You are on page 1of 44

CHEM231 Lab Manual 2013/14

Organic Chemistry -1 (Practical)


CHEM 231

________________________________________________________________________

Laboratory Manual 2013/2014

Pepared by

Dr. Khaid Shadid & Prof. Dr. Abdulfatah Haikal

Islamic University in Madinah

1
CHEM231 Lab Manual 2013/14

Melting Point Determination


Identity and Purity of Solid Organic Compounds

Objectives

To introduce the technique of melting point determination.


To use the concept of melting points for identification and characterization of organic
compounds.
Properly fill and use a capillary melting point tube.
Determine accurate melting point ranges for a wide variety of organic substances.

Introduction
The melting point of a solid can easily and accurately be determined using only a small
amount of material. In combination with other measurements, melting point information can
provide rapid confirmation of the identity of unknown substances.
The method of capillary melting point determination involves placing a small amount of
sample in the bottom of a narrow capillary tube that has been closed at one end. The melting
point is then determined using a melting point apparatus (as in Figure) that simultaneously
heats both the sample tube and a thermometer. The temperature range over which the
substance melts is recorded.

Melting is said to begin when the first indication of liquid is seen. The end of the melting
point range is the temperature at which all of the solid material has become a liquid.
Some pure materials possess a very narrow melting range, perhaps as little as 0.5-1.0 C,
while more typically a 2-3 C range will be observed. You will usually see data recorded as, for
example, mp 232-234 C. Though formally denoting the melting range, this piece of data is
almost universally referred to as the melting point (mp).

2
CHEM231 Lab Manual 2013/14

For the most accurate results, the rate of heating should be kept relatively low, especially for low-
melting samples, to ensure that the thermometer reading represents as accurately as possible
the true temperature experienced by the sample tube (since the transfer of heat within the
apparatus is relatively slow). With this fact in mind, it is sensible when recording a melting
point of an unknown material to perform a trial run where the temperature range is increased
relatively rapidly in order to ascertain a rough melting range. The determination is then
repeated by heating rapidly to within around twenty degrees of the expected melting point and
then very carefully increasing the temperature the remaining few degrees until the melting
point is reached.

Apparatus for manual melting point determination.


"Mel-temp" device (left) and hot oil method (right).

Melting Points as Criteria of Purity. Thermodynamics tells us that the freezing point of a pure
material falls as the amount of an impurity is increased. The presence of an impurity in a
sample will both lower the observed melting point and cause melting to occur over a broader
range of temperature. Generally, a melting temperature range of 0.5-1.0 C is indicative of a
relatively high level of purity. It follows that for a material whose identity is known, an estimate
of the degree of purity can be made by comparing melting characteristics with those of a pure
sample.

3
CHEM231 Lab Manual 2013/14

Melting Points of Urea and Succinic Acid


1. Determine the melting point range of urea, then the melting point range of succinic acid .
2. The capillary melting point tubes should be filled by crushing the sample to a fine
powder on a watch glass with the end of a glass rod and then introducing this powder to
the tube by pressing the open end into the powdered sample.
3. Once a 4-5 mm depth of powder has been introduced, the tube should be turned over.
Tap the sealed end of the tube on a hard surface to compact all of the material at the
bottom.
4. Assemble the melting point apparatus as in the figure above.
5. Record the melting point range of both urea and cinnamic acid.

Compound Melting Range, C

Acetanilide 113.5-114

Salicylic Acid 158.5-159

Sulfanilamide 165-166

Benzoic Acid 121.5-122

p-Terphenyl 210-211

4
CHEM231 Lab Manual 2013/14

Melting Point Determination


Report Sheet Name:----------------------

Melting Points of Urea and Succinic acid

Melting Range Melting Range


Trial 1 Trial 2 (if needed)

Succinic acid ---------------- ---------------

Urea ---------------- ---------------

Note

See How to fill the melting point capillaries

1. Pack the capillary tube by pressing the open end gently into a sample of compound to be analyzed.
Crystals will stick in the open end of the tube.
The solid should fill the tube to a depth of 2-3 mm. Tap the bottom of the capillary on a hard surface so that the
crystals pack down into the bottom

5
CHEM231 Lab Manual 2013/14

Recrystallization

A purification Technique for Solids


Objectives

1. To understand the concept behind recrystallization and its usefulness in organic synthesis.
2. Learn how to choose the correct solvent for recrystallization.

Introduction

Crystallization may be defined as the process in which a solid compound precipitates from a
saturated solution in the form of crystals. Saturation is usually effected through cooling or
evaporation. - Purification by recrystallization depends on the following facts:

1. Different solids have different solubilities in a given solvent.


2. Most solids are more soluble in hot than in cold solvents.

a crystallized compound

When the impure solid is dissolved in a minimum volume of a suitable hot solvent and the
resulting solution is gradually cooled, saturation and eventual crystallization of the pure
compound occurs.

Impurities in a solid are of two kinds: soluble and insoluble and recrystallization involves the
removal of both to purify a solid. Insoluble impurities are first removed by gravity filtration of
the hot solution while the soluble impurities remain dissolved in the cold saturated solution
(mother liquor) after precipitation of the desired compound. The pure crystals are separated from
the supernatant liquid by suction filtration.

6
CHEM231 Lab Manual 2013/14

Generalized Experimental Procedure:

1. Recrystallization involves the following sequence of steps:


2. Selection of a suitable solvent.
3. Preparation of the solution.
4. Hot Filtration (Gravity Filtration).
5. Cooling.
6. Collecting and Drying of Crystals.

Each step will now be discussed more fully.

