EXTRACTION OF CAFFEINE FROM TEA

A Project Report
Submitted in partial fulfillment of requirement for the award of Degree of
Bachelor of Engineering in Chemical Engineering

Submitted to

Rajiv Gandhi Proudyogiki Vishwavidhyalaya, Bhopal (M.P.)

Submitted By

ABHISHEK SINGH CHANDEL (0103CM151002)

SHUBHAM NARWARE (0103CM151044)
JOEL VARGHESE (0103CM151017)
PRAMOD JAIPRAKASH (0103CM151029)

Under The Guidance of

PROF. RANJANA SINGH YADAV

Department of Chemical Engineering

Lakshmi Narain College of Technology, Bhopal

Session 2015-2019

1
 Lakshmi Narain College of Technology, Bhopal

DEPARTMENT OF CHEMICAL ENGINEERING

CERTIFICATE
This is to certify that the work embodied in this entitled “EXTRACTION OF CAFFEINE
FROM TEA” has been satisfactorily completed by ABHISHEK SINGH CHANDEL,
SHUBHAM NARWARE, JOEL VARGHESE, PRAMOD JAIPRAKASH. It is a bona-fide
piece of work, carried out under our supervision and guidance in the Department of Chemical
Engineering, Lakshmi Narain College of Technology, Bhopal, for partial fulfillment of the
Bachelor of Engineering during the academic year 2015-2019.
Under the supervision of

PROF. RANJANA SINGH YADAV

Approved by

Prof. Ranjana Yadav

(Head of Department)

Forwarded By

Dr. Rakesh Mowar

(Principal)
Lakshmi Narain College of Technology, Bhopal

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 LAKSHMI NARAIN COLLEGE OF TECHNOLOGY, BHOPAL

DEPARTMENT OF CHEMICAL ENGINEERING

DECLARATION

We ABHISHEK SINGH CHANDEL, SHUBHAM NARWARE, JOEL VARGHESE,

PRAMOD JAIPRAKASH students of Bachelor of Engineering, Department of Chemical
Engineering, Lakshmi Narain College of Technology, Bhopal, hereby declare that the work
presented in this Minor Project is outcome of our own work, is bonafide correct to be best of our
knowledge and this work has been carried out taking care of engineering ethics. The presented
work does not integrate any patent work and has not been submitted to any University for the
award of any degree or any professional diploma.

ABHISHEK SINGH CHANDEL (0103CM151002)

SHUBHAM NARWARE (0103CM151044)
JOEL VARGHESE (0103CM151017)
PRAMOD JAIPRAKASH (0103CM151029)

DATE:

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 LAKSHMI NARAIN COLLEGE OF TECHNOLOGY, BHOPAL
DEPARTMENT OF CHEMICAL ENGINEERING

ACKNOWLEDGEMENT

We express our deep sense of gratitude to our project supervisor and Head of The Department
Prof. Ranjana Yadav whose kindness, valuable guidance and deep subjective knowledge
encouraged us to complete this project.
We also very much thankful to Dr. Rakesh Mowar (Principal of LNCT Bhopal), for his support
in guiding and selecting the right project and his valuable advice in completion of this project.
We would also thank our institution and all faculty members without whom this project work
would have been distant reality.

ABHISHEK SINGH CHANDEL (0103CM151002)

SHUBHAM NARWARE (0103CM151044)
JOEL VARGHESE (0103CM151017)
PRAMOD JAIPRAKASH (0103CM151029)

4
 ABSTRACT

Extraction is a method used for the separation of organic compound from a mixture of
compound. This technique selectively dissolves one or more compounds into an appropriate
solvent. The solution of these dissolved compounds is referred to as the extract. In the case of
Caffeine extraction from tea powder, the solubility of caffeine in water is 22mg/ml at 25°C,
180mg/ml at 80°C, and 670mg/ml at 100°C. Here the organic solvent Dichloromethane is used
to extract caffeine from aqueous extract of tea powder because caffeine is more soluble in
dichloromethane (140mg/ml) than it is in water (22mg/ml).The dichloromethane - caffeine
mixture can then be separated on the basis of the different densities of dichloromethane and
water because dichloromethane is much denser than water and insoluble in it. Residual water is
separated from dichloromethane by drain out the dichloromethane through separating funnel,
thus dichloromethane passed through the funnel while polar solvents such as water is still
remains in the funnel. Water and dichloromethane is slightly soluble in each other. So, after
separating the solvents, residual water will remain the organic layer. Mainly anhydrous sodium
sulfite is used for the removal of water from organic layer. Anhydrous sodium sulfite is an
insoluble inorganic solid which will absorb water, thus drying it.

Hence this project give us knowledge about caffeine,its sources ,extraction process including its
advantages and disadvantages.

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 INDEX
1. WHAT IS CAFFEINE…………………………………………………………...................8
2. LITERATURE REVIEW……………………………………………………………...…11
2.1. Discovery and spread of use 11
2.2. Chemical identification, isolation, and synthesis 14
3. NATURAL OCCURRENCE………………………………………………....…………....16
4. USE OF CAFFEINE………………………………………………..………….…………...18
4.1. Medical 18
4.2. Enhancing performance 18
4.3. Cognitive 18
4.4. Physical 18
4.5. Specific populations
4.5.1.1. Adults 19
4.5.1.2. Children 19
5. ADVERSE EFFECTS…………………………………………………………………..….20
5.1. Physical 20
5.2. Reinforcement disorders 21
5.3. Dependence and withdrawal 21
5.4. Risk of other diseases 22
6. OVERDOSE………………….…………………………………………………….…….…23
7. PRODUCTS…………………………………………………………………………….….25
7.1. Beverages 25
7.2. Coffee 25
7.3. Tea 25
7.4. Soft drinks and energy drinks 25
7.5. Other beverages 25
7.6. Chocolate 27
7.7. Tablets 27
7.8. Other oral products 27
8. COMBINATIONS WITH OTHER DRUGS…………………………………………….27
9. EXTRACTION OF CAFFIENE FROM TEA…………………………………………..28
9.1. Principle 29
9.2. Background to the Procedure 29
9.3. Procedure 30
10. REFERENCES…………………………………………………………………………… 32

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 LIST OF FIGURE

FIGURE TITLE PAGE NO.

1 Structure of Caffeine 8
2 Roasted coffee beans 16
3 Primary symptoms of caffeine intoxication 23

LIST OF TABLE

TABLE TABLE PAGE NO.

1 Chemical and Physical Properties of Caffeine 10
2 Daily caffeine intake 19
3 Caffeine content of selected common food and drugs 26

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WHAT IS CAFFEINE

Caffeine is a central nervous system (CNS) stimulant of the methylxanthine class. It is the
world's most widely consumed psychoactive drug. Unlike many other psychoactive substances, it
is legal and unregulated in nearly all parts of the world. There are several known mechanisms of
action to explain the effects of caffeine. The most prominent is that it reversibly blocks the action
of adenosine on its receptor and consequently prevents the onset of drowsiness induced by
adenosine. Caffeine also stimulates certain portions of the autonomic nervous system.

Caffeine is a bitter, white crystalline purine, a methylxanthine alkaloid, and is chemically related
to the adenine and guanine bases of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). It
is found in the seeds, nuts, or leaves of a number of plants native to South America and East Asia
and helps to protect them against predator insects and to prevent germination of nearby seeds.
The most well-known source of caffeine is the coffee bean, a misnomer for the seed of Coffea
plants. Beverages containing caffeine are ingested to relieve or prevent drowsiness and to
improve performance. To make these drinks, caffeine is extracted by steeping the plant product
in water, a process called infusion. Caffeine-containing drinks, such as coffee, tea, and cola, are
very popular; as of 2014, 85% of American adults consumed some form of caffeine daily,
consuming 164 mg on average.

