Int J Med Res.

2010; 1(2): 94-98 ISSN: 0976-8971

IJMBR
International Journal of Medicobiological
Research
(An International peer review journal)
Journal homepage: ijmedres.com

Research article

Phytochemical screening and Antibacterial activity of Sida cordifolia L. (Malvaceae)

leaf extract
A. Kalaiarasan, S. Ahmed John
PG and Research Department of Botany, Jamal Mohamed College, Tiruchirappalli-620020, Tamil Nadu, India.

Corresponding author: [email protected]

Article information ABSTRACT

Keywords: Phytochemical screening and antibacterial investigation was made in leaf extracts of Sida
Sida Cordifolia L. cordifolia L. The plant materials were collected and shade dried. The ethanol and methanol
Antibacterial activity extracts were prepared using soxhlet apparatus. Ethanol and methanol extracts revealed the
Pathogenic strain presence of alkaloids, glycosides, carbohydrates, flavonoids, phenols, saponins and tannins. The
antibacterial activity was studied by using agar–disc diffusion method. The results showed that
ethanol and methanol extracts inhibit the growth, in selected organisms. The higher number of
zone inhibition were found on the ethanol extract showed activity against virulent bacteria such
Received on: 24.09.2010 as Escherichia fecalis, Pseudomonas aeroginosa. The 11 mm of zone of inhibition found at 75
Revised on: 30.09.2010 mL/disc concentration, and methanol extract showed that the high inhibitory zone towards,
Accepted on: 26.11.2010 Escherichia coli (8.53 mm) at 75 mL/disc concentration no inhibitions is other two extracts.

1. INTRODUCTION extract of Sida cordifolia L. can be effectively used in curing the

bacterial diseases such as treat as variety of health disorders like
The plant Sida cordifolia L., the plant grows well throughout the bronchial asthma, cold and flu, chills, lack or perspiration,
plains of India., especially is damp climates. The shrub grows up headache, nasal congestion–aching joints and bones, cough,
to 0.75–1.5 m in height. The roots and the stem being stout and wheezing, edema and keeping the good health of the people.
strong. The leaves are 2.5–7 cm and 2.5–5 cm broad with 7–9
veins. They are heart shaped, serrate and truncate. The flowers 2. MATERIALS AND METHODS
are small. Yellow or white in colour, solitary and auxiliary. The
fruits are a moon–sized, 6–8 mm in diameter. The seeds are 2.1. Plant collection, authentication and garbling process
called as bijabanda in Ayurveda, are greyish black in colour and
smooth. The flowers blossoming from August to December and The plant material is leaf of Sida cordifolia L. were collected in
fruiting occurs from October to January. It was observed that sufficient quantities from the natural habitats from the riverbank
ethanol and methanol extract of the leaves of Sida cordifolia L. of Cauvery and roadside Tiruchirappalli. The collected leaves
The both extracts were showed that the presence of alkaloids, were shade–dried at room temperature into powder form for
glycolysides, carbohydrates, flavonoids, phenols, saponins and preparing different solvents extract. The plant Voucher No. 12.
tannins. It was observed that ethanol and methanol extract of The plant authenticated by Dr. S. Ahmed John. Associate
good inhibitory action against all the selected bacteria except Professor and Head of Department of Botany, Jamal Mohamed
Escherichia coli. The present findings showed that ethanol College (Autonomous), Tiruchirappalli – 620 020, Tamilnadu,
extract was more effective than methanol extract against all the India.
selected bacteria. It is concluded that ethanol and methanol

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 A. Kalaiarasan, et al.,/Int J Med Res. 2010; 1(2): 94-98 95

2.2. Preliminary phytochemical analysis[1-3] and aldehyde. The reduced ninhydrin then reacts with the
liberated ammonia forming blue complex proteins and hydroxyl
The preliminary phytochemical screening was followed proteins produce a yellow rather than a purple colour with
by medicinal chemistry, Horborne 1973. The antibacterial ninhydrin.
studies were performed by following methods as given in Indian
pharmacopoeia. The collected fresh leaves of Sida cordifolia L. c) Millon’s Test
used for the extraction purposes 450 g of powdered were evenly
packed in the soxhlet apparatus. It was then extracted with It is a detecting reagent for proteins. Dissolve 1 g of mercury is 9
ethanol and methanol solvent. mL of foaming nitric acid. Keeping the mixture well cooled
during the reaction when the action is complete add equal
2.2.1. Test for fixed oils and fats volume of distilled water.

