CHEMISTRY PROJECT REPORT ON COMPARE THE

POLYPHENOL CONTENT IN THE VARIOUS SAMPLES

OF TEA LEAVES

Submitted by

Hariharasudhan.A ,

CLASS: XII

PROJECT GUIDE

Dr. S.SIVA, M.Sc; M.PHIL; B.Ed; Ph.D.

MADURAI PUBLIC SCHOOL

MADURAI – 625017.

MARCH-2024
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 MADURAI PUBLIC SCHOOL (CBSE)
MADURAI – 626123.

BONAFIDE CERTIFICATE

This is to certify that HARIHARASUDHAN.A Reg. No. of

Grade XII of Madurai Public School, Sivakasi has

successfully completed his project work under my guidance for

the academic year 2020-21. He has taken proper care in

completing this project. I certify that this project is up to my

expectation and as per the CBSE guidelines.

INTERNAL EXAMINER EXTERNAL EXAMINER

PRINCIPAL

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 MARCH 2024

DECLARATION

I declare that the Project entitled CHEMISTRY

PROJECT REPORT ON COMPARE THE POLYPHENOL

CONTENT IN THE VARIOUS SAMPLES OF TEA

LEAVES is the result of a study originally carried out by

me under the guidance of Dr. S. SIVA, MADURAI

PUBLIC SCHOOL (CBSE), MADURAI.

Signature of the Candidate

(HARIHARASUDHAN.A)

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 ACKNOWLEDGEMENT

“Thankfulness is to God immense who gives us the triumph”

Management, Principal and Staff Members, MADURAI

PUBLIC SCHOOL (CBSE), MADURAI for their valuable

help rendered to me in providing me the facilities.

It is impossible to express my indebtedness to my

Guide Dr. S.SIVA, MADURAI PUBLIC SCHOOL (CBSE),

MADURAI for the dynamic guidance, constant help,

sincere and compassionate advices, patience and

insightful discussion and parental care for completion of

this project work.

Last but not the least, I express my gratitude to

Almighty for giving me strength and health to finish

this work successfully.

(HARIHARASUDHAN.A)

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 CONTENTS

CHAPTER NO PAGE NO.

1. Introduction 5

2. Materials and Methods 20

3. Results and Discussion 23

4. Conclusion 25

5. Reference and Bibliography 2

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 CHAPTER-I

INTRODUCTION

Introduction

Caffeine in tea is dependent on many conditions such as type of cultivar, soil

conditions, temperature, etc. Studies show that the levels can change from plantation

to plantation, from tea to tea. Research has shown that some black teas actually have

less caffeine than some green teas. Some studies are not rigorously controlled or they

have examined certain teas with a specific water temperature and steep time, but not at

any other temperature or steep time. One study examined caffeine in a black tea after

a seven minute steep but not at the usual five minute steep. Suffice it to say, tea has

less caffeine than coffee. If you want to reduce caffeine intake, just make weaker

cups of tea and discard the first rinse, which may or may not lower the caffeine level. I

have reservations about the stated amount of caffeine in white tea, which is the top

bud and leaves. It should at least have the same amount as some greens, even taking

into account the volume of leaf, since most caffeine is at the top of the bush to repel

insects.

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Coffee is an excitant and caffeine reaches the brain in less than 5 minutes, 2-5 hours

later the effect has gone. Tea is a stimulant and caffeine is released gradually over 10

hours.

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 There are approximately 600 traces of aroma compounds in tea leaves, some of

which are lost and some that are released during the manufacturing process. Once

plucked, tea leaves begin to wither, their cell walls begin to break down and chemical

compounds begin to form new chemical compounds, not all of which are

watersoluble.

Carbohydrates, which add to the sweetness of tea, help to fuel the enzymatic

reactions that take place during oxidation and are also responsible for the creation of

polyphenols in young tea leaves.

The term polyphenol refers to a categorization of compounds composed of many

phenolic groups – up to 30,000. These compounds are plant metabolites produced as

a defense against insects and other animals; such as the alkaloid caffeine and two

similar compounds: theobromine and theophylline, which add to the bitterness of tea.

Alkaloid levels depend on the varietal and cultivar of camellia sinensis used, along

with climate, geology, geography, age of the leaves and the propagation method used.

The bud and first leaf have the highest concentration of polyphenols, 2-3 times more

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caffeine, and polyphenol levels decrease in each leaf moving down the plant.

One compound, flavanoids (also known as tannins) are responsible for the health

claims of tea. The major flavanols in tea are: catechin (C), epicatechin (EC),

epicatechin gallate (ECG), gallocatechin (GC), epigallocatechin (EGC), and

epigallocatechin gallate (EGCG), EGCG is the most active of these catechins and is

often the subject of studies regarding tea antioxidants especially as it relates to cancer

research.

