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Synthetic Experiment 1: Isolation and Purification of Caffeine from Tea

Riddhi Patel
Madison Salvitti
Chemistry 213 W 001
June 25, 2016

Introduction: Caffeine is an alkaloid, nitrogen containing basic compound, found in

plants that is an active ingredient in coffee beans, tea leaves, cocoa and more. It acts as a stimulant

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in humans, stimulating the heart, respiration, and the central nervous system.3,4 It can be physically
addictive, potentially causing nervousness, irritability and headaches.2 It is also a well-known
diuretic.3 Some benefits include improved alertness, learning capacity and exercise performance.4
It can be useful in treating migraines, relieving asthma attacks and increasing the potency of
analgesics. Sometimes it can be used as a pesticide or an aromatic flavor4
Of the four main types of teas (white, green, oolong and black), black tea tends to
contain the highest amount of caffeine1, guaranteeing a relatively substantial yield, which is why
it is the chosen starting material. There are several different ways of extracting caffeine from tea
leaves; solid liquid extraction followed by liquid-liquid extraction and sublimation are just two
examples. Once extracted, the caffeine needs to be purified as many of the tannins in the tea leaves
could have been extracted along with the caffeine.

The isolated compound, Caffeine

Figure 1. The isolation of 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione or the compound
more commonly known as caffeine, from English Teat Time Black Tea leaves.
Solid-Liquid and Liquid-Liquid extraction are very commonly used techniques to
separate compounds in a mixture by using acid base chemistry to pull certain compounds into the

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organic or aqueous layer.6 High solubility of caffeine in hot water is the reason it is the chosen
extractant to dissolve all of the caffeine in it from the tea bag. Dichloromethane is an organic
solvent thus adding it to the aqueous tea results in the formation of an aqueous layer followed by
an organic layer (since it is denser than water).7 Since caffeine is an organic compound,
dichloromethane is used as the extractant to pull the desired compound, caffeine, into the organic
layer, separating it from the water and any other inorganic compounds/ salts residing in the
aqueous layer as a result of their polarity.
Thin Layer Chromatography (TLC) is an important technique in organic chemistry that
can be used to monitor the progress of a reaction and to verify a substances identity based on
molecular interactions and the use of a standard.5 Column Chromatography (CC) involves the
same principles as TLC except it can be applied to separate mixtures, both solids and liquids on
small or large scale. The experiment aimed to collect the purified caffeine and CC allows for the
collection of the separated components, which is why CC was the chosen purification method.5
Tea contains many other tannins, large acidic molecules, and pigments which can be separated
from caffeine based on difference in their polarities/affinity to the polar adsorbent alumina versus
the mobile ethyl acetate. This is another reason the column was run to purify caffeine.
Caffeine is a very important and most common psychoactive analgesic/stimulant used by
majority of the population, with many health benefits as well as risks. The purpose of this
experiment was to extract caffeine from black tea and to purify it using column chromatography.
The product was isolated and characterized by IR, 1HNMR (60 MHz and 400 MHz), 13CNMR and
melting point determination (Caffeine M.P.: 236 C 237.5 C).3
Experimental: Caffeine: English Teatime Black tea (2 tea bags, 45 mg caffeine/bag) were steeped
in hot water for 30 minutes. The aqueous tea solution was extracted with dichloromethane (3 x 20

