Lab 3.

Isolation of caffeine from coffee

Purpose: Isolation of caffeine from coffee
Introduction
Caffeine is stimulant. The most common sources of caffeine are: tales, tea, cocoa, mate, and kola
nuts. On the dry weight basis, tea leaves contain more caffeine than coffee powder. However, a
cup of coffee actually contains more caffeine because more coffee than tea is used in making a cup
of “drinking strength” brew.

Because caffeine has stimulant effect on the central nervous system, it has been used as an
antidote to counteract the depressant effects of morphine poisoning. The lethal dose for caffeine
in humans is estimated to be about 10g (equivalent to ~100 cups of coffee). There is no report
about deaths that caused by coffee.
We can extract caffeine from your beverage with dichloromethane (DCM). Caffeine is slightly
soluble in water but very soluble in DCM. Thus, mixing these two insoluble liquids together will
cause most of the caffeine to dissolve preferentially in the organic DCM layer. The base that is
added reacts to form salts with many of the non-caffeine substances (e.g. tannic acid). This
prevents them from also dissolving in the DCM thus contaminating the caffeine.
Because water and DCM are not miscible, the two liquid layers are then separated by filtration.
Firstly, a filter paper cone will be wetted with water, and the DCM layer with a small amount of
the water later, will then be poured into the funnel. Once wetted with water, the pores of a filter
paper will only allow water to drain through them. When the two liquids are poured into this wet
paper the DCM layer will stay trapped inside the filter cone while the water layer drains through.
After evaporation of DCM, you can obtain a product consisting of brown to white caffeine crystals.
The question then arises “how do you know that these crystals are indeed caffeine?”
In this experiment, we will use two methods to identify your product: Melting point
measurement and infrared spectrum. Melting point is a physical constant property for a particular
substance (like density and boiling point, etc). For example, all samples of ice melt at 0 C.
Similarly, all samples of pure caffeine melt at 238 C. By comparing the observed melting point
of product you obtained from your experiment with recorded melting point of pure caffeine,
evidence for identity can be obtained. Usually impurity can cause a sample to begin melting at a
temperature lower than melting point. So if there is impurities in your caffeine sample, it will begin
melt at a temperature lower than 238 C (sometimes at ~190 to 220 C). Please note that caffeine
can sublime (go directly from a solid to a gas phase) and may cause difficulty in observing the
melting point. So rapid heating of your sample is necessary.

Infrared (IR) spectroscopy is also very useful for the quick identification of functional group(s)
in a molecule. Like other characteristic properties, the response of a molecule to IR light is also
characteristic. Light in the IR wavelength range (700 nm – 1 mm) can cause molecules to vibrate
along bonds. Thus, IR spectroscopy is most often used technique for characterization and
identification, since the number and type of chemical bonds gives a specific response. Here we
will also use IR spectroscopy to identify our caffeine samples. The IR spectrum for pure caffeine
is provided for you to use as a comparison to the spectrum obtained for your sample. Directions
for collecting an IR spectrum will be provided by your instructor.

Minimum safety standards for this experiment

As in all laboratory procedures, you should use proper PPE and look up any potential hazards
associated with the substances you will be using:
(a) Hot glass; (b) Odor chemicals should be handled in fume hood; (c) Look up the MSDS for each
reagent used; (d) be careful to avoid spillage, especially during the extraction portion.

Equipment/Materials
250 mL Erlenmeyer flask, beaker, filter paper, long stem funnel, instant/ground coffee, sodium
carbonate, Melting point apparatus

Procedure
Step 1. (a) Weigh out at least 1g of instant coffee.
(b) Dissolve the solid in 100 mL of cold water in a clean, 250 mL Erlenmeyer flask.

Step 2. Extraction
1. Dissolve 2 g of Na2CO3 in your sample in the Erlenmeyer flask as prepared from Step 1
above.
2. Add 25 mL of CH2Cl2 (DCM) and agitate by “aggressively swirling” for about 10 minutes.
Attention: Do not shake or a persistent liquid/liquid emulsion will likely form (a frothy
layer of water and DCM mixed that does not separate easily).
3. Let the flask contents stand undisturbed for several minutes while you suspend a long stem
funnel in an iron ring and place a collection beaker or flask underneath (size unimportant).
Fit the funnel with a cone made from a Filter paper and wet the paper thoroughly with
water. Your lab instructor or TA can show you how to fold the filter paper.
4. The upper water layer is possibly dark brown. By decantation, slowly and carefully pour
off and discard the upper layer as much as possible. Leave behind in the flask all of the
DCM layer (higher density), together with a little remaining water.
5. Slowly pour the entire contents of your Erlenmeyer flask (the DCM solution + upper layer
of remaining water) into the wetted filter paper cone. Keep the liquid level up near the top
of the paper cone until all of the flask contents have been thus transferred. The upper water
layer will wick into the paper cone and drain through it. Left behind trapped in the filter
paper will be the clear DCM solution, which may be yellow to dark brown. This solution
contains your caffeine.
6. While the last bit of water is draining through the filter cone, take a clean and dry 50 mL
(or 100 mL) beaker and put your group number or your name on it. Weigh this beaker to
the nearest milligram (0.001g) using an electronic balance and record data on the report
 sheet. Write down the weight!
7. Using a medicine dropper, transfer only the clear DCM layer (possibly yellow or dark
brown) remaining in your filter cone to the weighed dry beaker.
8. Go to the fume hood and place your beaker on the hot plates. Watch the liquid contents
closely and do not leave the beaker unattended. Avoid breathing the DCM vapors. When
the liquid level in the beaker gets down to about 1 mm, immediately remove the beaker
from the hot plate and set it aside in the hood to cool. Because DCM has low boiling point
at only 40 C, the small remainder of DCM liquid evaporates very quickly. It is not
necessarily to hold the beaker with tongs as the DCM boils at only 40 C. So you can safely
grab the beaker with your fingers. Record your observations on the report. Reweigh the
beaker to the nearest 0.001g. Now we are ready to try to find out if what we have isolated
is really caffeine.

Data Sheet
A. Preparation

Appearance and source of original coffee sample:

Weight of coffee sample _________ g

B. Extraction

Appearance of solid residue in beaker after evaporation of DCM

Weight of beaker (empty and dry) __________ g

Weight of beaker + dry residue of caffeine __________ g
Total weight of residue (caffeine) __________ g
The extraction yield (%) of raw caffeine __________

C. Identification

Melting point of residue __________

Melting point of pure caffeine (consult the Merck Index) ___238C___

QUESTIONS
1. In the caffeine isolation steps:
(a) Which layer in the actual extraction step was the DCM (methylene chloride) the top or the
bottom?
(b) What physical property of DCM would cause it to be above or below the water layer? Is this
property intensity or extensive property?

2. The symptoms of withdrawal from an addictive drug are irritability, desire for the addicting
drug and physical discomfort. All of these symptoms are relieved by a dose of the addicting drug.
Using these criteria, is caffeine an addicting drug?