Tafline Grace Borromeo Sia 29C

Extraction of Caffeine from Tea 26 November 2019

I. Data and Results [10.0 pts]

Table 1. Gravimetric data for the extraction of caffeine from tea

Mass of tea bags 9.02 g

Mass of extracted caffeine crystals 140 mg

Appearance of caffeine crystals light brown needles

Caffeine content per tea bag [1] 44.8 mg

Total amount of caffeine present in tea bags 179.2 mg

% yield 78.1 %

Table 2. Comparison of retention factors for the caffeine-containing samples

Rf

Substance Spot 1 Spot 2 Spot 3

Caffeine standard 0.30 - -

Tea 0.30 0.13 0.39

Pepsi 0.30 - -

Figure 1. TLC visualisation plate. From left to right: caffeine standard, Lipton tea, Pepsi
Cola. The plate was developed using a 20:10:3:0.2 benzene/diethyl ether/acetic acid/methanol
solvent system in a scintillating vial, and the spots were visualised with a UV lamp. TLC
analysis indicated that the tea extract contained two components in addition to caffeine.

II. Discussion [20.0 pts]

Tea leaves contain several components that are extracted during the brewing process.
One of these is caffeine, which is a bitter, slightly polar alkaloid that is soluble in both water
and organic solvents like ethyl acetate. Caffeine is a powerful central nervous system
stimulant and vasodilator that is found in coffee, tea, and energy drinks. There are three main
methods of decaffeination, but the most common and least costly method is the direct solvent
method, which utilises an organic solvent such as dichloromethane or ethyl acetate to extract
caffeine from beverages [2]. In this experiment, the direct solvent method was used to isolate

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caffeine from both Lipton Yellow Label tea and Pepsi Cola. Ethyl acetate was used as the
organic solvent because it readily dissolves caffeine and is less toxic and hazardous than
dichloromethane [3]. The presence of caffeine in the two extracts was then verified using thin
layer chromatography by comparing the calculated retention factor values of each sample to
that of caffeine standard.
To extract caffeine from tea, four tea bags were steeped in hot water with added
sodium carbonate. The basicity of the solution not only ensured that the caffeine would be
readily solvated in organic solvents and less attracted to water, but also that the tannins would
not co-dissolve with caffeine in ethyl acetate [3]. A selective liquid-liquid extraction using
multiple portions of ethyl acetate was performed to extract caffeine from the mixture. This
was done in order to maximise the extractive power of the solvent [4]. Afterwards, the
harvested caffeine was purified through simple distillation and dried with sodium sulfate. A
similar procedure was utilised in order to extract caffeine from Pepsi Cola.
Using the direct solvent method, a total of 140 mg of caffeine was harvested from the
four tea bags of Lipton Yellow Label Tea. This amounts to 78.1% of the maximum amount of
caffeine that could be extracted from the tea bags (Table 1), a moderate yield that was due to
the use of multiple extractions. f a single liquid-liquid extraction had been performed using
30 mL ethyl acetate, only 82% of the caffeine would be extracted into the organic layer. On
the other hand, the same amount of solvent used in five successive extractions would result to
96.3 mg caffeine being extracted from the aqueous layer, resulting in a percent yield of
96.3%. This shows that performing multiple extractions maximises the solvent’s extracting
ability and thus results in maximal recovery or yield of the material <AtQ#1> [4]. In
comparison, while the caffeine extracted from Pepsi Cola was neither recrystallised nor
measured, the amount was approximated using the nutrition facts provided on the company
website. One 16 oz. serving of Pepsi contains only 50 mg of caffeine [5]. This means that a
total of 1.06 mg of caffeine was extracted from 5-mL portion of Pepsi <AtQ#2>.
After the amount of harvested caffeine for both the tea and Pepsi extracts was
quantitated, the presence of caffeine in both samples was verified using a 20:10:3:0.2
benzene/diethyl ether/acetic acid/methanol solvent system to develop the TLC plate. Based
on the calculated retention factors, caffeine was indeed present not only in the tea extract but
also in the Pepsi extract (Table 2). However, while pure caffeine is white in color [6], the
caffeine extract from tea was discolored to a light yellow to beige color (Table 1) <AtQ#3a>.
Furthermore, instead of only exhibiting one spot corresponding to that of extracted caffeine,
the caffeine extract from tea also contained two more spots that were not present in the
caffeine standard (Table 2; Figure 1). This, along with the light brown color of the sample,
indicated that there were other compounds present in the sample, such as theobromine and
theophylline [7] <AtQ#4>. In contrast to this, only one spot was visualised for the Pepsi
sample (Table 2; Figure 1). This indicated that the caffeine extracted from Pepsi was pure.
As indicated by the spots of caffeine along the path of each sample and the retention
factor value of 0.30 for the three samples (Figure 1), the direct-solvent method using ethyl
acetate was successful in extracting caffeine from both tea and Pepsi Cola. A good yield of
caffeine was obtained because caffeine was extracted using a multiple extraction procedure.
However, although the caffeine extract from Pepsi Cola was pure, the caffeine extract from
tea contained impurities, such as theobromine and theophylline, which may not have been
properly removed from the sample, or else may have been extracted together with caffeine.
For future experiments, the purity of caffeine extracts may be improved by modifying the
experimental design to include vacuum sublimation and recrystallization as additional steps
to ensure the removal of contaminants <AtQ#3b>. The yield of caffeine may also be

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increased by using more solvent in the solid-liquid extraction phase so as to be able to extract
a greater amount of caffeine from the tea bags.