8
 Fig 1 structure of Caffeine

Caffeine can have both positive and negative health effects. It can treat and prevent the
premature infant breathing disorders bronchopulmonary dysplasia of prematurity and apnea of
prematurity. Caffeine citrate is on the WHO Model List of Essential Medicines. It may confer a
modest protective effect against some diseases, including Parkinson's disease. Some people
experience sleep disruption or anxiety if they consume caffeine, but others show little
disturbance. Evidence of a risk during pregnancy is equivocal; some authorities recommend that
pregnant women limit consumption to the equivalent of two cups of coffee per day or less.
Caffeine can produce a mild form of drug dependence – associated with withdrawal symptoms
such as sleepiness, headache, and irritability – when an individual stops using caffeine after
repeated daily intake. Tolerance to the autonomic effects of increased blood pressure and heart
rate, and increased urine output, develops with chronic use (i.e., these symptoms become less
pronounced or do not occur following consistent use).

Caffeine is classified by the US Food and Drug Administration as "generally recognized as safe"
(GRAS). Toxic doses, over 10 grams per day for an adult, are much higher than typical doses of
under 500 milligrams per day. A cup of coffee contains 80–175 mg of caffeine, depending on
what "bean" (seed) is used and how it is prepared (e.g., drip, percolation, or espresso). Thus it
requires roughly 50–100 ordinary cups of coffee to reach a lethal dose. However, pure powdered
caffeine, which is available as a dietary supplement, can be lethal in tablespoon-sized amounts.
Caffeine and other purine alkaloids, including theobromine and theophylline, have played a major
role in the long-standing popularity of non-alcoholic beverages and foods such as coffee, tea, cocoa,
mate, chocolate and a wide range of soft drinks (Asahihara 2008). Caffeine is a naturally
9
occurring chemical stimulant found in the leaves, seeds and fruits of a numerous plant species of a
group of compounds called trimethylxanthine. Its chemical formula is C 8H10N4O2. In its pure form,
caffeine is a white crystalline powder that tastes very bitter. It is medically useful to stimulate the
heart and also serves as increasing the rate of urine excretion. It is one of the most studied ingredients
in the food supply. The most commonly known sources of caffeine are coffee and cocoa beans,
guarana, and tea leaves. The amount of caffeine in food and beverage products varies depending on
the serving size, the type of product and preparation method. Tea which we generally drink is made
from the leaves of an Asian evergreen known as Camellia sinensis. White tea, green tea, red tea, and
black tea all come from this plant, and all contain caffeine. The presence of caffeine in plants helps to
prevent them from insects and other herbivores with the compound’s bitter taste and stimulating
qualities. The growing buds and young leaves of tea plants manufacture the highest amounts of
caffeine. Caffeine can be isolated from teas with liquid-liquid extraction and quantitated by gas
chromatography with nitrogen phosphorus detection. The decaffeinated teas contained less than 12
mg of caffeine per serving, and caffeine was not detected in the herbal tea varieties. The steep time
affect the caffeine concentration of the tea. Most brewed teas contain less caffeine per serving than
brewed coffee. (Jenna et al., 2008). A special ultrasound method with high-frequency, well
penetrating power and the sound wave can be used to extract the caffeine. The quantity of caffeine is
increased with temperature raised

Chemical and Physical Properties of Caffeine

General
Systematic
1,3,7-trimethyl-1H-purine-2,6(3H,7H)-dione
name
1,3,7-trimethylxanthine, trimethylxanthine,
Other names theine, mateine, guaranine,
methyltheobromine

Molecular
C8H10N4O2
formula

SMILES O=C1C2=C(N=CN2C)N(C(=O)N1C)C

Molar mass 194.19 g mol−1

Appearance Odorless, white needles or powder

CAS number [58-08-2]

Properties
Density and 1.2 g/cm³, solid
phase

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 Solubility in Slightly soluble
water
Soluble in ethyl acetate, chloroform, pyrimidine, pyrrole, tetrahydrofuran
Other
solution; moderately soluble in alcohol, acetone; slightly soluble in petroleum
solvents
ether, ether, benzene.

Melting point 237 °C

Boiling point 178 °C (sublimes)

Acidity (pKa) 10.4 (40 °C)

Hazards
MSDS External MSDS

Main hazards May be fatal if inhaled, swallowed

or absorbed through the skin.

Flash point N/A

RTECS
EV6475000
number
REF- Malenka RC, Nestler EJ, Hyman SE (2009). "Chapter 15”

LITERATURE REVIEW

Discovery and spread of use

According to Chinese legend, the Chinese emperor Shennong, reputed to have reigned in about
3000 BCE, accidentally discovered tea when he noted that when certain leaves fell into boiling
water, a fragrant and restorative drink resulted. Shennong is also mentioned in Lu Yu's Cha Jing,
a famous early work on the subject of tea.

The earliest credible evidence of either coffee drinking or knowledge of the coffee tree appears
in the middle of the fifteenth century, in the Sufi monasteries of the Yemenin southern Arabia.
From Mocha, coffee spread to Egypt and North Africa, and by the 16th century, it had reached
the rest of the Middle East, Persia and Turkey. From the Middle East, coffee drinking spread to
Italy, then to the rest of Europe, and coffee plants were transported by the Dutch to the East
Indies and to the Americas.

Kola nut use appears to have ancient origins. It is chewed in many West African cultures,
individually or in a social setting, to restore vitality and ease hunger pangs.

The earliest evidence of cocoa bean use comes from residue found in an ancient Mayan pot dated
to 600 BCE. Also, chocolate was consumed in a bitter and spicy drink called xocolatl, often
seasoned with vanilla, chile pepper, and achiote. Xocolatl was believed to fight fatigue, a belief
11
probably attributable to the theobromine and caffeine content. Chocolate was an important luxury
good throughout pre-Columbian Mesoamerica, and cocoa beans were often used as currency.

Xocolatl was introduced to Europe by the Spaniards, and became a popular beverage by 1700.
The Spaniards also introduced the cacao tree into the West Indies and the Philippines. It was
used in alchemical processes, where it was known as "black bean"

The leaves and stems of the yaupon holly (Ilex vomitoria) were used by Native Americans to
brew a tea called asi or the "black drink". Archaeologists have found evidence of this use far into
antiquity, possibly dating to Late Archaic times.

Caffeine is a plant alkaloid, found in numerous plant species, where it acts as a natural pesticide
that paralyzes and kills certain insects feeding upon them. The most commonly used caffeine-
containing plants are coffee, tea, and to some extent cocoa. Other, less commonly used, sources
of caffeine include the yerba mate[4] and guaranà plants, which are sometimes used in the
preparation of teas and energy drinks. Two of caffeine's alternative names, mateine and
guaranine, are derived from the names of these plants.

The world's primary source of caffeine is the coffee bean (the seed of the coffee plant), from
which coffee is brewed. Caffeine content in coffee varies widely depending on the type of coffee
bean and the method of preparation used; even beans within a given bush can show variations in
concentration. In general one serving of coffee ranges from about 40 milligrams for a single shot

(30 milliliters) of arabica-variety espresso to about 100 milligrams for strong drip coffee.
Generally, dark-roast coffee has less caffeine than lighter roasts because the roasting process
reduces the bean's caffeine content. Arabica coffee normally contains less caffeine than the
robusta variety. Coffee also contains trace amounts of theophylline, but no theobromine.

