Department of P.

G Studies and Research in Applied Botany

Janna Sahyadri Shankaraghatta – 577-451
PROJECT DEFENDING ON

“Investigation of Preliminary Phytochemical and Antimicrobial properties

of Alangium salviifolium(Linn.f.) Wang. And Mimusops elengi Linn.”

Under the Guidence of : Presented by:

Dr. M.B.SHIVANNA. Mr. Veeranna B Shetter,
Professor, M.Sc. II year,
Dept of Applied Botany Dept of Applied Botany.

2011
 The medicinal plants have always been the principle
source of medicine in throughout world.
 Every culture on the earth, through written or oral
traditional, as relied on the natural chemistry found
in healing plants for their therapeutic properties.
 In Ayurveda, more than 2000 plants are find in
medicine use among them only 600 species are most
commonly exploited by the traditional practitioners
in the formulation of drugs(Schultes,1978).
 Plant possess various types of chemical compounds
and these are classified as primary and secondary
metabolites like amino acids, carbohydrates,
proteins, fats, oils, alkaloids, steroids and phenolics
etc.
 Many secondary metabolites serve as antimicrobials. An anti
microbial is a substance that kills or inhibits the growth of
microorganisms such as bacteria, fungi and viruses.

 Screening of medicinal plants for anti microbial agents has

gained much importance because lately World Health
Organization(WHO) is keenly interacted in the development
and utilization of medicinal plant resource in the traditional
system of medicine.

 This will be a major step in the development and

standardization of botanical medicines for sustainable global
health care.

 The present study will be testing the efficacy of two medicinal

plants Alangium salviifolium (Linn.f.) Wang. and Mimusops
elengi Linn.
 Alangium salviifolium (Linn.f) wang.
  Family: Alangiaceae (Alangium family)
 The plant is commonly known as : Sage Leaved Alangium in English
• Hindi: Ankol • Kannada: Ankolamara .
 Sage Leaved Alangium is a tall thorny tree native to India. It grows to a
height of about 3 to 10 meters. The bark is ash colored, rough and
faintly fissured. The leaves are elliptic oblong, elliptic lanceolate or
oblong lanceolate. The flowers are greenish white, fascicled, axillary or
on old wood. The berries are ovoid, ellipsoid or nearly globose.
glabrous, smooth and violet to purple. The flowering season is February
to June.
 Medicinal Uses: In Ayurveda, the roots and the fruits are used for
treatment of rheumatism, and hemorrhoid. Externally it is used for the
treatment of bites of rabbits, rats and dogs.
 Mimusops elengi Linn.
Family: Sapotaceae
 Vernacular Names: Engish: Spanish cherry, Kannada: Ranjal ,San: Bakulah, Hin: Bakul,
Maulsiri .
 Spanish cherry is a lovely green small tree of the Indian subcontinent It is an evergreen
tree with dark grey fissured bark and densely spreading crown. Leaves are oblong,
glabrous and leathery with wavy margins5-16 cm and 3-7 cm wide.. Flowers are small,
star-shaped, yellowish white in color, with a crown rising from the center. are white,
fragrant, axillary, solitary or fascicled. Fruits are ovoid or ellipsoid berries. it is a prized
oranamental specimen because it provides a dense shade and during the months from
March to July fills the night air with the delicious heady aroma of its tiny cream colored
flowers about In the morning the fragrant flowers which so graciously scented their
surroundings with their deep, rich, fragrance during the evening hours, fall to the
ground.
 Medicinal Uses: It is used in the treatment and maintenance of oral hygiene. Rinsing
mouth with water solution made with bakul helps in strengthening the teeth. It also
prevents bad breath and helps keep the gums healthy.
 The earliest known medical document is a 4000-year-old Sumerian clay tablet
that recorded plant remedies for various illnesses.
 The ancients possessed an extensive pharmacopoeia. Hippocrates (in the late
fifth century B.C.) mentioned 300 to 400 medicinal plants. Dioscorides (78
A.D.) is among the earliest generation of famous botanist who recognized the
aromatic mint and umbel as a natural taxon, and he started the systematic
survey of plant chemicals.
 He documented characters and properties of numerous drugs of the old world,
in the famous work “Materia Medica”, published in first century.
 About 80% of the population in the developing countries still depends
traditional medicine for their health care. Modern pharmacopeias contain at
least 25% of drugs derived from plants (Kameshwara Rao, 2000).
 In 1742 – 1786, the phytochemical studies were at its peak and isolated several
organic acids such as citric acid, gallic acid, oxalic acid, tartaric acid, etc.
(Latha, 1997).
 Plant secondary metabolites are a diverse group of molecules that are involved
in the adaptation of plants to their environment but are not part of the primary
biochemical pathways of cell growth and reproduction.
 The discovery of antimicrobials like penicillin and tetracycline proved the way
for better health for millions around the world.
 Before 1941, the year penicillin was discovered; no true cure for gonorrhea,
strep throat, or pneumonia existed. Patients with infected wounds often had to
have a wounded limb removed, or face death from infection. Now, most of
these infections can be cured easily with a short course of antimicrobials.
 Among ancient civilization, India has been known to be rich repository of
medicinal plants. The forest in India is the principal repository of large number
of medicinal and aromatic plants, which are largely collected as raw materials
for manufacture of drugs and perfumery products.
 About 8,000 herbal remedies have been codified in Ayurveda. The Rigveda
(5000 BC) has recorded 67 medicinal plants, Yajurveda 81 species,
Atharvaveda (4500-2500) 290 species, Charak Samhita (700 BC) and Sushrut
Samhita (200 BC) had described properties and uses of 1100 and 1270 species
respectively.
 Traditional Chinese Medicine being an important type of alternative medicine
needs a special attention for the study of antimicrobial properties. However
Janovská et al. (2003), screened some medicinal plants species of TCM.
 There is also significant literature available saying that the many plant species
are used for burns, cholera, fever, rheumatism, pneumonia, skin diseases and
wounds, astringent, itch, leprosy, inflammation, ulcers, oil-alopecia, acne
(Nadkarni, 1982; Joshi, 2000; Kameshwara Rao, 2000).
 The extracts of Mimusops elengi L. (family: Sapotaceae) bark, fruit and seed
were evaluated for antibacterial activity (Durre Shahwar and Muhammad Asam
Raza et al.,2010).
 Different solvent extracts of bark, fruits (fleshy portion) and leaves of
Mimusops elengi were screened for their antibacterial and antifungal activities
against some pathogenic bacteria and fungi. The activities of the extracts were
not significantly enough against most of the tested organisms. Fruit extracts
were less potent against most of the tested organisms (M. Abbas Ali et.al,2010).
 The effect of aqueous bark extract of Mimusops elengi Linn. in alloxan induced
diabetic rats (JERLINE, M.et.al, 2010).
 evaluate the antimicrobial and antioxidant activities of the alcoholic and
aqueous extracts from the root of Alangium salvifolium (L.f) Wang(Vineet C.
Jain et.al,2010).
 However, there are no works reported upon the
preliminary phytochemical analysis of secondary
metabolites of the soxhlet extracts of Alangium
salviifolium (stem bark) and Mimusops
elengi(stem bark). Neither there is any studies
conducted strictly upon the antibacterial
properties of the Alangium salviifolium and
Mimusops elengi has been analyzed for the
antimicrobial properties, hence the present study
aims to prove that Alangium salviifolium and
Mimusops elengi has the antibacterial property.
 Aims and Objectives

