S. Selva Kumar et al.

/BioMedRx 2013,1(4),371-373
Research Article
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Preliminary phytochemical analysis of Excoecaria agallocha and Avicennia

S. Selva Kumar*, Prajeet Sen and S. Anbuselvi
Department of Industrial Biotechnology, Bharath University, Chennai-600073, India

Received on:13-01-2013; Revised on: 11-02-2013; Accepted on:21-03-2013

ABSTRACT
In the present study, the mangroves, Avicennia and Excoecaria agallocha (Euphorbiaceae), were selected for this study. The crude extracts
from dried leaf of the mangroves were extracted using soxhlet extraction method and obtained extracts were concentrated by using rotary
evaporator. Two solvents Methanol and Ethyl acetate were used to extract the leaf material. Extracts from mangroves and mangrove-
dependent species have proven effective against human, animal and plant pathogens, but only limited investigations have been carried out
to identify the metabolites responsible for their bioactivities. Hence it is of interest to investigate the phytochemical analysis of mangrove
plants such as Avicennia and Excoecaria Agallocha.

Key words: Avicennia, Excoecaria Agallocha, Saponins, Tannins, Mangroves

INTRODUCTION
The mangrove plant, Avicennia is a genus of flowering plants cur- Excoecaria agallocha L. (Euphorbiaceae) is a small mangrove tree
rently placed in the bear’s breeches family, Acanthaceae. It contains found extensively in the tidal forests and swamps of the Sundarbans
mangrove trees, which occur in the intertidal zones of estuarine areas and other coastal areas of Bangladesh (1). This plant is also well-
and are characterized by aerial roots. Species of Avicennia occur distributed in a number of other countries of temperate and tropical
worldwide south of the Cancer. The taxonomic placement of Avicennia Asia, Australasia and Southwestern Pacific (2). However, the milky
is contentious. In some classifications it has been placed in the family sap of this tree can cause temporary blindness if it enters the eyes.
Verbenaceae, but more recently has been placed by some botanists in The sap can also cause skin blisters and irritation. Clinical trials car-
the monogeneric family. ried out on this plant have shown its potential anti-HIV, anticancer,
antibacterial and antiviral properties (3).

Fig.1. Plant Avicennia

*Corresponding author.
Dr. S Selva Kumar, Ph.D.
Associate Professor,
Dept. Of Industrial Biotechnology,
Bharath University,
Fig.2. Plant Excoecaria agallocha
Chennai-600073,India.

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Exploration of the chemical constituents of mangrove plants is nec- 2.3 e. Test for flavonoids
essary to find new therapeutic agents and this information is very 2 ml of extract solution was treated with 1.5 ml of 50% methanol
important to the community. There are two important reasons for the solution. The solution was warmed and metal magnesium was added.
need to study the chemical constituents of mangrove plants. First, To this solutionfew drops of conc. Hydrochloric acid was added and
the mangrove is one of the tropical forest that is easy to grow and has the red colour was observed for flavonoides and orange colour for
not been widely utilized. Second, chemical aspects of mangrove plants flavons. [9]
is very important because potential to develop compounds agro-
chemical and medical value. 2.3 f. Test for anthraquinones
About 0.5 g of extract was taken in a dry test tube and 5 ml of chloro-
In the recent years, screening of Mangrove plants for a variety of form was added and shaken for 5 min. The extract was filtered and the
biological activities , further attention should be paid to develop the filtrate shaken with equal volume of 10% of ammonia solution. A pink
novel drug from natural product is prerequisite. Hence it is of interest violet or red colour in the ammonical layer indicates the presence of
to investigate the phytochemical analysis of mangrove plants. anthraquinones.

MATERIALSAND METHODS 2.3 g. Test for cardiac glycosides

0.2 g of extract was dissolved in 1 ml of glacial acetic acid containing
2.1. Collection of samples 1 drop of ferric hloride solution. This was then under layered with 1ml
The medicinal plants used for the experiment were leaves of of concentrated sulphuric acid. A brown ring obtained at the interface
Excoecaria agallocha, and Avicennia. indicated the presence of a deoxysugar characteristic of cardioids.