1. Selection of a suitable solvent

A suitable solvent for recrystallization should possess the following important properties:

1. Dissolve a large amount of the solid to be purified at high temperature.


2. Dissolve impurities readily at low temperatures (= soluble impurities) or not at all even at the
boiling point (= insoluble impurities).

Experimentally: the suitable solvent is determined through solubility tests.

This is done by shaking about 0.1 g of the powdered solid with ~2 mL of the given solvent in a
dry test tube. If the entire solid has nearly dissolved in the cold solvent, the solvent considered
unsuitable. If not, the mixture is heated gently to the boiling point with stirring. If most of the
solid did not dissolve, the solvent is also unsuitable.

If a substance is found to be too soluble in one solvent and insoluble in another, then a mixture
of both solvents (solvent pair) may be used. In such cases, the solvents must be completely
miscible. The compound to be recrystallized is dissolved in the solvent in which it is very
soluble and then the other solvent is added gradually at the boiling point until a slight turbidity
occurs. The solution is then allowed to stand at room temperature to effect slow crystallization
before chilling in ice.

7
CHEM231 Lab Manual 2013/14

Table: Common solvents for Recrystallization

Solvent b.p., C Particulars of Solvents

Water 100 To be used whenever suitable

Methanol 65 Flammable, toxic

Ethanol 78 Flammable

Acetone 56 Flammable

Chloroform 61 Non-flammable, vapor toxic

Cyclohexane 81 Flammable

Ethyl acetate 78 Flammable

2. Preparation of the solution

To prepare the hot solution, the finely divided solid is placed in an Erlenmeyer (conical) flask
and the selected solvent is added in small portions. The mixture is stirred and heated to boiling
after each addition, until the solid dissolves completely. A slight excess of the solvent is usually
added to compensate for any losses (through evaporation) during hot filtration.

NOTE: Decolorizing charcoal may added at this stage if the solution is colored due to colored
impurities.

The flask should be removed from the heat source before adding charcoal to it, otherwise
bumping will occur.

8
CHEM231 Lab Manual 2013/14

3. Hot Filtration (Gravity Filtration)

Filtration of the hot solution is necessary to remove insoluble impurities. Upon addition to the
funnel, the solution will cool rapidly and this can cause unwanted precipitation. This can be
minimized, or avoided, by using a fluted filter paper and a stemless funnel placed on top of a
beaker on a hotplate containing a few milliliters of the recrystallization solvent.

4. Cooling

Cooling the filtered solution will allow crystals to form. The rate of cooling plays a role in
determining the size of the crystals that form: fast cooling will tend to generate more crystals of
small size, slow cooling can allow largest crystals to form. The best compromise of speed,
convenience, and crystal quality, is simply to let the solution cool to room temperature on the
lab bench. To ensure maximum recovery of material, the solution should be cooled in an ice-
water bath after the solution has reached room temperature.

NOTE: Scratching of the inner surface of the glass can help in crystal formation.

9
CHEM231 Lab Manual 2013/14

5. Collecting and Drying of Crystals

When the crystallization is complete, the crystals need to be collected by suction filtration using
a Buchner funnel to ensure rapid and complete removal of the solvent. Transfer the crystalline
material as a suspension to the filter, being careful to never fill the funnel over half full. Most of
the time, it will be difficult to completely transfer all of the crystals to the funnel, quantitatively,
so it will be necessary to add a small amount of ice-cold solvent to the flask to help facilitate the
transfer of all of the crystals. The crystals are then washed with a little more ice-cold solvent to
remove any final impurities that may remain on the surface of the crystals. This solvent should
be as cold as possible to keep the crystals from redissolving.

The crystals are finally dried in an oven or allowed to air-dry, in case the melting point is low,
by spreading them over a sheet of paper.

10
CHEM231 Lab Manual 2013/14

Experimental

Selection of Solvent

Perform solubility tests on anthracene, salicylic acid, benzoic acid, and sodium benzoate in
water, alcohol, and ligroin as follows:

With a spatula take about 0.1 g of the powdered solid and place in a dry test tube. Start by
dissolving it in about 2 mL of solvent with stirring. If insoluble, heat the mixture to boiling (in
water bath) and observe the solubility. The results should allow the selection of a suitable
solvent for each compound.

Recrystallization of an impure unknown

Weigh about 1 g of the impure unknown and recrystallize it from the solvent you have
selected.(By making a solubility tests on pure unknown to choose the suitable solvent).

Make sure you use the minimum volume of solvent; otherwise the amount of recovered
product will be small. Determine the weight and melting point of the purified compound.

11
CHEM231 Lab Manual 2013/14

Recrystallization

Report Sheet
Name:---------------------

Selection of Recrystallization Solvent

Solubility
Suitable
Compound Water Alcohol Ligroin
Solvent
Cold Hot Cold Hot Cold Hot

Sodium benzoate

Salicylic acid

Anthracene

Benzoic Acid

12
CHEM231 Lab Manual 2013/14

Procedure for Crystallization

The photos below illustrate the process of crystallization.

Heat some solvent to boiling. Place Pour a small amount of the hot Swirl the flask to
solvent into the flask containing the dissolve the solid.
the solid to be recrystallized in an
solid.
Erlenmeyer flask.

Place the flask on the steam bath to If the solid is still not dissolved, add When the solid is all in
keep the solution warm. a tiny amount more solvent and solution, set it on the bench
swirl again. top. Do not disturb it!

13
CHEM231 Lab Manual 2013/14

After a while, crystals should You can now place the flask in an ice
appear in the flask. bath to finish the crystallization process.

You are now ready to filter the solution to isolate the crystals. Please see the section on vacuum
filtration. After your crystals are filtered from the solution, put them on a watch glass as shown below.

Here is the filter paper with crystals Carefully scrape the crystals onto
on it. the watch glass.