III. Sample Calculations [3.0 pts]

Maximum amount of caffeine that can be extracted (AtQ#1)

Single extraction
(𝑥/30𝑚𝐿)
7.8 = (100𝑚𝑔 − 𝑥)/50𝑚𝐿
3 𝑥 50
7.8( 5 ) = ( 30 )( 0.100𝑔−𝑥 )
0.468 - 4.68x = x
5.68x = 0.468 g
x = 82.4 mg

Multiple extractions

First extraction 𝑥 50
7.8 = ( 6 )( 0.0267𝑔−𝑥 )
(𝑥/6𝑚𝐿)
7.8 = (0.100𝑔 − 𝑥)/50𝑚𝐿 0.0249912 = 1.936x
𝑥
7.8 = ( )(
50
) x = 12.91 mg
6 0.100𝑔−𝑥
0.468 - 4.68x = x
Fourth extraction
1.936x = 0.0936 g (𝑥/6𝑚𝐿)
x = 48.3 mg 7.8 = (0.0267𝑔 − 𝑥)/50𝑚𝐿
𝑥 50
7.8 = ( 6 )( 0.01379𝑔−𝑥 )
Second extraction
(𝑥/6𝑚𝐿) 0.01290744 = 1.936x
7.8 = (0.0517𝑔 − 𝑥)/50𝑚𝐿 x = 6.67 mg
𝑥 50
7.8 = ( 6 )( 0.0517𝑔−𝑥 )
Fifth extraction
0.0483912 = 1.936x (𝑥/6𝑚𝐿)
x = 25.0 mg 7.8 = (0.00712𝑔 − 𝑥)/50𝑚𝐿
𝑥 50
7.8 = ( 6 )( 0.00712𝑔−𝑥 )
Third extraction
(𝑥/6𝑚𝐿) 0.00664= 1.936x
7.8 = (0.0267𝑔 − 𝑥)/50𝑚𝐿 x = 3.44 mg

Total amount of caffeine that can be extracted = (48.3+25.0+12.91+6.67+3.44)mg =96.32 mg

Maximum amount of caffeine that can be extracted for this experiment (AtQ#2)
Caffeine content of Lipton Yellow Label tea
224mg caffeine/1000mL water * 200mL water = 44.8 mg
% yield = actual/theoretical * 100 % = (140mg/179.2mg)*100% = 79.1%

Rf values
Substance: caffeine standard
Rf = distance travelled by spot/distance travelled by solvent
= 0.30

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IV. References [2.0 pts]
[1] Astill C, Birch M, Dacombe C, Humphrey P, Martin P. Factors Affecting the Caffeine and
Polyphenol Contents of Black and Green Tea Infusions. Journal of Agricultural and Food
Chemistry [Internet]. 2001 [cited 14 October 2019]; 49(11):5340-5347. Available from:
https://pubs.acs.org/doi/abs/10.1021/jf010759%2B
[2] Farah A. Chapter 15: coffee as a speciality and functional beverage. In: Paquin, P (ed).
Functional and Speciality Beverage Technology. Cambridge, UK: Elsevier Science; 2009.
370-295.
[3] Cammack J. Extraction of Caffeine from Tea [Internet]. Hayesville [Oregon]: Chemeketa
Community College; [cited 2019 November 24]. Available from
http://faculty.chemeketa.edu/jcammack/CH241-3B%20Lab/CH241B%20Labs/CH241%206
%20Caffeine%20Extraction%20F14.pdf
[4] Nichols L. Organic Chemistry Laboratory Techniques. Oroville, CA: Butte Community
College; 2016.
[5] The facts about your favorite beverages: caffeine [Internet]. Pepsico. 2019 [cited 14
October 2019]. Available from: https://www.pepsicobeveragefacts.com/home/caffeine
[6] National Center for Biotechnology Information. Pubchem. [Internet]. Bethesda
[Maryland]: National Library of Science; [cited 2019 November 23]. Available from:
https://pubchem.ncbi.nlm.nih.gov/
[7] Bispo MS, Veloso MCC, Pinheiro HLC, De Oliveira RFS, Reis JON, De Andrade JB.
Simultaneous Determination of Caffeine, Theobromine, and Theophylline by High-
Performance Liquid Chromatography. Journal of Chromatographic Science [Internet]. 2002
[cited 2019Sep25];40:45–8. Available from: https://pdfs.semanticscholar.org/4aff/84257a3ed
1ddac7ca35a2622d3e0b0819b33.pdf