P.Jayanthi et al.

/ Journal of Pharmacy Research 2011,4(5),1405-1406

Research Article Available online through
ISSN: 0974-6943 www.jpronline.info
Phytochemical investigation of the extracts of
Eichhornia crassipes and its solvent fractionates
P.Jayanthi, P.Lalitha*, and Shubashini.K.Sripathi
Department of Chemistry,Avinashilingam Deemed University for Women,Coimbatore,Tamilnadu,India
Received on: 05-12-2010; Revised on: 14-01-2011; Accepted on:09-03-2011
ABSTRACT
Petroleum ether, acetone, ethyl acetate, aqueous, acid extracts and the chloroform and ethanol fractionates of aqueous extracts of Eichhornia crassipes were screened for the presence of
phytochemicals by standard procedures. Presence of sterols was observed in acetone and aqueous extract. Petroleum ether and aqueous extracts tested positive for flavonoids. Terpenoids were
present in the acetone extract. Proteins were present in aqueous extract and carbohydrates in acid extract. Presence of alkaloids, quinones and anthocyanins was noted in aqueous and acid extract.
Anthraquinones were found to be present in all extracts except petroleum ether and phenols were present in ethyl acetate extract. Ethanol fractionate showed the presence of proteins and terpenoids
which separated from aqueous extract. The results obtained provide a motivation that this plant which poses major threat to the environment and economy could be utilised in an efficient means.

Key words: Eichhornia crassipes, water hyacinth, solvent fractionation

INTRODUCTION
Plants synthesize a wide variety of chemical compounds, which can be sorted by their chemical extracts and fractionates of aqueous extract were screened for the presence of different
class, biosynthetic origin and functional groups into primary and secondary metabolites. phytochemicals [6, 9, 20].
Primary metabolites make up the physical integrity of the plant cell and are involved with the
primary metabolic process of building and maintaining of living cells. Secondary metabolites Phytochemical tests:
do not seem to be vital to the immediate survival of the organism that produces them and are
not an essential part of the process of building and maintaining living cells [18] whereas these Test for Alkaloids
carry out a number of protective functions in the human body. Plant secondary metabolites can Mayer’s test
boost the immune system, protect the body from free radicals, kill pathogenic germs and much A fraction of extract was treated with Mayer’s test reagent (1.36 g of mercuric chloride and 5 g
more. Knowledge of the chemical constituents of plants is desirable, not only for the discovery of potassium iodide in 100 ml of water) and observed for the formation of cream coloured
of therapeutic agents, but also because such information may be of value in disclosing new precipitate.
sources of such economic materials as tannins, oils, gums, precursors for the synthesis of Wagner’s test
complex chemical substances, etc. In addition, the knowledge of the chemical constituents of A fraction of extract was treated with Wagner’s reagent (1.27 g of iodine and 2 g of potassium
plants would further be valuable in discovering the actual value of folkloric remedies [4]. iodide in 100 ml water) and observed for the formation of reddish brown colour precipitate.
Hager’s test
The water hyacinth, Eichhornia crassipes (Mart.) Solms, is a tropical species belonging to the A few ml of extract was treated with Hager’s reagent (saturated aqueous solution of picric acid)
pickerelweed family (Ponte-deriaceae) [2]. It is a free floating aquatic plant well known for its and observed for the formation of prominent yellow precipitate.
production abilities and removal of pollutants from water. It can quickly grow to very high
densities (over 60kg m-2); thereby completely clogging water bodies, which in turn may have Test for Flavonoids
negative effects on the environment, human health and economic development [3,5]. NaoH test
A small amount of extract was treated with aqueous NaOH and HCl, observed for the formation
On the other hand, when looked from a resource angle, it appears to be a valuable resource with of yellow orange colour.
several unique properties. As a result, research activity concerning control (especially biologi- H2SO4 test
cal control) and utilization (especially wastewater treatment or phytoremediation) of water A fraction of the extract was treated with concentrated H 2SO4 and observed for the formation of
hyacinth has boomed up in the last few decades. This study focuses on the preliminary orange colour.
screening of the phytochemicals present in various extracts of water hyacinth which may aid in
the identification and isolation of compounds which may be of use to the public. Test for Sterols
Liebermann-Burchard test
MATERIALS AND METHODS: Extract (1ml) was treated with chloroform, acetic anhydride and drops of H 2SO4 was added and
observed for the formation of dark pink or red colour.
Plant collection:
Six tonnes of fresh water hyacinth was collected from Singanallur boat house, Coimbatore, Test for Terpenoids
Tamilnadu. The root portion was cut off and the plant was washed thoroughly to free from Liebermann-Burchard test
debris. The leaves and shoot portion were shade dried for 20 days. The dried plant material was Extract (1ml) was treated with chloroform, acetic anhydride and drops of H 2SO4 was added and
sliced, ground coarsely and stored for further use. observed for the formation of dark green colour.

