Research Paper Volume 3 Issue 3 November 2015

International Journal of Informative & Futuristic Research ISSN: 2347-1697

Preliminary Phytochemical Analysis Of Hydrilla
Verticillata (L.F.) Royle. Collected From Polluted
And Unpolluted Water Sources
Paper ID IJIFR/ V3/ E3/ 082 Page No. 1125-1128 Subject Area Botany
Keywords Preliminary Phytochemical, Hydrilla Verticillata, Polarity, Water pollution

Assistant Professor ,
1st Dr. Mary KensaV. Department of Botany and Research Centre,
S.T. Hindu College, Kanyakumari, Tamil Nadu
Associate Professor ,
2nd Dr. Neelamegam R. Department of Botany and Research Centre,
S.T. Hindu College, Kanyakumari, Tamil Nadu

Abstract
Preliminary phytochemical studies in whole plant dry sample extracts of H.
Verticillata grown in unpolluted and polluted water sources were carried out to
determine the presence or absence of possible secondary metabolites using various
solvents (petroleum ether, chloroform, ethyl alcohol, methanol, and water).
Among the phytochemicals tested, flavonoids and phenols are not reported in all
the extracts of H. Verticillata from unpolluted and polluted water sources. The
steroids and tannins are present in the petroleum ether and chloroform extracts of
plant samples from unpolluted and polluted water sources. Presence of alkaloids
in water, chloroform, ethyl acetate and methanol extract (except petroleum ether)
of both unpolluted and polluted plant samples was noted. Saponins are present
only in water and petroleum ether extracts of both unpolluted and polluted plant
samples. The results of the qualitative preliminary phytochemical screening of
whole plant extracts of H. Verticillata plants from unpolluted and polluted water
sources exhibited the presence of alkaloids, flavonoids, phenols, saponins, steroids
and tannins depend upon the polarity of solvent and solubility level of
phytocomponents.

1. Introduction
Phytochemicals are defined as bioactive, non-nutrient plant compounds found in fruits, vegetables,
grains and other plant foods that have been linked to reducing the risk of major chronic diseases The
medicinal values of plants exists in their phytocomponents which produce a definite physiological
action on the human body. Preliminary phytochemical screening reports, generally revealed the
presence of alkaloids, tannins, cardiac glycosides and saponins in the methanol leaf extracts of

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Published Online On: November 30, 2015
Copyright©IJIFR 2015 1125
 ISSN: 2347-1697
International Journal of Informative & Futuristic Research (IJIFR)
Volume - 3, Issue -3, November 2015
Continuous 27th Edition, Page No.:1125-1128

Mitracarpus scaber (Abere et al. 2007); presence of alkaloid, catachin, coumarin, steroid, phenols,
amino acid, sugar, protein and xanthoprotein in the ethanolic extract of Alpinu calcarata and
Hiptage benghalensis leaves (Thirumurugan et al., 2008); presence of glycosides, mucilage,
carbohydrates, tannins, alkaloids and reducing sugars in the hexane, methanol and the aqueous
extracts of Tridax procumbens leaf (Kiran et al. (2008); and presence of iridiods, flavonoids and
triterpenes in methanol extract of Mussanda pupescens leaf (Vidyalakshmi et al., 2008).
Ethanol extract of Rubia cordifolia leaf showed positive results for alkaloids, cardiac
glycosides, tannins, flavonoids, and phenols and negative for saponins, volatile oils, anthraquinones
and cyanogenic glycosides and very trace for steroids (Kannan et al., 2009); the ethanolic extracts of
Costus mexicanus leaf showed the presence of phytosterols, saponins, glycosides and phenolic
compounds (Sheba et al., 2009). The occurrence of coumarines, flavonoids, sterols and triterpenoids
was reported by Kalaskar et al. (2010) in petroleum ether, ethanol, methanol and aqueous extract of
Ficus carica leaf samples. Ethanolic leaf extract of Pistia stratiotes (aquatic weed) revealed the
presence of anthraquinone, glycosides, cardiac glycosides, flavonoids, steroids and tannins (Khan et
al., 2011). The present study was carried out to determine the phytocomponents present in the H.
verticillata plant grown in polluted and unpolluted water conditions.