2.2. Preparation of extracts 2.3 h. Test for Proteins

500 grams of dried powder of Excoecaria Agallocha and Avicennia To 2ml of protein solution 1ml of 40% NaOH solution and 1 to 2 drops
was packed in separate round bottom flask for sample extraction us- of 1% CuSO4 solution was added. A violet color indicated the pres-
ing different solvents namely ethanol, methanol, chloroform, ethyl ence of peptide linkage of the molecule.
acetate and water. The extraction was conducted with 750 ml of each
solvent for a period of 24 hours. At the end of the extraction the 2.3 i. Test for Amino Acids
respective solvents were concentrated under reduced pressure and To 2ml of sample was added to 2ml of Ninhydrin reagent and kept in
the crude extracts were stored in refrigerator. water bath for 20 minutes. Appearance of purple color indicated the
presence of amino acids in the sample.
2.3 Phytochemical analysis
The extracts prepared were analyzed for the presence of alkaloids, 2.3 j. Test for Tri-Terpenoids
saponin,tannins, steroids, flavonoids, anthraquinone, cardiac glyco- 5ml of each extract was added to 2ml of chloroform and 3ml of con.
sides and reducingsugars based on the protocols available in the H2SO4 to form a monolayer of reddish brown coloration of the inter-
literature. [4, 5, 6] face was showed to form positive result for the terpenoids.

2.3 a. Test for alkaloids 2.3 k. Test for Triple Sugar

The extract of the crude dry powder of each solvent was evaporated To 2 ml of extract 2drops of Molisch’s reagent was added and shaken
to dryness in boiling water bath. The residues were dissolved in 2 well.2ml of con. H2SO4 was added on the sides of the test tube. A
NHydrocholoric acid. The mixture was filtered and the filtrate was reddish violet ring appeared at the junction of two layers immediately
divided intothree equal portions. One portion was treated with a few indicated the presence of carbohydrates.
drops of Mayer’sreagent; one portion was treated with equal amount
of Dragondroff’s reagentand the third portion was treated with equal RESULTS AND DISCUSSION
amount of Wagner’s reagent respectively. The creamish precipitate,
the orange precipitate and brownprecipitate, indicated the presence
of respective alkaloids. [7] Table 1 Phytochemical analysis of avicennia

2.3 b.Test for saponins S.no. Phytochemical Ethyl Ethanol Chloroform Water
About 0.5 g of the plant extract was shaken with water in a test tube Constituents acetate
and then heated to boil. Frothing was observed which was taken as a 01 Flavinoids + + + +
preliminary evidence for the presence of the saponin. 02 Alkaloids - + + -
03 Tri terepenoids - - + -
04 Saponins + + + +
2.3 c. Test for tannins 05 Tannins - - + +
About 0.5 g of extract was added was in 10 ml of water in a test tube 06 Triple sugar - - - -
and filtered. A few drops of 0.1% ferric chloride was added and ob- 07 Amino acid - - - -
served forbrownish green or blue-black coloration. [8] 08 Anthroquinones - - + +
09 Steroids + + + +
10 Proteins - - - -
2.3 d. Test for steroids
11 Cardiac glycosides - - + -
2 ml of acetic anhydride was added to 0.5 g of methanol extract of
each sample with 2 ml sulphuric acid. The colour changed from violet
to blue or green in some samples indicating the presence of steroids. “+” – Presence, “-“ – Absence.

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Table 2. Phytochemical analysis of Excoecaria agallocha chloroform and aqueous extracts of Excoecaria Agallocha. Phy-
S.no. Phytochemical Ethyl Ethanol Chloroform Water tochemical screening of the crude extracts revealed the presence of
Constituents acetate flavinoids, saponins and alkaloids in all the four extracts. Tri-terpinoids
are present in ethyl acetate and ethanolic extracts where as absent in
01 Flavinoids + + + + chloroform and aqueous extracts. In case of tannins, tannins are
02 Alkaloids + + + +
03 Tri terepenoids + + - - present in ethyl acetate, ethanolic and chloroform extracts where as
04 Saponins + + + + absent in aqueous extract. Triple sugar, amino acid, anthroquinones
05 Tannins + + + - and proteins are absent in all the four extracts. In case of steriods,
06 Triple sugar - - - -
steriods are present in ethyl acetate, ethanolic and chloroform extract
07 Amino acid - - - -
08 Anthroquinones - - - - where as absent in aqueous extract. Cardaic glycosides are present in
09 Steroids + + + - ethyl acetae, chloroform and aqueous extract where as absent in
10 Proteins - - - - ethanolic extarct.
11 Cardiac glycosides + - + +

“+” – Presence, “-“ – Absence. REFERENCES

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Source of support: Nil, Conflict of interest: None Declared

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