Preparation of extracts: Test for Anthraquinones

Water hyacinth (1500g) was defatted twice with petroleum ether (5L) for 6 hours and then Borntrager’s test
refluxed twice with ethanolic KOH (4.5L) for 6 hours. The extract was desolvetised under About 50 mg of powdered extract was heated with 10% ferric chloride solution and 1ml
reduced pressure and the residue was extracted thrice with acetone under reflux for 1 hour. The concentrated HCl. The extract was cooled, filtered and the filtrate was shaken with diethyl
acetone extracts were pooled and concentrated to get acetone extract. ether. The ether extract was further extracted with strong ammonia; pink or deep red colourations
of aqueous layer indicate the presence of anthraquinone.
Water hyacinth (140kg) was extracted successively with ethyl acetate (800L) to get the ethyl
acetate extract . The plant residue was then refluxed with water (800L) twice for 6 hours to get Test for Anthocyanin
the aqueous extract. A small portion (1500g) of the plant residue was extracted with 1:1 NaOH test
hydrochloric acid (3L) for 6 hours to get the acid extract. A small amount of extract was treated with 2M NaOH and observed for the formation of blue
green colour.
The aqueous extract was fractionated with chloroform and ethanol. The various solvent
Proteins
*Corresponding author. Ninhydrin test (Aqueous)
P.Lalitha The extract was treated with aqueous ninhydrin and observed for the presence of blue colour,
Department of Chemistry, indicating the presence of amino acid or purple colour indicating the presence of protein.
Avinashilingam Deemed University for Women, Ninhydrin (acetone)
Coimbatore,Tamilnadu,India Ninhydrin was dissolved in acetone; the extract was treated with ninhydrin and observed for
Tel.: 098422 92614 the formation of purple colour.
E-mail:goldenlalitha@gmail.com

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 P.Jayanthi et al. / Journal of Pharmacy Research 2011,4(5),1405-1406
Biuret test Triterpenoids are a large class of natural isoprenoids present in higher plants, which exhibit
The extract was heated in distilled water and filtered. The filtrate is treated with 2% copper a wide range of biological activities [7]. Steroids (anabolic steroids) have been observed to
sulphate solution, to this added 95% ethanol and potassium hydroxide and observed for the promote nitrogen retention in osteoporosis and in animals with wasting illness [1]. An-
formation of pink ethanolic layer. thraquinones are a main source of natural dye, which are gaining importance due to the
Test for Phenols environmental pollution caused by synthetic dyes [11]. Carbohydrate is reported to have
Ferric chloride test numerous roles in living things, such as the storage and transport of energy (starch,
The fraction of extract was treated with 5% ferric chloride and observed for the formation of Glycogen) and structural components (cellulose in plants, chitin in animals). Additionally
deep blue or black colour carbohydrates and their derivatives play major roles in the working process of the immune
Liebermann’s test system, fertilization, pathogenesis, blood clotting and development [14].
The extract was heated with sodium nitrite, added H2SO4 solution diluted with water and
excess of dilute NaOH was added and observed for the formation of deep red or green or blue Lata et al. [12] demonstrated the presence of few secondary metabolites in Eichhornia
colour. crassipes except saponins in aqueous extract which was prepared by soaking the dried,
powdered plant material for 12 hours. Kandukuri et al. [8] reported the presence of alkaloids,
Test for Quinones phenols, steroids, tannins and triterpenoids in the methanol extract which was prepared by
A small amount of extract was treated with concentrated HCl and observed for the formation soxhlet extraction whereas the authors have reported the absence of flavonoids in water
of yellow colour precipitate. hyacinth. Ndubuisi et al. [16] revealed the presence of saponin, glycoside and anthraquinone
but absence of alkaloid in the chloroform extract of water hyacinth. Dubey et al. [13] reported
Test for Carbohydrates the presence of alkaloids in the aqueous extract of water hyacinth.
Molisch’s test for Carbohydrates
Few drops of Molisch’s reagent was added to each of the portion dissolved in distilled water, Our present study involves a detailed phytochemical investigation of bulk quantities of water
this was then followed by addition of 1 ml of conc. H2SO4 by the side of the test tube. The hyacinth. As a preliminary step, phytochemical screening of different extracts was carried out.
mixture was then allowed to stand for two minutes and then diluted with 5 ml of distilled Fractionation makes it possible to isolate more than two components in a mixture in a single
water. Formation of a red or dull violet colour at the interphase of the two layers was a run. This property sets it apart from other separation techniques. Hence solvent fractionation
was employed for the aqueous extract. The ethanol fractionate of aqueous extract showed that
positive test.
flavonoids, terpenoids and proteins which were present in aqueous extract, after fractionation
Fehling’s test for free reducing sugar
was found in ethanol fractionate.
About 0.5 g of each extract was dissolved in distilled water and filtered. The filtrate was
heated with 5 ml of equal volumes of Fehling’s solution A and B. Formation of a red Bulk isolation of compounds based on the results of phytochemical screening is under
precipitate of cuprous oxide was an indication of the presence of reducing sugars. progress in our laboratory. Preliminary screening tests may be useful in the detection of the
bioactive principles and subsequently may lead to the drug discovery and development.
RESULTS AND DISCUSSION: Further, these tests facilitate their quantitative estimation and qualitative separation of pharma-
The results obtained in the present investigation for the extracts viz. petroleum ether(PEE), cologically active chemical compounds.
acetone(AcE), ethyl acetate(EAE), aqueous(AqE) and acid (AcidE) and the fractionates of
aqueous extract i.e chloroform(CHCl3F), ethanol (EtOHF), and aqueous(AqF) are summarised ACKNOWLEDGMENT:
in table 1. The preliminary phytochemical tests result indicates the presence of phenol, The financial support of DRDO-LSRB is acknowledged and the authors thank Avinashilingam
flavonoids, alkaloids, terpenoids, sterols, tannins, carbohydrates, anthroquinone, anthocya- Deemed University for providing necessary facilities to carry out this work.
nins, and proteins in different extracts. The presence of wide range of phytochemical
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Source of support: DRDO-LSRB, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.4.Issue 5. May 2011 1405-1406