INTRODUCTION

Now a day’s guava leaves and any other part of guava tree is useful in
medical field or industrial field. It has 100 species of tropical shrubs. It
is cultivated all over the world, including India, Native to Mexico,
Central America, and Northern South America. It is a frequently eaten
species. The two guava leaves grows in an opposite arrangement, it
have short petioles or stalk that join the leaf to the stems.

CLASSIFICATION OF GUAVA

1. Kingdom:- Plantae
2. Sub-kingdom:-Tracheobionata
3. Super-division:-Spermatophyta
4. Division:-Magnoliophyta
5. Class:-Magnoliopsida
6. Sub-class:-Rosidae
7. Order :-Myrtales
8. Family :-Myrtaceae
9. Genus:-Psidiumguajava

The guava leaves is obtained from guavas tree, they belong to other
species or genera. The colour of guava leaves is dull green to dark
green pink in colour. It has characteristic odour and the size of guava
leaves is 7 to 15 cm in length and 3 to 5 cm in width. The leave of
guava leave may be oval, oblong, and ovate-elliptic and they have hairy
undersides entire margin. The chemical constituent of Guava contains
phenolic compound, isofalvonoside, Gallic acid, catechin, rutin,
tannins, saponins, glycoside, starch etc.The guava leaves is used in the

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treatment of colic, diarrhoea, diabetes, cough, pain, a high cholesterol,
Heart disease, pain, weight loss, and cancer. It is also used as
antioxidant, stimulant and as dysentery. The properties of guava leaves
are anti spasmodic and anti microbial properties. The other important
properties of guava leave are following Hepatoprotective, anti –allergy,
anti cough, anti diabetic and anti inflammatory etc. (santoshmajumder,
dr. sudhankarkokate et al).The extract of guava leaves also have
antimicrobial and antioxidant activity. Psidiumguajava also contain
ascorbic acid beneficial for skin and effective against oxidation and act
as antioxidant and shows free radical scavenging activity.
Psidiumguajava parts like roots, stem, bark, fruits, leaves, flowers and
juices also shows therapeutic effects against several diseases, it is the
most common and popular traditional remedy for GIT disorders like
indigestion, stomachic, diarrhoea, dysentery. The present study reveals
the phytochemical study of different leaves extract of Psidiumguajava
and also shows their therapeutic importance.

1.1.Synonyms Psidiumguajava L., Psidiumguajavavahl

1.2.Biological source It is obtained from a small trees in the related
species of guavas. The belong to other species or genera.
1.3.Family Myrtaceae
1.4.Morphological characteristics

Colour: - dull green to dark green pink colour

Odour: - characteristic

Shape: - Ovate-elliptic or oblong elliptic, they have hairy undersides

entire margin

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Size 7-15 cm in length and 3-5 cm in width.

Plate (i):- Guava leafs shape Plate (ii):- size of guava leafs

Plate (iii):- Stem of Guava tree Plate (iv) :- Psidium gujava tree

1.5.Cultivation and collection :- Guava is cultivated maximum in

tropical and subtropical regions up to 1500m above from sea levels
.so it is found in our country (India) in every state.
i. Soil:- The seed of guava is hardy, so it is not choosy soil.
It can be grow in slightly alkaline and poor soil. For well
growth of Psidium guajava well drainage system require
with sandy loams to clay loams and PH varies from 6.5 to
8.5.
ii. Climate:-It takes mild rainy season during fruit maturity. It
requires dry climate condition at the time of flowering.