Tea is another common source of caffeine. Tea usually contains about half as much caffeine per
serving as coffee, depending on the strength of the brew. Certain types of tea, such as black and
oolong, contain somewhat more caffeine than most other teas. Tea contains small amounts of
theobromine and slightly higher levels of theophylline than coffee. Preparation has a significant
impact on tea, and color is a very poor indicator of caffeine content.[8] Teas like the green
Japanese gyokuro, for example, contain far more caffeine than much darker teas like lapsang
souchong, which has very little.---Chocolate derived from cocoa contains a small amount of
caffeine. Chocolate is a weak stimulant, which is mostly due to its content of theobromine and
theophylline. It contains too little of these compounds for a reasonable serving to create effects in
humans that are on par with coffee. A typical 28-gram serving of a milk chocolate bar has about
as much caffeine as a cup of decaffeinated coffee.

Caffeine is also a common ingredient of soft drinks such as cola, originally prepared from kola
nuts. Soft drinks typically contain about 10 to 50 milligrams of caffeine per serving. By contrast,
energy drinks such as Red Bull contain as much as 80 milligrams of caffeine per serving. The
caffeine in these drinks either originates from the ingredients used or is an additive derived from
the product of decaffeination or from chemical synthesis. Guarana, a prime ingredient of energy

12
drinks, contains large amounts of caffeine with small amounts of theobromine and theophylline
in a naturally occurring slow-release excipient.

Humans have consumed caffeine since the Stone Age.[11] Early peoples found that chewing the
seeds, bark, or leaves of certain plants had the effects of easing fatigue, stimulating awareness,
and elevating mood. Only much later was it found that the effect of caffeine was increased by
steeping such plants in hot water. Many cultures have legends that attribute the discovery of such
plants to people living many thousands of years ago.

According to one popular Mongolian legend, the Emperor of China Shennong, reputed to have
reigned in about 3,000 BC, accidentally discovered that when some leaves fell into boiling water,
a fragrant and restorative drink resulted Shennong is also mentioned in Lu Yu's Cha Jing, a
famous early work on the subject of tea.

The early history of coffee is obscure, but a popular myth traces its discovery to Ethiopia, where
Coffea arabica originates. According to this myth, a goatherder named Kaldi observed goats that
became elated and sleepless at night after browsing on coffee shrubs and, upon trying the berries
that the goats had been eating, experienced the same vitality. The earliest literary mention of
coffee may be a reference to Bunchum in the works of the 9th century Persian physician al-Razi.
In 1587, Malaye Jaziri compiled a work tracing the history and legal controversies of coffee,
entitled "Umdat al safwa fi hill al-qahwa". In this work, Jaziri recorded that one Sheikh, Jamal-
al-Din al-Dhabhani, mufti of Aden, was the first to adopt the use of coffee in 1454, and that in
the 15th century the Sufis of Yemen routinely used coffee to stay awake during prayers.
Towards the close of the 16th century, the use of coffee was recorded by a European resident in
Egypt, and about this time it came into general use in the Near East. The appreciation of coffee
as a beverage in Europe, where it was first known as "Arabian wine," dates from the 17th
century. During this time "coffee houses" were established, the first being opened in
Constantinople and Venice. In Britain, the first coffee houses were opened in London in 1652, at
St Michael's Alley, Cornhill. They soon became popular throughout Western Europe, and played
a significant role in social relations in the 17th and 18th centuries.

The kola nut, like the coffee berry and tea leaf, appears to have ancient origins. It is chewed in
many West African cultures, individually or in a social setting, to restore vitality and ease hunger
pangs. In 1911, kola became the focus of one of the earliest documented health scares when the
US government seized 40 barrels and 20 kegs of Coca-Cola syrup in Chattanooga, Tennessee,
alleging that the caffeine in its drink was "injurious to health".[15] On March 13, 1911, the
government initiated The United States vs. Forty Barrels and Twenty Kegs of Coca-Cola, hoping
to force Coca-Cola to remove caffeine from its formula by making exaggerated claims, such as
that the excessive use of Coca-Cola at one girls' school led to "wild nocturnal freaks, violations
of college rules and female proprieties, and even immoralities."[16] Although the judge ruled in
favor of Coca-Cola, two bills were introduced to the U.S. House of Representatives in 1912 to
amend the Pure Food and Drug Act, adding caffeine to the list of "habit-forming" and
"deleterious" substances which must be listed on a product's label.

The earliest evidence of cocoa use comes from residue found in an ancient Mayan pot dated to 600
BC. In the New World, chocolate was consumed in a bitter and spicy drink called xocoatl, often

13
seasoned with vanilla, chile pepper, and achiote. Xocoatl was believed to fight fatigue, a belief
that is probably attributable to the theobromine and caffeine content. Chocolate was an important
luxury good throughout pre-Columbian Mesoamerica, and cocoa beans were often used as
currency.

Chocolate was introduced to Europe by the Spaniards and became a popular beverage by 1700.
They also introduced the cacao tree into the West Indies and the Philippines. It was used in
alchemical processes, where it was known as Black Bean.

The first coffee house in Europe was opened Paris in the 1800s by an French-Armenian named
Pascal. Armenian merchants played in role in the more modern history of coffee and this is the
reason why the coffee growing region in is named the Armenia Region of Columbia.In 1819, the
German chemist Friedrich Ferdinand Runge isolated relatively pure caffeine for the first time.
According to a legend, he did this at the behest of Johann Wolfgang von Goethe.[17]Today,
global consumption of caffeine has been estimated at 120,000 tons per annum,[18] making it the
world's most popular psychoactive substance. This number equates to one serving of a caffeinic
beverage for every person, per day. In North America, 90% of adults consume some amount of
caffeine daily.

Caffeine is a central nervous system and metabolic stimulant,[19] and is used both recreationally
and medically to reduce physical fatigue and restore mental alertness when unusual weakness or
drowsiness occurs. Caffeine stimulates the central nervous system first at the higher levels,
resulting in increased alertness and wakefulness, faster and clearer flow of thought, increased

focus, and better general body coordination, and later at the spinal cord level at higher doses.[20]
Once inside the body, it has a complex chemistry, and acts through several mechanisms as
described below.

Caffeine is metabolized in the liver into three primary metabolites: paraxanthine (84%),
theobromine (12%), and theophylline (4%) Caffeine is completely absorbed by the stomach and
small intestine within 45 minutes of ingestion. After ingestion it is distributed throughout all
tissues of the body and is eliminated by first-order kinetics.

The half-life of caffeine , the time required for the body to eliminate one-half of the total amount
of caffeine consumed at a given time varies widely among individuals according to such factors
as age, liver function, pregnancy, some concurrent medications, and the level of enzymes in the
liver needed for caffeine metabolism. In healthy adults, caffeine's half-life is approximately 3-4
hours. In women taking oral contraceptives this is increased to 5-10 hours, and in pregnant
women the half-life is roughly 9-11 hours.Caffeine can accumulate in individuals with severe
liver disease when its half-life can increase to 96 hours. In infants and young children, the half-
life may be longer than in adults; half-life in a newborn baby may be as long as 30 hours. Other
factors such as smoking can shorten caffeine's half-life.