Let the crystal finish drying on the


watch glass.

14
CHEM231 Lab Manual 2013/14

Boiling Point and Distillation

Identify and Purify of Liquid Organic Compounds


Distillation as a method for the Separation of Liquids

Objectives

1. Distillating a pure liquid (acetone) and determining its boiling point.


2. Separating a mixture of acetone and water by simple distillation.
3. Separating a mixture of acetone and water by fractional distillation.
4. Compare the efficiency of each type of distillation.

Introduction

The boiling point of a pure organic liquid is a physical property of that liquid. It is
defined as the temperature at which the total vapor pressure of the liquid is equal to the
external (atmospheric) pressure.
Boiling points can be determined using the technique of simple distillation.
Simple Distillation is the condensation of vapors from a boiling liquid and collection of
the condensed vapors in a receiving vessel.
Fractional Distillation is equivalent to several repeated simple distillations.
Distillation is a technique that is used to:
- obtain a boiling point of a pure liquid.
- purify a mixture of liquids based on boiling point differences among compounds.

15
CHEM231 Lab Manual 2013/14

Background

Boiling Points of Pure Liquids


The boiling point of a pure liquid depends on the following variables:

1- Nature of intermolecular attractive forces: H-bonding, dipole-dipole, or London forces.


2- Molar masses: boiling point increases with molar mass.
3- Shape of molecules: among isomers having the same functional group, straight chain molecules
have higher boiling points than corresponding branched ones (less molecular surface area).

Boiling Points of Solutions


The effect of any solute, A, on the boiling point of a liquid B, depends on the nature of A:
If A is less volatile than B:
the total vapor pressure of the solution is lower at any given temperature.

the boiling point of the solution is higher than that of pure B.

Example: a solution of sugar in water.

If solute A is more volatile than B:


the total vapor pressure of the solution is higher at any given temperature.

the boiling point of the solution is lower than that of pure B.

Example: a solution of acetone and water.

Simple and Fractional Distillation


- In practice, separation of a liquid mixture into its components by a single distillation
(simple distillation) is possible when the boiling points of the components are 80 degrees or
more apart.
- For mixtures of liquids having boiling points much less than 80 degrees apart, separation
can be achieved only by fractional distillation. The basic idea behind fractional
distillation is the same as simple distillation only the process is repeated many times. It
uses a fractionating column which provides a large surface area for continuous heat
exchange between the hot ascending vapor and the cooler descending liquid, thus
resulting in a series of evaporations and condensations leading to separation of the two
components.

16
CHEM231 Lab Manual 2013/14

The Apparatus

A typical set-up for simple distillation is given in Figure 2 below.

Simple Distillation Apparatus

1: Heat source, 2: Round bottom flask, 3: Still head, 4: Thermometer, 5: Condenser, 6: Cooling water in

7: Cooling water out , 8: Distillate/receiving flask, 9: Vacuum/gas inlet, 10: Still receiver, 11: Heat control

12: Stirrer speed control , 13: Stirrer/hot plate, 14: Heating (Oil/sand) bath, 15: Stirrer bar/anti-bumping
granules , 16: Cooling bath.

Observe the following practical points:


1- The boiling flask should not be more than half full.
2- Boiling stones are added to the liquid to prevent bumping.
3- Each ground joint should be greased to ensure a completely sealed system.
4- Cooling water in the condenser should enter at the lower end and exit at the upper end. This
ensures that the condenser jacket is always full of water.
5- The bulb of the thermometer should be below the opening of the side arm so as to measure the
temperature at which liquid and vapor are in equilibrium.

17
CHEM231 Lab Manual 2013/14

The photo below is for a fractional distillation set-up. The only difference between this set-up and that of
a simple distillation set-up is the inclusion of a fractionating column (Figure 4) between the round
bottom flask and the Y-adaptor (still head).

Fractional Distillation Apparatus Fractionating Columns

Experimental
Determination of Boiling Point (bp) of Pure Acetone
Arrange a simple distillation apparatus. Introduce about 20 mL of acetone and a few boiling
stones in a 50 mL round-bottomed flask. Heat gently so that the distillate collects in the receiver
drop by drop. Make sure that there is a drop of liquid hanging from the bulb of the thermometer
to ensure that the thermometer is reading the correct bp. Absence of this drop indicates
superheating. Wait until 1-2 mL of the distillate have been collected before recording the
temperature. Continue the distillation until about 2 mL of residue are left in the distillation flask,
and record the temperature again. Keep the acetone for the following part.

Separation of a Mixture
(Simple Distillation)

Make a mixture of the two liquids (acetone-water) 20 mL each and pour it into a 100 mL round
bottomed flask. Carry out a simple distillation as before and collect five fractions in the following
boiling ranges: 50-62, 62-72, 72-82, 82-95 plus the fifth fraction which is the residue. Measure the
volume of each fraction and record the results.

18
CHEM231 Lab Manual 2013/14

(Fractional Distillation)

Combine the five fractions and pour into a 100 ml round-bottomed flask, attach the fractionating
column and proceed as for simple distillation. Measure the volume of each fraction as before and
record your results.

19
CHEM231 Lab Manual 2013/14

Boiling Point and Distillation

Report Sheet Name:----------------------

Simple Distillation of Pure Acetone

Boiling point of pure acetone: ---------

Separation of a Mixture of Acetone and Water

1.

Fraction Boiling Range, C Volume of Distillate, mL Composition


Simple Fractional
I 50-62
II 62-72
III 72-82
IV 82-95
V residue

2. Plot the boiling point versus the volume of distillate for the acetone-water mixtures using
simple and fractional distillation.