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 iii. Raising of rootstock:- Propagation of uniform material for
planting is very important. For the vegetative propagation
of guava, rootstock raised from the seed extract. The seed
is extract from the ripe fruits. The seed is sown in raise bed
closely immediately after extraction in august or in
February. Seedling of 10 to 15 cm height are uprooted from
these beds and replanted in well prepare field for patch
budding.
iv. Rooting of cutting:-The guava also can propagate through
cuttings. Cutting of 20 to 25 cm length are prepare from
one year shoots. The planted cuttings should be applied
light irrigation, keep the soil medium at field capacity by
spreading trash for conservation of soil moisture.
v. Vegetative propagation:-Guava is usually propagated
from seed can also be propagated vagetatively.
vi. Harvesting:- The quality of guava fruits and leaves depend
on the season of the cropping. For example winter cropping
gives good quality then rainy cropping. The guava can be
picked when mature, light green in colour.
1.6.MICROSCOPIC CHARACTER OF GUAVA LEAVE:- The
T.S of guava leafs contain upper epidermis,chlorenchyma,
palisade, air globules, lower epidermis, phloem, xylem,
sclerenchyma; cuticle and tricomes. They have different function
which help to growth the guava tree. Xylem tissue is used mostly
for transporting water from roots to stems but leaves also transport
other dissolved compound and phloem is responsible for
transporting food produced from photosynthesis from leave to non
photosynthesis parts of a plant such as roots and stems.

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  The microscopic structure of T.S of guava leaves is shown in
plate (v).

Plate (v):- T.S of guava leave

Palisade absorb the light energy used by the leaf. Trichomes has
several function, it absorb water and minerals, they reflect radiation,
lower plant temperature and reduce water loss. The epidermis forms a
boundary between the plant and external environment, it protect
against water loss, gas exchange and secretes metabolic compound.

1.7.Chemical constituent:-The chief constituents like essential oil,

poly phenols, tannin, pinene, gallic acid, isoflavonoid, catechin,
rutin, ascorbic acid, beta carotene, terpenoid and triterpene shows
pharmacological importance against several disorders like
antibacterial, anti-inflammatory, antispasmodic, diarrhoea,
diabetes, colic, cough, pain, high cholesterol, heart disease, cancer
and weight loss. The extract of guava leaves also have
antimicrobial and antioxidant activity. The constituents of guava
leaves are shown in Table(i).

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 a) Phenolic compound
b) Isoflavonoid
c) Gallic acid
d) Rutin
e) Kaemoferol
f) Ascorbic acid
g) B-carotene
h) Carotenoid
i) Terpenoid and triterpene
j) Epicathechin

Table (i):- Chemical constituents

But the main constituents of guava leave are phenolic compound like
stenophylaninA, ellagic acid, beta isomers, deoxyhexoside and
catechin.

1.8.USES OF GUAVA LEAVES

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  The guava leaves are used in the treatment of following
problem, they are shown in table (ii).
I. colic
II. diarrhoea

III. Diabetes

IV. Cough

V. Pain

VI. Cataracts

VII. High cholesterol

VIII. Heart disease

IX. Weight loss

X. Cancer

Table (ii):- Uses

Pharmacological Importance
Sr. No. Pharmacological Importance
1. Antispasmodic
2. Anti- malarial
3. Antioxidant
4. Hepatoprotective
5. Antidiabetics
6. Anticough
7. Anti-inflammatory, anti-allergics
8. Antidiarroea and cytotoxic

Table (iii):- Pharmacological Importance

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1.9. SIDE EFFECTS
It is not recommended for the pregnant women and breast feeding
women, but guava is safe when eaten as food.

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CHAPTER II

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2. REVIEW OF LITERATURE

Now a day’s guava leaves and any other part of guava tree is useful in
medical field or industrial field. The extract of guava leaves also have
antimicrobial and antioxidant activity. Psidium guajava also contain
ascorbic acid beneficial for skin and effective against oxidation and act
as antioxidant and shows free radical scavenging activity. The other
important properties of guava leave are following Hepatoprotective,
anti –allergy, anti-cough, anti diabetic and anti inflammatory etc.
(Santosh Majumder, Dr. Sudhankarkokate et al).

Today’s guava leaves extract are very essential for health which
decreases cholesterol level and glucose concentration which can be
used as anti-diabetic and also used as anti-diarrhoeal. Antidiabetic and
antidiarrhoeal effect on ethanolic extract of Psidium guajava leaves Bat.
Leaves in Wister rats (Santosh Majumder et al.)

