MSc practical

ISOLATION OF CAFFEINE FROM TEA

Introduction

The principle xanthines used in medicine are caffeine and theophylline. This group of drugs is
of value for their effects on the kidney, smooth muscle of the bronchi and the central nervous
system. (Theophylline is widely used as a bronchodilator in asthma).
Caffeine and its salts are most active in their stimulation of the central nervous system and are
principally used for this purpose.

Caffeine is administered in powder or tablet form and is frequently included in analgesic

preparations with aspirin, paracetamol, or codeine. Caffeine is claimed to enhance the activity
of analgesics and also ergotamine, and is often added to “tonics” for tiredness. It may be
obtained from tea or coffee or prepared synthetically.

The object of this exercise is to isolate caffeine from tea. Identification of the caffeine should
be made by IR and TLC

1) Method of isolation of methylxanthines

1. Tea (~10g weighed to 4 decimal places) is extracted with water (200ml) plus 0.05N
sulphuric acid (50ml) in a beaker using a Bunsen bring to boil then remove from heat.

2. Leave to cool for a few minutes and filter using Buchner system.

3. Add to the filtrate 20 ml of 10% sulphuric acid and stir.

Sulphuric acid converts the tannins to their salts, thus rendering them insoluble in chloroform
but they are soluble in water

4. The filtrate should now be extracted by 3 successive washes (in a separating funnel) of
chloroform (50ml each) .With each wash collect the organic layer (Bottom)

caffeine is more soluble in chloroform than the tannins.

5. Combine all the extracts.

6. Dry by adding large spatulas of anhydrous sodium sulphate until water is removed. It is dry
when you get a fine powder.

7. Clean Buchner system and dry using acetone in the fume cupboard.

8. Remove sodium sulphate by filtering with Buchner system.

9. Pour the filtrate into a pre-weighed (to 4 decimal places) round-bottomed flask.
10. Evaporate all the chloroform using a rotary evaporator (see demonstrators) you will be left
with methylxanthine crystals.

2) Thin Layer Chromatography

After obtaining a yield for your methylxanthines re-dissolve your crystals in 2ml of
chloroform. Examine 1ml of your solution, as well as reference solutions and mother liquor by
TLC.

Methylanthines absorb UV light and therefore the inclusion of a fluorescent agent in the
stationary phase is particularly useful since visualisation in the UV at 254nm can then be used.

Plates: Silica gel GF254

Solvent: Chloroform: Acetone: N-Butanol: 30% Ammonia, (30:30:40:10)
Reference solutions: Caffeine, Theobromine, Theophylline, and Mother Liquor

3. Infra Red Spectroscopy

Using the rest of your methylxanthine solution obtain an IR spectrum