The amino acid L-theanine found in tea has been shown to reduce cognitive stress

and used in conjunction with caffeine, show a possible improvement in memory and

learning tasks.

The volatile substances in tea leaves are largely responsible for a tea’s flavor and

aroma, and many are derived from the actual manufacturing process itself, e.g.: The

differences of varieties, the environmental effects, various methods of processing and

modes of propagations cause the change of chemical composition of tea leaves. As

shown in the figure on the right, the composition of fresh tea flush contains various

components, such as polyphenol (include catechins),caffeine,amino acids,

vitamins,flavonoids, polysaccharides and fluorine. Structural formulae of catechins,

caffeine, theanine, saponins as main green tea componets are drawn in the figure

below. Polyphenols and caffeine are the most important chemicals of tea, considerable

pharmacological significance. Polyhenols are present to the extent of 30-35 % in the

dry tea leaf matter and their content determines the quality of the beverage.

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Polyphenols

Plant-derived polyphenol, tannic acid, formed by esterification of ten

equivalents of the phenylpropanoid-derived gallic acid to a monosaccharide (glucose)

core from primary metabolism

Phenol-phenolate equilibrium, and resonance structures giving rise

to phenol aromatic reactivity

Polyphenols also known as polyhydroxyphenols are a structural class of

mainly natural, but also synthetic or semisynthetic, organic chemicals characterized by

the presence of large multiples of phenol structural units. The number and

characteristics of these phenol structures underlie the unique physical, chemical, and

biological (metabolic, toxic, therapeutic, etc.) properties of particular members of the

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class. Examples include tannic acid (image at right) and ellagitannin(image below).

The historically important chemical class of tannins is a subset of the polyphenols.

The name derives from the Ancient Greek word πολύς (polus, meaning "many,

much") and the word phenol which refers to a chemical structure formed by attaching

to an aromatic benzenoid (phenyl) ring, an hydroxyl (-OH) group akin to that found in

alcohols (hence the -ol suffix). The term polyphenol appears to have been in use since

1894.

Definition of the term polyphenol

Ellagic acid, a core-type 'component' of polyphenols

Raspberry ellagitannin, a tannin composed of 14 gallic acid units around a core

of three units of glucose, with two gallic acids as simple esters, and the remaining 12

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appearing in 6 ellagic acid-type units. Ester, ether, and biaryl linkages are present,

see below.

Original "WBSSH" definition of polyphenols

The earliest widely accepted definition of polyphenols, the White–Bate-Smith–

Swain–Haslam (WBSSH) definition,was offered and justified by natural product and

organic chemist Edwin Haslam and co-workers, based on the earlier natural products

research of Edgar Charles Bate-Smith, Anthony Swain, and Theodore White that

characterized specific structural characteristics common to plant phenolics used

in tanning (i.e., the tannins).The WBSSH describes the polyphenol class as:

• generally moderately water-soluble compounds

• with molecular weight of 500–4000 Da

• with >12 phenolic hydroxyl groups

• with 5–7 aromatic rings per 1000 Da

where the limits to these ranges are somewhat flexible.[1][7] The definition further

states that polyphenols display unique physical and chemical behaviors related to their

high molecular weights and profusion of phenolic substructures—precipitation of

proteins and particular amine-containing organics (e.g., particular alkaloid natural

products), and formation of particular metal complexes (e.g., intense blue-black

iron(III) complexes).

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Proposed Quideau definition of polyphenols

The need to clarify the definition of 'polyphenols' in the light of the extensive

research into this large substance class and of increasingly ambiguous use of the

polyphenol term led Stéphane Quideau, Bordeaux 1 University, France, to offer a

definition not given formal status by IUPAC: The term "polyphenol" should be used

to define compounds exclusively derived from the shikimate/phenylpropanoid and/or

the polyketide pathway, featuring more than one phenolic unit and deprived of

nitrogen-based functions. Structurally, this definition continues to steer the definition

away from exclusively man-made structures without corresponding natural products,

and explicitly excludes monophenolic structures (man-made or naturally occurring)

and their derivatives, e.g., phenyl esters, methyl phenyl ethers and O-phenyl

glycosides. This definition departs from the WBSSH definition in terms of

physicochemical behavior, with its lack of reference to solubility, precipitation, and

complexation phenomena.

Ellagic acid (M.W. 302, right), a molecule at the core of naturally occurring

phenolic compounds of varying sizes, is itself not a polyphenol by the WBSSH

definition, but is by the Quideau definition. The raspberry ellagitannin (M.W.

~2450),on the other hand, with its 14 gallic acid moieties (most in ellagic acid-type

components), and more than 40 phenolic hydroxyl groups, meets the criteria of both

definitions of a polyphenol. Other examples of compounds that fall under both the

WBSSH and Quideau definitions include the black teaantioxidant theaflavin-3-

gallate shown below, and the hydrolyzable tannin, tannic acid, shown above.