20
CHEM231 Lab Manual 2013/14

Chromatography
A Separation and Purification Technique
Introduction

Chromatography is a technique that may be used:

1. to separate the components of a mixture as well as


2. to identify organic substances and
3. examine their purity.

Chromatography encompasses several techniques such as column, thin-layer, paper, gas


liquid, etc.

Two principles are basically involved in Chromatography: adsorption (as in thin-


layer Chromatography) and partition (as in paper Chromatography), and certain terms are
common to both types of Chromatography.

In adsorption Chromatography, separation depends on the selective desorption of the


components of a mixture by the eluent (mobile phase) from the surface of a solid
adsorbent (stationary phase). The adsorbent may be packed in a column (Column
Chromatography) or spread as a thin layer on a glass plate as in thin-layer
Chromatography.

In partition Chromatography, separation depends on partition of the components of a


mixture between the stationary and mobile phases. The mobile phase may be a
liquid (liquid-liquid partition Chromatography) or a gas (gas-liquid partition
Chromatography).

21
CHEM231 Lab Manual 2013/14

ANALYSIS OF CHROMATOGRAMS:
In thin layer and paper Chromatography, substances are characterized by their Rf values
(retardation factor). The Rf-value is a number (less than one) which is characteristic of a
compound for a given adsorbent and developing solvent.

The retention factor, or Rf, is defined as the distance traveled by the compound divided by
the distance traveled by the solvent.

For example, if a compound travels 2.1 cm and the solvent front travels 2.8 cm, the Rf is 0.75:

The Rf for a compound is a constant from one experiment to the next only if the
chromatography conditions below are also constant:
solvent system
adsorbent
thickness of the adsorbent
amount of material spotted
temperature
Since these factors are difficult to keep constant from experiment to experiment, relative Rf
values are generally considered. Relative Rf means that the values are reported relative to a
standard, or it means that you compare the Rf values of compounds run on the same plate at the
same time.
The larger an Rf of a compound, the larger the distance it travels on the TLC plate. When
comparing two different compounds run under identical chromatography conditions, the

22
CHEM231 Lab Manual 2013/14

compound with the larger Rf is less polar because it interacts less strongly with the polar
adsorbent on the TLC plate. Conversely, if you know the structures of the compounds in a
mixture, you can predict that a compound of low polarity will have a larger R f value than a
polar compound run on the same plate.
The Rf can provide corroborative evidence as to the identity of a compound. If the identity of a
compound is suspected but not yet proven, an authentic sample of the compound, or standard,
is spotted and run on a TLC plate side by side (or on top of each other) with the compound in
question. If two substances have the same Rf value, they are likely (but not necessarily) the same
compound. If they have different Rf values, they are definitely different compounds. Note that
this identity check must be performed on a single plate, because it is difficult to duplicate all the
factors which influence Rf exactly from experiment to experiment.
In gas-liquid chromatography, compounds are characterized by their retention times.
THIN-LAYER CHROMATOGRAPHY (TLC):

This is one application of adsorption chromatography in which an adsorbent, usually silica gel
or alumina, is spread out as a thin layer on an inert surface, such as a glass plate or microscope
slide. The mixture is applied at one end of the coated plate and, as the mobile phase (a liquid)
moves up the solid adsorbent by capillary action, the adsorbed components of the mixture get
desorbed and carried along at different rates by the moving solvent. Adsorption of the
components of the mixture, on the surface of the adsorbent, occurs to differing extents
depending on their structural features and polarity. The more strongly adsorbed a given
compound is, the slower it is transported by the mobile phase, and conversely, the more weakly
adsorbed the compound is, the faster it is transported up the stationary phase. The result is that
the components of the mixture are separated into different zones or spots.

Separation by thin-layer chromatography

23
CHEM231 Lab Manual 2013/14

Separation by thin-layer chromatography depends on:

1. the kind and activity of the adsorbent (stationary phase).


2. the polarity of the eluent (mobile phase).
3. the chemical nature of the components of the mixture.

The most common adsorbents employed in TLC are silica (SiO2.xH2O) and alumina (Al2O3.
xH2O), and the activity of these adsorbents is largely determined by their water content. For a
given adsorbent and compound, the greater the polarity of the eluent, the greater is its ability to
dislodge a compound from the surface of the adsorbent, and therefore the higher the R f-value.
Eluting power of solvents:

Acetic acid > Ethyl alcohol > Acetone > Diethyl ether > Dichloromethane > Hexane.

GENERALIZED EXPERIMENTAL PROCEDURE


Preparation of TLC Plates. Large glass plates (20x20 cm) are commonly used for
quantitative separations, while microscope slides are usually used for qualitative
purposes. A homogeneous slurry of the adsorbent in a volatile organic solvent
(chloroform or dichloromethane) is poured over the glass plates and allowed to air-dry
at room temperature. Microscope slides can be coated, two at a time, by dipping them
into the slurry for sometime then holding them vertically to air-dry. The jar of adsorbent
must be shaken thoroughly before each use to homogenize the slurry.

Spotting. The mixture to be analyzed is dissolved in a suitable solvent (1% solution).


With a drawn capillary tube, a small amount of this solution is spotted on the TLC plate
about 1 cm from the bottom. The spots should have a diameter not larger than 1-2 mm,
since larger spots result in "tailing" and overlapping of close spots. Once the solvent
evaporates from the spots, the plate is ready for developing.

Development of the Chromatogram. The eluent, also called developing solvent, is


chosen on the basis of the nature and polarity of the compounds being studied. It is best
to choose the solvent that will give a satisfactory separation within the range of 0.2-0.8 Rf
values. The plate is placed in a developing chamber (e.g. a covered beaker) containing the
solvent and lined with filter paper soaked in the solvent to help saturate the atmosphere
with solvent vapors. When the solvent front reaches the finish line, the plate is removed
from the beaker and placed on the bench top to air-dry.

