Extraction Of Caffeine From Tea Leaves

Principle of the experiment:

The technique used to separate an organic compound from a mixture of compounds is called Extraction.
Extraction process selectively dissolves one or more of the mixture compounds into an appropriate
solvent. The solution of these dissolved compounds is referred as the Extract. Caffeine is a naturally
occurring alkaloid (a class of naturally occurring compounds containing nitrogen and having the
properties of an organic amine base) found in coffee, tea, Cocoa, Guarana (Paullinia cupana) and kola
nuts.

Caffeine belongs to a family of naturally occurring compounds known as xanthines. Caffeine is

chemically 1,3,7-trimethylxanthine (C8H10N4O2). Caffeine acts as a stimulant. It stimulates the heart,
respiration, the central nervous system, and is a vasodilator (relaxes the blood vessels) as well as a
diuretic (increases urination). It is a beverage produced by steeping in freshly boiled water the young
leaves and leaf buds of the tea plant, Camellia sinensis. Two principal varieties of tea are used, the small-
leaved China plant (C. sinensis) and the large-leaved Assam plant (C. assamica). The leaves may be
fermented or left unfermented. Fermented teas are referred to as black tea, unfermented teas as green
tea, and partially fermented teas as oolong tea. Tea leaves consist mostly of cellulose (a water-insoluble
polymer of glucose), caffeine, tannins (phenolic compounds, compounds that have an -OH directly
bonded to an aromatic ring) and a small amount of chlorophyll. The solubility of caffeine in water is 22
mg/ml at 25·C, 180 mg/ml at 80·C, and 670 mg/ml at 100·C. Here the organic solvent dichloromethane is
used to extract caffeine from aqueous extract of tea leaves because caffeine is more soluble in
dichloromethane (140 mg/ml) than it is in water (22 mg/ml). However, the tannins that are slightly soluble
in the dichloromethane can be eliminated by converting it to their salts (phenolic anions by adding sodium
carbonate) (tannins are phenolic compounds of high molecular weight and being acidic in nature can be
converted to salts by deprotonation of the -OH group) which remain in the water.

Procedure

Place 30 g of the tea leaves in a 500 ml beaker. Add 250 mL of distilled water and 5 g of sodium
carbonate and stir the contents of the beaker with a glass rod. Boil the contents of the beaker on a hot
plate/water bath for 10 minutes. Cool the tea solution to room temperature using an ice-water bath. Filter
the cooled solution using glass wool or muslin cloth or cheese cloth. Transfer the filtrate into a separating
funnel and extract 3-4 times (4 x 10-25 mL) using dichloromethane. Take care to avoid formation of
emulsion for each extraction. Don’t shake the separating funnel vigorously, but gently swirl the two
immiscible layers for 5 minutes. After each extraction, remove the lower organic layer into 250 mL beaker,
leaving any emulsion layer behind. Add anhydrous sodium sulphate to the combined extracts. The
sodium sulfate will remove any water and water soluble salts that are retained in the dichloromethane
(organic layer) or accidentally transferred during decantation. Filter to remove the solid sodium sulphate
and transfer the dry solution to a pre-weighed 250 mL beaker. Evaporate it to dryness by boiling it on a
water bath. (It can also be evaporated under vacuum or by blowing dry air or nitrogen gas on the surface
of the liquid). The residue will be crude caffeine (usually slight green in color) (determine its weight-the
yield will be 0.5 g approx for 25 g of tea leaves). Purify the crude caffeine either by sublimation or by
recrystallization using methanol. Caffeine is reported to sublime at 170°C. Pure caffeine appears as white
glistering needles. It is bitter in taste. Determine the melting point using a sealed capillary. The melting
point of caffeine is 238°C.

Identification test

Murexide test

Add few crystals of caffeine with 3-4 drops of conc. nitric acid in a porcelain dish and evaporate to
dryness. Addition of 2 drops of ammonium hydroxide to the residue gives purple color.

Thin Layer Chromatography

To assess the purity of caffeine extracted perform TLC on both for your caffeine sample and an authentic
sample of caffeine. Add a few drops of ethanol to caffeine on your watch glass. This will not dissolve all of
it, but enough will dissolve to allow you to do TLC on the solution. Spot the TLC plate with the isolated
caffeine and with authentic caffeine from a solution of caffeine in ethanol. Label both the sample and
authentic sample on your plate. Develop the chromatogram using a mixture of ethyl acetate (95%) and
acetic acid (5%) as mobile phase. After the development air dry the plate and examine the plate under UV
light to observe the spots. Outline the spots with a pencil and confirm the purity of caffeine from Rf value
(It is the ratio of distance traveled by sample spot from the origin to the solvent front from the origin) of
both the spots.

The IR Spectrum of Caffeine in a Nujol Mull