The potential of natural substances to improve the immune system has

long been the subject of investigation. Guava leaves has been used
traditionally in the treatment of various diseases. Guava leaves is
commonly used to treat diarrhea, gastroenteritis and other digestive
complaints, while the guava fruit has been used to increase platelets in
patient with dengue fever (NoerLaily* ,Retnowindyakusumaningtyas,
Lim Sukarti, Maria Rosari Devi KartikaRini/Procedia chemistry 14
(2015) page no. 301-307)
Medicinal plant have been used in the treatment and improvement of
human disease (Gutierrez et al. 2008; Nyirenda et al. 2012).
Guava leaves has antioxidant abilities can be used as natural medicines
for preventing aging and chronic diseases (Kahonen et al. 1999).

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These plants have various physiologically active substances with
anticancer and antimicrobial abilities (Bhanot et al. 2011; Miyake and
Hiramitsu et al.2011).
The free radical scavenging abilities of plant has been evaluated
(Brand-Williams et al.1995; Jayanthi and Lalitha 2011).
Psidium guajava leaves are used in diarrhoea and dehydration and also
used in the treatment of gastroenteritis, dysentery, stomach pain,
diabetes mellitus and wound. It also has antioxidant and antibacterial
and anti-inflammatory properties (Qian and Nihorimbere 2004; Cheng
et al.2009; Han et al.2011a).
The main active substances in guava leaves are tannins (Ocuda et al.
1987), ethanol and hydro ethanol (Qian Nihorimbere 2004), methanol
(Chah et al.2006) and water (Moreno et al. 2000).
Therefore the studies of guava leaves are analyse and evaluated with
regard to anti-inflammatory, antibacterial, anti-oxidant, anti-microbial
and anti-diabetes properties.

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CHAPTER III

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 MATERIAL AND METHOD

3. PHYSICAL EVALUATION:-For physical evaluation firstly have

to collect the leaves of guava, then wash properly with tap water or
free flowing water, after washing the guava leaves keep for drying for
5 to 7 days, also you can dry it artificially by using hot air oven etc.

Plate (vi):- wet guava leafs Plate (vii):- Dry guava leafs
The physical evaluation can be done by using following method:-

3.1. LOSS ON DRYING:-

Procedure:-
Beaker Beaker + leafs( B+ leafs
L)
30.6g 31.6g 1g
29.6g 30.1g 1g
28.7g 29.7g 1g

Then keep it for 1 hour at 60°C.After that you can calculate the
final value and % yield of moisture content or LOD (loss on
drying).

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 Initial value( B+ L) Final value Moisture content or
LOD
31.6g 31.4g 0.2g
30.1g 30.4g 0.2g
29.7g 29.5g 0.2g

Table (iv):- LOD value

% yield of moisture content = 0.2/1×100
=20%
Result: - The moisture content of guava leaves is 20%.

3.2. ASH VALUE:- the object of ashing drugs is to remove traces

organic matter which can be interfere during determination process.

Procedure:-

Weigh (3g) dry guava leafs in crucible

↓
Then keep it in Muffle furnace for 5 to 6 hour
↓
After 5 to 6 hour switch off the instrument
↓
Then after 15 minute crucible is cool and weight

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 Plate (viii):- Muffle furnace
Crucible weight ( C ) C + L (leafs) initial Leafs (L)
wt
18.5g 21.5g 3g
21.3g 24.3g 3g
20.2g 23.2g 3g

Plate (ix):- ash of guava leafs

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The final weight and % yield are:-
Initial weight Final weight Ash value
21.5g 18.78g (2.8) 0.24g( 3-2.8)
24.3g 21.52g(2.8) 0.25g(3-2.8)
23.2g 20.39g(2.8) 0.21g(3-2.8)

Table (v):- Ash Value

% yield of ash value = 0.23/3×100

= 7.6%

The total ash value is 7.6 % and acid insoluble ash value is 6.66%.

3.3. SWEELING INDEX:-

Procedure:-

Measure 20ml of water 3g of dry guava leaves in measuring cylinder

↓
Then keep it for 24 hour
↓
After that Observe the swelling value of guava leaves

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 Plate (x):- swelling index
Result:-There are no swelling it means there are absence of
saponin.

