Screening of Phyto-chemical compounds from hydro-ethanolic and ethanolic leaf and bark extracts of Terminalia arjuna and Syzygium cumini

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056
Volume: 04 Issue: 06 | June -2017 www.irjet.net p-ISSN: 2395-0072
© 2017, IRJET | Impact Factor value: 5.181 | ISO 9001:2008 Certified Journal | Page 1934
Screening of Phyto-chemical compounds from hydro-ethanolic and
ethanolic leaf and bark extracts of Terminalia arjuna and Syzygium
cumini
Dr. Swaati Sharma , Priya Vishnoi
Department of Agriculture, Food and Biotechnology,
Jayoti Vidyapeeth Women’s University, Jaipur, Rajasthan, India.
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Abstract: Medicinal plants have many phyto-chemicals or secondary metabolites which are used to treat many human
diseases (oxidative stress diseases and neurodegenerative diseases). Neurodegenerative diseases include stroke, Parkin-son’s
disease and Alzheimer’s disease and oxidative stress diseases such as inflammatory bowel diseases, retinal ischemia and
cardiovascular diseases. In this study, different parts of Terminalia arjuna and Syzygium cumini plants (mainly leaves and
bark) are used for screening of bioactive compounds. Plant parts are extracted with ethanolic and hydro-ethanolic (50%)
solvents by soxhlet apparatus extraction method and screened for the presence of secondary metabolites namely alkaloids,
flavonoids, rasins, saponins, steroids, tannins, glycosides, phenol, terpenoids and fatty acids.
In this research work, We found that selected plants are good source of various bioactive compounds. However, both leaves
and bark of Terminalia arjuna contain high amount of phyto-chemicals than Syzygium cumini. From this study, We concluded
that both plants are good source of secondary metabolites and can be used as medicinal source for the treatment of various
diseases.
KEYWORDS: Terminalia arjuna, Syzygium cumini, secondary metabolites, phyto-chemicals, bioactive compounds.
1. INTRODUCTION:
Since ancient times, secondary metabolites have ability to prevent chronic diseases far away their role as dietary antioxidants
(1). According to World Health Organisation (WHO), medicinal plants can be good source to obtain variety of drugs (2).
Generally herbal drugs are less expensive, safe, and easily available and rarely have side effects. These are widely used in
human therapy, veterinary, scientific research and agriculture etc. A large number of phyto-chemicals belong to various
chemical classes have prohibitory effects on all types of microorganism invitro. Medicinal plants contain many bioactive
compounds and are concentrated in the endangered parts of the plants such as leaves, bark, flowers, fruit and skin, used for
various medicinal purposes (3,4, 5). Secondary metabolites have their role in several health benefits and provide a source of
medicine since historic times. Traditional plants are helpful in world’s pharmaceuticals and serve as starting material in drug
development. It has been reported that these natural secondary metabolites may reduce the risk of cardiovascular diseases,
oxidative stress diseases and neurodegenerative diseases (1).
The plants chosen for this study are Terminelia arjuna (T.arjuna) and Syzygium cumini (S. cumini). T.arjuna is large sized tree
to 30 m tall, belongs to Combretaceae family. This large an evergreen tree appeared in hot season (February to April), found in
South Asian region having wide spectrum of biological activity. The bark of T. arjuna have been reported for anti-dysentric,
antipyretic, astringent, cardiotonic, lithotriptic, anticoagulant, hypolipidemi, antimicrobial (6) and antiuremic (7) activities.
Many useful phytochemicals have been isolated from T.arjuna including terpenoids for cardiovascular properties, tannins and
flavonoids for its antimicrobial and anticancer properties etc (8).
On the other hand, S.cumini is commonly known as jambolan or java plum, belongs to family Myrtaceae. Plants of this family
are rich in volatile oils which are used in medicines (9). Fruits of this family are used as edibles and as traditional medicines in
divergent ethno-botanical practices throughout the tropical and subtropical world (10). The fruits are berries (edible), 1.5 to
3.5 centimeters long, dark purple and nearby black color contain single large seed (11, 12). The fruit is sweet, mildly sour and
astringent in taste and contains raffinose, glucose, fructose (13), citric acid, mallic acid (14), gallic acid, anthicyanins (15).