Chemical identification, isolation, and synthesis

14
In 1819, the German chemist Friedlieb Ferdinand Runge isolated relatively pure caffeine for the
first time; he called it "Kaffebase" (i.e., a base that exists in coffee). According to Runge, he did
this at the behest of Johann Wolfgang von Goethe. In 1821, caffeine was isolated both by the
French chemist Pierre Jean Robiquet and by another pair of French chemists, Pierre-Joseph
Pelletier and Joseph Bienaimé Caventou, according to Swedish chemist Jöns Jacob Berzelius in
his yearly journal. Furthermore, Berzelius stated that the French chemists had made their
discoveries independently of any knowledge of Runge's or each other's work. However,
Berzelius later acknowledged Runge's priority in the extraction of caffeine, stating: "However, at
this point, it should not remain unmentioned that Runge specified the same method and
described caffeine under the name Caffeebase a year earlier than Robiquet, to whom the
discovery of this substance is usually attributed, having made the first oral announcement about
it at a meeting of the Pharmacy Society in Paris."

Pelletier's article on caffeine was the first to use the term in print (in the French form Caféine
from the French word for coffee: café). It corroborates Berzelius's account:

Caffeine, noun (feminine). Crystallizable substance discovered in coffee in 1821 by Mr.

Robiquet. During the same period – while they were searching for quinine in coffee because
coffee is considered by several doctors to be a medicine that reduces fevers and because coffee
belongs to the same family as the cinchona [quinine] tree – on their part, Messrs. Pelletier and
Caventou obtained caffeine; but because their research had a different goal and because their
research had not been finished, they left priority on this subject to Mr. Robiquet. We do not
know why Mr. Robiquet has not published the analysis of coffee which he read to the Pharmacy
Society. Its publication would have allowed us to make caffeine better known and give us
accurate ideas of coffee's composition.

Robiquet was one of the first to isolate and describe the properties of pure caffeine, whereas
Pelletier was the first to perform an elemental analysis.

In 1827, M. Oudry isolated "théine" from tea but it was later proved by Mulder [ and by Carl
Jobst] that theine was actually caffeine.

In 1895, German chemist Hermann Emil Fischer (1852–1919) first synthesized caffeine from its
chemical components (i.e. a "total synthesis"), and two years later, he also derived the structural
formula of the compound. This was part of the work for which Fischer was awarded the Nobel
Prize in 1902.

Caffeine has been a part of our global history for thousands of years. Each country has its own story
and source of caffeine. One of the most eccentric caffeine findings was in Ethiopia. The folk stories
passed between generations says that a farmer, who had recently moved his goats to a new pasture
found them to be restless and ancy. For the next few days he watched them, and noted that they were
grazing on small berries. These berries were later dried and called “coffee beans.”

15
NATURAL OCCURRENCE

Around sixty plant species are known to contain caffeine. Common sources are the "beans"
(seeds) of the two cultivated coffee plants, Coffea arabica and Coffea canephora (the quantity
varies, but 1.3% is a typical value); in the leaves of the tea plant; and in kola nuts. Other sources
include yaupon holly leaves, South American holly yerba mate leaves, seeds from Amazonian
maple guarana berries, and Amazonian holly guayusa leaves. Temperate climates around the
world have produced unrelated caffeine-containing plants.

Caffeine in plants acts as a natural pesticide: it can paralyze and kill predator insects feeding on
the plant High caffeine levels are found in coffee seedlings when they are developing foliage and
lack mechanical protectionIn addition, high caffeine levels are found in the surrounding soil of
coffee seedlings, which inhibits seed germination of nearby coffee seedlings, thus giving
seedlings with the highest caffeine levels fewer competitors for existing resources for survival
Caffeine is stored in tea leaves in two places. Firstly, in the cell vacuoles where it is complexed
with polyphenols. This caffeine probably is released into the mouth parts of insects, to
discourage herbivory. Secondly, around the vascular bundles, where it probably inhibits
pathogenic fungi from entering and colonizing the vascular bundles. Caffeine in nectar may
improve the reproductive success of the pollen producing plants by enhancing the reward
memory of pollinators such as honeybees.

16
 Fig 2 Roasted coffee beans

The differing perceptions in the effects of ingesting beverages made from various plants
containing caffeine could be explained by the fact that these beverages also contain varying
mixtures of other methylxanthine alkaloids, including the cardiac stimulants theophylline and
theobromine, and polyphenols that can form insoluble complexes with caffeine.

The most important source of caffeine worldwide is the coffee bean. This is the seed of the
coffee plant from which coffee is brewed. The exact amount of caffeine contained in coffee
beans varies greatly and depends on several factors such as climate and soil type.

One serving of coffee usually contains around 40 mg of caffeine in a single 30 ml shot. Drip coffee
contains around 100 mg in a 120 ml cup. Arabica coffee normally contains less caffeine than the
robusta variety. In general, dark-roast coffee has less caffeine than lighter roasts because roasting
reduces the caffeine content. As far as tea is concerned, pale teas such as the Japanese green tea
gyokuro contains a higher amount of caffeine than darker teas such as lapsang souchong.
Chocolates made from the cocoa bean contain a small amount of caffeine. A 28 mg serving of
milk chocolate contains as much caffeine as a cup of decaffeinated coffee.
Caffeine is also used in soft drinks such as Coca-Cola. Typically, a serving of Coca-Cola contains
around 10 to 50 mg of caffeine, while energy drinks such as Red Bull can contain 80 mg or more

17
of caffeine. One main ingredient of energy drinks is guarana, which contains large amounts of
caffeine.

USE OF CAFFEINE

Medical

Caffeine is used in:

• Bronchopulmonary dysplasia in premature infants for both prevention and treatment. It

may improve weight gain during therapy and reduce the incidence of cerebral palsy as
well as reduce language and cognitive delay. On the other hand, subtle long-term side
effects are possible.

• Apnea of prematurity as a primary treatment, but not prevention.

• Orthostatic hypotension treatment.

• In moderate doses, caffeine may reduce symptoms of depression and lower suicide risk.

Enhancing performance

18
Cognitive

Caffeine is a central nervous system stimulant that reduces fatigue and drowsiness. At normal
doses, caffeine has variable effects on learning and memory, but it generally improves reaction
time, wakefulness, concentration, and motor coordination. The amount of caffeine needed to
produce these effects varies from person to person, depending on body size and degree of
tolerance. The desired effects arise approximately one hour after consumption, and the desired
effects of a moderate dose usually subside after about three or four hours.

Caffeine can delay or prevent sleep and improves task performance during sleep
deprivation.Shift workers who use caffeine make fewer mistakes due to drowsiness.

A systematic review and meta-analysis from 2014 found that concurrent caffeine and L-theanine
use has synergistic psychoactive effects that promote alertness, attention, and task switching;
these effects are most pronounced during the first hour post-dose.

Physical

Caffeine is a proven ergogenic aid in humans. Caffeine improves athletic performance in

aerobic (especially endurance sports) and anaerobic conditions. Moderate doses of caffeine
(around 5 mg/kg) can improve sprint performance, cycling and running time trial performance,
endurance (i.e., it delays the onset of muscle fatigue and central fatigue), and cycling power
output. Caffeine increases basal metabolic rate in adults.

Specific populations

Adults

For the general population of healthy adults, Health Canada advises a daily intake of no more
than 400 mg.

Children

In healthy children, caffeine intake produces effects that are "modest and typically innocuous".
There is no evidence that coffee stunts a child's growth. For children age 12 and under, Health
Canada recommends a maximum daily caffeine intake of no more than 2.5 milligrams per
kilogram of body weight. Based on average body weights of children, this translates to the
following age-based intake limits.

