered crystals

Evaporation of CH2Cl2

- Residue

10
Table 1.8 Data on % recovery.
Mass, g % Recovery
Mixture 1, 2 and 3

- Container + sample ---

- Empty container ---

- Sample ---

Component 1

- Container + sample ---

- Empty container ---

- Sample

Component 2

- Container + sample ---

- Empty container ---

- Sample

Component 3

- Container + sample ---

- Empty container ---

- Sample

Answer to Questions

Q1: What happens to the unit particles of a solute when it dissolves in a

liquid?
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q2: A true solution is homogeneous and said to have only one phase. What
is a “phase”?

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___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: How do you explain the difference in the solubility of the Set A
compounds in the organic solvent? How about the Set B compounds?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: How do you explain the difference in the solubility of the Set A
compounds in water? How about the Set B compounds?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q5: How do you explain the difference in the solvent power of water and
the organic solvent?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q6: What is meant by the salting-out effect? Illustrate.

Q7: Reports of exact solubility figures in g solute per 100 mL solvent are
accompanied by a notation on the temperature measurement. Why is this
necessary? How would you explain the effect of temperature on solubility?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________

12
 Exercise #2
Volatility: Distillation and Determination of Boiling Point

Objectives

1. To compare the volatilities of different organic compounds.

2. To demonstrate simple distillation set up.
3. To separate the components of a mixture of 2 liquids.

Introduction

Volatility is the relative ease with which a substance passes from the
liquid to the gaseous (vapor) state. To escape from the liquid into the
gaseous state, an individual molecule requires energy – to break away from
other liquid molecules, and to maintain the vigorous movement
characteristic of a gaseous particle. The amount of a substance that is
present as gas in the space above the surface of the liquid contained in a
closed vessel is measured by its vapor pressure, which increases with
temperature.

Evaporation is the slow volatilization by the escape of surface

molecules from the liquid into the air. Boiling is the fast volatilization of a
liquid; the turbulence observed is due to the formation of vapor even in the
body of the liquid. Boiling occurs when the vapor pressure is equal to the
atmospheric pressure, and the temperature required for this quality is
called the boiling point. The less volatile a liquid is, the higher is its boiling
point.

Distillation is the process of vaporizing a liquid and converting the

vapor to the liquid state by cooling. The sample mixture (generally a
solution) is heated to sustain boiling, and the vapor is allowed to pass
through a cooling tube where it condenses to form the distillate. Thus,
distillation serves to separate the volatile from the non-volatile component
of a mixture.

If the original liquid sample contains only one volatile component, the
vapor produced will consist of this component alone, and its temperature
will be near the boiling point of the latter. (At a given pressure, the boiling
point of a pure liquid is a characteristic constant and serves as a clue to its
identity, just as the melting point does for a pure solid). A mixture
containing two or more main components that are volatile will give a mixed

13
vapor, the composition of which depends on the relative volatility of these
components and their respective concentrations in the liquid phase. As
distillation proceeds, the vapor composition changes towards increasing
concentration of the less volatile component(s) as indicated by the variation
of the vapor temperature.

Procedure

A. Comparison of Volatility by Rate of Evaporation

Arrange 5 small, clean, dry watch glass in a row on the bench top.
Near each, place a piece of paper as label to indicate the compound
under examination: a) acetone, b) methanol, c) ethylacetate, d) 1-
butanol and e) water. When ready, place 2 drops of the compound to be
tested in the center of the watch glass, and immediately note the time
on the label. When the liquid has completely disappeared, note down
the time again.

B. Distillation and Boiling Point Determination

Place 30 mL ordinary tap water in a 50 mL distilling flask, taking
care that no liquid enters the side arm. Add a piece of porcelain or boiling
chips to prevent bumping, and assemble the rest of the distillation
apparatus. The tip of the thermometer bulb should be in the center of
the flask neck, about 5 mm below the side arm. Check all the
connections for tightness and turn the water on the condenser. Heat the
flask contents to boiling then adjust the flame so the condensate falls
from the condenser at the rate of 1 drop in 5 seconds. Record the
thermometer reading when the volume of the distillate is about 3 mL.
Record again the thermometer reading at each succeeding 3 mL interval
until a total of 15 mL distillate has been collected. The boiling range of
water is the temperature indicated at the first drop and at the last drop
collected, provided that the distillation has proceeded at a steady rate.

Note:
1. Bumping is the term given to irregular boiling, whereby the liquid
alternates between relative calm and violent motion, which may lead
to spillage.
2. Rate of heating must be controlled to avoid superheating of the
vapor.

14
 Exercise #2
Volatility: Distillation and Determination of Boiling Point

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 2.1 Volatilities of different substances.

Compounds Time Exposed Time Completed Minutes Elapsed
Acetone

Methanol

Ethylacetate

1-butanol

Water

Table 2.2 Simple distillation and boiling point of water.

Descriptions:
Original tap water:
___________________________________________________________
Residue after distillation:
___________________________________________________________
Distillate:
___________________________________________________________

Volume of distillate 1 drop 3 mL 6 mL 9 mL 12 mL 15 mL

T of vapour, °C

Boiling Point range of water:

15
 Exercise #3
Extraction through Immiscible Liquids

Objectives

1. To extract caffeine from tea samples using liquid-liquid extraction.

2. To compute for the percent caffeine in the tea sample.

Introduction

When 2 liquids which are very sparingly soluble in each other are
shaken together, they form 2 layers after some time: the more dense liquid
below and less dense above. Their immiscibility may be traced to the wide
difference in their polarities. Separation of 2 immiscible liquids maybe
effective by the use of separatory funnel.

If there is a solute in any of the immiscible liquids, the solute will

distribute itself between the 2 layers, in a definite proportion, depending
on its solubilities in the 2 solvents. The distribution may be expressed thus:

C1
K=
C2

where: K is distribution coefficient, C1 is concentration in solvent 1 and C2

is concentration in solvent 2. The distribution coefficient may be obtained
from literature values or calculated from solubility values of the solute in
the 2 solvents. This process is called liquid-liquid extraction.

Procedure

1. Place 15 g of tea, 150 mL of distilled water, 7 g of CaCO 3, and a

boiling chip in a 250-mL beaker.
Note: Calcium carbonate reacts with tannins to form insoluble
precipitate subsequently removed by filtration.
2. Boil the mixture gently over a hot plate for 15 – 20 min with
occasional stirring. Allow the mixture to settle and cool to room
temperature.
3. Filter the mixture through a cotton plug in a glass funnel. Transfer
the filtrate into a separatory funnel and add 25 mL chloroform.

16
4. Stopper and shake vigorously until a fine emulsion is formed. Allow
the liquids to separate into clear layers, then draw off the chloroform
layer.
Note: The tea-chloroform emulsion sometimes takes a long time to
stratify. If work is interrupted at this point, drain the emulsion into a
250-mL flask stopper and store it inside the locker for stratification.
In the next laboratory period, the layers formed may be separated.
5. Extract the aqueous layer once more with 25 mL chloroform.
6. Dry the combined chloroform extracts by adding 0.5 – 1.0 g of
anhydrous sodium sulfate.
Note: The most commonly used drying agents are anhydrous
magnesium sulfate and calcium chloride. They absorb water by
forming hydrates.
7. Remove the sodium sulfate by filtering the solution through a cotton
plug in a dry glass funnel and into a previously dried and weighed 25-
mL beaker.
8. Evaporate the concentrate to dryness in a steam bath.
9. Determine the weight of crude caffeine and compute for % caffeine
in the tea sample.

17
 Exercise #3
Extraction through Immiscible Liquids

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 3.1 Extraction of caffeine from tea leaves.

Sample/ Actions Taken Description
Solid sample (type, brand)

Mixture after boiling, cooled to RT

Filtrate

Extraction with chloroform:

- Organic layer

- Aqueous layer

Extract after addition of sodium

sulphate
Chloroform concentrate

Crude caffeine

Table 3.2 Determination of % caffeine in the sample.

Mass of crude caffeine + container, g

Mass of container, g

Mass of crude caffeine, g

Mass of sample, g

% caffeine in sample

18
Answer to Questions

Q1: Discuss briefly the role of the following reagents in the extraction of
caffeine.
a. Calcium carbonate
___________________________________________________________
___________________________________________________________
b. Anhydrous sodium sulfate
___________________________________________________________
___________________________________________________________
Q2: What are the characteristics of chloroform that make it a good
extracting solvent for caffeine?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: What are emulsions? Why do they form during extractions? How is the
formation of an emulsion minimized?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: Why is it necessary to remove a stopper from a separatory funnel when
liquid is being drained from it through a stopcock?
___________________________________________________________
___________________________________________________________
___________________________________________________________

19
 Exercise #4
Chromatography

Objectives

1. To learn the techniques of paper chromatography and thin layer

chromatography.
2. To apply chromatographic methods in the separation of the
components of a mixture.
3. To identify an unknown sample by comparing its R f value and other
characteristics with those of a standard.

Introduction

Chromatography includes a variety of techniques for separating

individual compounds or compound types from a mixture. Separation is
effected by the distribution of the components of the mixtures between a
stationary phase and a mobile phase. Various types of chromatography are
possible depending on the physical states of the stationary and mobile
phases involved. Adsorption chromatography uses a solid stationary phase
and a liquid mobile phase. Separation using adsorption chromatography is
governed by surface adsorption phenomena. On the other hand, partition
chromatography uses a liquid stationary phase supported on the surface of
a solid and a liquid or gas mobile phase which is insoluble in the stationary
phase. Partition chromatographic separations may be due to differences in
the solubility of the sample in the stationary and mobile phases.

In adsorption chromatography, the mixture to be separated is

adsorbed on the solid stationary phase over which the liquid mobile phase
is allowed to flow. The transfer of the adsorbed compound between the
stationary phase and the mobile phase is an equilibrium process.

𝐾
compound in stationary phase ↔ compound in mobile phase

The extent of adsorption of a single component depends on the

polarity of the molecule, the activity of the adsorbent, and the polarity of
the liquid mobile phase. In general, the more polar a functional group in
the compound is, the more strongly it will be adsorbed on the surface of
the polar stationary phase. The actual separation of the components in a
mixture is dependent on the relative values of the adsorption-desorption
equilibrium constant, K, for each of the components. The individual

20
components will move with the mobile phase at different rates, resulting in
their separation into different regions (as bands or spots) in the stationary
phase.

Thin layer chromatography (TLC) is a form of solid-liquid adsorption

chromatography that uses a thin layer of adsorbent (usually alumina or
silica gel) supported on a flat surface (usually glass) is stationary phase.
TLC is very useful in monitoring the progress of reactions, detecting
intermediates in reactions, analyzing crude products or unknown mixtures,
determining the number of components in a mixture and evaluating the
efficiency of purification processes.

Paper chromatography bears a resemblance to TLC but is highly

different in principle. Filter paper which is made of highly purified cellulose
absorbs and retains water molecules strongly. This is because cellulose is
a polyhydroxy compound. The paper (cellulose) and the bound water which
forms part of its structure constitute the stationary phase in paper
chromatography. Paper chromatography is an example of liquid-liquid
partition chromatography. Like TLC, paper chromatography is used in the
rapid analysis of the components of reaction mixtures and as a tentative
means of identification.