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Theaflavin-3-gallate, a plant-derived polyphenol, an ester of gallic acid and

a theaflavin core. There are 11 phenolic hydroxyl groups if one counts two that

are engaged in ether linkages.

Defining chemical reactions of the polyphenol class

Individual polyphenols engage in reactions related to both their core phenolic

structures, their linkages, and types of glycosides they form. Standard phenolic

reactions include ionization (which contributes to solubility and complexation),

oxidations to ortho- and para-quinones (which contributes to antioxidant

characteristics), and underlying aromatic transformations related to the presence of

the phenolic hydroxyl (see phenol image above); reactions related to their linkages

include nucleophilic additions, and oxidative and hydrolytic bond cleavages.[10]In

addition, as noted above, a traditional feature of polyphenols was their ability to

form particular, characteristic metal complexes.

Chemical Structure and its synthesis

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 As opposed to smaller phenols, polyphenols are often larger molecules

(macromolecules) deposited in cell vacuoles. The upper molecular weight limit for

small molecules is about 800 Daltons, which allows for the possibility to rapidly

diffuse across cell membranes so that they can reach intracellular sites of action or

remain as pigments once the cell senesces. Hence, many larger polyphenols are

biosynthesized in-situ from smaller polyphenols to nonhydrolyzable tannins and

remain undiscovered in the plant matrix. Most polyphenols contain repeating

phenolic moieties of pyrocatechol, resorcinol, pyrogallol, and phloroglucinol

connected by esters (hydrolyzable tannins) or more stable C-C bonds

(nonhydrolyzable condensed tannins). Proanthocyanidins are mostly polymeric

units of catechin and epicatechin. Catechol and resorcinol (benzenediol) types of

polyphenols have two, and pyrogallol and phloroglucinol (benzenetriol) types

have three phenolic hydroxyl groups, respectively, though mixing of these types

within polyphenols is also possible. The phenolic substructures arise from various

biosynthetic pathways (WBSSH definition), especially phenylpropanoid and

polyketide branches aimed at plant and related secondary metabolites (both

definitions).

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Pyrocatechol Resorcinol Pyrogallol Phloroglucinol

The C-glucoside substructure of polyphenols is exemplified by the phenol-

saccharide conjugate puerarin, a midmolecular-weight plant natural product. The

attachment of the phenol to the saccharide is by a carbon-carbon bond.

The isoflavone and its 10-atom benzopyran "fused ring" system, also a structural

feature here, is common in polyphenols.

Polyphenols always have heteroatom substituents other than hydroxyl

groups; ether and ester linkages are common, as are various carboxylic

acid derivatives (see theaflavin gallate image); ester linkages are common in

the hydrolyzable tannins. Apart from simple heteroatom links, the carbon

frameworks can become complex, e.g., various carbon-carbon bond linkages join

hydrolytically labile esters and ethers as common in non-hydrolyzable condensed

tannins.

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 The biphenyl/biaryl substructure of polyphenols, here as prepared by synthetic

chemists using the copper-mediated Ullmann reaction. The carbon-carbon bond

in biaryls in nature is also synthesized though a metal-mediated coupling

reaction, often involving iron. The biaryl substructure can be seen in ellagic acid

above.

An example of the spiro-type substructure found in polyphenols—where two

rings are joined at a single shared point—with illustration of the two

stereoisomers that can arise, here labeled R (rectus) and S (sinister) based on

the CIP system to describe stereochemistry.

In these, diverse biosynthetic steps abound: the seven-atom ring (seven-

membered ring) appearing in theaflavin structure above is an example of a

"carbocycle" that is of a nonbenzenoid aromatic tropolone type. In addition, there

are periodic occurrences of:

benzopyrans and normal and C-glucoside derivatives (figure at right)—e.g.

in condensed, complex and hydrolyzable tannins such as in stenophyllanin A,

acutissimin B, mongolicain A, stenophynin A, mongolicanin, and mongolicin B,

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 True polyphenols from the tannin and other WBSSH types are routinely

biosynthesized in the natural sources from which they derive; their 'chemical'

syntheses (using standard "bench" organic chemical methods) were somewhat limited

until the first decade of the new millennium because these syntheses involve

challenging regioselectivity and stereoselectivity issues. Early work focused on the

achiral synthesis of phenolic-related components of polyphenols in the late 70's, and

the Nelson and Meyers synthesis of the permethyled derivative of the ubiquitous

diphenic acid core of ellagitannins in 1994 followed by stereoselective synthesis of

more complex permethylated structures such as a (+)-tellimagrandin II derivative by

Lipshutz and coworkers in the same year,[ and Itoh and coworker's synthesis of a

permethylated pedunculagin with particular attention to axial symmetry issues in

1996.The total synthesis of a fully unmasked polyphenol, that of the ellagitannin

tellimagrandin I, was a diastereoselective sequence reported in 1994 by Feldman,

Ensel and Minard.

