PHARMACOGNOSY & ORGANIC CHEMISTRY LAB CHEAT SHEET

Experiment 08: Extraction and Isolation of Caffeine from Camellia sinensis (Tea Leaf)

EXPERIMENT #08 (Date: 25-04-2026) APPLICATIONS & PRECAUTIONS

Object: Extraction and isolation of Caffeine from leaves of Camellia sinensis (Tea leaf). Applications:
Requirements: Tea leaf (sample), Beaker, Muslin cloth, Stand, Water, Filter paper, Funnel, • Use as OTC for cold and flu medication.
$H_2SO_4$ sol., Separating funnel, Chloroform, Ethanol, Lead acetate. • Use in dietary products.
• Used in pain-killer medicines.
THEORY: ALKALOIDS • Widely used in energy drinks to improve mental alertness.
Nitrogen-containing compounds present as physiologically active compounds, produced Precautions:
by a variety of organisms such as bacteria, fungi, plants, and animals. • Stomach ulcer due to increased gastric acid.
• Sleep disorder due to brain disturbances.
Alkaloid Obtained Plant Source
• Can worsen anxiety.
Nicotine Nicotiana tabacum (Leaves) • Should be avoided in high B.P.
Morphine Papaver somniferum (Seed)
Strychnine Nux-vomica (Seed) EXTRACTION PROCEDURE (STEPS 1–20)
1. Weight $10 ext{ g}$ of tea grains.
Cocaine Erythroxylum coca (Leaves)
2. Add in beaker with $50 ext{ ml}$ of water.
Types of Alkaloids: 3. Boil for $10 ext{ minutes}$ and filter it.
1. True Alkaloids: Nitrogen compound with a heterocyclic ring & derived from amino 4. Put residual grains again in water ($50 ext{ ml}$) for $10 ext{ mins}$ $ ightarrow$ repeat
acids. 3 times.
2. Proto Alkaloids: Naturally occurring nitrogen-containing compounds where the 5. Obtained filtrate contains caffeine, tannin, and other organic compounds.
nitrogen atom is derived from amino acids but is not a part of the heterocyclic 6. Add $10 ext{ ml}$ lead acetate to filtrate.
structure. For example: Mescaline, Ephedrine. 7. Lead acetate precipitates tannin $ ightarrow$ leaving pure caffeine.
3. Pseudo Alkaloids: Not derived from amino acids but from terpenoids. For example: 8. Allow mixture to set for $10 ext{ mins}$.
Caffeine, Theophylline. 9. Filter it to remove tannins.
10. Now add $2 ext{ ml}$ $H_2SO_4$ to filtrate for removing acid impurities.
CLASSIFICATION (BASED ON CHEMICAL STRUCTURE) 11. Transfer filtrate to separating funnel and add $10 ext{ ml}$ chloroform.
12. Set it on stand and shake vigorously until two different layers appear.
Tropane alkaloids Atropine, Cocaine
13. Open the stopper and collect bottom layer in beaker.
Quinoline alkaloids Quinine, Quinidine 14. Repeat procedure to obtain max. caffeine.
Isoquinoline alkaloids Morphine, Papaverine 15. Weight the empty beaker ($w_1$).
16. Add $10 ext{ ml}$ ethanol in chloroform to separate out caffeine.
Indole alkaloids Reserpine, Vincristine
17. Next day, shiny white crystals are obtained in beaker.
Purine alkaloids Caffeine, Theophylline 18. Weight beaker having shiny crystals ($w_2$).
Pyridine alkaloids Nicotine, Ricinine 19. Weight of caffeine: $w = w_2 - w_1$.
20. Find percentage yield of caffeine.

CAFFEINE: PROFILE & STRUCTURE

OBSERVATIONS & CALCULATIONS
Caffeine is a methyl-xanthine derivative with the molecular formula $$C_8H_{10}
N_4O_2$$. Weight of empty beaker ($w_1$) 36 grams
Structure: Fused ring system having a pyrimidine & imidazole ring $ ightarrow$ Weight of beaker + sample ($w_2$) 37 grams
xanthine core. Having two amide functional groups and three methyl groups at positions Weight of Caffeine ($w = w_2 - w_1$) $37 - 36 = 1 ext{ gram}$
1, 3, 7.
Weight of initial tea sample 10 grams
Physical Properties:
• White, odorless, crystalline powder. Formula:
• Has a distinctly bitter taste. \% ext{ Yield of Caffeine} = rac{ ext{Weight of Caffeine}}{ ext{Weight of Sample}} imes 100
• Moderately soluble in water. \% ext{ Yield} = rac{1}{10} imes 100 = \mathbf{10\%}
• Excellent solubility in Chloroform & Dichloromethane.
• Melting point of caffeine is 235°C. Result: We have successfully extracted caffeine from tea leaves of brand 10 grams and
percentage yield is 10%.
PHARMACEUTICAL ACTIONS & SIDE EFFECTS
Actions:
• CNS stimulant; relieves fatigue + drowsiness.
• Promotes lipolysis by increasing cAMP level.
• Acts as a diuretic and respiratory stimulant.
• Increases heart rate and Blood Pressure (B.P.).
• Increases gastric-acid secretion.
Side-Effects:
Anxiety, High B.P., Headache, Insomnia, Nervousness, Respiratory problems.