3.4.Water soluble and insoluble ash value:-

Procedure:-

Leaves (0.7g) with 25 ml of water in beaker

↓
Heat it for 5 min
↓
Then filter and warm the filter paper after filtration
↓
Weight the ash of filter paper which is remaining upper the filter
paper and
Are water insoluble extract
↓
The filtrate is water soluble extract
Result:-The water insoluble ash 74% and water soluble ash 25%

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3.5. FOAMING INDEX:-

Procedure: - Firstly take 10ml of water and 1 g extract

↓
Then shake it for few minute or 5 min
↓
If any foam is observed then there is the present of Sapponin.

Result: - In guava leave the small amount of Saponin was observed.

3.6. FOREIGN ORGANIC MATTER:-

The guava leaves was collected carefully and wash it in tab water and
inspect the leaves or dry leaves with unaided eye. After inspection
guava leaves visually there are no organic foreign matter.

Physical evaluation is help to determined Ash value, Foamingindex,

Swelling index, foreign organic matter and Loss on drying.

4. CHEMICAL EVALUATION

4.1. Extraction method:-

6g dried mashed of guava leaves was extracted with 600 ml of

boiling water at 100°C for 20 min
↓
The obtained of guava leafs extract was filter using a filter paper
↓
And evaporate to obtain the final extract volume 60ml.
↓
These extract is used for further analysis and determination.

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 Plate (xi):-Extract at 100°C

Plate (xii):- Extract value

4.2. CHEMICAL TEST:-The chemical test of Guava leaves extract

which determine the present of chemical constituent. They are shown
in table (vi).

TEST PROCEDURE RESULT

I. For Alkaloids
a) Hager’s Take 2ml of sample Alkaloid
reagent and add few drops of present
Test reagent. Alkaloid
gives yellow ppt with
Hager’s reagent.

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 II. For Amino acid
a) Million’s To the test solution Negative
test add 2ml of million’s
reagent white ppt
indicate the present of
amino acid.
b) Ninhydri- To the test solution Negative
ne test add few drops of
ninhydrin solution
then boil, Violet
colour indicate the
present of amino acid
III. For
Carbohydrate
a) Molisch’s test To the test sample add Carbohydra
molisch’s reagent, te was
then add drop wise present
sulphuric acid. If
violet ring is observed
then carbohydrate will
be present.
IV. For volatile oil To the thin section of Volatile oil
drug add present in
sudanIIIsolution. guava
Redcolour obtained by leaves.
globules indicates the
present of volatile oil.
V. For flavonoid

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 a) Alkaloid reagent To the test solution
test add few drops of Flavonoid
sodium hydroxide present.
solution intense
yellow colour is
formed which turn to
colourless on addition
of few drops of dilute
acid indicate the
present of flavonoid.
b) Shinoda test To the test solution Flavonoid
add few magnesium present.
and conc. Hcldrop
wise, pink scarlet,
crimson red or green
to blue colour appears
after few minutes.
c) Zinc HCL test To the test sol. add Present
mixture of zinc dust
and conc. Hclacid;
give Red colour after
few minute.
VI. For Cardiac
glycoside
 Baljet’s Treat the solution with Present of
test picric acid or sod. cardiac
Picrate, orange colour glycoside.
is formed.

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  Legal’s Treat the test solution Cardiac
test with hot pyridine and glycoside
add alkali sodium present.
nitroprusidesolution,
blood red colour
appears.
 Borntrag Boil the test material Negative
er’s test with 1ml of sulphuric means
for acid in a test tube for absence of
anthraqui 5 min.Filter while hot anthraquino
none ,cool the filtrate and ne
glycoside shake with equal vol. glycoside.
of dichloromethane or
chloroform separate
the layer of
chloroform or
dichloromethane and
shake it with half of
the vol. of dilute
ammonia. A rose pink
to red colour appears
in the amminiacal
layer.
VII. For steroid and Treat the extract with Yellow
triterpenoid few drops of sulphuric colour,
(Salkowski test) acid, red colour at which
lower layer indicate indicate the
present of steroid and present of

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 formation of yellow triterpenoid
colour lower layer .
indicate the present of
triterpenoid.
VIII. For tannin
Condensed
Ferric chloride Treat the extract with tannins are
test ferric chloride test, present
blue colour appears if
hydrolysable tannins
are present and green
colour appears if
condensed tannin are
present.