Further total syntheses of deprotected polyphenols that followed were led by the

Feldman group, for instance in Feldman and Lawlor's synthesis of the ellagitannin,

coriariin A and other tannin relatives. Khanbabaee and Grosser accomplished a

relatively efficient total synthesis of pedunculagin in 2003.

Work proceeded with focus on enantioselective total syntheses, e.g.,

on atroposelective syntheses of axially chiral biaryl polyphenols,with recent further

important work including controlled assembly of a variety of polyphenols according

to integrated strategies, such as in syntheses of extended series

of procyanidins (oligomeric catechins) by various groups and of resveratrol

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polyphenols by the Snyder group at Columbia that included the diverse carasiphenols

B and C, ampelopsins G and H, and nepalensinol B. A biomimetic synthesis, and the

first formal total synthesis 5-O-Desgalloyl-epi-punicacortein A, a

further ellagitannin in its C-glucosyl (C-glucoside subclass), has also recently been

accomplished. The novel strategies and methods referred to in these recent examples

helped to open the field of polyphenol chemical synthesis to an unprecedented degree.

Chemical properties and uses

Polyphenols are molecules owing their UV/Vis absorptivity to aromatic

structures with large conjugated systems of pi electron configurations; they also have

autofluorescence properties, especially lignin and the phenolic part of suberin.

They are reactive species toward oxidation. ABTS may be used to characterise

polyphenol oxidation products.

Polyphenols also characteristically possess a significant binding affinity for

proteins, which can lead to the formation of soluble and insoluble protein-polyphenol

complexes.

Chemical uses

Some polyphenols are traditionally used as dyes. For instance, in the Indian

subcontinent, the pomegranate peel, high in tannins and other polyphenols, or its juice,

is employed in the dyeing of non-synthetic fabrics.

Polyphenols, especially tannins, were used traditionally for tanning leather and

today also as precursors in green chemistry notably to produce plastics or resins by

polymerisation with or without the use of formaldehyde or adhesives for

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particleboards. The aims are generally to make use of plant residues from grape, olive

(called pomaces) or pecan shells left after processing.

Pyrogallol and pyrocatechin are among the oldest photographic developers.

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 CHAPTER-I

MATERIALS AND METHODS

Materials:

1. Beakers

2. Conical flask

3. Funnel

4. Heating arrangement

5. Tea Bags

6. Different brands of tea

Methods:

(i) Weigh exactly 5g of each of the different samples of tea and place them in tea

bags

(ii) Take three or four conical flasks and put 100 ml of hot water to each of them.

(iii) Put tea bags of different samples of tea in the different conical flaks. After

10 minutes remove the tea bags and dry them by placing in an oven.

(iv) When the tea bags become dry, reweigh them.

(v) Note the flavor of the tea solution obtained.

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22
 CHAPTER-III

RESULTS AND DISCUSSION

Table: 1 Corrosion inhibition by Polystyrene on Iron Metal

S.No Brand Initial Final Loss in % of water

weight weight weight soluble

[x-y/x (x)100]

1. Nestle 5 2.8 2.2 44%

2. Gold 5 2.5 2.5 50%

Cup

3. Jiyo tea 5 2.3 2.7 54%

(i) From the obtained data’s, we know that the Jiyo tea has more polyphenol

contents as compared to the other tea companies.

(ii) Hence, Jiyo tea has more flavor contents.

CONCLUSION

CONCLUSION

(i) % of Polyphenol in Nestle tea company : 44%

(ii) % of Polyphenol in Gold cup company: 50%

(iii) % of Polyphenol in Jiyo Tea Company: 54%

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 CHAPTER-V

REFERENCE AND BIBLIOGRAPHY

Reference and Bibliography

1. Quideau, S. P.;. (2011). "Plant Polyphenols: Chemical Properties, Biological

Activities, and Synthesis". Angewandte Chemie International

Edition. 50 (3): 586–621.

2. Quideau S (2011). "Why bother with polyphenols?". Groupe Polyphenols.

Retrieved 26 March 2014.

3. Polyphenol on www.merriam-webster.com online dictionary

4.Polyphenol definition and meaning | Collins English

Dictionary". www.collinsdictionary.com. Retrieved 2017-12-02.

5. Nonaka, G. (1989). "Isolation and structure elucidation of tannins" (PDF). Pure

Appl. Chem. 61 (3): 357–360. doi:10.1351/pac198961030357.

6. www.google.com

7. 11th and 12th NCERT Chemistry Laboratory Lab Manual.

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