24
CHEM231 Lab Manual 2013/14

Visualization of Spots. Compounds on the plate are located according to their


characteristics:
a). If the spots are colored, they can be observed in ordinary light.

b). If the compounds are colorless, they can be seen under UV-light where they appear as dark
spots on a white background.

c). Colorless spots may also be located with an indicator. Most organic compounds form
complexes with iodine giving dark brown spots when the plate is exposed to iodine vapor.
Sulfuric acid may also used to make colorless spots visible. Most organic compounds turn black
when sprayed with sulfuric acid.

COLUMN CHROMATOGRAPHY:

In column chromatography, the stationary phase, a solid adsorbent, is placed in a vertical


glass (usually) column and the mobile phase, a liquid, is added to the top and flows down
through the column (by either gravity or external pressure). Column chromatography is
generally used as a purification technique: it isolates desired compounds from a mixture.
The mixture to be analyzed by column chromatography is applied to the top of the column.
The liquid solvent (the eluent) is passed through the column by gravity or by the application of
air pressure. An equilibrium is established between the solute adsorbed on the adsorbent and
the eluting solvent flowing down through the column. Because the different components in the
mixture have different interactions with the stationary and mobile phases, they will be carried
along with the mobile phase to varying degrees and a separation will be achieved. The
individual components, or elutants, are collected as the solvent drips from the bottom of the
column.
Column chromatography is separated into two categories, depending on how the solvent
flows down the column. If the solvent is allowed to flow down the column by gravity, or
percolation, it is called gravity column chromatography. If the solvent is forced down the
column by positive air pressure, it is called flash chromatography, a "state of the art" method
currently used in organic chemistry research laboratories. The term "flash chromatography" was
coined by Professor W. Clark Still because it can be done in a flash."

25
CHEM231 Lab Manual 2013/14

Columns Packed Column Column Chromatography

The Adsorbent
Silica gel (SiO2) and alumina (Al2O3) are two adsorbents commonly used by the organic chemist
for column chromatography. These adsorbents are sold in different mesh sizes, as indicated by
a number on the bottle label: silica gel 60 or silica gel 230-400 are a couple examples. This
number refers to the mesh of the sieve used to size the silica, specifically, the number of holes in
the mesh or sieve through which the crude silica particle mixture is passed in the
manufacturing process. If there are more holes per unit area, those holes are smaller, thus
allowing only smaller silica particles go through the sieve. The relationship is: the larger the
mesh size, the smaller the adsorbent particles.
Adsorbent particle size affects how the solvent flows through the column. Smaller particles
(higher mesh values) are used for flash chromatography, larger particles (lower mesh values)
are used for gravity chromatography. For example, 70230 silica gel is used for gravity columns
and 230400 mesh for flash columns.
Alumina is used more frequently in column chromatography than it is in TLC. Alumina is quite
sensitive to the amount of water which is bound to it: the higher its water content, the less polar
sites it has to bind organic compounds, and thus the less sticky it is. This stickiness or activity
is designated as I, II, or III, with I being the most active. Alumina is usually purchased as
activity I and deactivated with water before use according to specific procedures. Alumina
comes in three forms: acidic, neutral, and basic. The neutral form of activity II or III, 150 mesh,
is most commonly employed.

26
CHEM231 Lab Manual 2013/14

The Solvent
The polarity of the solvent which is passed through the column affects the relative rates at
which compounds move through the column. Polar solvents can more effectively compete with
the polar molecules of a mixture for the polar sites on the adsorbent surface and will also better
solvate the polar constituents. Consequently, a highly polar solvent will move even highly polar
molecules rapidly through the column. If a solvent is too polar, movement becomes too rapid,
and little or no separation of the components of a mixture will result. If a solvent is not polar
enough, no compounds will elute from the column. Proper choice of an eluting solvent is thus
crucial to the successful application of column chromatography as a separation technique. TLC
is generally used to determine the system for a column chromatography separation. Often a
series of increasingly polar solvent systems are used to elute a column. A non-polar solvent is
first used to elute a less-polar compound. Once the less-polar compound is off the column, a
more-polar solvent is added to the column to elute the more-polar compound.
Interactions of the Compound and the Adsorbent
Compounds interact with the silica or alumina largely due to polar interactions.
Analysis of Column Eluents
If the compounds separated in a column chromatography procedure are colored, the progress
of the separation can simply be monitored visually. More commonly, the compounds to be
isolated from column chromatography are colorless. In this case, small fractions of the eluent
are collected sequentially in labeled tubes and the composition of each fraction is analyzed by
thin layer chromatography. (Other methods of analysis are available; this is the most common
method and the one used in the organic chemistry teaching labs.).
This Figures shows the use of column chromatography

1 2 3 4 5 ..
Column Chromatography

27
CHEM231 Lab Manual 2013/14

Alcohols

Classification and tests


Objective:

1. Determine chemical and physical properties of alcohols.


2. Classify an alcohol as primary, secondary, or tertiary.
3. Perform a chemical test to distinguish between the classes of alcohols.
4. Write the formulas of the oxidation products of alcohols

Structures of Alcohols

Alcohols are organic compounds that contain the hydroxyl group (OH). The simplest alcohol
is methanol. Ethanol is found in alcoholic beverages and preservatives, and is used as a solvent.
2-Propanol (isopropanol), also known as rubbing alcohol, is found in astringents and perfumes.

Classification of Alcohols

In a primary (1) alcohol, the carbon atom attached to the OH group is bonded to one other
carbon atom. In a secondary (2) alcohol, it is attached to two carbon atoms and in a tertiary (3)
alcohol to three carbon atoms.

