Pharmaceutical and Medicinal Organic
Chemistry (lab)
3. Hold the stopper in place firmly and invert
EXERCISE 1 - MACROSCALE EXTRACTION the separatory funnel. While the
OF CAFFEINE FROM AQUEOUS SOLUTION separatory funnel is inverted, release the
pressure by slowly opening the stopcock.
Extraction is one of the most important Continue inverting and venting until the
techniques for isolating and purifying organic “whoosh” is no longer audible. Note that
substances. In this method, a solution is the two layers must now be mixed
mixed thoroughly with a second solvent that is thoroughly so that as much caffeine as
possible is transferred from the aqueous
immiscible with the first solvent.
layer to the dichloromethane layer.
4. Shake the mixture for at least one minute.
MATERIALS, APPARATUS AND REAGENTS 5. Place the separatory funnel in the iron ring
NEEDED and let it stand until the layers separate
completely.
MATERIALS AND 6. Place an Erlenmeyer flask under the
REAGENTS
APPARATUS separatory funnel and remove the top
2% Aqueous caffeine 125 mL Separatory stopper on the funnel. Allow the bottom
solution funnel (organic) layer to drain slowly by partially
opening the stopcock.
Ammonia solution 10 mL Pipet 7. When the interface between the upper and
Anhydrous sodium lower phases just begins to enter the bore
Beaker of the stopcock, close the stopcock
sulfate
immediately.
Concentrated
Bulb aspirator 8. Repeat this extraction two more times
hydrochloric acid
using 5.0 mL of fresh dichloromethane
Dichloromethane Cork each time.
Distilled water Erlenmeyer flask 9. Combine the organic layer from each of
these extractions with the dichloromethane
Potassium chlorate Evaporating dish solution from the first extraction.
Glass rod
Hotplate
Drying the Organic Layer
Iron ring
Iron stand 1. Weigh four portions of anhydrous sodium
sulfate in a paper box, each weighing 0.50
Test tube
g.
2. Add one portion of anhydrous sodium
Watch glass
sulfate to the combined solution.
3. Swirl and stir the mixture.
PROCEDURES 4. Continue adding the remaining portions
while swirling and stirring the mixture
SAFETY AND SPECIAL INSTRUCTIONS until 1-3 mm layer of the solid forms at the
bottom of the Erlenmeyer flask.
Dichloromethane (CH2Cl2) is an acute 5. Note that the mixture is dry if it appears
inhalational hazard. Avoid breathing its clear. Otherwise, add more drying agent
fumes and avoid skin contact. and repeat the drying procedure.
Concentrated hydrochloric acid is 6. Filter the whole solution into a previously
corrosive. Avoid any skin contact. weighed evaporating dish.
Dispose all organic solvents in the
designated container.
Extraction of Caffeine from the Aqueous Evaporation of the Solvent
Solution
1. Under the hood, evaporate the
1. Pipet out 10 mL of the 2% aqueous caffeine dichloromethane by heating the
solution. Transfer the solution in a 125 mL evaporating dish in a hot water bath at
separatory funnel. Do not forget to close about 45°C.
the stopcock of the separatory funnel 2. When the solvent has evaporated, remove
before transferring. the evaporating dish from the bath and dry
2. Then add 5.0 mL dichloromethane to the the outside of the dish. Do not leave the
caffeine solution on the separatory funnel. evaporating dish in the water bath for a
Stopper the separatory funnel and hold it long time after the solvent has evaporated
as shown on Figure 1.2. because the caffeine may sublime.
3. Cool the evaporating dish at room 2. Anhydrous Sodium Sufate will start
temperature. absorbing the water from the solution
4. Weigh the evaporating dish with its 3. Gently stir the Erlenmeyer flask using a
content to determine the amount of
circular motion to make sure all the
caffeine.
water is absorbed.
Murexide Test for Caffeine - When the Anhydrous Sodium Sufate
swirls freely in the flask, the solution is
1. Place a pinch of the extracted caffeine on a now water-free.
watch glass. - Anhydrous Sodium Sufate – will
2. Add 3 drops of concentrated hydrochloric aggregate to the bottom of the flask.
acid. - It will stick mostly to the
3. Then add a pinch of potassium chlorate.
glass and form clumps,
Stir the mixture using a glass rod.
4. Gently heat watch glass over steam bath making it easy to
until the mixture is dry. separate.
5. Allow to cool at room temperature then 4. Get a clean beaker and pour the dry
moisten the dried mixture with ammonia liquid into it carefully.
solution. - Do not let the Anhydrous Sodium
6. Observe for any changes in the color of the Sufate be transferred to the clean
solution.
beaker.
Module 1 Pretask: - You can use glass funnel with filter
paper.