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mL). The organic layers were collected. The organic solution was washed with saturated sodium
chloride (20 mL) and dried over anhydrous sodium sulfate for 20 minutes. The resulting solution
was analyzed using TLC (100% ethyl acetate) and evaporated with nitrogen to yield a crude
product (0.07 g, 0.361 mmol). The crude caffeine sample was dissolved (0.4 mL dichloromethane
and 0.2 mL ethyl acetate). Column Chromatography (100% ethyl acetate) monitored by TLC
(100% ethyl acetate), afforded a pure white powder (0.04 g, 0.206 mmol, 44% yield). 1HNMR
(400 MHz, CDCl3): (ppm) 3.24 (s, 3H), 3.41 (s, 3H), 3.89 (s, 3H), 7.88 (s, 1H); .13CNMR (400
MHz, CDCl3): (ppm) 27.78, 29.65, 33.51, 107.33, 141.33, 148.48, 151.54, 155.26; IR (ATR)
max (cm-1): 3426.76, 3110.32, 2952.35, 1692.02, 1642.45, 1546.24, 1448.13, 1357.80 and
1232.78. M.P.: (237.1 C 241.1 C).
Results and Discussion:
Extraction of caffeine from tea leaves and coffee beans among other plants, is a very
important and widely practiced process known as decaffeination or removal of caffeine, regardless
of whether the goal of extracting the caffeine is to consume a less caffeinated product or a more
concentrated one. In this experiment, caffeine was isolated from black tea leaves by using solidliquid extraction followed by liquid-liquid extraction.
The method involved steeping the tea bags in hot water since the solubility of caffeine
increases significantly with temperature causing it to be extracted into the aqueous solution.
Caffeine is only minimally soluble in water at room temperature (2.2 mg/ml) 8 but its solubility
greatly increases at higher temperatures (670 mg/ml at 100C).8 Thus, using hot water (100C) as
an extractant in the solid-liquid extraction process allows the caffeine from the tea bags to
completely dissolve in it initially, and then upon cooling, the caffeines decreased solubility in

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water makes it very easy to extract from the other acidic tannins and by-products found in tea
leaves.
Tea leaves contain acidic tannins, cellulose, undecomposed chlorophyll, and pigments
along with caffeine.3 For a successful liquid-liquid extraction, caffeine must be present as free base
and acidic tannins must remain water soluble. Therefore, caffeine is extracted from the aqueous
tea solution by washing with dichloromethane (d = 1.325 g/mL)5 with which has proven to be the
highest percent yielding extractant in previous studies.6 Dichloromethane, an organic solvent, is
an alkyl halide and dissolves caffeine very well because of its polarity being similar to caffeine.
Dichloromethane also has a low boiling point (39.75 C)8, doesnt react with the solute or any
aqueous by-products, and is immiscible in water, making it an ideal solvent for separating the
homogeneous solution. Because it is denser than water, the caffeine containing organic layer lies
on the bottom and the aqueous layer floats on top. The organic solution was washed with brine, or
saturated sodium chloride, to remove any remaining water droplets or inorganic salts. A drying
agent, anhydrous sodium sulfate, was used to dry the solution and remove any traces of water by
absorbing the water molecules and assuming its hydrous state.
The crude caffeine product was then purified using Column Chromatography with
100% ethyl acetate as the mobile phase, considering the highly polar nature of caffeine evident
with the Rf value of 0.03 on average, followed by evaporation of the solvent. Ethyl Acetate is a
slightly polar organic solvent that is effective in running TLC analysis on highly polar molecules
such as caffeine with the two ketones and two amide functional groups contributing to its ability
to form hydrogen bonds, 6 electronegative atoms (4 nitrogen and 2 oxygen atoms), the big dipole
moment (difference in the electronegativities of atoms in C=O and C-N bonds) and high
polarizability along with its high molecular weight (size).5 And since column chromatography

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follows the same basic principles as TLC, ethyl acetate is also used as the mobile phase to elute
the caffeine.
The experiment resulted in 0.070 g crude product and 0.040 g of purified caffeine from
the two tea bags that contain 45 mg caffeine each on average.1 This meant the final product yield
was 44% and percent recovery of the pure caffeine from the crude was 58%. This is consistent
with the percent yield found in studies that involved extracting caffeine from tea leaves. In a study
conducted by Onami and Kanazawa that aimed to extract caffeine from black tea leaves using
dichloromethane-alkaline water as an extractant, the yield was shown to be 45% in the first several
minutes and 58% over time.3 Alkaline water seemed to be more effective than water when
combined with dichloromethane as an extractant.3
The IR spectra shows peaks at: 3426.76 cm-1, 3110.32 cm-1, 2952.35 cm-1, 1692.02 cm-1,
1642.45 cm-1, 1546.24 cm-1, 1448.13 cm-1, 1357.80 cm-1 and 1232.78 cm-1, indicating the presence
of alkene or aromatic hydrogens (attached to a carbon in N-C=N cyclic structure), aliphatic CH bonds, a C=O bond attached to a C=C bond, a carbonyl in an amide group, C=N bond, C-N
bond and C-C bond, respectively (Figure 1, Supplemental Information). 1HNMR (60 and 400
MHz) show three singlet peaks at 3.409 or 3.24 ppm, 3.585 or 3.41 ppm and 3.995 or 3.89 ppm
integrating very close to 3.00 hydrogens per peak; each of these three peaks are characteristic of
the three hydrogens in methyl groups attached to nitrogen atoms in caffeine. The fourth singlet
peak, observed at 7.518 or 7.88 ppm, integrating to about 1.00 hydrogen is indicative of the
hydrogen attached to the carbon double bonded to one nitrogen atom and single bonded to the
other. Since the starting material (tea leaves) consists of a lot of tannins which are huge molecules
and if present, would display plenty of supplemental peaks pointing to the extra hydrogens, but

