Name: Dan Erson B.

Talplacido Experiments Performed: February 21 & 28, 2017

Section: 2ChE-A Report Submitted: March 28, 2017

Group No.: 13

Experiment No. 3: Extraction of Caffeine from Tealeaves

I. Introduction

The active ingredient that makes tea and coffee valuable to humans is caffeine. Caffeine

is classified as an alkaloid due to its structure. Alkaloids are a highly diverse group of

compounds that contain a ring structure and a nitrogen atom. In most cases, the nitrogen

atom is located inside the heterocyclic ring structure [1]. A classification based on

biosynthetic pathways is mostly used to categorize different alkaloid [1]. Alkaloids have a

wide distribution in the plant kingdom and mainly exist in higher plants, such as those

belonging to Ranunculaceae, Leguminosae, Papaveraceae, Menispermaceae, and

Loganiaceae [1]. Moreover, several alkaloids exhibit significant biological activities, such as

the relieving action of ephedrine for asthma, the analgesic action of morphine, and the

anticancer effects of vinblastine [1]. In fact, alkaloids are among the most important active

components in natural herbs, and some of these compounds have already been

successfully developed into chemotherapeutic drugs, such as camptothecin (CPT), a

famous topoisomerase I (TopI) inhibitor, and vinblastine, which interacts with tubulin.

Caffeine (chemically, 1,2,3-trimethylxanthine) is a white crystalline xanthine alkaloid,

which is bitter in taste. It is an alkaloid compound naturally found in tea leaves, cocoa beans,

nuts such as bitter kola and kola nut, and fruits of more than sixty plants [2]. Industrially,

extract of caffeine is used in the production of soft drinks, coffee, and tea and in some

processed foods. In medicine, it is chiefly a psychoactive drug used for nervous system

stimulation. Nawrot et al. [3] noted that caffeine is probably the most frequently ingested

pharmacologically active substance in the world, found in common beverages (coffee, tea,
and soft drinks), products containing cocoa or chocolate, medications including headache or

pain remedies, and over-the-counter stimulants [2]. The wide use of caffeine in productions

and its increasing world consumption could be the reason why little or no effort has been

made to set aside the existing controversy surrounding the conflicting results generated by

the use of variable forms of caffeine and experimental methods towards the study of its

possible toxicity [2].

Extraction is the technique used to separate an organic compound from a mixture of

compounds. The theory of extraction lies in the concept of immiscibility between two phases

to separate a solute from the other phase. [3] There are two main types of extraction in

chemistry namely: liquid-liquid extraction (also known as solvent extraction or partitioning),

which is further divided into two types (single and multiple), and solid-liquid extraction. [4]

Liquid-liquid technique is a very common method in Organic Chemistry [4].

Among the materials to be utilized in this experiment, tea is central, being the source of

the component to be extracted: caffeine. This second-most popular drink in the world comes

in a variety of forms-green, black, oolong- but is scientifically noted for its high content of

polyphenols. This substance combats free-radicals and prevents cancer and other body

ailments. With the tea leaves tough cellulose insoluble in water, boiling has been an easy

separating method to get at the chemicals in store (e.g. caffeine, tannin, ascorbic acid, etc.).

To isolate one of these components is a different matter, however [5].

In order to extract caffeine from tea, several methods are used. First, a solid/liquid

extraction must take place in order to get the solid natural product into the liquid solvent. In

order to isolate the desired reaction compounds from the natural product, liquid/liquid

extractions are used.

Neutral and acid/base are two forms of liquid/liquid extractions [6]. Caffeine extraction

from tea leaves involves an acid/base liquid/liquid extraction [8]. The reaction involves a

homogenous mixture of an organic and aqueous layer. The ideal solvent in the extraction
 should have a low boiling point, not react with the solute or other solvents, not be toxic or

highly flammable, not miscible with water, be inexpensive, and should readily dissolve

caffeine at room temperature. A common liquid/liquid solvent pair for the extraction of

caffeine is water-dichloromethane [6]. Because water is present in the pairing, it possible to

separate inorganic compounds from organic compounds due to the fact that organic

substances are immiscible in water [9].

When mixing the liquid pairs, the density of the both solvents predict which solvent is the

top and which the bottom layer is. Caffeine, which was present in the organic layer, was

located below the aqueous layer [6]. The product that is collected after extraction still has

many impurities. Sublimation is one way to purify the sample, because caffeine has the

ability to pass directly from the solid to vapor and reverse to form a solid all without

undergoing the liquid phase. Caffeine has the ability to undergo sublimation under different

conditions than the impurities, and can thus be isolated [7].

II. Methodology

For the extraction of crude caffeine from teabags, the weight of 3 teabags were

measured after that it was boiled in a 100 mL of water (H 2O) for 5 minutes. The mixture that

has been boiled was filtered using a fluted filter paper and transferred in an erlenmeyer flask.