Small spots of the mixture to be separated are placed near the

bottom of a strip of filter paper or a TLC plate and a solvent (mobile phase)
is allowed to travel up by capillary action. Separation takes place due to
the different affinities of the components of the mixture for the polar
stationary phase and the mobile phase which is a relatively nonpolar
solvent or solvent system. There is a continuous back-and-forth exchange
of solutes between the two phases, but those which are more soluble in the
mobile phase spend more time in it and are carried up faster.

A compound will move up the TLC plate or paper strip at a rate

relative to that of the solvent front. The relative mobility is known as the R f
value of the compound and is defined by the equation,

distance travelled by compound from the origin

Rf =
distance travelled by the solvent from the origin

where the origin is the midpoint of the original spot. The distance traveled
by a compound is obtained by measuring the distance from the origin to
the point of greatest density (center of mass) of the spot corresponding to

21
the compound. Under a defined set of conditions (adsorbent, solvent,
temperature, and humidity) the Rf value is a characteristic property which
can be used for the identification of a compound.

Procedure

A. Separation of Plant Pigments by Paper Chromatography

1. Collect about 1 g of leaves from any one kind of plant in the
locality. Take note of the plant’s scientific name and the place
where it was collected.
2. Cut the leaves into small pieces.
3. Extract the leaf pigments with minimal amount of acetone using a
mortar and pestle. Filter the extract through a piece of cotton.
4. Draw a clear acetone extract into a capillary tube and use this
extract to make spot on a strip of chromatographic paper about 1
cm from the bottom. (To save time and materials, practice your
spotting technique on a piece of scratch paper before attempting
to use the chromatography strip). Make sure that the spot does
not exceed 2 mm in diameter.
5. Repeat step 4 five times, allowing the spot to dry each time.
6. Take another strip and do steps 4 and 5.
7. Pour 5 mL of 9:1:1 (v/v/v) hexane: ethanol: acetone and 9:1 (v/v)
hexane: acetone into two separate 50 mL test tubes.
8. Mount the test tubes in 250-mL Erlenmeyer flasks.
9. Attach the paper with the spotted side at the far end to a hook
mounted on a stopper and carefully insert the spotted end of the
strip into the test tube. Make sure that the paper does not touch
the side of the test tube. See to it that the spot is above the
level of the solvent.
10. Stopper the tube tightly and watch the solvent rise on the
paper.
11. When the solvent front is about 1 in from the top of the strip,
remove the paper from the test tube and mark the solvent front
with a pencil.
12. Allow the strip to dry.
13. Mark the outline of the individual spots with a pencil. Draw the
pattern produced on your data sheets and label each spot
according to its color and shape.
14. Compare the chromatograms produced by the two solvent
systems as to the extent or degree of separation of the spots.

22
 Strip of
chromatographic paper

Figure 4.1 Setup for paper chromatography.

(http://www.tanlam.com/science/biology/eoi/eoi.htm)

B. Analysis of the Component Dyes of Black Ink by TLC

CAUTION: Avoid inhaling silica gel in TLC plates!

1. Draw an aliquot of the ink sample into a capillary tube and spot it
on one TLC plate/chalk, approximately 1 cm from the bottom. Spot
a sample of another ink brand onto another TLC plate/chalk.
2. In a wide-mouth screw-cap bottle, pour about 20 mL of the solvent
system – 6:2:2 (v/v/v) 1-butanol: ethanol: 2N NH3. Line the sides
of the chamber with a piece of filter paper and allow the system
to equilibrate.
3. After about 2 min, place the TLC plate/chalk in the developing
chamber. Cover the chamber tightly.
4. Allow the chromatogram to develop (about 20 min).
5. Analyze and compare the chromatograms as in Part A.

23
 Figure 4.2 Setup for thin layer chromatography.
(http://www.chemguide.co.uk/analysis/chromatography/thinlayer.html)

C. Identification of Amino Acids by Paper Chromatography

1. Obtain a clean sheet of Whatman no. 1 filter paper, about 12 cm
by 18 cm. Handle it only at its corners.
2. Using a pencil, lightly draw a thin line parallel to one side,
approximately 1.5 cm from the edge of the paper.
3. Lightly place 8 X’s along the line at 2 cm intervals.
4. Under each X, place an identifying mark, two for each standard
(P=phenylalanine, T=tyrosine and A=aspartic acid) and two for
the unknown substance (U).
5. Spot a small amount of each standard and unknown solution on
its designated position on the paper. Do this five times allowing
the spot to dry each time.
6. Roll the paper into a cylinder and staple the ends. Make sure that
the edges of the paper do not touch each other.
7. Fill the developing chamber up to about 0.75 cm deep with the
solvent system 4:1:1 (v/v/v) n-butanol: glacial acetic acid: water.
8. Place the cylindrical paper in the chamber, observing the usual
precautions.
9. Cover the chamber tightly and allow the chromatogram to develop
(about 1 and ½ hours).
10. After the chromatogram has been developed, remove the paper
and open it. Mark the solvent front lightly with a pencil.
11. Allow the paper to dry.
12. Dip the paper in a 2% ninhydrin solution in acetone. CAUTION:
Ninhydrin is a neurotoxin. Avoid direct skin contact.
13. Allow the solvent to evaporate.
14. Place the paper in an oven at 100-110°C for about 10 min.
15. Encircle each spot with a pencil and calculate Rf values.
24
16. Compare the spots in terms of shape and color.
17. Draw the chromatogram.

Figure 4.3 Setup for paper chromatography of amino acids

(http://employees.oneonta.edu/helsertl/AAChrom.html)

25
 Exercise #4
Chromatography

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 4.1 Data on the plant sample.

Scientific Name:

Common Name:

Place and Date of Collection:

Description of Sample:

Description of Acetone Extract:

Table 4.2 Data on the analysis of plant pigments by paper chromatography.

Solvent System

Sketch of
chromatogram

26
Table 4.3 Data on the analysis of the ink samples by TLC.
Solvent system: ______________________________________________
Ink sample
(color and brand)

Sketch of
chromatogram

Table 4.4 Data on the analysis of amino acids by paper chromatography.

Solvent system: ____________________________________________

Distance travelled by solvent: _________________________________

Diagram of the chromatogram:

27
Table 4.5 Data on the Rf values of the standard amino acids.
Amino Acid Distance Traveled Average
Name Color and Trial 1 Trial 2 Average Rf
shape of spot values

Table 4.6 Data on the identification of the unknown amino acid.

Amino Acid Distance Traveled Average
Sample Code Color and Trial 1 Trial 2 Average Rf
shape of spot values

Identity of the Unknown Sample: Basis of identification:

Answer to Questions
Q1: What would be the effect of the following errors in chromatographic
work?
a. The solvent level in the developing chamber is higher than the
spotted sample.
______________________________________________________
______________________________________________________
______________________________________________________
b. Too much sample is applied to the paper.
______________________________________________________
______________________________________________________
______________________________________________________
c. The paper is allowed to remain in the chamber after the solvent front
has reached the top of the plate.
______________________________________________________
______________________________________________________
______________________________________________________
Q2: Why is it necessary to cover the developing chamber tightly during the
development of a chromatogram?
___________________________________________________________
___________________________________________________________
___________________________________________________________

28
Q3: Can TLC or paper chromatography be used to separate and identify
very volatile substances? Explain your answer.
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: In Part C, why were you required to handle the chromatographic paper
only at its corners?
___________________________________________________________
___________________________________________________________
___________________________________________________________

29
 Exercise #5
Stereochemistry

Objectives

1. To construct three-dimensional models of molecules and translate

them to two-dimensional representations (drawings) and vice-versa.
2. To determine chirality of molecules in terms of
a. Superimposability/non-superimposability with corresponding
mirror image.
b. Presence/absence of a simple element of symmetry such as a
plane of symmetry.
c. Presence/absence of stereocenters.
d. Presence/absence of optical activity.
3. To predict the existence of enantiomerism and diastereomerism in
certain compounds.
4. To differentiate between:
a. Enantiomers and racemic mixtures.
b. Enantiomers and diastereomers.
5. To recognize that
a. Free rotation about C–C single bonds gives rise to different
conformations.
b. Conformations are continuously and rapidly interconverting.
c. There is a preferred conformation.
6. To differentiate between conformational and configurational
stereoisomers.

Introduction

Stereochemistry is concerned with the shapes of molecules and how

the differences in shape can affect the properties and reactions of
compounds. Subtle differences in molecular shape have far-reaching
consequences and it is probably in the field of molecular biology that a full
awareness of the importance of molecular shape has emerged. For
example, the shape of the complex DNA molecule, which has been
described as the key to life itself, is involved in passing the genetic code
from parents to offspring. Knowledge of the molecular shapes of such vital
compounds as sugars, amino acids and proteins is essential in
understanding their reactions and properties.

30
 Molecular structures are so frequently represented in two dimensions
that thinking about molecules in three dimensions can be considerably
difficult. The purpose of this exercise is to help organic chemistry students
think in three dimensions. The exercise is designed in a manner that would
give step-by-step introduction to the basic concepts of stereochemistry. A
student, however, must first be able to:

a. Write and interpret molecular and structural formulas and

b. Recognize that a compound with a particular Lewis structure can
exist in a number of forms called stereochemical isomers that
differ only in the arrangement in space of the bonded atoms.

Throughout the exercise, students should actively participate in the

learning process by answering questions and constructing and inspecting
molecular models.

Procedure

Read through the exercise and perform the required activities

(constructing models, drawing, answering questions) as they are asked.
When an instruction does not seem to be clear, or when you have difficulty
in constructing or analyzing your model, consult with your laboratory
instructor.

Materials:
Each group will be provided with a kit containing the following:
Spheres: assign a color for each element [7 carbon, 3 oxygen, 1
chlorine, 1 bromine, and 1 fluorine]
1 short white sticks
7 long white sticks
2 coils of springs
2 forks
2 pentagonal pieces of cardboard
1 mirror

A. Chirality, Enantiomerism, Diastereomerism, Optical Activity

Objects that are IDENTICAL are SUPERIMPOSABLE – that is,
when we imagine the two objects “fused” together, all the corresponding
parts match exactly. Thus, a sphere is superimposable on another
sphere of the same diameter. A sphere, however, is not superimposable
on a cube or on a sphere of a different diameter.

31
1. Determine whether each of the following pairs of objects are identical
with each other, i.e., superimposable or not.
a) Two forks b) your left hand and your right hand

2. View your right hand in front of the mirror.

a) Is the reflection of your right hand superimposable on your left
hand?
b) Is your right hand the mirror image of your left hand?

3. Using a mirror, determine whether the following objects are

superimposable on their mirror images or not.
a) a fork b) a pentagon c) your left hand

So far we have examined certain simple relationships between the

shapes of pairs of objects. Two objects may be IDENTICAL or DISTINCT.
Distinct objects may be related as mirror images of each other or not
related as mirror images at all.

IDENTICAL: superimposable mirror images

DISTINCT: non-superimposable

mirror images not mirror images

Molecules can be classified in a similar way. Many molecules are

superimposable on their mirror images, others are not. Those which are
related as non-superimposable mirror images are particularly important
in biological systems.

4. A PLANE OF SYMMETRY is an imaginary plane or sheet that divides

the object so that one half is the exact reflection of the other half.
Determine whether each of the following has at least one plane of
symmetry or none:
a) a sphere b) a regular tetrahedron c) a shoe

5. Determine whether the objects in no. 4 are superimposable on their

mirror images or not.

