INTRODUCTION
Medicinal plants compose the most important resource of novel pharmaceuticals
and health care products (Ivanova et al., 2005).
Curative plants are natural source of medicine molecules since they have active
compounds, properties of which are useful to humans. Since, primeval times,
medicinal flora has attracted researchers for detection of active biomolecules
having significant biological actions (Aiyelaagbe and Osamudiamen,
2009).
Phytochemical studies have concerned the awareness of plant scientists suitable
to the development of novel and sophisticated techniques. These techniques played
a momentous role in giving the resolution to systematic problems on the one
hand and in the search for supplementary resources of raw materials for pharmaceutical
industry on the other hand. Plants synthesises number of chemical compounds
with different chemical and pharmacological property. Thus the awareness of
the chemical constituents of plant origin is desirable for discovery of curative
agents (Pandith, 2012). For the identification of different
chemical groups and chemical compounds several researchers has reported Phytochemical
screening of various plants (Parekh and Chanda, 2007;
2008).
According to World Health Organization (WHO), more than 80% of the world’s
population relies on traditional medicines for their most important health care
requirements. The medicinal value of plants lies in few chemical substances
responsible for several definite physiological actions in human. Such chemical
group includes alkaloid, flavonoids, tannins and phenolic compounds (Singh,
2012). Awareness of the chemical constituents of plants is pleasing, not
only for the invention of curative agents but also for the renaissance of the
lost information about these valuable plants (Mojab et
al., 2003).
Presently 120 active compounds have been isolated from the higher plants out
of which 80% show an optimistic correlation between their traditional use and
therapeutic uses (Fabricant and Farnsworth, 2001). The
Phytochemical interaction and trace components may change the drug response
in ways that cannot presently be replicated with a grouping of few purative
active ingredients. Pharmaceutical researchers noted that the clinical trials
may be used to evaluate the effectiveness of a particular herbal preparation
(Izhaki, 2002).
So the present study was undertaken to evaluate phytochemical Comparison between
Pet ether and Ethanolic extracts of Bacopa monnieri, Evolvulus alsinoides
and Tinospora cordifolia.
MATERIALS AND METHODS
Selection, collection and authentication of plants: Plants were selected
on the basis of the literature survey, information collected from standard books
and also from traditional medicine system practitioners. Plants were collected
from the hills of Solan region of Himachal Pradesh (HP), India. These plants
were authenticated in department of forestry Dr. Y.S. Parmar University, Solan,
Himachal Pradesh (HP), India. The samples of plants were linked to UHF-Herbarium
with Field book number 12547, 12548 and 12549 for Bacopa monnieri, Evolvulus
alsinoides and Tinospora cordifolia, respectively.
Extraction and preparation of combinations: The plants materials were
processed and dried in shade. Dried plant materials were crushed and were extracted
with ethanol using soxhlet apparatus after defating with pet ether (40:60).
Ethanolic extracts were dried at 40°C using rotary vacuum evaporator and
kept in air tight container till any further use.
Phytochemical screening: Detailed phytochemical testing was performed
to identify presence (+) or absence (-) of different phytoconstituents (Kokate
et al., 2006).
Test for carbohydrates
Molish test: About 2 mL of aqueous extract was treated with 2 drops
of alcoholic α-naphthol solution in a test tube and then 1 mL of concentrated
sulphuric acid was added carefully along the sides of the test tube. Formations
of violet ring at the junction indicate the presence of carbohydrates.
Test for glycosides
General test: Add Extract 200 mg+5 mL dilute H2SO4
by warming on a water bath then filter it. Then neutralize the acid extract
with 5% solution of NaOH and add 0.1 mL of Fehling solution A and B unit, it
becomes alkaline (test with pH paper) and heat in a water bath for 2 min. Note
the quantity of red precipitate formed and compare with that of formed in test
B.
Keller-killiani test: To 2 mL of test solution, 3 mL of glacial acetic
acid and 1 drop of 5% ferric chloride were added in a test tube. Add carefully
0.5 mL of concentrated sulphuric acid by the side of the test tube. Formation
of blue color in the acetic acid layer indicates the presence of Cardiac glycosides.
Test for saponins
Froth test: The extract was diluted with distilled water and shaken
in graduated cylinder for 15 min. The formation of layer of foam indicates the
presence of saponins.
Test for alkaloids: To the extract, dilute hydrochloric acid was added,
shake it well and filtered. With the filtrate, the following tests were performed.
Hager’s test: To 1-2
mL of filtrate, few drops of Hager’s
reagent were added in a test tube. Formation of yellow color precipitate indicates
the presence of alkaloids.
Tannic acid test: Alkaloids give buff colour precipitate with tannic
acid solution.
