ISOLATION OF CAFFEINE

FROM TEA LEAVES

Botanical: Camellia thea

Synonyms---Thea sinensis
(Sims). Thea veridis. Thea bohea.
Thea stricta Jassamica. Camellia
theifera (Griff.).

Green tea, black tea, and oolong tea are all derived from
http://www.nlm.nih.gov/medlineplus/druginfo/natural/patientthe same plant.
black_tea.html
White teas? These are less processed tea and contain more

antioxidants than black teas and oolongs, which fall midway

between green and black teas. TIME December 3, 2007 p 41 India Brews A Stronger Cup
---Constituents---Caffeine (theine), tannin (10 to 20 per cent
gallotannic acid), boheic acid, volatile oil, aqueous extract,
protein wax, resin, ash and theophylline.
http://www.botanical.com/botanical/mgmh/t/tea--08.html$$

Black tea

is a source of caffeine, a methylxanthine

O
H3C
O

CH3
N

N
N
CH3

N
1,3,7-trimethyl-2,6-dioxopurine

It stimulates the central nervous system,

relaxes smooth muscle in the airways to the lungs
(bronchioles), stimulates the heart,
and acts on the kidney as a diuretic (increasing urine).

Green Tea:

Chinese Tea, Unfermented Tea, Sencha

Active Ingredients:
1.Polyphenols (Catechins) (10-25%):
epigallocatechin-3-gallate, epicatechin,
epicatechin-3-gallate, epigallocatechin
2.Purine Alkaloids (Methyl Xanthines): caffeine((2.94.2%)
3.Inorganic Ions: fluoride (130-160 mg/kg),
potassium, aluminum
4.Other Ingredients: flavonoids, caffeic acid
derivatives, triterpene saponins, volatile oils

Oolong Tea,

the tea that represents China, is

produced in the province of Fujian. The tea consists of
several kinds of leaves that goes through different
production processes giving different flavours and aromas.
http://www.oolongtea.org/e/welcome/04.h
tml

Caffeine is not only in coffee, tea, cola, even in chocolates!

Caffeine-infused gum, lip balm, mints, beer, candy, sunflower seeds,

How does it work?

Caffeine crosses the blood-brain barrier, dampening
neurotransmitters that would normally make you sleepy,
improving temporarily, cognitive function and even athletic
performance.
A typical cup of coffee has 80 to 150mg caffeine;
Softdrinks have 34 mg to 50 mg per can;
Energy drinks may pack from 75 mg to 300 mg per serving.

TIME October 1, 2007 p.38 The Caffeine Habit

One cup

of tea contains approximately 50

milligrams of caffeine, depending on the strength
and size of cup (as compared to coffee which
contains 65 to 175 milligrams of caffeine per cup).

Tea also contains polyphenols

(catechins, anthocyanins, phenolic
acids), tannin, trace elements, and
vitamins.

Caffeine is a stimulant of the central

nervous system, and may cause
insomnia in adults, children, and
infants (including nursing infants of
mothers taking caffeine). Caffeine
acts on the kidneys as a diuretic
(increasing urine and urine
sodium/potassium levels, and
potentially decreasing blood
sodium/potassium levels), and may
worsen urge incontinence. Caffeinecontaining beverages may increase
the production of stomach acid, and
may worsen ulcer symptoms. Tannin
in tea can cause constipation.
Caffeine in doses of 250 to 350
milligrams can increase heart rate
and blood pressure, although people
who consume caffeine regularly do
not seem to experience these effects

TECHNIQUES
1. Extraction
1.1 Solid-Liquid Extraction
1.2 Liquid-Liquid Extraction
o Single Extraction
o Multiple Extraction
2. Sublimation/Recrystallization
3. Characterization by Melting Point Determination

SOLID-LIQUID EXTRACTION
Solvents for extraction:
1. Non-toxic
2. Easily removed
3. Desired constituent is soluble
4. Non-reactive

Extract with Na2CO3 in

BOILING WATER

MARC
Na-Tannate

HOT WATER EXTRACT

Extract is BROWN in color
due to flavonoids, tannins and
its oxidation products.

EXTRACTION
-

the removal of a desired compound from a

solid or liquid mixture by a solvent.

transferring a solute from one solvent to an

Types:
SOLID-LIQUID EXTRACTION
LIQUID-LIQUID EXTRACTION

SOLID-LIQUID EXTRACTION

DECOCTION the act or process of boiling anything in a

watery fluid to extract its virtues; an extract from a body by
boiling it in water
http://naturalhealthcare.ca/medical_terms.phtml?term=decoction.
Taken 02 Dec 2007

INFUSION is the flavour thats extracted from an

ingredient such as tea leaves, herbs or fruit by steeping
(soaking) them in a liquid (usually hot), such as water, for
tea.
http://www.epicurious.com/tools/fooddictionary/entry?id=3033. Taken 02 Dec

CONTINUOUS EXTRACTION PROCESS

Used when the organic compound is more
soluble in water than in the organic solvent

BATCH EXTRACTION PROCESS

Most frequently used.
Separation of a neutral organic
compound in an aqueous medium, by
shaking with an organic solvent in
which the compound is soluble but is
immiscible with water.