2. MATERIALS AND METHODS:-
2.1COLLECTION OF PLANT MATERIAL:
The leaves and bark of T.arjuna and S.cumini were procured from the botanical garden of jayoti vidhyapeeth women’s
university.
2.2EXTRACTION OF PLANT MATERIAL:
Fresh plant materials were washed with running tap water and dried at room temperature. After washing and drying, leaves
and barks of the plants were grinded using grinder to make powder. The powder was then subjected to successive extraction
with different solvents (50% hydro-ethanol and ethanol) using soxhlet apparatus method. The hydro-ethanolic and ethanolic
extracts were prepared by taking 25 grams of each powdered sample and soaked in 250 ml of 50% hydro-ethanol or ethanol
solvent system. The collected extracts were stored in calcium carbonate at very low temperature and then taken up for further
research work.
2.3PHYTOCHEMICAL ANALYSIS:
Chemical test were carried out on 50% hydro-ethanolic and ethanolic extracts of aerial parts of T.arjuna and S.cumini using
standard procedures to identify the constituents (16,17,18).
Test for Glycosides: For the estimation of glycosides, extract was diluted with solvent by which extract was extracted from
plant materials. 2ml of diluted plant extract was mixed with 2ml of chloroform and 2ml of sulphuric acid then shaken well.
Reddish brown color indicated the presence of steroidal ring (glycone portion of glycosides).
Test for Fatty acids: 0.5gm of plant extract was mixed with 5ml of ether and then extract was evaporated on filter paper
and dried the filter paper. The transparency appeared on filter paper indicated the presence of fatty acids.
Test for Flavonoids: Alcoholic solution of extract was mixed with few drops of 10% lead acetate. Yellow precipitate
indicated the presence of flavonoids.
Test for Phenol: Diluted extract was mixed with few drops of ferric chloride solution. Appearance of bluish black color
showed the presence of phenol.
Test for Resins: 0.5gm of plant extract was diluted with 10 ml of water and then shaken for 5 min. Turbidity formed in the
solution indicated the presence of resins.
Test for Saponins: Diluted 5ml extract was stirred with 5ml of distilled water and then warmed in a test tube. Stable
formation of foam indicated the presence of saponins.
Test for Steroids: Diluted 2ml of extract was stirred with 2ml of chloroform and 2ml of conc. Sulphuric acid. Red color
produced in the lower chloroform layer indicated the presence of steroids.
Test for Tanins: 2ml of extract was mixed with 2ml of DW and then 2 or 3 drops of ferric chloride was mixed to the solution.
Green precipitate indicated the presence of tannins.
Test for Terpenoids: To identify the presence of terpenoids, Salkwoski method was used. 0.5 gm plant extract was mixed
with 2ml of chloroform and 3ml of conc. Sulphuric acid to form a layer. Reddish brown color indicated the presence of
terpenoids.

QUALITATIVE ESTIMATION OF PHYTOCHEMICALS:-
TABLE NO:- 1
NAME OF
PHYTO-
CHEMICALS
NAME OF MEDICINAL PLANTS
TERMINALIA ARJUNA SYZYGIUM CUMINI
LEAVES
AND BARK EXTRACTED
WITH
50%
HYDRO-ETHANOL
LEAVES AND BARK
EXTRACTED WITH
ETHNOL
LEAVE AND BARK
EXTRACTED
WITH 50%
HYDRO-
ETHANOL
LEAVES AND
BARK
EXTRACTED
WITH ETHANOL
Leaves Bark Leaves Bark Leaves Bark Leaves Bark
Saponins ++ +++ ++ ++ ++ ++ +++ ++
Steroids ++ +++ ++ + +++ ++ ++ +
Flavonoids +++ ++ +++ ++ ++ +++ +++ ++
Phenols +++ ++ +++ ++ +++ +++ +++ ++
Resins +++ + ++ + +++ +++ + ++
Tanins +++ ++ +++ ++ ++ ++ ++ ++
Terpenoids +++ ++ +++ ++ ++ + ++ ++
Fatty acids +++ - +++ - - - - -
Glycosides +++ +++ +++ +++ +++ ++ +++ ++
+++ High concentration, ++ Moderate concentration, + Low concentration, - Absence.
3.RESULT AND DISCUSSION :
The qualitative estimation of phyto-chemicals of plants T.arjuna and S. cumini exhibited the presence of various bioactive
components such as glycosides, saponins, flavonoids, phenol, resins, tannins, terpenoids, fatty acids. From this study, I found
that T.arjuna contains high amount of phyto-chemicals than S.cumini. Different plant parts with different solvents were
screened to check the presence of various secondary metabolites. Results have been shown in table no.1.
Leaves of T.arjuna extracted with 50% hydro-ethanol contain high (+++) amount of flavonoids, phenols, resins, tannins,
terpenoids, fatty acids and glycosides and moderate (++) amount of saponins and steroids, whereas, the bark extracted with
the same solvent contains high (+++) amount of saponins, steroids and glycosides, moderate (++) amount of flavonoids,
tannins, terpenoids and phenols, low (+) amount of resins and fatty acids are absent(-).
In case of leaves extracted with ethanol contain high (+++) flavonoids, phenol, tannins, terpenoids, fatty acids and glycosides,
moderate (++) saponins, steroids and resins, whereas, the bark extracted with the same solvent contains high (+++)
glycosides, moderate (++) saponins flavonoids, phenols, tannins and terpenoids, low (+) steroids and resins, fatty acids are
absent(-). The bark of T.arjuna functions as a diuretic in cirrhosis of liver and gives relief in symptomatic hypertension (19). Its
leaves have been reported to have analgesic and anti-inflammatory properties (20). T.arjuna is also known as cardiotonic
since historic times (21). The bark of T.arjuna and juice of it is used as antacid by the people live in malkangiri district of orrisa