Test for flavonoids
Alkaline reagent test: The extract was treated with few drops of
sodium hydroxide separately in a test tube. Formation of intense yellow color
which becomes color less on addition of few drops of dilute acid, indicate presence
of flavonoids.
Shinoda test: To the extract, 5 mL (95%) of ethanol was added. The mixture
was treated with few fragments of magnesium turning, followed by drop wise addition
of concentrated hydrochloric acid. Formations of pink color indicate presence
of flavonoids.
Zinc hydrochloride test: Add a mixture zinc dust and concentrated HCl.
It gives red colour after few min.
Test for triterpenoids and steroids
Salkowski’s test: The extract was treated with chloroform and
filtered. The filtrate was added with few drops of concentrated sulphuric acid,
shaken and allowed to stand. If the lower layers turns red, sterol are present.
Presence of golden yellow layer at bottom indicates the presence of triterpenes.
Test for tannin and phenolic compounds
Lead acetate test: Some amount of extract was dissolved in distilled water.
To this solution few drops of lead acetate solution was added. Formation of
white precipitate indicates presence of phenolic compounds (Kokate
et al., 2006).
RESULTS
Phytochemical screening of Bacopa monnieri: Phytochemical screening
revealed the presence of different phytoconstituents both Pet ether and Ethanolic
extracts.
| Table 1: |
Phytochemical screening of pet ether and ethanolic extracts
of Bacopa monnieri |
 |
| +: Present, -: Absent |
| Table 2: |
Phytochemical screening of pet ether and ethanolic extracts
of Evolvulus alsinoides |
 |
| +: Present, -: Absent |
In pet ether extracts; Bacopa monnieri showed different phytoconstituents
like Carbohydrates, Phenols, Glycosides and Anthraquinones (Table
1). And in Ethanolic extracts; Bacopa monnieri showed different phytoconstituents
like Carbohydrates, Phenols, Glycosides, Tannins, Flavonoids and Saponins (Table
1). When comparison between Pet ether and Ethanolic extracts; Ethanolic
extract showed more phytoconstituents as compared to Pet ether extract.
Phytochemical screening of Evolvulus alsinoides: Phytochemical
screening revealed the presence of different phytoconstituents both Pet ether
and Ethanolic extracts. In Pet ether extracts; Evolvulus alsinoides showed
different phytoconstituents like Carbohydrates, Phenols, Glycosides and Flavonoids
(Table 2). And in Ethanolic extracts; Evolvulus alsinoides
showed different phytoconstituents like Carbohydrates, Phenols, Glycosides,
Alkaloids, Tannins, Terpenoids, Steroids, Amino acids and Anthraquinones (Table
2). When comparison between Pet ether and Ethanolic extracts; Ethanolic
extract showed more phytoconstituents as compared to Pet ether extract.
Phytochemical screening of Tinospora cordifolia: Phytochemical
screening revealed the presence of different phytoconstituents both Pet ether
and Ethanolic extracts. In pet ether extracts; Tinospora cordifolia showed
different phytoconstituents like Phenols, Glycosides, Alkaloids, Terpenoids,
Steroids, Saponins and Anthraquinones (Table 3).
| Table 3: |
Phytochemical screening of pet ether and ethanolic extracts
of Tinospora cordifolia |
 |
| +: Present, -: Absent |
And Ethanolic extracts; Tinospora cordifolia showed different phytoconstituents
like Carbohydrates, Phenols, Glycosides, Alkaloids, Terpenoids, Steroids, Saponins,
Amino acids and Anthraquinones (Table 3). When comparison
between Pet ether and Ethanolic extracts; Ethanolic extract showed more phytoconstituents
as compared to Pet ether extract.
DISCUSSION
Plants synthesises a broad range of primary and secondary metabolites with
different functional groups (Sharanabasappa et al.,
2007). The presence of phytochemicals is a marker that the plant can be
a prospective source of precursors in the formation of synthetic drugs (Ayoola
et al., 2008).
In present study it was observed that Pet ether and Ethanolic extracts of Bacopa
monnieri, Evolvulus alsinoides and Tinospora cordifolia showed
the presence of various chemical constituents (Table 1-3).
When comparison between Pet ether and Ethanolic extracts of Bacopa monnieri,
Evolvulus alsinoides and Tinospora cordifolia; Ethanolic extracts
of these plants showed more phytoconstituents as compared to Pet ether extracts
of these plants (Table 1-3).
CONCLUSION
From present investigation, it can be concluded that phytochemical comparison
is subsequently momentous and useful in finding chemical constituents in the
plant substances that may lead to their quantitative evaluation and also pharmacologically
active chemical compounds.
ACKNOWLEDGMENT
We are thankful to Dr. YS Parmar University Solan (HP), India to help us in
authenticating these plants and to PBRI Bhopal (MP), India for to provide us
all lab facilities.
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