6. Describe the relationship between the presence of a plane of

symmetry in an object and the object’s superimposability on its
mirror image.

32
 Objects which are not superimposable on their mirror images are
CHIRAL objects, while those which are superimposable on their mirror
images are ACHIRAL.

There are two alternative methods which may help in the

analysis of the shape of an object to decide whether it is chiral or
achiral:

a) Construct the mirror image of the object. If the mirror image is

identical to the object, that is, superimposable, then the object is
ACHIRAL. Otherwise, it is chiral.
b) Inspect the object for the presence of a plane of symmetry. If it has
a plane of symmetry, then the object is ACHIRAL. Otherwise, it is
chiral.

The two approaches can be summarized in the following scheme:

Given Model A

Construct model B, the Inspect for the presence of

mirror image of A a plane of symmetry

Are models A and B Does model A have a plane

identical? of symmetry?

NO NO
Model A is
CHIRAL
YES YES

Model A is ACHIRAL Model A is ACHIRAL

7. Construct a model of CH4. Is it chiral or achiral? Why?

8. Construct a model of CBrFClH. Is it chiral or achiral? Why?

33
 A chiral molecule such as molecule I below is distinct from its non-
superimposable mirror image, molecule II. Construct models for I and
II, and verify this relationship.

I II

(http://plaza.ufl.edu/tmullins/BCH3023/isomers.html)

Molecules such as I and II, which are related as non-

superimposable mirror images, are called ENANTIOMERS.

9. a) Are enantiomers chiral?

b) Do enantiomers have the same molecular formula?
c) Does one structural formula represent both members of a pair of
enantiomers?

Stereoisomers possess the same molecular and structural

formula, but have different spatial arrangements of their atoms.

10. Are enantiomers stereoisomers?

One of the most important properties of chiral molecules in

solution is their effect on plane-polarized light. Both enantiomers rotate
the plane of polarized light. Substances with this property are said to be
optically active. Chiral substances have molecules that are not
superimposable on their mirror images and are optically active.
Enantiomers are also known as optical isomers.

11. Determine whether each of the following is optically active or

not.
a) CH3CH(NH2)COOH
b) CH3CH2CHClCH3
c) CH3CH2CH2CH3

34
 The rotation of plane polarized light is used to observe
experimentally one of the main differences between a pair of
enantiomers. One enantiomer rotates plane-polarized light clockwise (as
seen by observer), the other enantiomer, counterclockwise by the same
magnitude. Conventions used to indicate direction of rotation of the
plane polarized light are:
Clockwise (+), dextrorotatory, d
Counterclockwise (–), levorotatory, l

Alanine, , is a chiral amino acid that has two

enantiomers: (+)-alanine and (-)-alanine. These two are optical
isomers.

12. Which enantiomer of alanine rotates the plane of polarized light

clockwise?

13. Substances which do not rotate the plane of polarized light are
said to be optically inactive. Determine whether each of the following
aqueous solutions are likely to be optically active or inactive.
a) a solution of ethanol, CH3CH2OH
b) a solution of (+)-alanine
c) a solution of (-)-alanine
d) a solution containing equimolar quantities of (+)-alanine and (-)-
alanine

A racemic mixture containing equimolar quantities of a pair of

enantiomers is called a RACEMIC MIXTURE. A racemic mixture is
represented as (±) or sometimes dl, e.g., (±)-glucose or dl-glucose. A
racemic mixture is optically inactive.

14. How is the racemic mixture of alanine represented?

In the preceding section, we have seen that if a substance can be

shown to be optically active in solution, then we know it is chiral.
However, absence of optical active does not prove that the substance is
achiral, since a racemic mixture may be present. It is therefore also
useful to be able to “detect” molecular chirality directly from molecular
shape, if necessary, with the aid of models.

35
 Two methods have been introduced so far. Another method that
allows the detection of chirality directly from molecular structure (and
shape) is described below. This method is useful for simple organic
compounds and easily works even without the aid of models.

15. Construct models of molecules with the following generalized

formulas using differently colored markers for the atoms or groups of
atoms W, X, Y and Z.
W Y X W

W Y Z Y
W Y X X
W W Y Z

model I model II model III model IV

16. Construct the mirror image of each model. Which are

superimposable on their mirror images?

17. Which model represents a chiral molecule?

18. How many different atoms or groups of atoms must be attached

to the central carbon atom in order to confer chirality to the molecule?

A carbon atom linked to four different groups is asymmetric and

known as a tetrahedral STEREOCENTER. The presence of at least one
carbon stereocenter renders chirality to a molecule.

19. Consider the following structural formulas:

OH Cl

COOH CH2Br

(a) (b)

Match these structural formulas with the generalized formulas

given in number (15). Which of the two contains a stereocenter? Draw
the structure and mark the stereocenter with an asterisk.

36
20. Which of the above is a chiral molecule? Explain your answer.

21. a) Which of the molecules in number (19) can exist as a pair of

enantiomers?
b) Draw the pair of enantiomers.

In the preceding section, you have learned to recognize that the

presence of a stereocenter can confer chirality to a molecule as a whole.

Lactic acid, , for example contains one stereocenter

(marked with *). It is a chiral molecule and therefore can exist as two
pure optical isomers, (+)-lactic acid and (-)-lactic acid.

Now we will consider molecules with more than one stereocenter.

Consider 2,3-dihydroxybutanoic acid.

22. a) How many stereocenters does it have?

b) Copy the structure and mark the first stereocenter with an
asterisk. Encircle the four groups attached to this stereocenter.
c) Copy the structure and mark the second stereocenter (if any)
with an asterisk. Encircle the four groups attached to this
stereocenter.

The molecule 2,3-dihydroxybutanoic acid is an example of a

substance with two different stereocenters.

The flying wedge representation is a two-dimensional

representation where a solid wedge ( ) indicates a bond projecting
up and out of the plane of the paper while a dashed wedge ( )
indicates a bond projecting into the paper. Thus, in I, –OH and –H are
projecting towards you, the observer, while the –COOH and –CH3 groups
are projecting into the paper. (Differentiate H, OH, COOH and CH 3 by
using differently colored markers.)

37
 COOH

H C OH

H C OH

CH3

23. a) Make a model of this molecule whose three dimensional

structure is given above in flying wedge representation. Copy
structure I.
b) Construct a second model, II. Draw a flying wedge
representation of II, related to I as mirror image.
c) Are I and II chiral?
d) What kind of stereoisomers do they represent?

A compound that contains two different stereocenters can exist as

four optically active stereoisomers. Construct the models of the other
two stereoisomers of 2,3-dihydroxybutanoic acid, III and IV. The flying
wedge representations of the four stereoisomers are given below:
COOH COOH COOH COOH

H C OH HO C H H C OH HO C H

H C OH HO C H HO C H H OH
C

CH3 CH3 CH3 CH3

I II III IV

All of the above stereoisomers of 2,3-dihydroxybutanoic acid are

optical isomers. Some pairs are related as non-superimposable mirror
images (enantiomers). Other, called DIASTEREOMERS, are also non-
superimposable and are not related as mirror images.

24. What is the relationship between

a) I and II b) III and IV c) I and IV d) II and IV

38
25. What is the relationship between
a) I and III b) II and III

26. The constitutional formula of tetrose is shown below

O
║
H–C–CH(OH)CH(OH)CH2OH

a) Using the flying wedge representation, draw all the stereoisomers

of tetrose. Label each stereoisomer I, II, and so on. Mark the
stereocenters using asterisks.
b) Identify enantiomers.
c) Identify diastereomers.

B. Conformational Isomers
Atoms within molecules can vibrate and are free to rotate about
single bonds. This rotation may result in the atom taking different
positions relative to the rest of the molecule. Molecular structures that
are interconvertible by simple bond formations are called
CONFORMATIONAL ISOMERS or CONFORMERS.

Open Chain Compounds

27. Construct a model of ethane CH3CH3. Rotate the C–C single

bond.
a) Is there a change in the relative positions of the different atoms?
b) Does the ethane molecule have more than one conformation?

Rotation about the C–C bond changes the shape of the molecule
while rotation about the C–H bond has no effect. Atoms can rotate
“freely” about single bonds as opposed to double bonds (e.g., C=C,
C=N), which have restricted rotation. The ethane molecule can adopt an
infinite number of conformations according to the relative positions of
the hydrogen atoms. These conformations can be interconverted by
rotating about the C–C single bond. The two extreme conformations of
ethane, the ECLIPSED and STAGGERED can be seen by viewing long
the C–C axis. Examine these conformations using your model.

28. In which of the two conformations are the hydrogen atoms

farthest apart from each other?

39
 Conformation Representation
Sawhorse projection Newman projection

Eclipsed

Staggered

The conformation in which the atoms are farthest apart normally

has the lowest potential energy and is referred to as the preferred
conformation. Molecules are continuously and rapidly changing from one
conformation to another, but they spend most of their time in the
preferred conformation.

The changes in potential energy of ethane as one methyl group

moves relative to the other about the carbon-carbon single bond can be
shown diagrammatically in the figure below. The dihedral angle (or
rotation angle, Φ) is illustrated as:

The difference in the potential energy of the staggered and eclipse

conformers of ethane is 3 kcal/mol. Because this energy is readily
attained at room temperature, different conformations of ethane cannot
be isolated. An energy barrier greater than 15-20 kcal/mol is required
to be able to isolate different conformations at room temperature.

29. Complete the diagram in your sheet by indicating changes in

relative potential energy which occur during a full 360° rotation of
one methyl group of ethane relative to the other methyl group.

40
 1.2

Relative potential energy

1

0.8

0.6

0.4

0.2

0
0 60 120 180 240 300 360
Dihedral angle, Φ

30. Draw the Newman and sawhorse representations of the

conformation of ethane with the highest potential energy.

31. Draw the Newman and sawhorse representations of the

conformation of ethane with the lowest potential energy.

32. Which of these conformations is the preferred conformation of

ethane?

33. Construct a model of chloroethane, CH3CH2Cl. Remember that

these models do not actually show the relative sizes of different
atoms. View the model along the carbon-carbon axis and draw the
sawhorse and Newman projections of the preferred conformation.

34. Do all chloroethane molecules spend all of their time in this

preferred conformation?

35. What is the preferred conformation called?

In ethane, energy differences between various conformations are

mainly due to electronic interactions between the electron pairs in the
C–H bonds. In chloroethane, there are electronic interactions between
the C–H bonds and with the C–Cl bonds. Also, interactions between the
larger chlorine atom and hydrogen atoms are greater than any
hydrogen-hydrogen interactions.

41
36. Construct a model of 1,2-dibromoethane, BrCH2CH2Br.
Examine possible conformations of this molecule. Which of the
following interactions will be the greatest?
a) hydrogen-hydrogen interactions
b) hydrogen-bromine interactions
c) bromine-bromine interactions

Ring Compounds

Many important organic molecules contain rings of atoms, for

example, sucrose, chlorophyll, nicotine, etc. Rings range in size from
three-membered like cyclopropane to those containing more than 30
atoms. However, most cyclic compounds are made up of 5- or 6-
membered rings. In this exercise, only cyclohexane and its
monosubstituted derivatives will be considered.

Construct a model of the cyclohexane molecule (C6H12).

Rotate the carbon atoms and examine different conformations.

37. Are there any conformations in which all carbon atoms of the
ring are in one plane? (Note: Be careful not to distort models.)