How to use a separatory Funnel
MACROSCALE EXTRACTION
Separatory funnel- used to separate two
immiscible liquids. EXTRACTION
Funnel – holds TWICE the volume of the Extraction is one of the most important
solution to be extracted and the extraction techniques for isolating and purifying organic
solvent. substances that involves transferring a solute
from one solvent into another. The solute is
Ring stand- for support extracted from one solvent into the other
because the solute is more soluble in the
1. Add the solution first followed by the second solvent than in the first. The two
extracting solvent. Stop cock should be solvents must be immiscible (mix freely), and
they must form two separate phases or layers,
closed when pouring the solvent.
for this procedure to work. Therefore, the
2. Stopper the funnel while holding the difference in between the polarities of two
stopper in place, gently invert and swirl selected solvents play a very important role
the liquids. Do not shake the funnel using this liquid-liquid extraction procedure.
vigorously for an emulsion may form.
Extraction is used in many ways in organic
3. Open the stop cock to release pressure
chemistry. Many natural products are present
and close it to continue to swirl. Repeat
in animal and plant tissues having high water
steps for a few times.
content. Extracting these tissues with a water-
4. After mixing, place back in the ring
immiscible organic solvent is useful for
stand and remove the stopper.
isolating the natural products. Organic
5. Wait for the 2 liquids to separate.
solvents such as diethyl ether, hexane, and
6. Open the stop cock to remove the
dichloromethane (slightly polar to non-polar)
bottom layer. Close it before the top
are commonly used. For instance, caffeine, a
layer reaches the bottom of the funnel.
natural product, can be extracted from an
DRYING THE EXTRACT USING aqueous tea solution by shaking the solution
ANHYDROUS SODIUM SULFATE successively with several portions of
dichloromethane). Another example, vanilla
Anhydrous Sodium Sufate – used to remove extract, the popular flavoring agent, was
water from an organic material. originally extracted from vanilla beans using
alcohol ethanol (slightly polar to non-polar ) as
- Typically in powder form
the organic solvent.
1. Scoop a small amount of Anhydrous
Sodium Sufate to the Erlenmeyer flask.
- TOO MUCH – difficult to separate from
the organic solution.
4. The dehydrated solution is filtered and
the extract is collected in a pre-weighed
THE EXTRACTION PROCESS evaporating dish.
5. Dichloromethane is evaporated by
THE EXTRACTION PROCESS
steam bath. Too much heat should be
avoided because the caffeine may
A generalized extraction process, using a
specialized piece of glassware called a sublime.
separatory funnel, is illustrated in the Figure 6. The dried caffeine is weighed and its
1.3. One of the most common extraction percent yield is computed. After that, a
procedures involves using an organic solvent pinch of the sample is placed in a
(nonpolar or slightly polar) to extract an watch glass for the murexide test.
organic compound from an aqueous solution.
7. Murexide test is performed. The
Because water is highly polar, the mixture will
separate into two layers or phases: an extracted caffeine is added with
aqueous layer and an organic (nonpolar) layer. concentrated hydrochloric acid and
potassium chlorate. The mixture turns
Many substances are soluble in both water yellow after steam bath.
and organic solvents. Water can be used to
8. The dried mixture is moistened with
extract, or “wash,” water-soluble impurities
from an organic reaction mixture. To carry out ammonia solution. The mixture then
a “washing” operation, water and an turns pink to purple in color. Any
immiscible organic solvent must be added to purine alkaloids (such as caffeine)
the reaction mixture contained in a separatory produce a pink to purple color using
funnel. After stoppering the funnel and this test due to formation of murexide
shaking it, allow the organic layer and the or ammonium purpurate.
aqueous (water) layer to separate. A water
wash removes highly polar and water-soluble
materials, such as sulfuric acid, hydrochloric
acid, and sodium hydroxide, from the organic CAFFEINE
layer. The washing operation helps to purify
the desired organic compound present in the The following are the physicochemical
original reaction mixture. properties of caffeine:
During the swirling and shaking procedure,
emulsion may form (the organic layer may be IUPAC name: 1,3,7-trimethylpurine-2,6-
suspended within the aqueous solution). To dione
prevent this phenomena, a small volume of synonym: 1,3,7-trimethylxanthine
saturated sodium chloride may be added, or molecular formula: C8H10N4O2
the mixture inside the funnel may be gently physical description: odorless white powder
stirred using a glass rod to break off the with bitter taste
emulsion. After the two layers are formed, the boiling point: 178°C (sublimes)
solution can be separated. To identify which melting point: 235°C
layer is organic, get a sample from each solubility: 2.17 g per 100 mL water
layer then add a drop of water. If the drop density: 1.23 g/cm3
mixes with the sample, the layer is chemical safety: irritant
aqueous, if not, then the layer is organic. uses: stimulant
structure: three methyl groups attached in
MACROSCALE EXTRACTION OF CAFFEINE a purine ring
1. The caffeine from an aqueous solution
is extracted using dichloromethane
(CH2Cl2). Because dichloromethane is
denser than water, the organic layer
(dichloromethane) is the bottom layer
while the water is the upper layer.
2. The solution is allowed to settle. The
lower layer is collected, while the upper
layer is retained. This extraction
process using dichloromethane is
repeated three times
3. After extraction, the organic layer is
removed and dried over anhydrous
sodium sulfate.