 To extract the phytochemicals from two medicinal plants

such as Alangium salviifolium and Mimusops elengi in
different solvents.

 To know the primary and secondary metabolites present

in selected medicinal plants.

 To screening the crude solvent extracts for antifungal

an antibacterial activities.
 Methods and Materials
Plant collection:
  The plants used in the study were Alangium salviifolium(Linn.f.) Wang. And
Mimusops elengi Linn. and the plant materials were collected from Haniya, Hosanagar
taluk, Shimoga district during September 2010 .The plants were identified by using
standard manual “Flora of Madras Presidency” by Gamble.
Plants materials of each plant were air dried for about 15-30 days, ground and made
into powder. The powdered plant materials were subjected into successive extraction by
soxhlet apparatus.
  Preparation of crude extracts :
100 gram of the air-dried and coarsely powdered plants materials were exhaustively
extracted for about 40 cycles (≈18 hour) with petroleum ether in soxhlet apparatus. The
petroleum ether extract was separated and evaporated under a fan. The extracted plant
material was then air-dried, repacked in soxhlet apparatus and exhaustively extracted with
Chloroform about 20 hour (40 cycles) and the above procedure is repeated for the methanol
(take ≈24 hour to complete 40 cycles) .
Thus obtained extracts were collected separately in clean and dry screw capped
bottles, labeled and stored. After each successive solvent extraction the extract yield was
recorded. Thus obtained solvent extracts were used to carry out the preliminary
phytochemical analysis and antimicrobial assay.
Preliminary phytochemical Group Test
The preliminary phytochemical group test of solvents extracts of the two plants
was performed by the standard methods (Goering & Van Soet et al.,
1975;Krishnaswamy, 2003; Pederson, 2006; Adeloye et al., 2007; Arunachalam et
al.,2009).
1.Primary metabolites:
1.Test for Carbohydrates:
•Molisch’s Test: To the 2ml of sample 2 drops of alpha- napthol and mixed
carefully. Rundown the concentrated sulfuric acid along the walls of the test tube.
The purple violet ring at the junction of the two layers which indicates the
presence of carbohydrates.
2.Test for Polysaccharides:
To 1 ml of test solution, 2 drops of iodine added. Blue coloured solution
indicates the presence of polysaccharides.
3.Test for Proteins:
•Biuret reaction: Biuret reaction can be made to 2ml of test solution and 2ml of
10% sodium hydroxide mix, add 2 drops of 0.1% copper sulphate solution. If
violet or pink colour then compounds with two or more peptide bonds give a violet
colour with alkaline copper sulphate solution.
•Ninhydrin Test: To 4ml of solution which should bear neutral pH add 1ml of
0.1% freshly prepared ninhydrin solution mix the concentration and boil a couple
of minutes. Observation of violet or purple colour indicates the presence of
proteins.
4.Test for Lipids:
To 2ml of test sample iodine solution was added dropwise. Original iodine colour
disappears which indicates the presence of lipids.
5.Test for Oils:
The one drop of sample placed on filter paper and allowed to dry. Clear greasy
spot is observed which indicates the presence of oils.