28
CHEM231 Lab Manual 2013/14

1. Properties of Alcohols
Materials: 4 test tubes, pH paper, stirring rod, methanol, ethanol, 2-propanol, t-butyl
alcohol (2-methyl-2-propanol).
Odor: Place 5 drops of each of the alcohols, phenol to four separate test tubes. Carefully
detect the odor of each.
Solubility in water Add about 2 mL of water (40 drops) to each test tube. Shake and
determine whether each alcohol is soluble or not. If the substance is soluble in water, you
will see a clear solution with no separate layers. If it is insoluble, a cloudy mixture or
separate layer will form. Record your observations.
Acidity Obtain a container of pH paper. Place a stirring rod in one of the alcohols and touch
a drop to the pH paper. Compare the color of the paper with the chart on the container to
determine the pH of the solution. Record your observations.
Note: DISPOSE OF ORGANIC SUBSTANCES IN DESIGNATED WASTE CONTAINERS!

2. Oxidation of Alcohols
The Bordwell-Wellman test solution contains potassium dichromate dissolved in sulfuric
acid. It is an orange-yellow solution. The orange-yellow color is due to the Cr2O7 2- ion. The
oxidation number of chromium is +6. This reagent will oxidize primary and secondary
alcohols and, in turn, becomes reduced. The chromium in dichromate is reduced to chromic
ion, Cr+++.
A greenish colored solution results. This color change from orange-yellow to green serves
as an indicator for the presence of a primary or secondary alcohol. A primary alcohol is
oxidized first to an aldehyde, which will be further oxidized to an acid.

Materials: 4 test tubes, methanol, ethanol, 2-propanol, t-butyl alcohol (2-methyl-2-


propanol), 1% potassium dichromate solution, conc. H2SO4

Procedure: Place 3 mL of 1 % of potassium dichromate and 2 drops of conc. H2SO4 in each


of four test tubes. Mix thoroughly and add 2 drops of the alcohols. Shake the tube and Look
for a color change.
If the orange color turns to green in 12 minutes, oxidation of the alcohol has taken place. If
the color remains orange, no reaction has occurred. If a test tube becomes hot, place it in a
beaker of ice-cold water. Record your observations.
Caution: Concentrated H2SO4 is corrosive.

29
CHEM231 Lab Manual 2013/14

General reaction equation:

Experimental equation:

K2Cr2O7
Methanol: CH3OH HCOOH Cr 3+
H2SO4 green

K2Cr2O7
CH3CH2OH CH3COOH Cr 3+
Ethanol: H2SO4 green

OH O
K2Cr2O7 Cr 3+
Isopropanol: CH3CHCH3 C
H2SO4 H3C CH3 green

CH3
K2Cr2O7
2-methyl-2-propanol:
H3C C OH No Reaction
H2SO4
CH3

30
CHEM231 Lab Manual 2013/14

3. Lucas Test: for Primary, Secondary and Tertiary Alcohols.


The Lucas reagent is a solution of zinc chloride in concentrated HCl. This solution must
be made freshly to get proper results. The test depends on a difference in the rate of
reaction of these alcohols. The general equation for the reaction is:

Tertiary alcohols react IMMEDIATELY. The test tube will get hot, and because
the chloride is insoluble two layers may be apparent, or a cloudy dispersion
forms.
Secondary alcohols will become cloudy in 5 to 10 minutes. If cloudiness does not
appear place test tube in a hot water bath and observe.
Primary alcohols give no reaction in a reasonable length of time.

CAUTION! Lucas reagent contains concentrated hydrochloric acid - Handle It With Care

Procedure
1. Place 1 ml of Lucas reagent in each of three clean test tubes.
2. Add 4 drops of ethanol to one test tube. Shake the test tube to mix the reagents and
notice whether the mixture gets cloudy and how long it takes.
3. In a second test tube place 4 drops of Isopropanol, shake and note how long it takes the
tube to get cloudy.
4. In a third test tube place 4 drops of 2-methyl-2-propanol, shake and note how long it
takes the tube to get cloudy.
5. Record observations on the Report Sheet.
Dispose of these reagents in the "Halogenated Organic Liquid Waste" container in the hood.

31
CHEM231 Lab Manual 2013/14

4. Iodoform Test
The iodoform test is used to identify secondary alcohols that have a methyl group on the
alcohol carbon. This type of alcohol will react with I2 in NaOH to give a yellow
precipitate of iodoform, CHI3. The reaction is shown below.

The formation of a yellow precipitate in the test solution is taken as a positive reaction,
and it means that the reactant alcohol was a secondary alcohol with a methyl group on
the alcohol carbon.

Procedure: To test methanol, ethanol and isopropanol, Place about 1 mL of water (20 drops)
in each of 3 test tubes. Add 5 drops of the alcohols to be tested. Add 10 drops of 10% NaOH
and mix by shaking each tube side to side. Add 10 drops of KI/iodine solution and shake to
mix. A yellow precipitate indicates a positive reaction. Record observations on the Report
Sheet.

32
CHEM231 Lab Manual 2013/14

5. Methyl Salicylate (Wintergreen Oil)


1. Transfer about 0.5 g of salicylic acid to clean and DRY large test tube and add 1 mL of
methanol.
2. Holding the top of the tube, tap the bottom with your finger to stir the mixture.
Dissolve the salicylic acid completely in the methanol.
3. Carefully add 3 to 4 drops of concentrated H2SO4 to the mixture (in the hood) and mix
again by tapping the bottom of the tube with your finger.
4. Place this mixture in the hot water bath for 10 min to complete the reaction.
5. After the reaction is complete, remove the test tube from the water bath and allow it to
cool. Turn off the hot plate under the water bath.
6. After it has cooled, carefully waft the fumes by passing your hand over the top of the
tube toward your nose. Can you smell the minty aroma of wintergreen?
7. If you have difficulty smelling it, you can pour hot water from the water bath into the
tube. The aroma of the methyl salicylate should be apparent after it has mixed with
warm water.
8. Record your observations and answer the questions on the Report Sheet.