The two extreme conformations of cyclohexane are referred to as

the CHAIR and BOAT conformations.

chair boat

42
 Manipulate your models carefully to get the chair and boat
conformations. Be sure you can differentiate the two extreme
conformations. If in doubt, consult your laboratory instructor.

Put your models carefully in the chair conformation. Look at the

relative positions of the hydrogen atoms along the axes, C2–C3 and C5–
C6. Hold carbons 2, 3, 5 and 6 firmly in one hand, then without moving
carbon 1, flip carbon 4 up (or down) without breaking any bond. Again
look at the relative positions of the hydrogen atoms along C2–C3 and C5–
C6 bonds.

38. Does the flipping of carbon 4 result in a change in

conformation?

39. In which conformation are the hydrogen atoms farthest apart?

40. Which is the preferred conformation? Give reasons for your

answer. Compare your models with the Newman projections below
by looking through C2–C3 and C5–C6 bonds at the same time.

I II

41. Which is the eclipsed conformation? The staggered

conformation? Which is the boat conformation? The chair
conformation?

In the chair conformation, notice that there are two orientations

for the C–H bonds. Bonds that are oriented vertically are known as
AXIAL bonds; the rest are known as EQUATORIAL bonds. The axial
bonds alternate above and below the ring carbons while the equatorial
bonds point away from the ring. The drawings below are meant to show
the general orientations of equatorial and axial bonds. They do not
represent specific compounds.

43
equatorial bonds only axial bonds only axial and equatorial bonds

Use white sticks for the axial bonds. Let these be the marked
bonds. Hold carbon atoms 1, 2, 3, 5 and 6 firmly and flip carbon 4 up.

42. What conformation results?

43. Then holding carbons 2, 3, 4, 5 and 6 firmly, flip carbon 1 down.

What conformation results?

44. Did you break any bonds?

45. What is the orientation of the marked bonds now?

46. Make a sketch of the final conformation (include marked

bonds).

Now, with the equatorial bonds marked by the white sticks,

reverse the process and rotate the bonds to reform the chair
conformation such that the axial positions are now marked. This overall
process of one chair conformation changing to another chair
conformation is known as RING INVERSION or ring flipping and is a very
rapid process. The cyclohexane ring inverts approximately 10 times a
second at room temperature.

47. Represent ring inversion and indicate that the two chair
conformations are in dynamic equilibrium. The symbol is the
usual way of indicating that a dynamic equilibrium exists, in this case,
between the two conformations.

48. Consider methylcyclohexane. Construct the model of

methylcyclohexane with the methyl attached axially. Make the ring
undergo inversion. Complete the following:

44
49. Methylcyclohexane can exist in two chair conformations, one
with the methyl group on an axial position (a-methylcyclohexane)
and the other with the methyl group on an equatorial position (e-
methylcyclohexane).
a) In which chair conformation, a or e, is the methyl group farthest
away from the neighboring hydrogen atoms?
b) In which chair conformation, a or e, does the methyl group have
greater room?
c) Which is the preferred chair conformation of methylcyclohexane, a
or e?

50. Make a general statement about the preferred conformation of

monosubstituted cyclohexane.

45
 Exercise #5
Stereochemistry

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

A. Chirality, Enantiomerism, Diastereomerism, and Optical Activity

1. a. _______________________________
b. _______________________________

2. a. _______________________________
b. _______________________________

3. a. _______________________________
b. _______________________________
c. _______________________________

4. a. _______________________________
b. _______________________________
c. _______________________________

5. a. _______________________________
b. _______________________________
c. _______________________________

6. ________________________________________________________
________________________________________________________

7. ________________________________________________________
________________________________________________________

46
8. ________________________________________________________
________________________________________________________

9. a. _______________________________
b. _______________________________
c. _______________________________

10. _______________________________

11. a. _______________________________
b. _______________________________
c. _______________________________

12. _______________________________

13. a. _______________________________
b. _______________________________
c. _______________________________
d. _______________________________

14. _______________________________

15. ---

16. Model I ________________________________

Model II _______________________________
Model III _______________________________
Model IV _______________________________

17. _______________________________

18. _______________________________

47
19. _______________________________

20. ______________________________________________________
________________________________________________________
21. a. ______________________________
b.

22. a. ______________________________
b.

48
 c.

23. a. b.

c. _____________________________
d. _____________________________

24. a. I and II: ___________________________

b. III and IV: _________________________
c. I and IV: ___________________________
d. II and IV: __________________________

25. a. I and III: ___________________________

b. II and III: __________________________

26. Tetrose
a. Flying wedge representations of tetrose, labeled as I, II, etc.

49
 b. ______________________________________________________
c. ______________________________________________________

B. Conformational Isomers
27. a. ______________________________
b. ______________________________
28. ________________________________
29. .

1.2

1
Relative potential energy

0.8

0.6

0.4

0.2

0
0 60 120 180 240 300 360
Dihedral angle, Φ

50
30. .

Newman projection Sawhorse projection

31. .

Newman projection Sawhorse projection

32. _____________________________________

33. Preferred conformation of chloroethane

Newman projection Sawhorse projection

34. _______________________________

35. _______________________________

36. _______________________________

37. _______________________________

51
38. _______________________________

39. _______________________________

40. ______________________________________________________
______________________________________________________

41. Eclipsed conformation: __________

Staggered conformation: ________
Boat conformation: _____________
Chair conformation: ____________

42. _______________________________

43. _______________________________

44. _______________________________

45. _______________________________

46. Final conformation (include marked bonds)

47. .

52
48. .

49. a. ____________________________
b. ____________________________
c. ____________________________

50. ______________________________________________________
______________________________________________________

53
 Exercise #6
Hydrocarbons

Objectives

1. To develop the ability to detect various signs of chemical change.

2. To observe how hydrocarbon types may be detected and
differentiated from each other by means of simple chemical tests
(test tube reactions).
3. To compare the reactivity of alkanes, alkenes, alkynes and aromatic
hydrocarbons towards selected chemical reagents.
4. To carry out the laboratory preparation of acetylene.

Introduction

Hydrocarbons are compounds containing only hydrogen and carbon

atoms. They are classified as alkanes, alkenes, alkynes or aromatic
hydrocarbons depending on the presence of certain structural features:
alkane (no functional group); alkene (C=C); alkyne (C≡C); and aromatic
(cyclic pi system of electrons).

Alkanes and their cyclic analogs have only two types of bonds in their
molecules: C–C and C–H sigma bonds. Both types of bonds may also be
present in other hydrocarbon types. All carbon atoms in alkanes are sp 3
hybridized. Also, since both C–C and C–H bonds are non-polar, ionic
reactions are unusual for alkanes under ordinary conditions. Instead,
alkanes typically undergo reactions involving free radicals.

An example of a reaction undergone by alkanes is halogenation,

which is a free radical substitution reaction involving C–H bonds. The
reaction of a halogen with an alkane is initiated by heat or light and is
represented by the equation:
UV or high T
R−H+X−X → R−X+H−X

When the halogen used is bromine (Br2), the reaction is accompanied by

loss of the red-orange color of molecular bromine and evolution of a gas,
HBr. These are signs indicating that a free radical substitution reaction has
occurred.

Alkenes and their cyclic analogs contain at least one C=C in their
molecular structures. The double bond consists of a pi bond, which arises

54
from the overlap of the p orbital of each carbon atom (Cp–Cp), and a sigma
bond, which is due to Csp2–Csp2 orbital overlap. Since the double bond is a
site of high electron density (i.e., it is electron rich), alkenes react with
electrophilic reagents and oxidizing agents.

Alkenes react with halogens in a reaction known as electrophilic

addition. This reaction takes place at the double bond and does not require
the presence of heat or light. Free radical addition also occurs but at a
slower rate. The general reaction for electrophilic addition of halogen is:

X–X

X X
alkene alkyl halide

The presence of a C=C may be detected using bromine. As in the

halogenation of alkanes, the loss of the red-orange color of bromine (even
at room temperature or in the dark may indicate the presence of a C=C);
HBr is not evolved.

Another test for the presence of a C=C is the reaction with Baeyer’s
reagent (cold, dilute, neutral aqueous potassium permanganate). The
general reaction is:

KMnO4 + H2O + MnO2 + KOH

OH OH
alkene glycol brown ppt

In a positive test, the purple color of the reagent disappears completely as

the permanganate ion is reduced and a brown precipitate (MnO2) is formed.

Alkyne possess at least one C≡C in their molecular structures. The

triple bond consists of two pi bonds and one sigma bond. Like the C=C, the
C≡C is electron rich. Consequently, alkynes undergo the same type of
reactions as alkenes.

55
Halogenation:
X X

+ 2 X–X

X X
alkyne tetrahalide

Oxidation:

R R + KMnO4 + H2O 2 + MnO2 + KOH

alkyne carboxylic acids brown ppt

Whether the C≡C of an alkyne is found at the end of the carbon chain
(terminal alkyne) or not (internal/nonterminal alkyne) can be determined
by the test with alcoholic or ammoniacal silver nitrate. The test involves
the reaction of the of the acidic terminal H (attached to C≡C) resulting in
an acetylide ion that forms an insoluble precipitate with Ag+ ions.
ethanol
R − C ≡ C − H + AgNO3 → R − C ≡ C− Ag + + HNO3
terminal alkyne silver acetylide

ethanol
R − C ≡ C − H + Ag(NH3 )+
2 → R − C ≡ C− Ag + + NH3 + NH4+
terminal alkyne silver acetylide

Aromatic hydrocarbons are characterized by the presence of a cyclic

pi system of electrons. Although the pi system is also a region of high
electron density, aromatic hydrocarbons do not undergo the same types of
reactions as alkenes or alkynes.

Procedure

The following representative compounds will be used in this exercise:

cyclohexane cyclohexene benzene toluene acetylene

56
A. Investigation of Hydrocarbons
Note down the color, clarity and physical state of the above
representative compounds. Use cyclohexane, cyclohexene, benzene and
toluene for the tests below.

1. Solubility Behavior
a. Place 5 drops of CH2Cl2 in a test tube.
b. Add 3 drops of the sample.
c. Shake to mix.
d. Examine the mixture.
e. Repeat steps a-d using distilled water, 10% NaOH and then
concentrated H2SO4 as solvent.

2. Halogenation: Reaction with Bromine (Br2)

BROMINE CAUSES PAINFUL BURNS. IF SPILLED ON THE SKIN, FLUSH
WITH PLENTY OF WATER WITHOUT RUBBING THE SKIN AND TREAT
IMMEDIATELY WITH GLYCERINE.
a. Place 5 drops of the sample in each of two test tubes, one of which
is wrapped in carbon paper.
b. Add 3 drops of the reagent (Br2 in CH2Cl2). Shake to mix.
c. Expose the unwrapped test tube to light for 10 min. Observe the
appearance of the solution.
d. Unwrap the other test tube and compare its contents with that of
the tube exposed to light.

3. Oxidation: Reaction with Baeyer’s Reagent

a. Place 5 drops of Baeyer’s reagent (cold, dilute, neutral aqueous
potassium permanganate) in a test tube.
b. Add 3 drops of the sample. Shake to mix.
c. Observe. Take note of any color change. (Partial reduction of
KMnO4 can be due to impurities. Therefore, if the purple color of
the reagent is not completely destroyed, the test is to be
considered negative.)