2.Secondary metabolites:
1.Test for alkaloids:
1.Mayer’s test: Small quantities of the plant extracts of both the plants were
treated with few drops of dilute hydrochloric acid (HCl) and then they were treated
with Mayer’s reagent. The formation of yellowish buff coloured precipitate
indicated positive test for alkaloids.
2. Wagner’s test: Small quantities of plant extracts were separately treated with
few drops of dilute HCl and then with Wagner’s reagent, a reddish brown
precipitate developed, suggesting the presence of alkaloids.
3. Dragendroff’s test: In this test addition of 2ml of Dragendroff’s reagent and
1ml of diluted hydrochloric acid to the plant extract produced orange precipitate
indicating presence of alkaloids.
2.Test for flavonoids
1.Ferric chloride test: Test solution with few drops of ferric chloride
solution shows intense green colour indicating the presence of flavonoids.
2. Shinoda test: For the positive test the test solution with few fragments
of magnesium ribbon and concentrated HCl shows pink to magenta red
colour.
3. Zinc-Hydrochloric acid reduction test: Test solution with zinc dust and
few drops of hydrochloric acid shows magenta red colour.
4. Alkaline reagent test: Test solution taken when treated with sodium
hydroxide solution, shows increase in the intensity of yellow colour which
becomes colourless on addition of few drops of dilute acid.
5. Lead acetate solution test: Test solution with few drops of lead acetate
(10%) solution gives yellow precipitate.
6. Flavonoid test: in this test the plant extract a few magnesium turning
when added with concentrated sulphuric acid through sides of test tube
produced magenta colour indicating flavones or deep cherry red colour
indicating flavonoids
3.Test for glycosides
1. Keller – Killiani test: The test solution was treated with few drops of ferric chloride
solution and mixed. When concentrated sulphuric acid containing ferric chloride solution
was added, it forms two layers, lower layer reddish brown and upper acetic acid layer turns
bluish green.
2. Bromine water test: Test solution when dissolved in bromine water gives yellow
precipitate.
3. Raymond’s test: The test solution when treated with dinitrobenzene in hot ethanolic
alkali gives violet colour

4. Molisch’s test: In this test 1ml of molisch’s reagent was added to the plant extract and
when 1ml concentrated sulphuric acid was poured through the sides of the test tube, a
reddish violet ring was formed at the junction of two layers indicating the presence of
glycosides.

4.Test for tannins:

1.Ferricchloride test: Test solution is treated with 1ml of 5% ferric chloride solution;
formation of greenish black col.ouration demonstrated the presence of tannins.

2. Gelatin test: Test solution when treated with 1% gelatin solution gives white precipitate.

3. Sodium chloride test: The formation of precipitate with the addition of few drops
sodium solution to the extract indicated the presence of tannins.
5.Test for triterpinoids

Salkowaski test: A few drops of concentrated sulphuric acid was added to the test
solution, shaken and allowed to stand, lower layer turns yellow indicating the
presence of triterpinoids.

6.Test for saponins:

Foam test: Test solution is treated with water, shaken well for 15 minute,
formation of stable foam suggested the presence of saponins.

7.Test for lignins:

Furfuraldehyde test: when extract was treated with furfuraldehyde solution(2%),
formation of red colour indicated the presence of lignins.
8.Test for phenols
 1. Phenol test: Test was done by adding 0.5ml of ferric chloride solution
to the plant extract. The presence of phenolics was indicated by the
formation of the intense colour in the solution.
 2. Ellagic acid test: This test was yielding muddy yellow, olive brown,
niger brown or deep chocholate upon reaction of plant extract with
few drops of 5% mixture containing glacial acetic acid and 5% (V/V)
sodium nitrate solution.
 3. Hot water test: In this test, leaf and stem pieces when partially dipped
in hot water produced intense colour at the junction.

9.Test for steroids

Salkowaski test: A few drops of concentrated sulphuric acid was added
to the test solution, shaken and allowed to stand, lower layer turns to red
indication.

Further work will be carried out over on

antimicrobial activity ( bacteria and fungal species) by the
plants extract of Alangium salviifolium and Mimusops elengi.
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