COOH COOCH3
H2SO4
CH3OH H2O

OH OH

33
CHEM231 Lab Manual 2013/14

Alcohols
Report Sheet Name:----------------------

1. Solubility in water
Alcohol Structure Solubility

2. Oxidation of Alcohol
Alcohol Reaction equation Observations

3. Lucas Test
Alcohol Structure of Product Observations

4. Iodoform Test
Alcohol Structure of Product Observation

5. Formation of Methyl Salicylate


Alcohol Reaction equation Observation

34
CHEM231 Lab Manual 2013/14

Aldehydes and Ketones

Classification and tests


Objective:

5. Determine chemical and physical properties of aldehydes and ketones.


6. Perform a chemical test to distinguish between aldehydes and ketones

Structures of Aldehydes and Ketones


The major similarity between an aldehyde and a ketone is the carbonyl group. A
carbonyl group is a carbon atom doubly bonded to an oxygen atom.

O
C
Both molecules have a carbonyl group, the difference the number of carbons bonded to
the carbonyl carbon. An aldehyde will have none or one and a ketone will have two
carbons.

All aldehydes, except formaldehyde, will have a hydrogen atom on one side of the
carbonyl carbon and at least on carbon on the other side.
O
R C H

O O O

H C H CH3 C H C H

formaldehyde acetaldehyde benzaldehyde

All ketones have a carbon on each side of the carbonyl carbon.


O
R C R

O O O
O
CH3 C CH3 CH3CH2 C CH3 CH3CH2 C CH2CH3
Acetone butanone 3-pentanone cyclohexanone

35
CHEM231 Lab Manual 2013/14

1. The Bordwell-Wellman test solution contains potassium dichromate dissolved in sulfuric


acid: This test was used earlier when testing alcohols. Aldehydes are more rapidly oxidized
than ketones due to the hydrogen atom bonded to the -carbon. Aldehydes are oxidized to
carboxylic acids.
O [O] O

R H strong oxidizing agent R OH

aldehyde carboxylic acid


Ketones are not readily oxidized, which makes the two functional groups easily
distinguishable. Only under extreme conditions (strong reagents and high temperature) can
ketones be oxidized since the reaction requires the cleavage of a carbon-carbon bond.

Procedure: Add two drops of the carbonyl compound (Acetone, Acetaldehyde, and
Benzaldehyde) to 0.5 mL of the orange reagent (K2Cr2O7) in a test tube. Shake the tube vigorously
and heat in a water bath for about 3 - 4 minutes. Good mixing is essential, especially with water-
insoluble compounds. A few aldehydes may require longer heating; -- aromatic aldehydes are
more difficult to oxidize than aliphatic ones.

Test Tube Rxn:

O O
CH3 C H KH


2Cr2O7
+
CH3 C OH

Test Tube Rxn:

O
CH3 C CH3 HK NR
2Cr2O7
+

36
CHEM231 Lab Manual 2013/14

2. Tollens Test: Commonly known as the Silver Mirror Test, this distinctive qualitative test
involves the oxidation of aldehydes to their corresponding carboxylic acid. The oxidizing agent
is a silver complex ion [Ag(NH3)+2] which is reduced to a metallic silver which remains on the
walls of the test tube as a mirror. Otherwise, the silver is deposited as a black precipitate.
Tollens reagent is prepared by dissolving silver oxide in ammonia:

2 AgNO3 + 2 NaOH Ag2O (s) + H2O + 2 NaNO3


Grey Precipitate

Ag2O (s) + 4 NH3 + H2O 2 Ag(NH3)2 OH


Tollen's Reagent

O O
H + 2 Ag(NH3)2 OH + 2 Ag + 3NH3 + H2O
R R O NH4
silver

Procedure: add 3 to 4 drops of (Acetaldehyde, Benzaldehyde, and Acetone) to a basic solution of


AgNO3, NH3, ammonia is normally the base, if the test tube shows a mirrored surface, this is a
positive test for an aldehyde, gentle heating may be necessary. Ordinary ketones do not react with
the reagent.

To prepare Tollen's reagent: add 2 mL of 5% silver nitrate, and ONE DROP of 10% (3M) NaOH.
Mix the ingredients THOROUGHLY by STIRRING. If necessary addition of dropwise 2%
ammonia solution.

CAUTION!! Discard any unused Tollen's reagent after you have completed the tests. EXPLOSIVE
PRECIPITATES DEPOSIT ON STANDING!!

37
CHEM231 Lab Manual 2013/14

3. Iodoform Test: The Iodoform test was performed earlier on alcohols. The yellow ppt may be
collected on Buchner, washed with 10% NaOH, and dried. If there is doubt, its m.p. will con-
firm its identity. Since the procedure here is identical with that of the alcohols, this test will be
omitted here.

Iodoform Test for Methyl Ketones

Ketones containing a methyl group attached to the carbonyl give a reaction with iodine (I2) in a
NaOH solution. The reaction produces solid, yellow iodoform, CHI3.
Iodoform, which has a strong medicinal odor, is used as an antiseptic.