19
 Table 2 daily caffeine intake

Adolescents

Health Canada has not developed advice for adolescents because of insufficient data. However,
they suggest that daily caffeine intake for this age group be no more than 2.5 mg/kg body
weight. This is because the maximum adult caffeine dose may not be appropriate for light
weight adolescents or for younger adolescents who are still growing. The daily dose of 2.5
mg/kg body weight would not cause adverse health effects in the majority of adolescent caffeine
consumers. This is a conservative suggestion since older and heavier weight adolescents may be
able to consume adult doses of caffeine without suffering adverse effects.

Pregnancy and breastfeeding

The UK Food Standards Agency has recommended that pregnant women should limit their caffeine
intake, out of prudence, to less than 200 mg of caffeine a day – the equivalent of two cups of instant
coffee, or one and a half to two cups of fresh coffee. The American Congress of Obstetricians and
Gynecologists (ACOG) concluded in 2010 that caffeine consumption is safe up to 200 mg per day in
pregnant women. For women who breastfeed, are pregnant, or may become

Age range Maximum recommended daily caffeine intake

4–6 45 mg (slightly more than in 12 oz of a typical caffeinated soft drink)

7–9 62.5 mg
10–12 85 mg (about ½ cup of coffee)

REF- Mayo Clinic staff. "Pregnancy Nutrition: Foods to avoid during

pregnancy". Mayo Clinic. Retrieved 15 April 2012.

pregnant, Health Canada recommends a maximum daily caffeine intake of no more than 300
mg, or a little over two 8 oz (237 mL) cups of coffee.

There are conflicting reports in the scientific literature about caffeine use during pregnancy. A
2011 review found that caffeine during pregnancy does not appear to increase the risk of
congenital malformations, miscarriage or growth retardation even when consumed in moderate
to high amounts. Other reviews, however, concluded that there is some evidence that higher
caffeine intake by pregnant women may be associated with a higher risk of giving birth to a low
birth weight baby. and may be associated with a higher risk of pregnancy loss. A systematic
review, analyzing the results of observational studies, suggests that women who consume large
amounts of caffeine (greater than 300 mg/day) prior to becoming pregnant may have a higher
risk of experiencing pregnancy loss.

Adverse effects

Physical
Caffeine can increase blood pressure and cause vasoconstriction. Coffee and caffeine can affect
gastrointestinal motility and gastric acid secretion. Caffeine in low doses may cause weak
20
Broncho dilation for up to four hours in asthmatics. In postmenopausal women, high caffeine
consumption can accelerate bone loss.

Doses of caffeine equivalent to the amount normally found in standard servings of tea, coffee
and carbonated soft drinks appear to have no diuretic action. However, acute ingestion of
caffeine in large doses (at least 250–300 mg, equivalent to the amount found in 2–3 cups of
coffee or 5–8 cups of tea) results in a short-term stimulation of urine output in individuals who
have been deprived of caffeine for a period of days or weeks. This increase is due to both a
diuresis (increase in water excretion) and a natriuresis (increase in saline excretion); it is
mediated via proximal tubular adenosine receptor blockade. The acute increase in urinary output
may increase the risk of dehydration. However, chronic users of caffeine develop a tolerance to
this effect and experience no increase in urinary output.

Psychological

Minor undesired symptoms from caffeine ingestion not sufficiently severe to warrant a
psychiatric diagnosis are common and include mild anxiety, jitteriness, insomnia, increased
sleep latency, and reduced coordination. Caffeine can have negative effects on anxiety disorders.
According to a 2011 literature review, caffeine use is positively associated with anxiety and
panic disorders. At high doses, typically greater than 300 mg, caffeine can both cause and
worsen anxiety. For some people, discontinuing caffeine use can significantly reduce anxiety.

Some textbooks state that caffeine is a mild euphoriant, others state that it is not a euphoriant,
and one states that it is and is not a euphoriant.

Reinforcement disorders

Whether or not caffeine can result in an addictive disorder depends on how addiction is defined.
Some diagnostic models, such as the ICDM-9 and ICD-10, include a classification of caffeine
addiction under a broader diagnostic model. Some state that certain users can become addicted
and therefore unable to decrease use even though they know there are negative health effects.

Caffeine does not appear to be a reinforcing stimulus, and some degree of aversion may actually
occur, with people preferring placebo over caffeine in a study on drug abuse liability published
in an NIDA research monograph. Some state that research does not provide support for an
underlying biochemical mechanism for caffeine addiction. Other research states it can affect the
reward system.

"Caffeine addiction" was added to the ICDM-9 and ICD-10. However, its addition was
contested with claims that this diagnostic model of caffeine addiction is not supported by
evidence. The American Psychiatric Association's DSM-5 does not include the diagnosis of a
caffeine addiction but proposes criteria for the disorder for more study.
21
Dependence and withdrawal

Withdrawal can cause mild to clinically significant distress or impairment in daily functioning.
The frequency at which this occurs is self reported at 11%, but in lab tests only half of the
people who report withdrawal actually experience it, casting doubt on many claims of
dependence. Mild to increasingly severe physical dependence and withdrawal symptoms may
occur upon abstinence, with greater than 100 mg caffeine per day; some symptoms associated
with psychological dependence may also occur during withdrawal. Caffeine dependence can
involve withdrawal symptoms such as fatigue, headache, irritability, depressed mood, reduced
contentedness, inability to concentrate, sleepiness or drowsiness, stomach pain, and joint pain.
Withdrawal headaches are experienced by roughly half of those who stop consuming caffeine
for two days following an average daily intake of 235 mg.

The ICD-10 includes a diagnostic model for caffeine dependence, but the DSM-5 does not. The
APA, which published the DSM-5, acknowledged that there was sufficient evidence in order to
create a diagnostic model of caffeine dependence for the DSM-5, but they noted that the clinical
significance of this disorder is unclear.[3] The DSM-5 instead lists "caffeine use disorder" in the
emerging models section of the manual.

Tolerance varies for daily, regular caffeine users and high caffeine users. High doses of caffeine
(750 to 1200 mg/day spread throughout the day) have been shown to produce complete
tolerance to some, but not all of the effects of caffeine. Doses as low as 100 mg/day, such as a 6
oz cup of coffee or two to three 12 oz servings of caffeinated soft-drink, may continue to cause
sleep disruption, among other intolerances. Non-regular caffeine users have the least caffeine
tolerance for sleep disruption. Some coffee drinkers develop tolerance to its undesired sleep-
disrupting effects, but others apparently do not.

Risk of other diseases

A protective effect of caffeine against Alzheimer's disease is possible, but the evidence is
inconclusive. Caffeine increases intraocular pressure in those with glaucoma but does not appear
to affect normal individuals. It may protect people from liver cirrhosis. Caffeine may lessen the
severity of acute mountain sickness if taken a few hours prior to attaining a high altitude.

22
Overdose

Consumption of 1–1.5 grams (0.035–0.053 oz) per day is associated with a condition known as
caffeinism. Caffeinism usually combines caffeine dependency with a wide range of unpleasant symptoms
including nervousness, irritability, restlessness, insomnia, headaches, and palpitations after caffeine use.

Caffeine overdose can result in a state of central nervous system over-stimulation called caffeine
intoxication (DSM-IV 305.90). This syndrome typically occurs only after ingestion of large amounts of
caffeine, well over the amounts found in typical caffeinated beverages and caffeine tablets (e.g., more
than 400–500 mg at a time). The symptoms of caffeine intoxication are comparable to the symptoms of
overdoses of other stimulants: they may include restlessness, fidgeting, anxiety, excitement, insomnia,
flushing of the face, increased urination, gastrointestinal disturbance, muscle twitching, a rambling flow
of thought and speech, irritability, irregular or rapid heartbeat, and psychomotor agitation. In cases of
much larger overdoses, mania, depression, lapses in judgment, disorientation, disinhibition, delusions,
hallucinations, or psychosis may occur, and rhabdomyolysis (breakdown of skeletal muscle tissue) can
be provoked.