2. Materials And Method:

2.1 Study Area And Plant Materials Used

The selected plant H. verticillata belongs to the family Hydrocharitaceae was collected from
polluted and unpolluted water bodies in Asaripallam, Agastheeswaram Taluk, Kanyakumari
District, Tamil Nadu, India (Elevation about 460 meters (Mean Sea Level) and used for preliminary
phytochemical screening.

2.2 Preparation of whole plant dry powder

The selected plant was collected and dried separately at room temperature (30±2o C) for
about two weeks to get a constant weight. The dried plant materials (as whole plant) were ground to
powder by mechanical device and stored for further biochemical analysis.

2.3 Preparation Of Plant Extracts

The whole plant dry powder samples were extracted with different solvents such as
chloroform, methanol, ethyl acetate, petroleum ether and water at 20 % (w/v) level in a soxhlet
apparatus. The extracts were concentrated and used for qualitative phytochemical analysis.

3. Qualitative Phytochemical Analysis

Preliminary phytochemical screening of different solvent extracts of H. verticillata was
carried out separately following the methods of Herborne (1984), Trease and Evans (1987) and
Kolkate et al. (1995).
 Test for alkaloids (Wagner’s reagent test): To 1 ml of the extract, a few drops of Wagner’s
reagent were added and the formation of a reddish brown precipitate indicates the presence of
alkaloids.
 Test for flavonoids (Shinoda test): To 1 ml of the extract, magnesium turnings and 1-2 drops of
concentrated hydrochloric acid were added. Formation of pink colour indicates the presence of
flavonoids.

Dr. Mary KensaV. , Dr. Neelam egam R. :: Prelim inary Phy tochem ical
Analysis Of Hydrilla Verticillata (L.F.) Royle. Collected From Polluted And
1126
Unpolluted Water Sources
 ISSN: 2347-1697
International Journal of Informative & Futuristic Research (IJIFR)
Volume - 3, Issue -3, November 2015
Continuous 27th Edition, Page No.:1125-1128

 Test forcarbohydrates (Fehling’s test): 5 ml of Fehling’s solution was added to 2 ml of extract

and boiled in a water bath. The formation of yellow or red precipitate indicates the presence of
reducing sugars.
 Test for tannins and phenols (Lead acetate test): In a test tube containing about 5 ml of
extracts, 1 ml of 10 % lead acetate solution was added. Formation of yellow precipitate shows
the presence of tannins.
 Test for steroids (Salkowski’s test): To 2 ml of the extract, equal volume of conc. H 2 SO4 was
added carefully along the sides of the tubes. The upper layer turned red and the lower layer
turned yellow with green fluorescence, indicating the presence of steroids in the extract.
 Test for glycosides: To 1 ml of the test extract, 1 ml of sodium picrate solution was added. The
colour change from yellow to orange reveals the presence of glycosides.
 Test of saponins: To 5 ml of the filtrate, a drop of 5 % sodium bicarbonate solution was added.
The mixture was shaken vigorously and kept for 3 min. A honey comb like froth was formed
which shows the presence of saponins.
Preliminary phytochemical studies in different whole plant dry sample extracts of H.
verticillata grown in unpolluted and polluted water sources were carried out and the results are
presented in Table-1. Among the phytochemicals tested, flavonoids and phenols are absent (not
detected) in all the extracts of H. verticillata whole plant samples. The phytocomponents steroids
and tannins are noted only in the petroleum ether and chloroform extracts of H. verticillata whole
plant samples. But alkaloids found in all extracts (water, chloroform, ethyl acetate and methanol
extract) of H. verticillata. Similarly Saponins is present only in water and petroleum ether extracts of
H. verticillata samples.
Table –1: Preliminary phytochemical screening in the extract of Hydrilla verticillata whole plant
samples collected from unpolluted and polluted water sources.
Habitat of Phytochemicals screened in H. verticillata
Whole plant
whole plant
Alkal

Tann
Stero
solvent extracts
Phen

Sapo
Flav
onoi

nins
oids

samples ids

ins
ols

used
ds

collected
1. Petroleum ether UPWS - - - + + +
PWS - - - + + +
2. Chloroform UPWS + - - - + +
PWS + - - - + +
3. Ethyl acetate UPWS + - - - - -
PWS + - - - - -
4. Methanol UPWS + - - - - -
PWS + - - - - -
5. Water UPWS + - - + - -
PWS + - - + - -
UPWS-Unpolluted Water Source; PWS-Polluted Water Source;

The plant produced secondary metabolites in response to stress (Keeling and Bohlmann
2006). These plant based secondary metabolites, have been subsequently exploited by human beings
for their beneficial role in a diverse array of applications (Balandrin et al., 1985). Often, plants
secondary metabolites may be referred to as plant natural products, which have illicit effects on other
organisms.