Table (vi):- Chemical tests

Plate (xii):- Chemical test evaluation

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From the chemical evaluation or chemical test of guava leaves found
the present of Alkaloid, carbohydrate, Volatile oil, Flavonoid, Cardiac
glycoside, tannin and triterpenoid.

5. RESULTS AND DISCUSION

The solvent used for extraction of phenolic compounds of guava

leaves are water and others like methanol, ethanol, acetone, propenol,
ethyl acetate and combination of the solvents. The time required to
extract the active compound depend on active compounds to be
extracted and the used of the solvents. The physiochemical parameters
of leaves of Psidium guajava are as follows:-

Physiochemical Parameters Results

Loss on drying 20%
Ash value 7.6%
Acid insoluble ash value 6.66%
Water soluble 25%
Swelling index Nil
Foaming index Present

Table (vii):- physiochemical parameters of leaves of Psidium

guajava.

Results of Phytochemical screening of Psidium guajava leaves extract

are shown in Table no. (viii).

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Chemical Test Result
Alkaloids Present
Amino acid Nil
Volatile oil Present
Flavonoid Present
Cardiac glycoside Present
Triterpenoid Present
Tannin Present

Table (viii):- Phytochemical screening

DISCUSSION:- Physicochemical evaluation provide various

important parameter like Moisture content, Ash Value, acid insoluble
ash value, Foamingindex, Swelling index and foreign organic matter.
The ash values determined in the present study may be useful in
stabilizing standards of purity and quality. The low total ash value
indicates the low amount of inorganic salts. The extractive values give
an idea about the nature of chemical composition present in the leaves
and pants. And which is useful to determined specific constituents
soluble in that particular solvent used for the extraction.

Phytochemical screening was done to identify the presents of active

constituents of leaves. It revels the presence of flavonoids,
carbohydrates, cardiac glycosides, volatile oil, amino acids, alkaloids,
triterpenoid and tannin.

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 6. CONCLUSION

Guava leaves extract are herbal extract that can reduce glucose
concentration, insulin concentration, Triglycerides, total cholesterol,
LDL cholesterol, HDL cholesterol and LDL/HDL Ratio which
improved glucose and lipid metabolism in plasma. It also decreases
vascular reactivity in aorta which improved endothelial dysfunction.It
has pharmacological importance against several disorders like
antibacterial, anti-inflammatory, antispasmodic, diarrhoea, diabetics,
colic, cough, pain, high cholesterol, heart disease, cancer and weight
loss. The extract of guava leaves also have antimicrobial and
antioxidant activity. The project was successfully done.

7.REFERENCE

i. NoerLaily* ,Retnowindyakusumaningtyas, Lim Sukarti, Maria

Rosari Devi KartikaRini/Procedia chemistry 14 (2015) page
no. 301-307.

26
 ii. Santosh Majumder* Debashishtalukder, volume5, page no. 10-
14.
iii. JongkwonSeo, Soojung Lee, Marcus L Elam, Sarah A Johnson
et al. Food science and nutrition 2014 page no. 174-180
iv. S. Yosof, in Encyclopaedia of Food sciences and Nutrition
(second Edition), 2003
v. S.P. Singh, in Postharvest Biology and Technology of Tropical
and Subtropical fruits; Cocona to mango, 2011
vi. Dr. Sudhankarkokate et al.
vii. Dr. C.K. KokatePharmacognosy book etc.
viii. Gutierrez et al. 2008; Nyirenda et al. 2012.
ix. Kahonen et al. 1999
x. Brand-Williams et al.1995; Jayanthi and Lalitha 2011.
xi. Bhanot et al. 2011; Miyake and Hiramitsu et al.2011.
xii. Qian and Nihorimbere 2004; Cheng et al.2009; Han et
al.2011a.
xiii. Moreno et al. 2000

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