Tea Leaves
1

CELLULOSE, 2CAFFEINE, 3TANNINS

CELLULOSE
1

Glucose polymer, not soluble in water

CAFFEINE

2 Alkaloid compound; soluble in water but more

soluble in dichloromethane (DCM, CH2Cl2)
O
H3C
O

CH3
N

N
N
CH3

TANNIN
S
3 Phenolic compounds; all are potentially
3

soluble in water and some are hydrolyzed

in
water. refers to a class of compounds which
TANNINS
are PHENOLICS
hydrolyzable tannins found in tea
O
generally yield
glucose
and gallic acid
O
H

OH

OH

H
H

RO

OH

O C

CH2O C

OH

OH
CH2OH

RO

RO

OR

undergo hydrolysis in
RO
water

OH

H
+

RO

RO

H
H

ESTER OF GALLIC ACID AND GLUCOSE

HOOC

O
H

OR

OH
OH

Non-hydrolyzable tannins are condensation polymers

of catechins; these polymers are not uniform in structures
OH
OH
HO

O
OH
OH

catechin

Overview
Tannins that are hydrolyzed when mixed
with H2O produce glucose + gallic acid.
Adding Na
CO33 makes the acidic solution
Na2CO
basic & converts tannins & gallic acid to
H2O-soluble sodium salts.
This opens the door for a separation of
caffeine from the other components via
extraction by adding CH2Cl2 to the
aqueous solution.
Crude caffeine can then be isolated by
evaporating off the CH2Cl2 solvent and
further purified by sublimation or
crystallization.

LIQUID-LIQUID EXTRACTION
Hot WATER EXTRACT
Cool
Extract with 20 mL dichloromethane ( 3x)

AQUEOUS E XTRAC T

DCM EXTRAC T ( Organic)

Choice of solvent for liquid-liquid extraction:

1. Solvent must be able to dissolve compound of
interest
2. Solvent must not be very soluble in water
3. Solvent must be low boiling so that it can be
easily removed by distillation or evaporation
4. Solvent must have a density that is different
from water

LIQUID-LIQUID EXTRACTION is use for separation of complex mixtures

by selective partitioning between two phases, between two immiscible liq

Extraction are carried out in two immiscible solvents, usually one aqueous
one organic.

DC M EXTRAC T
Wash with 6M NaOH

AQUEOUS EXTRAC T

DCM EXTRAC T

AQUEOUS EXTRACT

DCM EXTRACT

Hydolyzable tannins

Caffeine

Non-hydrolyzable
tannins

Chlorophylls

Sugars, minerals, acids

Some flavonoids,waxes

Sodium salts of tannins

Condensed tannins
(non-hydrolyzable)

DISTRIBUTION COEFFICIENT (Partition

Coefficient)

Ratio of the concentrations of the solute in

each solvent at a particular temperature is
constant

It is independent of the total concentration and t

actual amounts of the two solvents mixed.

K = concentration in solvent 2
concentration in solvent 1

It is ALWAYS better to use several small amounts of the second s

than to make a single extraction with a large portion.

For example a compound A is soluble in diethylether to the

extent of 20g/100mL at 200C and soluble in water to the
extent of 5.0g/100mL at the same temperature.

What is the distribution coefficient of compound A?

We will calculate how much compound will be extracted by

single extraction.
If for example we have a solution containing 5.0 g of
compound A in 100 mL water. If we shake this with 100 mL
diethylether, how much a will be extracted by the ether?

Concentration of A in diethyl ether = X gms

100mL
Concentration of A in water = (5.0 X) gms
100 mL

Substitution,

K = concentration in diethyl ether

concentration in water

X=

Because a solute distributes itself between two solvent, a

single extraction may not be very efficient.
The 100 mL of ether maybe divided into three portions of
approximately 33 mL each.
X after three extractions =

EMULSION

A colloidal suspension of one liquid in another caused by vigorous shak

How to breakup emulsions?

(1) if one of the solvents is water, adding a saturated aqueous

sodium chloride solution
(2) gentle swirling of the liquid in the flask and gentle stirring

MELTING POINT DETERMINATION

MELTING POINT is the temperature at which solid changes
to liquid
At 1 atmosphere of pressure.
MELTING RANGE, the melting point of a solid is reported in
range

Effect of Impurities

a PURE ORGANIC COMPOUND usually has a SHARP melting po

SHARP MELTING POINT melts within a range of 1-2o

an IMPURE ORGANIC COMPOUND exhibits a broader range an

depressed melting point from that of pure compound

OTHER MELTING POINT BEHAVIORS:

1.Decomposition observed as darkening. All organic compounds decomp

when heated at sufficient temperature.

2. Polymorphs different crystalline forms of the same compound causes

different intermolecular attractions.

3.Hydrates some compounds can crystallize with water or other solvents

in their crystal lattice in a definite proportion by weight. Hy
would have different temperatures from anhydrous.

4.Racemic Mixtures a pair of nonsuperposable mirror image isomers cal

enantiomers would behave like a pure compou

5.Liquid Crystals intermediate between solid phase and liquid phase form
by heat causes some pure compounds not to
exhibit sharp melting point