23
 Fig 3 Primary symptoms of caffeine intoxication

Caffeine is a drug that in large amounts, especially over an extended period of time, can lead to a condition
termed "caffeinism." Caffeinism usually combines physical addiction with a wide range of unpleasant physical
and mental conditions including nervousness, irritability, anxiety, tremulousness, muscle twitching
(hyperreflexia), insomnia, and heart palpitations. (Under a rigid definition of addiction, meaning a process of
escalating use, "caffeine dependency" would be a more descriptive term. However, under the widely accepted
definition "chronic pattern of behavior that is perceived to be difficult to quit," caffeine may be said to be
addictive.) Furthermore, because caffeine increases the production of stomach acid, high usage over time can
lead to peptic ulcers, erosive esophagitis, and gastroesophageal reflux disease. However, since both "regular"
and decaffeinated coffees have also been shown to stimulate the gastric mucosa and increase stomach acid
secretion, caffeine is probably not the only component of coffee responsible.
There are four caffeine-induced psychiatric disorders recognized by the Diagnostic and Statistical Manual
of Mental Disorders, Fourth Edition: caffeine intoxication, caffeine-induced anxiety disorder, caffeine-
induced sleep disorder, and caffeine-related disorder not otherwise specified (NOS)

Massive overdose can result in death. The LD50 of caffeine in humans is dependent on individual sensitivity,
but is estimated to be 150–200 milligrams per kilogram of body mass (75–100 cups of coffee for a 70 kilogram
adult). A number of fatalities have been caused by overdoses of readily available powdered caffeine
supplements, for which the estimated lethal amount is less than a tablespoon. The lethal dose is lower in
individuals whose ability to metabolize caffeine is impaired due to genetics or chronic liver disease A death
was reported in a man with liver cirrhosis who overdosed on caffeinated mints.

Treatment of mild caffeine intoxication is directed toward symptom relief; severe intoxication may
require peritoneal dialysis

24
Products

Beverages

Coffee

The world's primary source of caffeine is the coffee "bean" (the seed of the coffee plant), from
which coffee is brewed. Caffeine content in coffee varies widely depending on the type of coffee
bean and the method of preparation used; even beans within a given bush can show variations in
concentration. In general, one serving of coffee ranges from 80 to 100 milligrams, for a single shot
(30 milliliters) of arabica-variety espresso, to approximately 100–125 milligrams for a cup (120
milliliters) of drip coffee. Arabica coffee typically contains half the caffeine of the robusta variety.
In general, dark-roast coffee has very slightly less caffeine than lighter roasts because the roasting
process reduces caffeine content of the bean by a small amount.

Tea

25
Tea contains more caffeine than coffee by dry weight. A typical serving, however, contains
much less, since tea is normally brewed more weakly than coffee. Also contributing to caffeine
content are growing conditions, processing techniques, and other variables. Thus, teas contain
varying amounts of caffeine.

Tea contains small amounts of theobromine and slightly higher levels of theophylline than
coffee. Preparation and many other factors have a significant impact on tea, and color is a very
poor indicator of caffeine content. Teas like the pale Japanese green tea, gyokuro, for example,
contain far more caffeine than much darker teas like lapsang souchong, which has very little.

Soft drinks and energy drinks

Caffeine is also a common ingredient of soft drinks, such as cola, originally prepared from kola
nuts. Soft drinks typically contain 0 to 55 milligrams of caffeine per 12 ounce serving. By
contrast, energy drinks, such as Red Bull, can start at 80 milligrams of caffeine per serving. The
caffeine in these drinks either originates from the ingredients used or is an additive derived from
the product of decaffeination or from chemical synthesis. Guarana, a prime ingredient of energy
drinks, contains large amounts of caffeine with small amounts of theobromine and theophylline
in a naturally occurring slow-release excipient.

Other beverages

Mate is a drink popular in many parts of South America. Its preparation consists of filling a
gourd with the leaves of the South American holly yerba mate, pouring hot but not boiling water
over the leaves, and drinking with a straw, the bombilla, which acts as a filter so as to draw only
the liquid and not the yerba leaves
Guaraná seeds ("beans") are used in making the commercially sold beverage Guaraná
Antarctica, which originated in Brazil and is currently the fifteenth most popular soft drink in
the world.
The leaves of Ilex guayusa, the Ecuadorian holly tree, are placed in boiling water to make a
guayusa tea, which is both brewed locally and sold commercially throughout the world.

Caffeine content of select common food and drugs

Caffeine per
Product Serving size
serving (mg)

Caffeine tablet 1 tablet 200

Excedrin tablet 1 tablet 65

Coffee, brewed 240 mL (8 US fl oz) 135*

Coffee, decaffeinated 240 mL (8 US fl oz) 5*

Coffee, espresso 57 mL (2 US fl oz) 100*

26
Chocolate, Dark (Hershey's Special Dark) 1 bar (43 g; 1.5 oz) 31

Chocolate, Milk (Hershey Bar) 1 bar (43 g; 1.5 oz) 10

Red Bull 240 mL (8.2 US fl oz) 80

Cocaine Energy Drink 250 mL (8.4 US fl oz) 280

Buckfast Tonic Wine 750 ml (1 bottle) 280

Bawls Guarana 296 mL (10 US fl oz) 67

Foosh Energy Mints 1 mint 100

Buzz Bites Chocolate Energy Chews 1 chocolate 100

Soft drink, Mountain Dew "Dew Fuel" 355 mL (12 US fl oz) 54.5

Soft drink, Coca-Cola Classic 355 mL (12 US fl oz) 34

Atomic Rush 255 mL (7 US fl oz) 100

Tea, green 240 mL (8 US fl oz) 15

Tea, leaf or bag 240 mL (8 US fl oz) 50

table 3 Caffeine content of select common food and drugs

REF- "Caffeine". Pubchem Compound. NCBI. Retrieved 16 October 2014.

Chocolate

Chocolate derived from cocoa beans contains a small amount of caffeine. The weak stimulant
effect of chocolate may be due to a combination of theobromine and theophylline, as well as
caffeine. A typical 28-gram serving of a milk chocolate bar has about as much caffeine as a cup
of decaffeinated coffee. By weight, dark chocolate has one to two times the amount of caffeine
as coffee: 80–160 mg per 100 g. Higher percentages of cocoa such as 90% amount to 200 mg
per 100 g approximately and thus, a 100-gram 85% cocoa chocolate bar contains about 195 mg
caffeine.

Tablets

Tablets offer the advantages over coffee and tea of convenience, known dosage, and avoiding
concomitant sugar, acid and fluid intake. Manufacturers of caffeine tablets claim that using
caffeine of pharmaceutical quality improves mental alertness. These tablets are commonly used
by students studying for their exams and by people who work or drive for long hours.

27
Other oral products

Another intake route is SpazzStick, a caffeinated lip balm. Alert Energy Caffeine Gum was
introduced in the United States in 2013, but was voluntarily withdrawn after an announcement
of an investigation by the FDA of the health effects of added caffeine in foods. Inhalants

There are several products being marketed that offer inhalers that deliver proprietary blends of
supplements, with caffeine being a key ingredient. In 2012, the FDA sent a warning letter to one
of the companies marketing these inhalers, expressing concerns for the lack of safety
information available about inhaled caffeine.

