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 the same plant.
http://www.nlm.nih.gov/medlineplus/druginfo/natural/patient-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 H3 C O N N CH3 CH3 N 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-3gallate, epicatechin, epicatechin-3-gallate, epigallocatechin 2. Purine Alkaloids (Methyl Xanthines): caffeine((2.9-4.2%) 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.html

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

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

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 in the long-term.
http://www.mayoclinic.com/health/green-tea/NS_patient-green_tea

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.

Types:

transferring a solute from one solvent to another

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 2007

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
1CELLULOSE

1CELLULOSE, 2CAFFEINE, 3TANNINS

1 Glucose polymer, not soluble in water

2CAFFEINE

2 Alkaloid compound; soluble in water but more soluble in

dichloromethane (DCM, CH2Cl2)
O H3 C O N N CH3 CH3 N N

3TANNINS

3 Phenolic compounds; all are potentially soluble in water and some are hydrolyzed in water. TANNINS refers to a class of compounds which are PHENOLICS
hydrolyzable tannins found in tea generally yield glucose and gallic acid
O H H H RO RO H
H

O O C OH OH OH
H CH2 OH O H RO RO RO

CH2O C O

OH OH

HOOC
H
+

OH OH

RO

OR

undergo hydrolysis in water

OR

OH

ESTER OF GALLIC ACID AND GLUCOSE

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 Na2CO3 makes the acidic solution Na2CO 3 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 WAT ER EXT RACT Cool Extract with 20 mL dichloromethane ( 3x)

AQU EOUS E XTR AC T

D CM EX TR AC 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 liquids.

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

D C M EXTR AC T Wash with 6M NaOH

AQU EOU S EXTR AC T

D C M EXTR AC T

AQUEOUS EXTRACT Hydolyzable tannins Non-hydrolyzable tannins Sugars, minerals, acids Sodium salts of tannins

DCM EXTRACT Caffeine Chlorophylls Some flavonoids,waxes Condensed tannins (nonhydrolyzable)

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 the 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 solvent 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 shaking.
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 point SHARP MELTING POINT melts within a range of 1-2o

an IMPURE ORGANIC COMPOUND exhibits a broader range and depressed melting point from that of pure compound

OTHER MELTING POINT BEHAVIORS: 1. Decomposition observed as darkening. All organic compounds decompose 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. Hydrates would have different temperatures from anhydrous. 4. Racemic Mixtures a pair of nonsuperposable mirror image isomers called enantiomers would behave like a pure compound. 5. Liquid Crystals intermediate between solid phase and liquid phase formed by heat causes some pure compounds not to exhibit sharp melting point