38
CHEM231 Lab Manual 2013/14

Aldehydes and Ketones


Report Sheet Name:----------------------

1. Acetaldehyde:
Physical State: Color: Odor:
2. Benzaldehyde:
Physical State: Color: Odor:
3. Acetone:
Physical State: Color: Odor:
6. Oxidation of Aldehyde and Ketone
Reaction equation Observations

Acetaldehyde

Benzaldehyde

Acetone

7. Tollens Test
Reaction equation Observations
Acetaldehyde

Benzaldehyde

Acetone

8. Iodoform Test
Structure of Product Observation
Acetaldehyde

Benzaldehyde

Acetone

39
CHEM231 Lab Manual 2013/14

Carboxylic Acids

Classification and tests


Objective:

7. Determine chemical and physical properties of carboxylic acid.


8. Perform a chemical test to distinguish carboxylic acids.

Structures of Carboxylic acids


Carboxylic acids are organic compounds that have a carboxyl group attached to an alkyl
group (R-COOH) or to an aryl group (Ar-COOH). The 'R' may be a hydrogen and the
result is formic acid. They may be mono carboxylated, multi carboxylated, substituted (e.
g., hydroxyl groups), or they may be aromatic

Physical properties

1. Only formic acid, acetic acid, and lactic acid are liquids at room temperature. The
others are solids.
2. Low molecular weight carboxylic acids are soluble in water. Water insoluble acids
dissolve in both sodium hydroxide solution and sodium bicarbonate solution, they
evolve carbon dioxide gas. This is considered as a good simple indication of them.
3. Aromatic carboxylic acids burn with a yellow smoky flame whereas aliphatic ones
bum with a blue flame without smoke.
4. Their boiling points are generally high due to the association through hydrogen

40
CHEM231 Lab Manual 2013/14

bonds: two molecules of the carboxylic acid are held together by two hydrogen bonds
rather than one.

Chemical properties
The acidic properties of carboxylic acids are attributed to the proton of the carboxyl
group. Mono carboxylic acids are weak acids except formic acid, which is the strongest.
The tendency of the alkyl group to release electrons weakens the acid; thus formic acid is
the strongest. On the other hand presence of electron withdrawing groups (such as
halogens) especially on the alpha carbon increases the acidity.
Reactions of carboxylic acids are related to:
The proton as in salt formation reactions.
Removal of the hydroxyl group as in conversion to derivatives such as esters,
amides, or acid chlorides.
Substitution either in the alpha position of aliphatic acids or in the meta position
of aromatic ones.

1. Solubility Test:
All aliphatic carboxylic acid are water soluble and change litmus paper from blue
to red color.
All aromatic carboxylic acid are water insoluble. Soluble in 10% NaOH due to its
acicdity

2. General test (Ferric chloride FeCl3 test)


The acid solution should be made neutral before performing the test with ferric chloride
solution. This is achieved by adding very dilute ammonia solution drop by drop with
shaking to a solution of about 0.5 g of the solid acid or 2 drops of the liquid acid in 1 mL
water until the medium becomes basic. At this stage the solution is slightly basic. To
make the solution neutral the excess ammonia should be removed by gently heating the
test tube in a water bath with shaking from time to time until the odor of ammonia
disappears. (In case of oxalic, tartaric, citric and lactic acids keep a portion of their
neutral solution for use in calcium chloride test). Cool the solution and then add few
drops of ferric chloride solution to get different colors (solutions or precipitates) as
follows:

41
CHEM231 Lab Manual 2013/14
1.
2.
4.
3.
3. Special test for acetic acid (ester formation)

Acetic acid, on contrary to formic acid, neither can be oxidized by, nor can reduce any of
the reagents applied to formic acid. Instead, it undergoes ester formation reaction:

Mix 1 mL of acetic acid with 2 mL of ethanol in a test tube and add to this mixture 2-3
drops of concentrated sulphuric acid. Heat the test tube in a water bath for 10 minutes,
and then pour the mixture into another test tube containing 5 mL sodium bicarbonate
solution; the characteristic fruity odor of ethyl acetate can be smelt, which indicates the
formation of this ester.

4. Special test for salicylic acid (ester formation)


In addition to the characteristic violet colour obtained with ferric chloride, salicylic acid
can also be detected by ester formation test. In this test methyl salicylate ester separates
out as an organic phase having a characteristic odor.

Follow the same procedure and conditions used for esterification of acetic acid but use
methanol instead of ethanol. Not that methanol is toxic internally so never withdraw it
by mouth to avoid accidental ingestion.

5. Reaction with Na2CO3

Carboxylic acid + Na2CO3 Strong effervescence (CO2 )

6. Reaction with calcium chloride CaCl2

General Carboxylic acid + CaCl2 + NH4OH precipitate

1. Oxalic acid: white ppt will dissolve in dil HCl, will not dissolve in acetic acid
2. Tartaric acid: white ppt with test tube scratching. The ppt will dissolve in acetic
acid
3. Citric acid: white ppt after heating, will not dissolve in acetic acid

42
CHEM231 Lab Manual 2013/14

Carboxylic acid
Report Sheet Name:----------------------

9. Solubility in water:
Carboxylic acid Structure Solubility
Acetic acid
Oxalic acid

Citric acid

Tartaric acid

Benzoic acid

Salicylic acid

10. Ferric chloride FeCl3:


Carboxylic acid Observation Conclusion

Acetic acid

Oxalic acid

Citric acid

Tartaric acid

Benzoic acid

Salicylic acid

11. Ester forming:


Carboxylic acid Observation Conclusion
Acetic acid

Salicylic acid

43
CHEM231 Lab Manual 2013/14

12. Reaction with Na2CO3:


Carboxylic acid Observation Conclusion
Acetic acid
Oxalic acid

Citric acid

Tartaric acid

Benzoic acid

Salicylic acid

13. Reaction with calcium chloride CaCl2


Carboxylic acid Observation Conclusion
Oxalic acid

Citric acid

Tartaric acid

44