Combinations with other drugs

• Some beverages combine alcohol with caffeine to create a caffeinated alcoholic drink.
The stimulant effects of caffeine may mask the depressant effects of alcohol,
potentially reducing the user's awareness of their level of intoxication. Such beverages
have been the subject of bans due to safety concerns. In particular, United States Food
and Drug Administration has classified caffeine added to malt liquor beverages as an
"unsafe food additive".

• Ya ba contains a combination of methamphetamine and caffeine.

• Painkillers such as propyphenazone/paracetamol/caffeine combine caffeine with an

analgesic.

Extraction of caffeine from tea

Caffeine is a member of the class of compounds organic chemists call alkaloids. Alkaloids are
nitrogen containing basic compounds that are found in plants. They usually taste bitter and often
are physiologically active in humans. The names of some of these compounds are familiar to you
even if the structures aren’t: nicotine, morphine, strychnine, and cocaine. The role or roles these
compounds play in the life of the plants in which they are found is not well understood. In some
cases they may act as pesticides; nicotine is found in tobacco and has been sprayed onto other
plants, in which it is not found, to function as an insecticide. The structure of caffeine is shown to
the right. It can be considered to be constructed from the purine ring system, which is important
biologically, being found in nucleic acids and elsewhere. Caffeine is found in a number of things
ingested by people. The table to the right lists some of them. Caffeine acts as a stimulant. It
stimulates the heart, respiration, the central nervous system, and is a diuretic. Its use can cause
nervousness, insomnia and headaches. It is physically addictive. A person who drinks as few as 4
cups of coffee a day and who attempts to stop “cold turkey” may experience headache, insomnia,
and possibly nausea as the result of withdrawal. Tea has been consumed as a beverage for almost
2,000 years, starting in China. It is a beverage produced
28
by steeping in freshly boiled water the young leaves and leaf buds of the tea plant, Camellia
sinensis. Today, two principal varieties are used, the small-leaved China plant (C. sinensis
sinensis) and the large-leaved Assam plant (C. sinensis assamica). Hybrids of these two
varieties are also grown. 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. As trade routes opened to Asia in the 17th century, tea was imported to Europe.

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 a
suitable solvent. The solution of these dissolved compounds is referred to as the Extract. Here the
organic solvent dichloromethane is used to extract caffeine from an 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 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.

Beverages cover a vast variety of addictive drinks out of which Tea and Coffee are the most
popular acceptable drinks. Tea powder is extracted from tea leaves which contain tannins,
which are acidic in nature, a number of colored compounds and a small amount of
unrecompensed chlorophyll and an important stimulant called Caffeine. Because of the
presence of Caffeine, tea and coffee are gaining popularity as an addictive stimulant. An
average 30g of tea can contain 20-ll0 mg of caffeine thereby making tea a significant source
of caffeine compared to other beverages. Caffeine can stimulate nervous system and can
cause relaxation of respiratory and cardiac muscles. Caffeine is well known to increase both
the alertness level and attention span. But like all other addictive’s, tea also shows withdrawal
symptoms like headache, nervousness and insomnia for a regular consuming person.

Caffeine, 1,3,7 - trimethylxanthine, belongs to a wide class of compounds known as alkaloids.

These are plant derived compounds with complex structure containing nitrogen, and usually
have roles in physiological activity. The melting point of Caffeine is 238°C.

Principle

Extraction is a method used for the separation of organic compound from a mixture of compound.
This technique selectively dissolves one or more compounds into an appropriate solvent. The
solution of these dissolved compounds is referred to as the extract. In the case of Caffeine
extraction from tea powder, the solubility of caffeine in water is 22mg/ml at 25°C, 180mg/ml at
80°C, and 670mg/ml at 100°C. Here the organic solvent Dichloromethane is used to extract
caffeine from aqueous extract of tea powder because caffeine is more soluble in dichloromethane
(140mg/ml) than it is in water (22mg/ml).The dichloromethane - caffeine mixture can then be
separated on the basis of the different densities of dichloromethane and water because
dichloromethane is much denser than water and insoluble in it. Residual water is separated from
dichloromethane by drain out the dichloromethane through separating funnel, thus
dichloromethane passed through the funnel while polar solvents such as water is still

29
remains in the funnel. Water and dichloromethane is slightly soluble in each other. So, after
separating the solvents, residual water will remain the organic layer. Mainly anhydrous
sodium sulfite is used for the removal of water from organic layer. Anhydrous sodium sulfite
is an insoluble inorganic solid which will absorb water, thus drying it.

Background to the Procedure

Tea leaves consist mostly of cellulose, a water-insoluble polymer of glucose, which is a simple
sugar (a monosaccharide). Cellulose performs a function in plants similar to that of fibrous
proteins in animals: it is structure building material. Along with the cellulose are found a number
of other things including caffeine, tannins (phenolic compounds, compounds that have an -OH
directly bonded to an aromatic ring) and a small amount of chlorophyll. The idea in this
experiment is to extract the water soluble materials in the tea leaves into hot water. [The solubility
of caffeine in water is 22 mg/ml at 25oC, 180 mg/ml at 80oC, and 670 mg/ml at 100oC.] The hot
solution is allowed to cool and the caffeine is then extracted from the water with dichloromethane
(methylene chloride), which is an organic solvent that is insoluble in water. Since caffeine is more
soluble in dichloromethane (140 mg/ml) than it is in water (22 mg/ml), it readily dissolves in the
dichloromethane. However, the tannins are slightly soluble in the dichloromethane. But we want
to separate the caffeine from the tannins by having the caffeine dissolve in the dichloromethane
and the tannins remain in the water. We can do thi by taking advantage of the fact that phenols are
acidic enough to be converted to their salts (deprotonation of the -OH group) by reaction with
sodium carbonate. So, we will add sodium carbonate to the water and the tannins will be
converted to phenolic anions, which are not soluble in the dichloromethane but are soluble in
highly polar water. There is one practical disadvantage in converting the tannins to their salts –
they become anionic surfactants. Detergents and soap are surfactants. It is the purpose of
surfactants to cause materials that do not dissolve in water (like oil, grease and dichloromethane)
to form an emulsion with water. We want to be able to separate the aqueous phase from the
dichloromethane phase, so the las thing we want is an emulsion of the two. Consequently, as you
extract the caffeine from th water into the dichloromethane do notshake the separatory funnel
vigorously.

Procedure

1. Open two tea bags by removing the staple, trying to not tear the paper. Weigh the total
contents to the nearest milligram and record this weight. Place the tea back into the
bags and staple the bags shut.

2. Place the tea bags into the bottom of a clean, but not necessarily dry, 150 ml beaker. Add
30 ml of deionized water and 2.0 g of anhydrous sodium carbonate. Place the beaker on a
wire gauze supported by a ring on a ring stand. Place a watch glass on top of the beaker.
Gently boil the contents of the beaker for 10 minutes using a micro burner. If the tea bags
rise out of the liquid push them down from time to time using a glass stirring rod.

30
3. Decant the hot liquid into a 50 ml Erlenmeyer flask. Add 20 ml of deionized water to
the beaker and again bring the contents to a boil. Again decant the liquid in the beaker
into the 50 ml Erlenmeyer flask. Press the tea bags with a clean cork to express as
much liquid as possible without breaking the bag and transfer this liquid to the 50 ml
Erlenmeyer flask also. Cool the tea extract to room temperature. Discard the tea bags.