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the absence of any additional hydrogens on the 1H NMR suggests little
to no contamination (Figures 2 and 3, Supplemental Information).
.13CNMR (400 MHz) showed peaks at 27.78 ppm, 29.65 ppm, 33.51
ppm, 107.33 ppm, 141.33 ppm, 148.48 ppm, 151.54 ppm and 155.26
ppm each integrating to one carbon per peak as there are 8 unique carbons in caffeine altogether.
The only contaminated peaks observed around 77 ppm were characteristic of CDCl3 solvent used
in the preparation of sample for NMR analysis (Figure 4, Supplemental Information).
In conclusion, the process of extracting caffeine from black tea leaves and purifying it
through Column Chromatography proved to be relatively effective as evident by the 44% yield
of pure caffeine and the melting point range of the purified product, 237.1 C 241.1 C which is
extremely close to the actual melting point range of caffeine, 236 C 237.5 C.3 The spectral
data including 1HNMR, 13CNMR and IR analysis (Supplemental Information) all indicate
successful purification as the sample isnt lacking or showing any extra peaks and integrates very
closely to the expected number of atoms, thus proving that the product is not contaminated with
the starting material or any other known by-products. Moreover, the percent recovery of 58%
further indicates that the crude product was purified of the contaminants while still yielding a
substantial amount of the desired product. Sources of error could include failing to maintain the
high temperature of water throughout the 30 minutes of solid-liquid extraction, theoretical
amount of caffeine in the tea bag varying from the average assumed, not eluting all of the
caffeine when running the column due to some air bubbles in the column, not draining/collecting
the organic layer completely or perhaps draining some of the aqueous layer along with it.
Research studies comparing methods for picking up caffeine have discovered that blending
certain amounts of alkali sodium carbonate in tea dust helps improve caffeine yield, and

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strengthen extraction rate.7 So future studies could experiment with compounds like alkali
sodium carbonate, different extractants/solvents, vary polarity of the mobile phases for TLC/CC
to determine which combination would yield a greater percent yield of caffeine with the highest
purity.
References:
1. "Bigelow Tea - Caffeine Levels." Caffeine Informer. Bigelow Tea, n.d. Web. 21 June
2016.
2. Dews, P., O'brien, C., and Bergman, J. (2002) Caffeine: behavioral effects of withdrawal
and related issues. Food and Chemical Toxicology 40, 12571261.
3. Onami, T., and Kanazawa, H. (1996) A Simple Method for Isolation of Caffeine from
Black Tea Leaves: Use of a Dichloromethane-Alkaline Water Mixture as an Extractant. J.
Chem. Educ. Journal of Chemical Education 73, 556.
4. Robertson, D., and Curatolo, P. W. (1984) The Cardiovascular Effects of Caffeine.
Caffeine 7785.
5. Rummel, S.; Beiswenger, MK. Chemistry 213 Introductory Organic Chemistry
Laboratory, Hayden McNeil. 2015-16, pp 41-150.
6. Sharif, R., Ahmad, S. W., Anjum, H., Ramzan, N., and Malik, S. R. (2013) Effect of
Infusion Time and Temperature on Decaffeination of Tea Using Liquid-Liquid Extraction
Technique. Journal of Food Process Engineering J Food Process Eng 37, 4652.
7. Tang, W.-Q., Li, D.-C., Lv, Y.-X., and Jiang, J.-G. (2010) Extraction and Removal of
Caffeine from Green Tea by Ultrasonic-Enhanced Supercritical Fluid. Journal of Food
Science 75.
8. Williamson, K and Katherine Masters. Macroscale and Microscale Organic Experiments,
6th ed.; Brooks/Cole, 2011.