The mixture was cooled down first by running tap water for 2 minutes and then was put in an

ice bath. It was then transferred in a separatory funnel which contained 20 mL of

dichloromethane (CH2Cl2). The lower layer (CH2Cl2) was extracted and another 20 mL of

dichloromethane (CH2Cl2) was used to extract again from the remaining mixture in the

separatory funnel. The water layer was discarded and both the extracted dichloromethane

(CH2Cl2) was mixed. The combined mixture was then put in a separatory funnel and washed

using 20 mL of sodium hydroxide (NaOH). Sodium hydroxide (NaOH) was discarded and the

remaining dichloromethane (CH2Cl2) was transferred in a clean erlenmeyer flask containing

 half spatula of anhydrous sodium sulfate (Na2SO4). The extract was then concentrated to

dryness over a boiling water bath until it forms white solids which is the crude caffeine from

the tea bags.

For purification, the flask with crude caffeine inside was sealed using a filter tube with a

fitted inner test tube serving as a cold finger and also an aluminum foil for which the vapor

will not escape, then it was put in an air bath with the beaker under an intense Bunsen flame

for 35 minutes. The inner test tube has an ice water inside and the ice water was constantly

replaced into new ice water every 2 minutes. After 35 minutes, the caffeine that clang in the

in cold finger and also into the aluminum foil was scraped and then weighed.

For characterization of crystalline caffeine, the crystals that was acquired was grinded

until it was a very fine powder. One side of the micro capillary was sealed by burning one

side of it using a Bunsen burner. With a sealed micro capillary, the pulverized crystal powder

was scooped inside the micro capillary until the height is 1 cm and was packed by letting the

micro capillary fall inside a 1-meter long glass tubing. It was then the micro tube was

attached to a thermometer and then clamping the thermometer to an oil bath using a heated

beaker. It was used to determine the melting point of caffeine. The first point which the

caffeine melted was the initial temperature while the final temperature is when the caffeine

was fully melted.

III. Results and Discussion

Result:

Weight of tea leaves used, g 6.26g

Weight of evaporating dish, g 88.65g
Weight of evaporating dish with tea 94.91g

leaves, g
Weight of flask, g 114.05

g
 Weight of flask with caffeine, g 114.24

g
Weight of pure caffeine, g 0.14g

Table 1. Measured Weights of Tea Leaves,

Dish, Flask/Caffeine, Caffeine Product

The various weights of flasks and dish were obtained in order to calculate the final

amount of caffeine product in grams.

weight of caffeine 0.14 g

Percent Yield = weight of tealeaves x 100 = 6.26 g x 100 = 2.24 %

Discussion:

A B

Figure 1. Tea extract and dichloromethane (A) and dichloromethane and

sodium hydroxide (B)

 The present study was conducted to isolate pure caffeine from the tealeaves. The

tealeaves weighing 6.26g was boiled in a 100 mL of water. Caffeine was isolated using

dichloromethane as an extracting solvent. The reason why the teabags was dissolved in

water because dichloromethane is immiscible with water. The organic solvent

dichloromethane is used to extract caffeine from aqueous extract of tea powder because

Figure 2. Hot air bath set-up

with aluminum foil

caffeine is more soluble in dichloromethane (140mg/ml) than it is in water (22mg/ml).The

dichloromethane caffeine mixture can then be separated on the basis of the different

densities of dichloromethane and water because dichloromethane is much denser than

water and insoluble in it (Figure 1).

Residual water is separated from dichloromethane by drain out the dichloromethane

through separating funnel, thus dichloromethane passed through the funnel while polar

solvents such as water is still remains in the funnel. Water and dichloromethane is slightly

soluble in each other. So, after separating the solvents, residual water will remain the

organic layer. Mainly anhydrous sodium sulfite is used for the removal of water from organic

layer. Anhydrous sodium sulfite is an insoluble inorganic solid which will absorb water, thus
 drying it. [10] The crude caffeine was then purified by using a hot air bath (Figure 2) with a

stopper with an inner test tube serving as a cold finger.

The reason for the use of aluminum foil is to make sure that the caffeine will cling into the

test tube and will not escape the into the surroundings, by doing this, it limits the possible

loss of pure caffeine. Weight of flask with pure caffeine was taken and found 114.24g.

Amount of pure caffeine (0.14g) was calculated by subtracting weight of empty flask

(114.05g) from weight of flask with crude caffeine (114.24g).

In determining if the caffeine is pure, capillary melting points is used as a criteria for

purity. A few crystals of the compound are placed in a thin walled capillary tube 10-15 cm

long, about 1 mm in inside diameter, and closed at one end. The capillary, which contains

the sample, and a thermometer are then suspended so they can be heated slowly and

evenly. The temperature range over which the sample is observed to melt is taken as the

melting point. The presence of impurities in the sample can also cause the sample to melt

over a range of temperatures. Thus, the melting point will usually be reported as a melting

range, the temperatures between which the sample melted. The first bubble appeared at the

temperature of 98C indicates the boiling point of the caffeine acquired and the caffeine

disappeared completely at the temperature reading of 237C which indicates its melting

point.