a) Spot test 2.2.4. Test for gum and mucilages

a) Molisch’s test
5 mL of extract solution placed in an ordinary paper.
A translucent spot is visible. This indicates the presence of fat. Dissolve 1g of napthol is 60 mL of 95% alcohol. It is used for
the detection of carbohydrates.
b) Saponification Test
b) Ruthenium Test
Take 5 mL of the extract in a test tube. Add 20 drops of 40%
NaOH and 2 mL of glycerol to it. Gently boil for about 3 It is used as a staining reagent for mucilage. In 10 ml of 10%
minutes until complete saponification occurs. If oil globules are solution of leaded acetate, dissolve 0.008 g of ruthenium red.
visible, boiling must be continued, divide the solution into 3 Use freshly prepared solution.
parts to carry the following experiments in test tube 1, 2, 3. To
test tube No. 1 was added with a saturated of NaCl. Make that 2.2.5. Test for alkaloids
the soap separates out and floats on the surface (Salting out a) Dragendroff’s Reagent
process). To test tube No. 2 was added a few drops of con. HCl
and oily layer of the fatty acids rise to the surface. To test tube It is used for the detection of alkaloids. Boil 14 g of sodium
No. 3 was then added a few drops of CaCl2 solution the soluble, iodide with 5.2 g basic bismuth carbonate is 50 mL glacial acid
calcium soap is precipitated. for few minutes. Allow it to stand overnight and filter off the
precipitate of sodium acetate crystal. To 40 mL of the red brown
2.2.2. Test for tannins and phenolic compounds filtrate add 160 mL of ethyl acetate add 1 mL water. Preserve
a) Ferric chloride test the stock solution is amber–coloured bottle. When needed
added 20 mL acetic acid to 10 mL of this stock solution and
5% W/V solution of ferric chloride in 90% alcohol and used for make up to 100 mL with water.
detection of phenols.
b) Mayer’s Reagent
b) Lead acetate test
Dissolve 1.36 g of mercuric chloride is 60 mL distilled water a)
25% of basic lead acetate solution is used for the detection of Dissolve 5 g of potassium oxide in 50 mL distilled water b) Mix
flavonoids. c) Adjust the volume to 100 mL with distilled water.
2.2.3. Proteins & amino acids
a) Biuret Test c) Hager’s Reagent

To 2 mL of test solution was taken with equal value of 10% A saturated aqueous solution of picric acid used for detection of
NaOH and one drop of 10% CuSO4 solution. A violet colour alkaloids.
formation indicates that the presence of peptide linkage.
d) Wagner’s Reagent
b) Ninhydrin Test
Dissolved 1.27 g of iodine and 2 g of potassium iodine in 5 mL
To 1mL of ninhydrin solution, add 1 mL test solution and heat. of water and make up the value to 100 mL distilled water.
Formation of violet colour indicates the presence of X-amino
acids. 2.2.6. Test for carbohydrate
a) Molisch Reagent
Ninhydrin is a powerful oxidizing agent, which causes.
Oxidative decaxbohydation of X-amino acids yielding CO2, NH3 Aqueous or alcoholic solution of substance was added with 10%
aqueous solution of alpha napthol shaken and added

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 A. Kalaiarasan, et al.,/Int J Med Res. 2010; 1(2): 94-98 96

concentrated sulphuric acid along the side of the tube. Violet transferred aseptically. The samples were added to the each cup
ring at the junction of two liquids shows presence of aseptically and labeled accordingly. The reference standard also
carbohydrates. added to the separate cups. The plates were kept undisturbed in
an incubator at 37±1oc for 24 hours and the zone of inhibition of
b) Fehling’s Test the standard and test samples were observed.