4. Reaction with ammoniacal AgNO3

Perform this test on cyclohexane and cyclohexene.
a. Place 5 drops of the reagent in a test tube.
b. Add 3 drops of the sample. Shake to mix.
c. Observe.

57
 ANY SOLID PRECIPITATE FORMED IN THIS TEST MAY BE EXPLOSIVE
WHEN DRY. TRANSFER ANY PRECIPITATE INTO A DESIGNATED
VESSEL CONTAINING HNO3.

B. Preparation and Testing of Acetylene Gas

This part must be performed with the assistance of the laboratory
instructor.

1. Introduce about 5 grams of calcium carbide (CaC2) into a dry filter

flask.
2. Assemble the setup with the help of the laboratory instructor.
3. Add 100 mL of distilled water to the separatory funnel.
4. Allow the water to fall into the solid dropwise in order to generate
acetylene gas. Observe.
5. Perform tests 2 – 4 (Part A) on the acetylene gas by bubbling
acetylene into each reagent.
DISCARD THE CONTENTS OF THE FILTER FLASK INTO THE
DESIGNATED CONTAINER.

58
 Exercise #6
Hydrocarbons

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 6.1 Sample description.

Sample Description
(i.e., color, clarity, physical state, odor)
Cyclohexane

Cyclohexene

Acetylene

Benzene

Toluene

Table 6.2 Solubility tests.

Sample Observations
CH2Cl2 +/- Water +/- NaOH +/- H2SO4 +/-
Cyclohexane

Cyclohexene

Benzene

59
Toluene

Give a generalization on the properties of each class of hydrocarbons based

on the result of the solubility tests (i.e., polarity, acidity/basicity, etc.)
Alkanes:
___________________________________________________________
Alkenes:
___________________________________________________________
Alkynes:
___________________________________________________________
Alkylbenzenes:
___________________________________________________________
Aromatic hydrocarbons:
___________________________________________________________
Table 6.3 Halogenation: Bromination with and without light.
Description of reagent (Br2 in CH2Cl2):
___________________________________________________________

Sample Observations
Final appearance of +/- Final appearance of +/-
mixture exposed to mixture not exposed
light to light
Cyclohexane

Cyclohexene

Acetylene

Benzene

Toluene

Which class/classes of hydrocarbons is/are reactive to Br2 in CH2Cl2 only in

the presence of light?
___________________________________________________________

60
Which class/classes of hydrocarbons is/are reactive to Br2 in CH2Cl2 with or
without light?
___________________________________________________________
Which class/classes of hydrocarbons is/are reactive to Br2 in CH2Cl2 both in
the presence and in the absence of light?
___________________________________________________________

Table 6.4 Oxidation: Reaction with Baeyer’s reagent (cold, dilute, neutral,
aqueous KMnO4).
Description of Baeyer’s reagent:
___________________________________________________________

Sample Observations +/-

Cyclohexane

Cyclohexene

Acetylene

Benzene

Toluene

Enumerate the classes of hydrocarbons that are oxidizable by Baeyer’s

reagent.
___________________________________________________________
What is the structural feature of these classes of hydrocarbons?
___________________________________________________________

Table 6.5 Reaction with ammoniacal AgNO3.

Description of ammoniacal AgNO3:
___________________________________________________________

Sample Observations +/-

Cyclohexane

Cyclohexene

Acetylene

61
Based on the test result, what is the structural requirement for a substance
to react with the ammoniacal AgNO3?
___________________________________________________________

Table 6.6 Generation of acetylene.

Sample Description
Water

Reactants Calcium carbide

Mixture during reaction

Mixture after reaction

Acetylene gas produced

Chemical reaction involved in the preparation of acetylene:

___________________________________________________________

62
Table 6.7 Summary of Results.
Reagent Alkanes Alkenes Alkynes Benzene Alkylbenzene
Is there a
reaction?
What is the
sign of a
Bromine positive
(in CH2Cl2) reaction?
Is light
necessary?
What is the
mechanism
involved?
Baeyer’s Is there a
reagent reaction?
(cold, dilute, What is the
neutral, sign of a
aqueous positive
KMnO4) reaction?
Is there a
Ammoniacal reaction?
silver nitrate What is the
[Ag(NH3)2+] sign of a
positive
reaction?

63
Answer to Questions

Q1: What is the function of CH2Cl2 in the bromination reactions? Why can
it fulfill this role?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q2: Explain why terminal alkynes are acidic.
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: Explain the difference on the rate of bromination reactions of toluene
and cyclohexane.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: Give the reagent and the reaction conditions that would distinguish
between the following compounds. Write equations for the reactions
involved.
a. benzene and 2-butyne

______________________________________________________
b. 1-butyne and 2-butyne

______________________________________________________
c. 2-methylpentane and 2-methyl-2-pentene

______________________________________________________
d. toluene and 1-methylcyclohexene

______________________________________________________

64
 Exercise #7
Organic Derivatives of Water

Objectives

1. To be acquainted with the chemical properties of the organic

derivatives of water.
2. To observe the differences in chemical reactivity of primary,
secondary and tertiary alcohols, phenols and ethers towards selected
chemical reagents.

Introduction

In addition to carbon and hydrogen, the element oxygen is found in

many important organic compounds. Among the classes of oxygen-
containing compounds are alcohols, phenols and ethers. They are also
known as the organic derivatives of water because they may be seen to
arise from the replacement of one or both of the hydrogen atoms of water
by an organic group. In alcohols, one hydrogen atom is replaced by an alkyl
group; in phenols, it is replaced by an aryl group. In ethers, both hydrogen
atoms are replaced by alkyl or aryl groups.

R Ar R R’
water alcohol phenol ether
R = alkyl Ar = aryl R, R’ = alkyl or aryl

The hydroxyl group (–OH) is the functional group of alcohols and

phenols. The O–H bond is polar with greater electron density around the
oxygen atom. Under the proper conditions, the O–H bond can split
heterolytically to release a proton:
_
:
:

+ Base + H–Base+
:

R
:

At the same time, the oxygen atom acts as a Lewis base because of
its unshared pairs of electrons. Hence, the oxygen atom is a site for attack
by Lewis acids or electrophiles.
E
│+
:

R–O–H + E+ R–O–H
:
:

65
 The nature of the substituent bonded to the hydroxyl group greatly
affects the reactivity of alcohols and phenols. Thus, alcohols are classified
as primary (1°), secondary (2°), or tertiary (3°), based on the type of
carbon atom bearing the hydroxyl group. In phenols, the hydroxyl group is
directly bonded to an aromatic system. The aromatic ring makes the
hydroxyl group more acidic that it is in alcohols.

Ethers are relatively inert molecules. The ether linkage, C–O–C, is

quite stable and can be cleaved only under drastic conditions. The unshared
pairs of electrons on the oxygen atom, however, make ethers susceptible
to electrophilic attack.

Alcohols of the type undergo a unique reaction with

halogens in the presence of a base. This is called the haloform reaction.
The mixture of halogen and base produces a hypohalite which is an
oxidizing agent:

2OH − + X2 → OX − + X− + H2 O
hypohalite

The hypohalite first oxidizes the alcohol to a methyl carbonyl form,

which subsequently undergoes the haloform reaction:
OH O O
│ X2/OH -
║ X2/OH -
│
R–CH–CH3 R–C–CH3 R–C–O- + CHX3

The compound classes considered in this exercise can be further

differentiated through their reaction with oxidizing and reducing agents.
Below is a summary of functional group transformations undergone by
organic derivatives of water upon oxidation:

1° alcohol:
[O] [O]

aldehyde carboxylic acid

66
2° alcohol:
[O]

ketone

phenol:
[O]
mixture of products

[O]
3° alcohols and ether: R3COH / ROR’ no reaction

Potassium permanganate is a powerful oxidizing agent with a

characteristic purple color due to permanganate ion. When it reacts with an
oxidizable compound, the purple color is discharged and a brown to black
precipitate of MnO2 is formed together with hydroxide ions. In this exercise,
a small amount of acid is added to the reagent to neutralize and hydroxide
ions produced for the reaction with oxidizable impurities.

Tollens’ reagent is a mild oxidizing agent which consists of

ammoniacal silver nitrate prepared by dissolving silver nitrate in an excess
of ammonium hydroxide. The oxidizing agent is the silver ion which exists
as a stable complex, Ag(NH3)2+. Upon reaction, the silver ion is reduced to
elemental silver, Ag, which adheres to the clean glass walls of the container
to form a mirror. Easily oxidizable compounds give a positive test with
Tollen’s reagent.

Whether an alcohol is primary, secondary or tertiary is shown by the

Lucas test, which is based on the difference in reactivity of the alcohol
toward hydrogen halides. Alcohols of no more than six carbon atoms are
soluble in the Lucas reagent, a mixture of concentrated hydrochloric acid
and zinc chloride, but the alkyl chlorides formed in this test are insoluble.
Thus, the appearance of cloudliness indicates the formation of an alkyl
chloride from the alcohol. The reactions for 3° and 2° alcohols are shown
below:

ZnCl2
2° alcohol: R2CHOH + HCl R2CHCl + H2O

ZnCl2
3° alcohol: R3COH + HCl R3CCl + H2O

67
Note that tertiary alcohols are most reactive and undergo reaction even in
the absence of the ZnCl2 catalyst.

Procedure

The following representative compounds (or a suitable substitute) will

be used in this exercise:

1-butanol diisopropyl ether

(n-butyl alcohol)

phenol

2-methyl-2-propanol 2-butanol
(tert-butyl alcohol) (sec-butyl alcohol)

CAUTION:

Phenol crystals slowly liquefy in hot and humid air. The yellow color
of the sample is due to phenol oxidation products. The sample is suitable
for the chemical tests, except where anhydrous samples are required.
HANDLE PHENOL WITH CARE AS IT CAN CAUSE PAINFUL BURNS. IF
SPILLED ON THE SKIN WASH FREELY WITH 95% ETHANOL, THEN WITH
SOAP AND WATER.

Diisopropyl ether is highly volatile and flammable. Its vapor has a

high tendency to form explosive hydroxides. Fire is therefore a hazard to
be guarded against. STAY AWAY FROM ANY OPEN FLAME WHEN WORKING
WITH ETHER AND MINIMIZE ITS EXPOSURE TO AIR.

68
A. Solubility Behavior
1. Place 5 drops distilled water in a test tube.
2. Add 3 drops of the sample.
3. Shake vigorously.
4. Examine the mixture.
5. Repeat steps 1 – 5 using 10% NaOH as solvent.
6. For compounds insoluble in base, repeat steps 1 – 5 using
concentrated H2SO4 as solvent.

BE CAREFUL IN HANDLING CONCENTRATED SULFURIC ACID. IT IS

CORROSIVE!

B. Chemical Reactivity
1. Reaction with Potassium Permanganate
a. Place 5 drops dilute, slightly acidic aqueous potassium
permanganate in a test tube.
b. Add 3 drops of the sample. Observe.
c. Warm the tube gently in a water bath for 5 min.
d. Examine the mixture.

2. Reaction with Tollens’ Reagent (ammoniacal silver nitrate)

NOTE: This test should be done using a test tube that is scrupulously
clean and has smooth walls. Otherwise, the silver will not
deposit on the tube to form a mirror.