4. Transfer the tea extract from the 50 ml Erlenmeyer flask to a 125 ml separatory funnel
that is supported by a ring on a ring stand.

5. Add 5 milliliters of dichloromethane to the separatory funnel. Stopper the funnel and
grab the neck of the funnel with one hand, holding the stopper into the neck of the
funnel. With your other hand grasp the stopcock in such a way that you can turn the
plug in the barrel to open and close the stopcock. While holding the stopper tightly into
the neck of the funnel, invert the funnel so the liquid no longer is in contact with the
stopcock. Pointing the stem of the funnel away from everybody, open the stopcock to
release any pressure that may have built up inside the funnel. Close the stopcock and
agitate the material in the funnel without shaking it vigorously – you want the contents
to mix, but you do not want to generate an emulsion. Point the stem of the funnel
upward, and making sure no liquid is in contact with the stopcock open it to again
relieve any pressure build-up. Close the stopcock. Agitate again as before and reclamp
the separatory funnel to the ring stand.

6. Allow the contents of the separatory funnel to settle. There should be two distinct mostly
clear layers. If there is an emulsion (cloudy) layer between two clear layers it is sometimes
possible to break the emulsion by swirling the contents of the funnel or stirring the
contents using a glass rod. If the emulsion persists seek your instructors help.
7. Carefully drain the lower (dichloromethane) layer into a 25 ml Erlenmeyer flask. Try
to not include any of the aqueous (upper) layer. If there is a lot of emulsion, include it
in the Erlenmeyer flask.

8. Repeat steps 5 through 7 using a second 5 ml portion of dichloromethane.

9. Add 0.5 g of anhydrous sodium sulfate to the combined dichloromethane extracts in the
25 ml Erlenmeyer flask. Swirl the contents of the flask. The anhydrous sodium sulfate will
absorb the small amount of water that is dissolved in the dichloromethane and small
amounts of water from the aqueous layer that may have gotten into the flask by accident.

10. Decant the liquid from the flask into a 25 ml beaker. Place one Boileezer into the
beaker. [Boileezer boiling chips or stones help to prevent bumping by allowing bubbles
to form smoothly during boiling.] Place the beaker on a hot plate and when the volume
of material in the beaker is between 3 and 5 ml start adding petroleum ether by means
of a Pasteur pipet. When the solution in the beaker begins to get cloudy remove the
beaker from the heat and allow it to cool at your bench. With luck, crystals of caffeine
will form in the solution.

31
 11. Set up the Hirsh funnel from the microkit as shown in the illustration to the right. Be sure
to clamp the filter flask to a ring stand and to place a piece of filter paper in the funnel.
Connect a rubber hose from the side-arm on the flask to the side-arm (NOT THE
BOTTOM) of the aspirator. Turn on the aspirator. Pour the crystals and mother liquor from
the beaker into the Hirsh funnel. If some crystals stick to the beaker, you can scrape them
out with a spatula or wash them out with the mother liquor in the filter flask if you used a
clean flask. Allow air to be drawn through the crystals for 10 minutes.

References

1. Malenka RC, Nestler EJ, Hyman SE (2009). "Chapter 15: Reinforcement and
Addictive Disorders". In Sydor A, Brown RY. Molecular Neuropharmacology: A
Foundation for Clinical Neuroscience (2nd ed.). New York: McGraw-Hill Medical.
p. 375. ISBN 978-0-07-148127-4. Long-term caffeine use can lead to mild physical
dependence. A withdrawal syndrome characterized by drowsiness, irritability, and
headache typically lasts no longer than a day. True compulsive use of caffeine has
not been documented.
2. Karch SB (2009). Karch's pathology of drug abuse (4th ed.). Boca Raton: CRC Press.
pp. 229–230. ISBN 978-0-8493-7881-2. The suggestion has also been made that a
caffeine dependence syndrome exists ... In one controlled study, dependence was
diagnosed in 16 of 99 individuals who were evaluated. The median daily caffeine
32
 consumption of this group was only 357 mg per day (Strain et al., 1994).
Since this observation was first published, caffeine addiction has been added as an
official diagnosis in ICDM 9. This decision is disputed by many and is not supported by
any convincing body of experimental evidence. ... All of these observations strongly
suggest that caffeine does not act on the dopaminergic structures related to addiction,
nor does it improve performance by alleviating any symptoms of withdrawal
3. American Psychiatric Association (2013). "Substance-Related and Addictive
Disorders" (PDF). American Psychiatric Publishing. pp. 1–2. Archived from the
original (PDF) on 15 August 2015. Retrieved 10 July 2015. Substance use disorder in
DSM-5 combines the DSM-IV categories of substance abuse and substance
dependence into a single disorder measured on a continuum from mild to severe. ...
Additionally, the diagnosis of dependence caused much confusion. Most people link
dependence with "addiction" when in fact dependence can be a normal body
response to a substance. ... DSM-5 will not include caffeine use disorder, although
research shows that as little as two to three cups of coffee can trigger a withdrawal
effect marked by tiredness or sleepiness. There is sufficient evidence to support this
as a condition, however it is not yet clear to what extent it is a clinically significant
disorder.
4. Introduction to Pharmacology (third ed.). Abingdon: CRC Press. 2007. pp. 222–
223. ISBN 978-1-4200-4742-4.
5. Juliano LM, Griffiths RR (October 2004). "A critical review of caffeine withdrawal:
empirical validation of symptoms and signs, incidence, severity, and associated
features" (PDF). Psychopharmacology. 176 (1): 1–29. doi:10.1007/s00213-004-2000-x.
PMID 15448977. Archived from the original (PDF) on 29 January 2012. Results: Of 49
symptom categories identified, the following 10 fulfilled validity criteria: headache,
fatigue, decreased energy/ activeness, decreased alertness, drowsiness, decreased
contentedness, depressed mood, difficulty concentrating, irritability, and foggy/not
clearheaded. In addition, flu-like symptoms, nausea/vomiting, and muscle
pain/stiffness were judged likely to represent valid symptom categories. In
experimental studies, the incidence of headache was 50% and the incidence of
clinically significant distress or functional impairment was 13%. Typically, onset of
symptoms occurred 12–24 h after abstinence, with peak intensity at 20–51 h, and for a
duration of 2–9 days.
6. "Caffeine". DrugBank. University of Alberta. 16 September 2013. Retrieved 8 August
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7. Poleszak E, Szopa A, Wyska E, Kukuła-Koch W, Serefko A, Wośko S, Bogatko K,
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Boiling Point
178 °C (sublimes)
Melting Point
238 DEG C (ANHYD)

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9. "Caffeine". ChemSpider. Royal Society of Chemistry. Retrieved 16 October
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234–236 °C Alfa Aesar
237 °C Oxford University Chemical Safety Data
238 °C LKT Labs [C0221]
237 °C Jean-Claude Bradley Open Melting Point Dataset 14937
238 °C Jean-Claude Bradley Open Melting Point Dataset 17008, 17229,
22105, 27892, 27893, 27894, 27895
235.25 °C Jean-Claude Bradley Open Melting Point Dataset 27892, 27893, 27894,
27895
236 °C Jean-Claude Bradley Open Melting Point Dataset 27892, 27893, 27894, 27895
235 °C Jean-Claude Bradley Open Melting Point Dataset 6603
234–236 °C Alfa Aesar A10431,
39214 Experimental Boiling Point:
178 °C (Sublimes) Alfa Aesar
178 °C (Sublimes) Alfa Aesar 39214
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