Natural Product Chemistry: Isolation

of Caffeine from Tea

By:
Kirsten Holloway
(​Kirsten.Holloway@aol.com​)

Organic Chemistry 2125

Sec 012

Experiment 6

March 22, 2018

Abstract
With the use of a separatory funnel, caffeine was to be extracted from 13.482g of tea by isolation
and the use of thin layer chromatography. 20 mL of ethyl acetate was obtained to extract the
caffeine/other compounds from the water over several fractions. (20 mL ethyl acetate for each
individual fraction). After drying the ethyl acetate solution, the solvent was purified yielding
.094 of crude caffeine. The percent recovery was determined to be 35% of the weight of the dry
tea. Percent recovery was compared to Rf values. Rf values: Spot 1: .35, Spot 3: .35. The
identical Rf values suggested that my sample of crude caffeine was in fact very similar to pure
caffeine.

Introduction
In this report a combination of techniques are used including extraction, crystallization, rotary
evaporation, and thin layer chromatography. The objective was to ultimately extract as much
caffeine from the brewed tea as possible in order to see it in its purest form. The crude caffeine
will be compared to a sample of pure caffeine by TLC. Rf values are calculated along with
percent recovery to assess the purity of the crude caffeine sample.

Experimental
First, the experiment was carried out with the use of extraction. To begin, 120 mL of water was
brought to a boil on a hot plate. Once boiling, the tea bags were placed in the beaker in order to
release the caffeinated tea into the water. 0.60 g sodium carbonate was then mixed into the tea.
The aqueous layer (tea) and the organic layer (caffeine/ethyl acetate) are placed in the separatory
funnel to isolate themselves. The aqueous layer is drained three times to insure that the caffeine
is obtained in the organic layer. 20 mL of ethyl acetate solvent was used in a separatory funnel
over several fractions (20 mL of ethyl acetate for each additional fraction).The resulting solution
of caffeine is washed in a separatory funnel with 10 mL of aqueous sodium carbonate. The ethyl
acetate layer or organic layer is drained into a flask along with 2.975g sodium sulfate. By adding
2.975g of sodium sulfate, it allows any remaining water in the ethyl acetate layer to be removed.
Next, isolation of the caffeine occurs with the use of the rotary evaporator. The rotovap will
remove all of the ethyl acetate leaving behind only the crusty deposits of caffeine. To wash the
flask, 2 mL of acetone is used creating an acetone solution. The acetone solution is brought to a
boil on a hot plate in order to evaporate all of the acetone. After acetone is removed, spot 1 for
thin line chromatography should be completed using a capillary tube to spot the crude caffeine
sample. Next, 2 mL hexane/.5 mL acetone is heated with the caffeine until the caffeine is
dissolved. It is then placed in an ice bath to carry out the crystallization portion of the
experiment. The mass of the extracted caffeine is calculated and a percent recovery is
determined. To complete thin line chromatography, pure caffeine crystals are obtained to
perform spot 3 with the use of a capillary tube. The TLC plate is developed in 3:1 ratio of ethyl
acetate and methanol, then placed under UV light. Rf values are calculated to compare the purity
of the two spots.

Results and Discussion

In this report, the data was calculated with the intent to compare and observe the extracted
caffeine sample versus the pure caffeine sample. This comparison would likely show if the
extracted caffeine sample had any impurities in it by the Rf value. The data was collected by
measurements of progress each sample made from the original spot it began on the TLC plate.
Those measurements were then used in the equation to calculate the Rf value. If the spots were
considerably different then we could assume that the extracted caffeine was impure and not
likely to be pure caffeine. If other elements or compounds were subject to the caffeine product,
this would result in a difference in polarity. However, my Rf values are identical which
concluded that my extracted caffeine is in fact, pure in comparison to the pure caffeine sample.
After the Rf values were calculated, percent recovery was also determined to be 35%. The
percent recovery is found to assess the amount of caffeine is recovered after extraction takes
place. The extraction portion of the experiment played a vital role also with the purity of the
organic layer and the aqueous layer. It was important to make sure when using the separatory
funnel that the extractions were precise. By extracting over several fractions, it helped to make
sure the organic and aqueous layers were sufficiently separated. Otherwise the ending results and
the TLC plate would have shown the impurities.

Distance moved by compound

Rf value equation​: _________________________
Distance moved by solvent

Amount of substance collected

Percent recovery equation​: __________________________
​ ​Amount of substance expected

TABLE 1. Rf VALUES

Substance Distance from Distance Rf Value

Baseline (cm) Between

Baseline and

Solvent

Crude Caffeine (Spot 1) 1.6 4.5 0.35

Pure Caffeine (Spot 3) 1.6 4.5 0.35

 The uncertainty of the distance

of the spot is ±0.001 cm

Table 1 (shown above) concludes that the extracted crude caffeine sample is very pure being that
it has an identical Rf value to the known sample of pure caffeine.

TABLE 2. PERCENT RECOVERY

Weight of the 6 tea bags prior to brewing: 13.482 grams

Amount of expected caffeine (2% of total tea bag 0.269 grams

weight):

Weight of dry 100 mL round bottomed flask: 44.345 grams

Weight of flask after rotary evaporation: 44.439 grams

Weight of crude caffeine product: 0.094 grams

Percent Recovery: .065/.269 =.35 x 100 = 35%

Percent recovery is determined to be 35% according to Table 2 (shown above).

It is possible that the data suggests some pure caffeine may have been lost throughout the
experiment due to human error.
Discussion Questions
1. Prepare a flowchart for the experiment including appropriate structures and explicitly
state what compounds are present and in which layer/fraction for each step.
-picture attached in email.

4. Percent recovery and percent yield are occasionally used as interchangeable terms. They do,
however, have different meanings. Why is it more correct to use percent recovery here?
-In this case, it is more correct to use percent recovery because we are comparing a pure
compound to an initial compound. For example and extracted sample in comparison to a known
sample. Percent yield is determined by actual yield vs. theoretical yield and that is not fitting for
this extraction experiment.

5. Would the apparent percent recovery be higher, lower, or unchanged if a student did the
following:
a. Used room temperature water to brew the tea?
b. Did not use sodium sulfate when directed?

-5a: If a student used room temperature water instead of boiling water to brew the tea, the
percent recovery would ultimately be lower due to a decrease in solubility level. A higher water
temperature allows more caffeine to be released at the time of extraction.
-5b: If sodium sulfate was not used when directed, percent recovery would be higher due to an
abundance of unwanted water in the organic layer. The objective in adding the sodium sulfate is
to remove any remaining water in the solution to find the purest form of caffeine as possible.