Perform this test on 1-butanol, 2-butanol, and phenol only.

a. Place 5 drops Tollens’ reagent in a test tube.
b. Add 3 drops of the sample.
c. Shake then heat gently in a water bath for 10 min.
d. Examine the mixture.

3. Lucas Test: Reaction with HCl-ZnCl2

Perform this test on 1-butanol, 2-butanol, and tert-butanol only.
a. Place 5 drops Lucas reagent in a test tube.
b. Add 3 drops of the sample.
c. If no cloudiness is observed, heat the solution in a hot water bath.
d. Allow to stand.
e. Note the time required for the appearance of turbidity, an insoluble
layer or emulsion.

69
4. Reaction with Ferric Chloride (FeCl3)
Perform this test on 1-butanol and phenol only.
a. Place 1 drop of the sample in a test tube.
b. Add 3 drops 2.5% aqueous FeCl3 then add about 1 mL of distilled
water.
c. Examine the mixture.
d. Compare the results with a blank using water as sample.

5. Iodoform Test: Reaction with I2/KI, NaOH

Perform this test on 1-butanol and 2-butanol only.
a. Place 3 drops of the sample in a test tube.
b. Add 5 drops distilled water.
c. Add 3 drops I2/KI solution.
d. Add 10% NaOH dropwise with shaking until iodine color disappears
and the solution is faintly yellow.
e. Examine the contents of the tube and note the odor.
f. If no change is observed, shake the tube then heat gently in a
water bath for 1 – 2 min.
g. Examine the mixture again.

70
 Exercise #7
Organic Derivatives of Water

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 7.1 Sample description.

Sample Description
(i.e., color, clarity, physical state, odor)
1-butanol

2-butanol

tert-butanol

phenol

diisopropyl ether

Table 7.2 Solubility tests.

Sample Observations
Water +/- NaOH +/- H2SO4 +/-
1-butanol

2-butanol

tert-butanol

phenol

71
diisopropyl ether

For the representative alcohols, explain the solubility behavior in water as

a function of branching in the molecular structure.
___________________________________________________________
___________________________________________________________
For phenol and ether, explain the solubility behavior in water as a function
of the relative proportions of hydrophilic bonds and hydrophobic bonds.
___________________________________________________________
___________________________________________________________
Which among the representative compounds is/are soluble in NaOH? Write
the acid-base reaction involved.
___________________________________________________________

Acid-base reaction

Enumerate the classes of organic derivatives of water that are soluble in

concentrated H2SO4. What generalization can be made for a substance to
be soluble in concentrated H2SO4?
___________________________________________________________
___________________________________________________________
___________________________________________________________

Table 7.3 Potassium permanganate test.

Description of reagent (acidic KMnO4):
___________________________________________________________
Sample Observations +/-
Before Heating After Heating
1-butanol

2-butanol

72
tert-butanol

phenol

diisopropyl ether

Table 7.4 Tollens’ test.

Description of reagent (ammoniacal AgNO3):
___________________________________________________________
Sample Observations +/-
1-butanol

2-butanol

phenol

Based on the results of the KMnO4 and Tollens’ tests, categorize the classes
of organic derivatives of water as:
Easily oxidizable: _____________________________________________
Oxidizable: _________________________________________________
Resistant to oxidation: _________________________________________

Table 7.5 Lucas test.

Description of reagent (HCl-ZnCl2):
___________________________________________________________
Sample Observations +/-
1-butanol

2-butanol

tert-butanol

73
Based on the results of the KMnO4 and Lucas tests on the representative
alcohols, give a generalization on how the different types of alcohols can
be differentiated from each other.
Primary alcohol: _____________________________________________
Secondary alcohol: ___________________________________________
Tertiary alcohol: _____________________________________________

Table 7.6 Ferric chloride test.

Description of reagent (aqueous FeCl3):
___________________________________________________________
Sample Observations +/-
1-butanol

phenol

Water

What class of organic compound reacts with FeCl3 solution?

___________________________________________________________

Table 7.7 Iodoform Test.

Description of reagent (I2/KI):
___________________________________________________________
Sample Observations +/-
1-butanol

2-butanol

What is the structural requirement for an alcohol to react with the iodoform
reagent?
___________________________________________________________

74
Answer to Questions

Q1: Explain the solubility behavior of the representative compounds in

water as a function of:
a. Branching in the structure
___________________________________________________________
___________________________________________________________
b. Relative proportion of hydrophilic to hydrophobic bonds
___________________________________________________________
___________________________________________________________
Q2: Explain the acidity differences observed for the compounds used in this
exercise in terms of the stability of the corresponding conjugate bases.
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: Based on the results of the oxidation test, classify the compounds
tested according to the following categories: easily oxidizable, oxidizable,
and resistant to oxidation.
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: Discuss the reactivity differences of the alcohols towards the Lucas
reagent.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q5: Suggest a simple chemical test that will differentiate between the
following pairs of compounds. Write equations for the reactions involved.
a. phenol and isopentyl alcohol _______________________________

b. tert-butyl alcohol and isobutyl alcohol ________________________

75
c. neopentyl alcohol and ether ________________________________

d. sec-butyl alcohol and neopentyl alcohol _______________________

e. propene and 2-butanol ____________________________________

76
 Exercise #8
Carbonyl Compounds and Carbohydrates

Objectives

1. To be acquainted with the chemical properties of carbonyl compounds

and carbohydrates.
2. To observe the differences in the chemical reactivity of aldehydes and
ketones.
3. To perform chemical tests that characterize the different types of
carbohydrates.

Introduction

A. Carbonyl Compounds
An important group of oxygen-containing compounds are the
carbonyl compounds – those that contain the carbonyl group, C=O, in
their molecular structures. They are classified as aldehydes or ketones
depending on what groups are bonded to the C=O group. Aldehydes
have hydrogen atom attached to the C=O group.

Since oxygen is more electronegative than carbon, there is greater

pi electron density at the oxygen end of the C=O bond. The carbonyl
group is therefore polar, with carbon bearing the partial positive charge.

This property of the carbonyl group gives rise to a set of reactions

characteristic of aldehydes and ketones – nucleophilic addition. The ease
of addition of the nucleophile depends on the degree of crowding at the
carbonyl group. Ketones are thus less susceptible to nucleophilic
addition than aldehydes.

77
 The compound 2,4-dinitrophenylhydrazine (2,4-DNP) may add to
the carbonyl group to form solid or oily derivatives called 2,4-
dinitrophenylhydrazones:

This reaction is very useful for the identification of aldehydes and

ketones.

Carbonyl compounds possessing the structure can also

undergo the haloform reaction. The bond between the carbonyl and
methyl group is cleaved to give a carboxylate ion and a haloform:

Aldehydes and ketones can be differentiated from each other by

the Tollens’ test. Aldehydes are oxidized by Tollens’ reagent to yield the
corresponding carboxylic acid salt and a silver mirror.

B. Carbohydrates
Carbohydrates are polyhydroxy aldehydes or ketones and the
derivatives or compounds, which yield such on hydrolysis. Many of them
are represented by the general formula Cx(H2O)x, which led early
workers to regard them as hydrates of carbon, hence the term
“carbohydrate”.

Carbohydrates that cannot be hydrolyzed to simpler units are

called monosaccharides. These may be classified according to the type

78
of carbonyl group present (ketose or aldose) or the number of carbon
atoms in the molecule (triose, tetrose, hexose, etc.). Carbohydrates
may exist as hemiacetals or hemiketals due to an intramolecular
reaction involving the C=O group and one of the –OH groups in the
chain. This reaction is shown below for D-glucose. The atom marked
with an asterisk is the masked carbonyl carbon (hemiacetal carbon).

The most common ketose is a hexose, D-fructose. The cyclization

(hemiketal formation) of D-fructose is shown below:

In the cyclic form, monosaccharides cannot undergo the reactions

typical of the carbonyl group. However, in aqueous solutions, the cyclic
forms exist in equilibrium with the open-chain forms. The presence of
even a small amount of the open-chain form allows the reactions
associated with the carbonyl group to take place. Moreover, in the open-
chain forms, the interaction between the carbonyl group and the

79
adjacent hydroxyl group makes the carbonyl group more susceptible to
oxidation and more reactive to nucleophilic reagents.

Carbohydrates that can be hydrolyzed to two monosaccharides

units are called disaccharides. The monosaccharide units in
disaccharides are joined by a glycosidic linkage (specifically, an acetal
or ketal linkage) which is really an ether linkage. Lactose is the dominant
carbohydrate in milk and is made up of β-D-galactose and α-D-glucose
joined by a β-1,4 glycosidic bond.

hydrolysis

α-lactose β-D-galactose α-D-glucose

Sucrose is common table sugar. It is composed of α-D-glucose and β-

D-fructose.

hydrolysis

sucrose α-D-glucose β-D-fructose

The glycosidic linkage joins the masked carbonyl group

(hemiacetal or hemiketal site) on one cyclic unit to the second. If the
point of attachment of the second unit is not at its carbonyl group, i.e.,
there is a hemiacetal carbon, the cyclic disaccharide can exist in
equilibrium with the free carbonyl form:

hemiacetal C

However, if the potential carbonyl group of the second unit is also

tied up in the glycosidic linkage, no such equilibrium will exist. The
disaccharide will not undergo the reactions that are typical of a free
carbonyl group.

80
 Monosaccharides and disaccharides are sometimes referred to as
“simple sugars” or simply “sugars”.

Polysaccharides are high molecular weight polymeric

carbohydrates made up of many cyclic monosaccharide units. Those
found in nature serve either a structural or nutritional function. Cellulose
is the chief structural material of plants. It is a linear polymer of β-D-
glucose:

hydrolysis

Starch is the storage form of glucose in plants. It is also a polymer of

D-glucose but the type of glycosidic linkage differs from that in cellulose.
Starch is made up of two components: amylose, which has a linear
structure and amylopectin which has a highly branched structure. Amylose
and amylopectin are made up of α-D-glucose units.

hydrolysis

hydrolysis
α-D-glucose

There is a number of reactions used to characterize carbohydrates.

The Molisch reaction is a general test for carbohydrates either in the free
or combined form. In the presence of concentrated sulfuric acid,
glycosidic linkages are hydrolyzed to give monosaccharides.
H+
(C6 H10 O5 )n + nH2 O → nC6 H10 O6

81
 The rate of hydrolysis depends on the solubility of the
carbohydrate in water. The monosaccharides formed are then
dehydrated to furfural (or hydroxymethylfurfural) and other colored
decomposition products.

H2SO4

a hexose hydroxymethylfurfural (HMF)

Carbohydrates that readily react with mild oxidizing agents are
called reducing sugars. The reducing property can be observed by their
reaction with Benedict’s reagent, which consists of copper(II) hydroxide
in aqueous solution complexed with sodium citrate. The Cu2+ complex
ion has a blue color. A positive test is indicated by the reduction of the
Cu2+ ion to Cu+ (Cu2O), which appears as a brick red precipitate. The
carbonyl group of the reducing sugar is converted to carboxylate.