2 mL of Fehling’s solution A and 2 mL of Fehling’s solution B 3. RESULTS AND DISCUSSION

mixed together and added to 2 mL of the sample solution.
Boiled for 2 minutes and cooled. Yellow precipitate or brick red Phytochemicals were screened by using the two different solvent
precipitate shows the presence of reducing sugars. extracts. The bioactive constituents of the leaves of Sida
cordifolia L. have been extracted and subjected to preliminary
c) Barfoed’s Test colour tests to identify the nature of the compounds present.
Ethanol and methanol extracts revealed the presence of
2 mL of test solution & 2 mL of barford’s reagent. Boil on alkaloids, glycosides, carbohydrates, flavonoids, phenols,
water bath. Brick red precipitate at the bottom of the test tube saponins and tannins were confirmed by suitable chemical tests.
monosaccharides present.
The antibacterial activity of the plant ethanol extracts
c) Benedict’s Test were determined against selected bacteria showing activities
(Table. 2.). The above result revealed that the ethanolic leaf
5 mL of Benedict’s reagents and then 3 mL of test solution boil extract have is very large activity Coryne bacterium (8.4 mm) at
for 2 minutes and cooled. Green yellow or red precipitate 75 mL/disc concentration Escherichia coli, Klebsiella
showed the presence of reducing sugar present. pneumonia (9.5 mm) zone of inhibition at this 75 mL/disc
concentration. Excherchia fecalis, Pseudomonas aeruginosa (11
d) Borntrager’s Test mm) zone of inhibition at this 75 mL/disc concentration.

Anthraquinone derivatives are generally detected by Antibacterial activity of the plant extracts was
determined against selected bacteria showing activities (Table.
Borntrager’s test. In this test, the drug is powered, boiled with
3.). The above results revealed that the methanolic extract have
dilute sulphuric acid and filtered while hot. The filtrate further
extracted with ether or any water immiscible organic solvent. significant activity against Escherichia fecalis, Klebsiella
The organic solvent separated and equal volume of ammonia pneumonia, Staphybococcus aureus (8 mm) at 75 mL/ disc
was added. A pink colour slowly developed and finally become concentrations. Escherchia (Coli 8.5 mm), Pseudomonas
red is the ammonia layer, confirm the presence of aeruginosa and coryne bacterium (7 mm) respectively at 75 mL/
anthroquinones. disc concentration. Zone of inhibition was recorded against
corynebacterium (7.5 mm) respectively at 50 mL/disc
2.3. Biological studies concentration. Partially zone of inhibition was recorded against.
2.3.1. Antibacterial activity Escherchia coli, Klebsilla pneumonia and Pseudomonas
aeruginosa, (7 mm). There was no activity all the bacterial
The antibacterial activity was studied systematically against strains tested at 25 mL/disc.[4-9]
different strains of bacteria by agar diffusion method in
particularly cup–plate method. The extracts were evaluated for 4. CONCLUSION
their antibacterial activity against Coryne bacterium,
Escherichia coli, Escherichia fecalis, kelbsiella pneumoniae, In present findings both the ethanolic and methanolic extract
Pseudomonas aeruginosa, Stceptococcus aureus. The test leaves of Sida cordifolia L. significant inhibitory action against
organisms were sub cultured using nutrient agar medium. The all the selected bacteria, it is concluded that ethanolic extract of
media used for antibacterial test was nutrient Agar/Broth of the Sida cordifolia L.., can be effectively used for curing the
Himedia pvt. ltd., Mumbai, India. The bacteria were supplied by bacterial diseases such as health disorders like bronchial asthma,
the Department of Microbiology, Institute of Basic Medical cold and flu, chills, lack of perspiration, headache, nasal
Science, Chennai, India. congestion, aching joints and bones, cough, wheezing and
edama., also recommended keeping the good health of the
The sterilized medium was inoculated with respective people.
bacterial strains after incubation at 37±1oc for 24 hrs. They were
stored in refrigerator and thus stock cultures were maintained. ACKNOWLEDGEMENT
The nutrient agar medium was prepared and sterilized by using
autoclave at 121ºC (1516/ sq.inc) for 1 hour. The petri-dishes, My heartful thanks to Dr. S. Ahmed John, Associates Professor
tube and flask were sterilized in hot air oven at 150oc for an and Head. PG and Research Department of Botany, Jamal
hour. Each sterilized petri dishes 30 mL of mother nutrient agar Mohamed College, Tiruchirappalli-620 020, Tamilnadu, India.
medium isolated with respective strain of bacteria was

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 A. Kalaiarasan, et al.,/Int J Med Res. 2010; 1(2): 94-98 97

