Chromatography
Charmaine Almonte, Arnold Balais Jr., Jan Michael Eli Bautista*, and Arvin Raphael
Bayot
ABSTRACT
Tea leaves (Thea sinensis) contain about 15-35mg of alkaloid caffeine which is known to have a
mild stimulating effect to the central nervous system. In the experiment, the group extracted, isolated and
characterized the caffeine present in the leaves. The members applied numerous steps in order to obtain
the results. The calculations are done through following a formula which then led to a 0.72% percent yield
of crude caffeine. In addition, the group performed Thin Layer Chromatography in order confirm the purity
of the crude caffeine.
INTRODUCTION
However, caffeine is not only commercialized within products such as coffee, but
also in tea leaves. Teas contain about 15-35 mg worth of caffeine, in contrast to the
average of 135 mg worth in coffee [3]. In this experiment, our group aims to isolate,
purify, and characterize the caffeine extracted from Lipton tea leaves, calculate the
percentage yield of this product, and finally testing its purity via Thin Layer
Chromatography.
METHODOLOGY
Three bags of tea were distributed to each group. The bags were emptied and
placed in a tared 50 mL beaker then the weight of the tea leaves was obtained. The tea
leaves were placed back in the tea bags. The tea bags were put in a boiling 100 mL
distilled water and let it boiled for 5 minutes. Next, the hot tea extract was cooled to
room temperature and was placed in a separatory funnel with 20 mL DCM
(dicholoromethane) to produce DCM extract (a) and Tea extract.
The tea extract was extracted with fresh 20 mL DCM which will then produce
DCM extract (b) and tea extract. The DCM extract (b) was combined with DCM extract
(a) to give 40 mL DCM extract. While the tea extract was extracted with another fresh
20 mL DCM which will produce DCM extract (c) and another tea extract that will be
discarded. The 40 mL DCM extract was combined with DCM extract (c) to give 60 mL
DCM extract. The 60 mL DCM extract was transferred using a rinsed separatory funnel
with 20 mL 6M NaOH extract to make an aqueous and DCM layer. The DCM layer was
dried with anhydrous Na2SO4 (sodium sulfate) and decanted to an evaporating dish
until the next meeting. Next, the dried caffeine was collected in a vial by using a micro
spatula. The percent yield of the caffeine was computed and then, a pea sized portion
of the caffeine was placed inside a calibrated 2 mL test tube, which contained 1 mL of
distilled water. It was later blotted on a particular point (point B) on a TLC plate, and
point A was blotted with pure caffeine. This was done in order to test its purity by
comparing the number of bubbles formed on the side of the pure caffeine against the
crude caffeine obtained by the group.
RESULTS
Part A
TLC plate
DISCUSSION
The last concept of the experiment is the Thin Layer Chromatography. Thin-layer
chromatography involves the same principles as column chromatography, it also is a
form of solid-liquid adsorption chromatography. In this case, however, the solid
adsorbent is spread as a thin on a plate of glass or rigid plastic. A drop of the solution to
be separated is placed near one edge of the plate, and the plate is placed in a
container, called a developing chamber, with enough of the eluting solvent to come to a
level just below the "spot." The solvent migrates up the plate, carrying with it the
components of the mixture at different rates. The result may then be a series of spots
on the plate, which means that the caffeine was not pure. However, in the results, it is
seen that our crude caffeine is in fact, pure caffeine since its TLC result was equivalent
with one another.
CONCLUSION
Caffeine is indeed found in tea leaves. It is confirmed by making use of
Dichloromethane to extract the product of interest from the substrate, and it is left to
sublime on an evaporating dish. The caffeine extracts purity is also confirmed by
comparing the bubbles formed on a TLC plate, where in points A and B contained a
drop of pure and crude caffeine respectively.
REFERENCES
[1] Retrieved February 18, 2017 from the World Wide Web:
http://scholars.unh.edu/cgi/viewcontent.cgi?article=1102&context=honors
[2] Retrieved February 18, 2017 from the World Wide Web: http://health.howstuffworks.com/wellness/drugs-alcohol/caffeine-
awake.htm
[3] Laboratory Experiments in Organic Chemistry. Carlos P. Garcia. University Publishing House, Manila, 2005.