IV. Conclusions and Recommendations

The objectives of the experiment is to isolate, purify and characterize caffeine from tea

leaves and to calculate the percentage yield of caffeine that will be acquired in the

experiment. The acquired pure caffeine weighing 0.14g from the initial tealeaves weighing

6.26g. The percentage yield computed was 2.24%, it indicates how much caffeine content is

in the tea leaves, however this result may still have impurities because the substance

acquired was liquid and not solid. The melting point range was 98C - 237C. The
 appearance of the first bubble at 98 C indicates that the acquired caffeine is almost water-

like.

The group recommends to be more careful in choosing the tea to be used in the

experiment. One big factor which caused the group to attain a very small amount of caffeine

is by using an herbal tea. Herbal teas are claimed as caffeine-free. Another is in putting the

tea leaves back in its bag after weighing it, caution is needed since it is very tiny in

composition. A small change in the mass recorded and the actual mass of tea leaves used

will contribute to a significant change in computing the percentage yield of the caffeine at the

end of the experiment. During the extraction of dichloromethane (CH2Cl2) and sodium

hydroxide (NaOH), make sure that method of swirling the separatory funnel is followed.

Make sure that the Erlenmeyer flask does not touch the beaker during hot air bath to prevent

the beaker from breakage. During the oil bath of the pulverized caffeine extract stored in a

micro capillary tube, it is advisable to keep distant from the setup because the oil may

splatter anytime. The group members must be attentive enough in determining the minimum

temperature of the sample indicated by the thermometer when the first bubble inside the

micro capillary tube is produced. Lastly, every member of the group must be cautious in

handling and assembling the laboratory apparatuses used in the experiment.

V. References

[1] Jin-Jian Lu, Jiao-Lin Bao, Xiu-Ping Chen, Min Huang, and Yi-Tao Wang, Alkaloids

Isolated from Natural Herbs as the Anticancer Agents, Evidence-Based Complementary

and Alternative Medicine, vol. 2012, Article ID 485042, 12 pages, 2012.

doi:10.1155/2012/485042. Date visited: March 4, 2017

[2] Agomuo Emmanuel, Duru Majesty, Amadi Benjamin, Amadi Peter, and Ugwokaegbe

Princess, Effect of Caffeine on Some Selected Biochemical Parameters Using Rat

Model, Advances in Biology, vol. 2017, Article ID 9303276, 8 pages, 2017.

doi:10.1155/2017/9303276 Date visited: March 4, 2017

[3] P. Nawrot, S. Jordan, J. Eastwood, J. Rotstein, A. Hugenholtz, and M. Feeley, Effects of

caffeine on human health, Food Additives and Contaminants, vol. 20, no. 1, pp. 130, 2003.

Date visited: March 4, 2017

[4]Crystallization of Caffeine. Retrieved from: www.ou.edu/englhale/pronarmode.doc

Date visited: March 4, 2017

[5] Extraction and Purification of Caffeine. Retrieved from:

http://www.linfield.edu/assets/files/chem/Courses/CHEM%20321/2014-week3-4-caffeine-

chem321l-53ebfd7b679e5.pdf Date Visited: March 5, 2017

[6] Williamson, K and Katherine Masters. Macroscale and Microscale Organic Experiments,

6th ed.; Brooks/Cole, 2011. Date visited: March 5, 2017

[7] Tello, J. Extraction of caffeine from Robusta coffee (Coffea canephora var. Robusta)

husks using supercritical carbon dioxide. [Online] 2011, 53-60.

www.elservier.com/locate/supflu Date visited: March 6, 2017

[8] Oneota. Isolation of Caffeine from Tea Leaves. [Online] 2003. Retrieved from:

http://employees.oneonta.edu/ knauerbr/chem226/226expts/226_expt06_pro.pdf Date

visited: March 6, 2017

[9] Amrita University. Extraction of Caffeine from Tea. 2013 Retrieved from:

http://vlab.amrita.edu/?sub=3&brch=64&sim=169&cnt=1 Date visited: March 6, 2017

[10] AHMAD, Iftekhar et al. EXTRACTION OF CAFFEINE FROM TEA AND

DEVELOPMENT OF CAFFEINATED FRUIT JUICE. Journal of Food Chemistry and

Nutrition, [S.l.], v. 1, n. 1, p. 01-05, june 2013. ISSN 2307-4124. Retrieve from:

http://escijournals.net/index.php/JFCN/article/view/72/151. Date visited: March 6, 2017