Reducing sugars may also be detected and identified by their

reaction with phenylhydrazine. The reaction involves nucleophilic
addition of phenylhydrazine to the carbonyl group of the reducing sugar
as well as to the α-position. The product is called an osazone. Osazones
are yellow, crystalline solids with well-defined melting points. They are
useful derivatives in the identification of simple sugars.

sugar osazone

Procedure

The following representative substances will be used in this exercise:

Aldehydes: acetaldehyde, benzaldehyde
Ketones: acetone, cyclohexanone

82
 Monosaccharides: glucose, fructose
Disaccharides: lactose, suctose
Polysaccharides: starch, cellulose (cotton fiber)

A. Solubility Behavior
Perform this test on benzaldehyde, acetone, glucose and starch.
1. Place 1 mL distilled water in a test tube.
2. Add 3 drops of liquid or a pinch (spatula tip) of solid sample.
3. Shake to mix. Examine carefully for homogeneity.

B. Chemical Reactivity of Carbonyl Compounds

1. Reaction with 2,4-DNP
Perform using acetaldehyde and acetone as samples.
a. Place 5 drops of 95% ethanol in a test tube.
b. Add 3 drops of the sample.
c. Add 5 drops of the 2,4-DNP reagent.
d. Shake. Allow to stand for 5 min.
e. Examine the mixture.

2. Reaction with Tollens’ Reagent

Perform using acetaldehyde and acetone as samples.
a. Place 5 drops Tollens’ reagent in a test tube.
b. Add 3 drops of the sample.
c. Shake then heat gently in a water bath for 10 min.
d. Examine the mixture.

3. Iodoform Test
Perform using acetone and cyclohexanone as samples.
a. Place 3 drops of the sample in a test tube.
b. Add 5 drops of distilled water.
c. Add 3 drops I2/KI solution.
d. Add 10% NaOH dropwise with shaking until the iodine color
disappears and the solution is faintly yellow.
e. Examine the contents of the tube and note the color.
f. If no change is observed, shake the tube then heat gently in a
water bath for 1 – 2 min.
g. Examine the mixture.

C. Hydrolysis of Di- and Polysaccharides

Perform using sucrose, starch and cellulose as samples.
1. Place 0.25 g of the sample in a test tube.

83
2. Add 2.5 mL distilled water and 0.5 mL 1M HCl.
3. Heat the mixture in boiling water for 30 min. Cool.
4. Neutralize the hydrolysates with 10% NaOH using phenolphthalein as
indicator. (NOTE: Keep the hydrolysates for the Benedict’s test.)

D. Color Reactions of Carbohydrate

1. Molisch Test
Perform using cyclohexanone, glucose, sucrose and starch as
samples.
a. Place 3 drops of 1% aqueous sample in a test tube.
b. Add 5 drops distilled water.
c. Add 3 drops Molisch reagent (10% α-naphthol in ethanol).
d. Shake and then add 5 drops concentrated H2SO4 slowly along the
side of the tube. Do not agitate the contents of the tube.
e. Note the color at the interface of the layers.

2. Benedict’s Test
Perform using glucose, fructose, sucrose, lactose and the
hydrolysates fro Part C as samples.
a. Add 3 drops sample to 10 drops Benedict’s reagent in a test tube.
b. Heat the mixture in a boiling water bath for 10 min.
c. Note the final appearance of the mixture.

3. Osazone Test
Perform using glucose, fructose, sucrose and lactose as samples.
a. Place a pinch (spatula tip) of each of the sample in separate test
tubes.
b. Add 2 mL phenylhydrazine-HCl/NaCH3COO in each tube
simultaneously.
c. Place the tubes together in a boiling water bath.
d. Shake the tubes occasionally.
e. Note the time of immersion and the time of precipitation of each
osazone.
f. Remove the test tubes from the hot water bath after 20 min.
g. Cool to room temperature. Observe.

84
 Exercise #8
Carbonyl Compounds and Carbohydrates

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data and Observations

Table 8.1 Sample description.

Sample Description
(i.e., color, clarity, physical state, odor)
Acetaldehyde

Benzaldehyde

Acetone

Cyclohexanone

Glucose

Fructose

Lactose

Sucrose

Starch

Cellulose

Table 8.2 Solubility behavior of selected samples.

Sample Observations +/-
Benzaldehyde

85
Acetone

Glucose

Starch

Account for the solubility behavior of each of the samples based on their
chemical structures.
Benzaldehyde: _______________________________________________
Acetone: ___________________________________________________
Glucose: ___________________________________________________
Starch: ____________________________________________________

Table 8.3 Reaction with 2,4-DNP.

Description of 2,4-DNP reagent:
___________________________________________________________
Sample Observations +/-
Acetaldehyde

Acetone

2,4-DNP is used to test for ______________________________________

Table 8.4 Reaction with Tollens’ reagent.

Description of Tollens’ reagent:
___________________________________________________________
Sample Observations +/-
Acetaldehyde

Acetone

Tollens’ test detects the presence of ______________________________

86
Table 8.5 Iodoform test.
Description of Iodoform reagent: _________________________________
Sample Observations +/-
Acetone

Cyclohexanone

What is the structural requirement that makes a carbonyl compound react

positively with the Iodoform reagent?
___________________________________________________________

Table 8.6 Hydrolysis of di- and polysaccharides.

Sample Observations Rxn
Upon adding After heating Upon adding No.
HCl NaOH
Sucrose 1

Starch 2

Cellulose 3

Write the equations for the hydrolysis reactions of the compounds tested.
Rxn 1

Rxn 2

87
 Rxn 3

Table 8.7 Molisch test.

Description of Molisch reagent:
___________________________________________________________
Sample Observations +/-
Cyclohexanone

Glucose

Sucrose

Starch

What class/classes of organic compounds react(s) with the Molisch

reagent?
___________________________________________________________

Table 8.8 Reaction with Benedict’s reagent.

Description of Benedict’s reagent:
___________________________________________________________
Sample Observations +/-
Glucose

Fructose

Sucrose

Lactose

88
Sucrose hydrolysate

Starch hydrolysate

Cellulose hydrolysate

Which sugars are reducing? _____________________________________

Which are non-reducing? _______________________________________
What structural feature is responsible for the reducing property of
carbohydrates? ______________________________________________
Account for the results of the Benedict’s test of the hydrolysates.
Sucrose hydrolysate __________________________________________
Starch hydrolysate ___________________________________________
Cellulose hydrolysate _________________________________________

Table 8.9 Osazone test.

Description of phenylhydrazine-HCl:
___________________________________________________________
Sample Time Time Reaction Description of +/-
immersed precipitate time crystals
formed
Glucose

Fructose

Sucrose

Lactose

Answers to Questions

Q1: Give a simple chemical test to differentiate between the following

compounds. Show equations for the reactions.
a. butanone and butanal ____________________________________

89
b. 2-propanol and acetone ___________________________________

c. glucose and butanal ______________________________________

d. sucrose and lactose ______________________________________

e. glucose and 1-pentanol ___________________________________

90
 Exercise #9
Acyl Compounds, Soaps and Detergents

Objectives

1. To observe the general properties of carboxylic acids.

2. To compare the acidity of carboxylic acids and phenols.
3. To verify experimentally the interconversion among the acyl
compounds.
4. To become familiar with the physical and chemical properties of fats
and oils and to understand the chemical basis for these properties.
5. To learn how to prepare soap.
6. To observe and compare the properties of soap and synthetic
detergents.

Introduction

A. Acyl Compounds

Acyl compounds are of the type , where Z is a halogen or a

group bonded to the carbonyl group through oxygen or nitrogen. The
most widely distributed acyl compounds are the carboxylic acids.
Carboxylic acids are the most important of the acidic organic
compounds. They contain the carboxyl group as their functional group:

R = alkyl or aryl

In the carboxyl group, the hydroxyl group is directly attached to

the carbonyl group. This allows the resonance stabilization of the
conjugate base formed upon release of a proton:

Base
+ H–+Base

carboxylic acid carboxylate anion

Thus, carboxylic acids have a greater tendency to release a proton

than other acidic organic compound types.

91
 Generally, carboxylic acid functional derivatives contain the

acyl group, . The following compound types are functional

derivatives of carboxylic acids:

Nitriles, R–C≡N, though not acyl compounds, are considered as

carboxylic acid functional derivatives because they are readily
hydrolyzed to carboxylic acids.

Nucleophilic addition to the carbonyl group is less likely in acyl

compounds than in aldehydes and ketones. The characteristic reaction
of acyl compounds is nucleophilic acyl substitution:

Through this reaction carboxylic acids may be converted to the

functional derivatives and, in turn, the functional derivatives may be
converted to carboxylic acids.

The acyl compounds differ in their reactivity towards nucleophilic

acyl substitution and this is evident from their hydrolysis reactions. The
more reactive compounds are hydrolyzed by water alone. The less
reactive ones either require a strong nucleophile such as the hydroxide
ion or an acid catalyst to enhance the electron deficiency of the carbonyl
carbon, making it more attractive to a nucleophile. Alcohols, phenols,
ammonia, and amines react with acyl compounds in much the same way
as water does.

B. Soaps and Detergents

Esters of long-chain carboxylic acids and the trihydric alcohol,
glycerol, are known as triacylglycerols or triglycerides. The carboxylic
acids attached to glycerol may be identical (simple triglyceride), or
different (mixed triglyceride).

92
 glycerol triacylglycerol

Triacylglycerols undergo the same reactions as the low molecular

mass esters. Hydrolysis in a basic medium, using alkali bases such as
sodium hydroxide or potassium hydroxide, yields long-chain carboxylate
salts and glycerol. The reaction called saponification, is represented by
the equation below. The salts produced are soaps. Soap produced from
triacylglycerols is a mixture of long chain carboxylate salts. Soaps,
having a long nonpolar chain and small polar end, are surface active
chemical species. They are amphiphatic and are used as detergents.

Synthetic detergents, like sodium lauryl sulfate, are also

amphiphatic and are generally characterized by the presence of a
different polar end like a sulfate group. They are usually derived from
C12 to C18 alcohols.

93
Procedure

A. Solubility Behavior
Perform this test on acetic acid, benzoic acid and sodium benzoate.
1. Place 5 drops distilled water in a test tube.
2. Add a small amount of sample (3 drops or a small piece).
3. Shake to mix.
4. Examine the mixture. Test with red and blue litmus paper.

B. Relative Acidity of Carboxylic Acids and Phenols

Perform this test on benzoic acid from part A and phenol.
1. Place 5 drops distilled water in a test tube.
2. Add a small amount of sample (3 drops or a small piece).
3. Add 5 drops of 10% NaOH.
4. Shake and then stand for 5 min. Examine the mixture.
5. Repeat the test using 10% NaHCO3 instead of 10% NaOH.

C. Hydrolysis of Acyl Compounds

Perform this test on acetyl chloride, acetic anhydride, ethyl benzoate
and benzamide.
ACETYL CHLORIDE AND ACETIC ANHYDRIDE MUST BE HANDLED IN THE
FUME HOOD. KEEP THEIR CONTAINERS CLOSED WHEN NOT IN USE.
1. Place 3 drops or a spatula tip of the sample in a test tube.
2. Add 10 drops distilled water.
3. Observe for any sign of reaction (e.g. heat evolution, bubble
formation, etc.).
4. If no change is seen, heat the tube for 3 min without boiling in a hot
water bath.
5. Cool the mixture.
6. Test with red and blue litmus paper.

D. Saponification of Coconut Oil

In this exercise, soap will be prepared using the cold process.
1. Weigh 22-23 g coconut oil in a 250 mL beaker. Warm this in a water
bath to about 50 – 60°C. Remove the beaker when ready to saponify.
2. Weigh 0.50 g freshly prepared 30% NaOH solution for each gram of
coconut oil.
3. While stirring, add about 1 mL of the NaOH solution to the warm
coconut oil. Continue mixing until a cloudy mixture is observed.