(22). Powder of bark is boiled with water and inhaled to cure headache and kill bacteria in teeth (23). Bark ash is used to treat
snakebite and scorpion sting (21). The bark of T.arjuna is also used to treat fever and high blood pressure in south surguja
district of Madhya Pradesh (24). It has been proven that T.arjuna and its species have potential to treat HIV (25).
On the other hand, leaves of S.cumini extracted with 50% hydro-ethanol contain high (+++) steroids, phenols, resins and
glycosides, moderate (++) flavonoids, tannins, terpenoids and saponins, fatty acids are absent(-), whereas, bark extracted with
the same solvent contains high (+++) flavonoids, phenols and resins, moderate (++) saponins, steroids, tannins and glycosides,
low (+) terpenoids and fatty acids are absent (-).
However, In case of leaves extracted with ethanol contain high (+++) saponins, flavonoids, phenols and glycosides, moderate
(++) steroids, tannins and terpenoids, low (+) resins and fatty acids are absent (-), whereas the bark extracted with the same
solvent contains moderate (++) saponins, flavonoids, phenols, resins, tannins, terpenoids and glycosides, low (+) steroids and
fatty acids are absent (-). The fruits of S.cumini having medicinal properties, used to treat cough, diabetes, inflammation,
dysentery and ringworm (10). The fruit (mainly seeds) of S.cumini is also claimed to contain an alkaloid, jambosine, and a
glycoside, jambolin or antimellin, which stop the diastatic coversion of starch into sugar (26). They are also rich in flavonoids
(a well known antioxidant), which accounts for the scavenging of free radicals (27, 28). Jambolan also have phenolics with
antioxidant activity (29) and is rich in protein and calcium. Java plums are also rich in mineral salts, sugar, vitamin C, PP
which boosts the beneficial effects of vitamin anthocyanins and flavonoids (30). Different parts of the jambolan have been
reported for its antioxidant, anti-inflammatory, neuropsycho-pharmacological, anti-microbial, anti-bacterial, anti-HIV,
antileishmanial and antifungal, nitric oxide scavenging, free radical scavenging, anti-diarrheal, antifertility, anorexigenic,
gastroprotective and anti-ulcerogenic and radioprotective activities (31).
Phyto-chemicals in our diet promote the health (1). Phyto-nutrients are related with the prevention of recognized chronic
diseases including cardio vascular diseases (CVDs), cancer, osteoporosis, diabetes and vision diseases that are mainly terrible
in Western countries. These diseases are mostly attributed to a high-sugar and high-fat diet, smoking, lack of exercise and
other unhealthy lifestyle (32). Interaction of carcinogens or oxidants with DNA causes many types of cancers. The powerful
antioxidant ability of phyto-nutrients can reduce this damage (33). Different parts of plant are also used to treat various
diseases such as diabetes, blisters in mouth, cancer, colic, diarrhea, digestive complaints, dysentery, piles, pimples and
stomachache (34).
Phyto-chemicals, found in bark and leaves of T.arjuna and S.cumini, play important roles to treat various diseases. Tannins
have the ability to treat sore throat, wound healing, antidiarrhoea and antihaemorrhagic agent (35, 36). Tannins have also
been studied for the antimicrobial degradation of dietary proteins of semen (37). Flavonoids can inhibit the initiation,
promotion and progression of tumors. Phenols and flavanoids, both are strong water soluble antioxidants (38). These phenolic
compounds also have some biological properties such as anti-apoptosis, anti-septics, anti-carcinogen, anti-ageing, anti-
inflammation, anti-atherosclerosis, improvement of endothelial function and cardiovascular protection (36). These phenolic
compounds also have ability to inhibit angiogenesis and cell proliferation activities (39). Terpenoids have carboxylic acid
group which is responsible for the activity of organic extracts (40,16). Glycosides are present in all the hydro-ethanolic and
ethanolic extracts of T.arjuna and S.cumini which lower the blood pressure (41). Saponins, found in hydro-ethanolic and
ethanolic extracts of both the plants are known to posses reduction in cholesterol level and can manage cardiovascular
diseases in humans (37). Saponins also have been used as emulsifying agents and having anti-fungal (42), anti-inflammatory
activity (43), coagulating and precipitating property in red blood cells. Steroids also have been found in these medicinal plants,
previously reported for its antibacterial activity (44) and their association with such compounds as sex hormones (45). Fatty
acids, found in hydro-ethanolic and ethanolic extracts of T.arjuna leaves are previously reported for their anti-fungal and anti-
bacterial activities (46). Resins are also present in these traditional medicinal plants, used for the treatment of arthritis,
wound healing and have anti-inflammatory, anti-microbial, anti-tumor, anti-hyperlipidemia properties (47).
4. CONCLUSION:
From this study, it can be concluded that these species may be used for drug development in pharmaceutical companies.
Different parts (leaves and bark) of both plants are good source of phyto-chemicals which have anti-fungal and anti-microbial
properties.
ACKNOWLEDGEMENT:-
The authors are thankful to Dr. Panckaj Garg, Founder & Advisor and Department of Agriculture, Food and Biotechnology,
Jayoti Vidyapeeth Women’s University, Jaipur for providing the laboratory and technical facilities used in this study.

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