5. REFERENCES [6] Baqir S., Naqvi., Rafi Shaikh M., Maleka F.A., Dilnawaz
th Sheikh. 1994. Antibacterial activity of ethanolic extracts
[1] G.E. Trease and W.C. Evans. Pharmacognosy., 13 edition. from Nericum indicum and Hibiscus rosasinensis. Journal
247-248. of Islamic academy of Science., 7(3): 167-168.
[2] Mebrata B.N. 1990. Quality control requirements of
medicinal plants used in traditional medicines ethanobotany. [7] Mukhtar H.M., Ansari S.H., Ali M., and Klani F.A 2002.
2: 19–24. Antimicrobial activity of Betula pendula and Hamdard
Medicus. 45(1):41-43.
[3] Indian Pharmacopoeia. 1996. Itoh. A et al Isolated mono
terpenes and glycosides, Journal of natural products. 2: [8] Chowdhury D., Sayeed A., Islamd A., Bhuiyan M.S.A and
100–115. Khan. G.R., M.A.M. 2002. Antimicrobial activity and
[4] Aliero A.A., and Afolayan A.J., 2006. Antimicrobial cylotoxicity of Aerva lamata. Fitoterapia., 73(1): 92-94.
activity of solanum tomentosum. Afr j Biotechnology., 5: [9] Kannan S., and Venkatakrishnan V., 2002. Antibacterial
369-372. activity of Phyllanthus amarus. Bionotes., 4(2): 40.
[5] Alam M.T., Karim M.M., and Shakila N. Khan. 2009.
Antibacterial activity of different organic extracts of
Achyranthes Aspera and Cassia Alata. Sci Res., 1: 393-398.

Table. 1. Qualitative phytochemical tests on various extracts of leaf of Sida cordifolia L.

S. No. Constituents Tests Ethanolic extract Methanolic extract

1. Alkaloids Mayer’s reagent + -

Dragondraff’s Reagent + -
Hager’s Reagent - -
Wagner’s Reagent - -
2. Sterols Libermann’s Reagent + -
Salkowski Reagent + -
3. Carbohydrates & glycosides Molisch Reagent - -
Felings Reagent + -
Barfoeds Reagent + +
Borntrager’s Reagent - +
5% KOH Reagent - -
4. Fixed oil fats Spot Test - +
Saponification - -
5. Phenolic compounds Extract +Fecl3 + +
6. Test Tannins Gelatin Test + -
FeCl3 Test + -
7. Protein and amino Acids Biurette test + +
Ninhydrin Test - +
Xanthoprotein Test - +
Millon’s test + -
8. Triterpenoid & Saponin Tin + Thionyl Chloride - +
9. Gums & mucilage With 95% alcohol - -
Molisch test
10. Falvone & Flavonoids Con H2SO4 + -

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 A. Kalaiarasan, et al.,/Int J Med Res. 2010; 1(2): 94-98 98

Table. 2. Antibacterial activities of Ethanol extract of Sida cordifolia L. Leaves against the pathogenic organisms

S. No. Name of the Organism Control Gendamicin Zone of inhibition (Diameter in mm)
25 mL/Disc 50 mL/Disc 75 mL/Disc
1. Corynebacterium 14.0 7.0 7.0 8.4
2. Escherichia Coli 15.0 - 8.4 9.5
3. Escherichia Fecalis 16.0 7.0 7.0 11.0
4. Klebsilla Pneumonia 14.0 7.0 8.0 9.5
5. Pseudomonas aeruginosa 15.0 - 8.0 11.0
6. Staphylococcus aureus 14.0 - 7.0 7.0

Table. 3. Antibacterial activities of Ethanol extract of Sida Cardifolia L. Leaves against the pathogenic organisms

S. No. Name of the Organism Control Gendamicin Zone of inhibition (Diameter in mm)
25 mL/Disc 50 mL/Disc 75 mL/Disc
1. Corynebacterium 14.0 - 7.5 7.0
2. Escherichia Coli 14.0 - 7.0 8.5
3. Escherichia Fecalis 14.0 - - 8.0
4. Klebsilla Pneumonia 14.0 - 7.0 8.0
5. Pseudomonas aeruginosa 14.0 - 7.0 7.0
6. Staphylococcus aureus 14.0 - - 8.0

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