94
4. Add the remaining NaOH solution gradually while stirring. When the
mixture temperature drops, warm to about 45°C by dipping the
beaker in a warm water bath.
5. Continue mixing until the mixture thickens to a point where there is
no distinct oil and water layers. Stop mixing when the mixture
becomes a soft gel. Take out the stirrer. If the shape of a portion
from the stirrer is retained, the soap is ready to be poured into molds.
6. Pour the soap into 4 or 5 molds (one for each member of the group
and one for the laboratory instructor). Optional: At this stage, very
small amounts of additives like essential oil or colorants may be
added before pouring the mixture into the molds.
7. Set aside. Allow the saponification to continue until the next meeting.

E. Comparison of Soaps and Synthetic Detergents

Dissolve a small chip of soap sample in 5 mL of warm distilled
water. Do the same with a sample of synthetic detergent. The mixtures
should be cloudy and should form suds when mixed. Use the mixtures
for the following tests.

1. Hydrolysis
Test each mixture with red and blue litmus paper.

2. Reaction with Acid

a. Place 10 drops of the sample in a test tube.
b. Add 5 drops 0.05N HCl gradually.
c. Shake to mix. Observe.

3. Reaction with “Soft” and “Hard” Water

a. Place 10 drops 0.02M NaCl (soft water) in a test tube.
b. Add 5 drops of the sample.
c. Repeat with 0.02M CaCl2 (hard water).

4. Emulsifying Action
On kerosene
a. Place 3 drops of kerosene in a test tube.
b. Add 10 drops of the sample.
c. Shake the tube vigorously for 1 min.
d. Allow to stand for 5 min.
e. Run a blank consisting of kerosene and water.

95
On oil
a. Put a layer of an oily material (oil from your face, coconut oil,
etc.) on your finger.
b. Mix with water. Observe.
c. Repeat with soap and synthetic detergent solution instead of
water. Observe.

96
 Exercise #9
Acyl Compounds, Soaps and Detergents

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 9.1 Sample descriptions.

Sample Description
Acetic acid

Benzoic acid

Acetic anhydride

Benzamide

Sodium benzoate

Acetyl chloride

Ethyl benzoate

Table 9.2 Solubility of carboxylic acids in water.

Sample Observations +/-
Acetic acid

Benzoic acid

Sodium benzoate

97
Account for the solubility behavior of each of the samples based on their
chemical structures.
Acetic acid:
___________________________________________________________
Benzoic acid:
___________________________________________________________
Sodium benzoate:
___________________________________________________________

Table 9.3 Relative acidity of carboxylic acids and phenols.

Sample Observations
Distilled H2O +/- 10% NaOH +/- 10% NaHCO3 +/-
Benzoic
acid

Phenol

Based on the results above, which is more acidic, carboxylic acids or

phenols? How did you arrive at this conclusion?
___________________________________________________________
___________________________________________________________
___________________________________________________________
Arrange the following compound types in order of increasing acidity:
alcohols, phenols, carboxylic acids, and water.
___________________________________________________________

Table 9.4 Hydrolysis of acyl compounds.

Sample Observations Litmus Paper Test
Sample +H2O On heating On cooling Red Blue Inference
Acetyl
chloride

Acetic
anhydride

98
Ethyl
benzoate

Benzamide

Which among the acyl compounds are easily hydrolyzed (do not require
heating)?
___________________________________________________________
Which are not easily hydrolyzed (require heating)?
___________________________________________________________

Table 9.5 Saponification of coconut oil.

Sample Observations
Oil in beaker +
30% NaOH

Warmed mixture
(while mixing)

Mixture after
mixing

Soap after one

week

Write the chemical equation for the saponification of coconut oil.

99
Table 9.6 Comparison of soaps and synthetic detergents.
Test Observations
Soap Synthetic detergent
Hydrolysis

Reaction with acid

Reaction with soft

water

Reaction with hard

water

Emulsifying action

Explain the cleaning property of soaps and synthetic detergents based on

your observation of their emulsifying action.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Between soap and synthetic detergent, which is a better cleaning agent in
hard water? Explain.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________

Answer to Questions

Q1: Explain the difference in the solubility of benzoic acid and sodium
benzoate in water. Which of the two would you predict to be more soluble
in CHCl3? Explain.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________

100
Q2: Based on the results in Part C, arrange the following compound types
in order of decreasing ease of hydrolysis: acid halides, acid anhydrides,
esters and amides. Give the theoretical explanation for the observed
differences in reactivity.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: On the basis of the litmus paper tests, are the soap-water and
detergent-water mixtures acidic, basic or neutral? Explain the difference, if
any, in the reactions of the soap-water and synthetic detergent-water
mixtures to litmus paper.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________

101
 Exercise #10
Synthesis of Aspirin
(Acetylsalicylic Acid from Salicylic Acid)

Objectives

1. To explain the concept of organic sysnthesis.

2. To synthesize the acetylsalicylic acid from salicylic acid by nuclephilic
acyl substitution.
3. To describe and explain differences in the properties of acetylsalicylic
acid and salicylic acid by simple chemical tests.

Introduction

A very important aspect of organic chemistry is the synthesis of a

compound from available ones in the laboratory. This aspect of organic
chemistry has contributed much to the knowledge there is regarding
reactions undergone by specific families of compounds. There are
reactions that appear to be happening neatly from a theoretical
standpoint. In the actual preparation, however, side reactions are
observed.

The goal in any synthesis is to obtain a pure sample of the desired

product by the most efficient and convenient procedure possible. For this
reason, one usually strives to use reactions that can reliably be expected
to give only a single product and avoid reactions that will give a mixture
of products. It is also important to plan a synthesis that entails the
fewest possible steps. This is necessary both in terms of the amount of
time and resources consumed in an overly long route and in the ultimate
yield that may be realized.

By definition, organic synthesis is the preparation of a desired

organic compound from commercially available materials. It is an
important element of the organic chemical industry. A scientist who
wishes to study the physical, chemical or physiological properties of a
compound obviously must have a sample of the specific compound.
Since some organic compounds may not be commercially available from
chemical suppliers, or may be too expensive, the chemist must often
synthesize the desired material.

102
 Acetylsalicylic acid is one of the most popular analgesics in the
market. Felix Hofmann, a chemist for the Bayer Company in Germany,
found a practical way to synthesize acetylsalicylic acid from salicylic acid
in 1897. In 1899, Bayer Co. started to market the compound under the
name Aspirin.

acetylsalicylic acid

In the stomach, acetylsalicylic acid is hydrolyzed back to salicylic

acid. First, it was thought that salicylic acid was the medically active
form of the analgesic. But today, it is well known that acetylsalicylic acid
is more potent than salicylic acid as an analgesic. The mechanism of
action involves a complex biochemical reaction. Aspirin blocks the
production of prostaglandins which, in excess amounts, trigger pain,
fever and inflammation.

Aspirin also hinders blood clotting. This is actually a negative

feature of the compound but it is now considered beneficial since it can
reduce the occurrence of both heart attacks and strokes.

Procedure

A. Preparation of Acetylsalicylic Acid (Aspirin)

SALICYLIC ACID IS MILDLY IRRITATING. ACETIC ANHYDRIDE AND
PHOSPHORIC ACID ARE CORROSIVE AND TOXIC. AVOID SKIN CONTACT
AND INHALATION OF FUMES.

1. Place 1 g salicylic acid in a 125 mL Erlenmeyer flask.

2. Add carefully 3 mL acetic anhydride and 5 drops 85% phosphoric
acid.
3. Swirl to mix and heat the flask in a steam bath for 15 min.
4. Without cooling, slowly add 2 mL distilled water.
5. When the vigorous reaction has ended, add 20 mL ice-cold water.
6. Cool to room temperature and then place in an ice bath to complete
the crystallization.
7. Collect the product by suction filtration.
8. Wash the crystals several times with small portions of cold water.

103
9. Transfer the crystals to a pre-weighed watch glass and air dry.
10. Weigh the dried aspirin and calculate the % yield of crude
product.

B. Recrystallization of Aspirin
1. Set aside a small quantity of crude aspirin for melting point
determination.
2. Transfer the rest of the crude aspirin into a 125 mL Erlenmenyer
flask. Add 95% ethanol dropwise while swirling the flask until almost
all the crude aspirin dissolves.
3. Add cold distilled water dropwise until almost all crystals appear.
Complete the recrystallization by cooling the flask in a cool bath.
4. Collect the crystals by suction filtration.
5. Wash the crystals several times with small portions of cold water.
6. Transfer the crystals to a pre-weighed watch glass and air dry.
7. Weigh the dry aspirin and calculate the % recovery.
8. Transfer the aspirin to a clean and dry vial and label properly.
9. Determine the melting points of the crude and recrystallized aspirin
and compare. Account for the difference.

C. Characterization of Aspirin
1. Propose a method of differentiating your product from the starting
material. Check with your instructor before performing the test.
2. Differentiate the synthesized acetylsalicylic acid from commercially
available aspirin by dissolving a pinch of aspirin in 5 drops of water
and 5 drops of iodine solution. Observe the reaction.

104
 Exercise #10
Synthesis of Aspirin
(Acetylsalicylic Acid from Salicylic Acid)

Name ____________________________ Date Performed _____________

Lab Section ___________ Date Finished ______________
Group No. _________
Score _________

Data

Table 10.1 Description of reagents used.

Sample Description
Salicylic acid

Acetic anhydride

Table 10.2 Observations on the synthesis of aspirin.

Sample / Actions Taken Observations
Salicylic acid + acetic
anhydride+H3PO4
Heating in steam bath

After adding distilled water

After adding ice-cold water

Product

105
Table 10.3 Calculations of % yield and % recovery.
Synthesis of Aspirin

Mass of watch glass, g

Mass of watch glass + product, g

Mass of crude product, g

Theoretical mass of product, g

% yield

Recrystallization of Aspirin

Mass of watch glass, g

Mass of watch glass + product, g

Mass of recrystallized product, g

% recovery

Melting Point of Crude Aspirin: ___________________________________

Melting Point of Recrystallized Aspirin: ____________________________

Draw the schematic diagram of your proposed method in differentiating

your product and starting material. Write the reactions involved.

106
Table 10.4 Difference between product and commercially available aspirin.
Sample Observations
(+ 5 drops of water and 5 drops iodine solution)
Product

Commercially available

Answer to Questions

Q1: Write the mechanism for the reaction involved in the synthesis of
acetylsalicylic acid from salicylic acid.

Q2: What is the role of phosphoric acid in the synthesis of acetylsalicylic

acid using acetic anhydride? Is it necessary to use phosphoric acid when
acetyl chloride is used as electrophile?
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q3: Explain how the following can favor the synthesis of aspirin:
a. Heating the mixture
___________________________________________________________
___________________________________________________________
___________________________________________________________

107
 b. Addition of water after heating
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q4: Aspirin undergoes the typical reactions of esters. With this in mind, can
aspirin be recrystallized using hot water? Explain.
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q5: Account for the difference in the reaction of the synthesized and
commercial aspirin with iodine.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________
Q6: Based on the results of the chemical tests, was aspirin synthesized in
the exercise? Explain.
___________________________________________________________
___________________________________________________________
___________________________________________________________
___________________________________________________________

108