Dr. Mary KensaV. , Dr. Neelam egam R. :: Prelim inary Phy tochem ical
Analysis Of Hydrilla Verticillata (L.F.) Royle. Collected From Polluted And
1127
Unpolluted Water Sources
 ISSN: 2347-1697
International Journal of Informative & Futuristic Research (IJIFR)
Volume - 3, Issue -3, November 2015
Continuous 27th Edition, Page No.:1125-1128

4. Findings & Discussion

The results of present study showed that the plant parts of H. verticillata having rich primary and
secondary metabolites such as alkaloids, flavonoids, phenols, saponins, steroids, tannins can be used
as industrial raw materials. These metabolites are further used for biosynthesis of bioactive
compounds.
Preliminary phytochemical screening of plants is very useful for the determination of the
active constituents in different solvent extracts. Among the phytochemicals tested, flavonoids and
phenols are not detected in all the extracts of H. verticillatasamples while steroids and tannins are
observed in petroleum ether and chloroform extracts of plant samples present in water, chloroform,
ethyl acetate and methanol extract (except petroleum ether) were also noted and the saponins are
present only in water and petroleum ether extracts of H. verticillata samples.
The results of the qualitative preliminary phytochemical screening of whole plant extracts of
H. verticillata plants from unpolluted and polluted water sources exhibited the presence of
alkaloids, flavonoids, phenols, saponins, steroids and tannins depend upon the polarity of solvent
and solubility level of phytocomponents.

5. References
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11-17.
[2] Balandrin, M. F., Klocke, J.A., Wurtele, E.S. and Bollinger, W.H. 1985. Natural plant chemicals: Sources
of industrial and medicinal materials. Science, 228: 1154-1160.
[3] Herborne, J. B. 1984. Recent advances in chemical ecology, Natural Products Reports, 6: 85-109.
[4] Kalaskar, M.G., Shah, D.R., Raja, N.M., Surana, S.J. and Gond, N.Y. 2010.Pharmacognostic and
phytochemical investigation of Ficuscarica. Ethnobotanical Leaflets, 14: 599-609.
[5] Kannan, M., Ranjit Singh, A.J.A. and Naryanan, M. 2009.Phytochemistry and Ethanopharmacological
studies on Rubiacordifolia.Ethnobotanical Leaflets,13: 338-342.
[6] Keeling, C. I. and Bohlmann, J. 2006.Genes, enzymes, and chemicals of terpenoid diversity in the
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[7] Khan, M.D., Ahad, A., Prasanta, P., Islam, T., Biswas, N. and Sadhu, K. 2011.Cytotoxicity, antimicrobial
and neuropharmacological evaluation of ethanolic extract of Pistia stratiotes.Int. Research journal of
pharmacy., 2(2): 82-92.
[8] Kiran Prajapati, D., Singh, S.B., Mishra, P., Dubey, S. and Sangameswaran, B. 2008.Pharmacognostical
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[9] Kolkate, C.K., Purohit, A.P. and Gokhale, S.B. 1995. In: Pharmacognosy, 3rd edition, NiralinPrakashan,
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[10] Sheba, R., Kaur, R. and Arora, R. 2009.Studies on phytochemical constituents of Costus mexicanus.
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[11] Thirumurugan, N., Bhan, J. and Dhar, Y. 2008. Screening and optimization of Andrographis paniculata.
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[12] Trease, G.E. and Evans, W.C. 1987.A Text Book of Pharmacognosy. Oxford: ELSB Baillere Tindall,
London.
[13] Vidyalakshmi, K.S., Hannah, R., Vasanthi, S. and Rajamanickam, G.V. 2008. Ethnobotany,
phytochemistry and pharmacology of Mussaenda sps.(Rubiaceae). Ethnobotanical Leaflets, 12: 469-475.

Dr. Mary KensaV. , Dr. Neelam egam R. :: Prelim inary Phy tochem ical
Analysis Of Hydrilla Verticillata (L.F.) Royle. Collected From Polluted And
1128
Unpolluted Water Sources