Research J. Pharmacognosy and Phytochemistry 2010; 2(3): 103-108 P Arun et.al.

Screening Antibacterial Activity of Various

Extracts of Lawsonia inermis.
P Arun1*, KG Purushotham1, Johnsy Jayarani1, Vasantha
Kumari1 and D Chamundeeswari2
1
Dr.M.G.R.Educational and Research Institute, Dr. M. G. R
University, Maduravoyal. Chennai-600 095, Tamil Nadu, India.
2
Sri Ramachandra Medical College and Research Institute, Chennai,
Tamil Nadu, India.
ISSN 0975- 2331
Research Journal of Pharmacognosy
and Phytochemistry. 2(3): May-June ABSTRACT:
2010, 103-108 Lawsonia inermis known as Henna is a woody and flowering plant found
in North Africa and South west Asia. Its leaves extensively in the
treatmernt of urinary tract infection in Siddha system of medicine.
Lawsonia inermis was subjected to antibacterial analysis. A battery of
assays were performed on different extracts of Lawsonia inermis (Henna)
for antibacterial activities. Antibacterial effects of n-hexane, choloroform
and methanol extracts of the leaves extract of Lawsonia inermis exhibited
various degree of inhibition activity. It was observed that leaves extract
Review Article were promising against gram positive and gram negative bacteria viz.
Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas
aeruginosa, Escherichia coli and Proteus mirablis. This study showed
that Lawsonia inermis could inhibit certain bacteria.

KEYWORDS: Anti-bacterial activity, Lawsonia inermis, Henna and

Medicinal plants.

INTRODUCTION:
Medicinal plants are the most exclusive source of life saving drugs for the
majority of the world’s population. Bioactive compounds currently
extracted from plants are used as food additives, dyes, insecticides,
cosmetics, perfumes and fine chemicals. These compounds belong to a
group collectively known as secondary metabolites1. A monotypic genus,
*Corresponding Author: represented by Lawsonia inermis, native of North Africa and South- west
P.Arun, Research Scholar, Asia, widely cultivated as an ornamental and dye plant. Lawsonia
Department of Industrial inermis, syn. Lawsonia alba (Henna) is a flowering plant the sole species
biotechnology, Dr. M.G.R. in the genus in the family Lythraceae. A glabrous, much branched shrub
Educational and Research Institute, or small tree with greyish brown bark. Leaves opposite, Sub–sessile,
Dr.M.G.R. University, elliptic or broadly lanceolate, entire, acute or obtuse, often mucronulate,
Maduravoyal. Chennai-600 095, Flowers numerous, small, white or rose – coloured, fragrant, in large
Tamil Nadu, India. terminal pyramidal panicled cymes; capsule globose, about the size of a
E-mail ID: pea, with numerous, pyramidal, smooth seeds2.Henna has been found to
arunarunpalani@yahoo.co.in exhibit Antibacterial, Antifungal and Dermatological properties. It is
Ph. No. +919884288233 useful in coloring of skin, scalp and nails etc. Henna has also shown anti-
diarrhoel, diuretic, emmanagogue and abortifacient prophetically and is
found to be practically non-toxic3. Alcoholic extract of shade dried leaves
of Lawsonia inermis intraperitonially injected to rats showed anti-
inflammatory activity comparable to that of hydrocortisone4.Chloroform
extract of leaves exhibit promising antibacterial activity against Shigella
Received on 07.03.2010 and vibrio cholera5. Astringent properties6.
Accepted on 12.04.2010
© A&V Publication all right reserved

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Research J. Pharmacognosy and Phytochemistry 2010; 2(3): 103-108 P Arun et.al.

No antifungal activity was detected for henna aqueous Antibacterial Assay:

solutions7.Most of the medicinal plants have identified and
used for treatment of human diseases are well documented8.
Collection and Identification:
Lawsonia inermis (Henna) leaves were collected from
Botanical garden, madras university campus, Maduravoyal,
Chennai. The specimen thus obtained was identified and
authenticated by- Dr. K. Balakrishnan, Research Officer,
Central Research Institute for Ayurveda and Siddha (Central
Council for Ayurveda and Siddha), Arumbakkam, Chennai.
The voucher specimen has been also deposited in Herbarium
of Department of Botany, University of Chennai.
Extraction and Isolation:
Lawsonia inermis leaves were taken in different aspirator
bottles and successively extracted with solvents in the order
n-hexane, choloroform and methanol after each extraction,
the solvents were evaporated under reduced pressure using
high vaccum conditions. These extracts were used for
antibacterial assays.
Antibacterial Activity:
Sample preparation:
All the leaves extracts (viz., n-hexane, chloroform, and
methanol) prepared in Dimethyl sulfoxide (Dmso) (5 mg/
ml) and aliquots were used to test the antibacterial activity.
The antibacterial activity had been tested against five
different species of gram positive and gram negative
bacteria. Bacterial test cultures were maintained on the stock
culture Brain heart infusion agar (BHA). From the stock
culture a loopful of the fresh culture was inoculated in Brain
heart infusion broth. The seed broths were incubated at 37°C
±1oC for 24 hours. Inocula were prepared by diluting 24
hours old cultures in saline. A dilution of 1: 100 was used in
all the tests.
MATERIALS AND METHODS:
Materials:
• Mueller Hinton Agar (Himedia)
• Brain heart infusion agar (Himedia)
• Culture plates
• Sterile cork borer
• Chloramphenicol (Himedia)
• Autoclave
• Incubators
• Wireloop
• Bacterial Test Cultures
Antibacterial activity was performed by well diffusion
technique9. Petri plates (10x10 cm) were prepared with
Mueller Hinton agar. 0.1 ml of the diluted culture was
spread evenly over plate with loop or sterile glass spreader.
The plates were dried for 30 minutes at 37°C. Wells of 6
mm (approximate) diameter were cut with sterile cork borer
in the inoculated agar. The wells were filled with the plant
extract. Chloramphenicol (1 mg/ml) were used as control in
the other well. The plates were incubated for 72 hours at
37°C. At the end of incubation period, the clear zone of
inhibition around the wells was measured in millimeter
(mm). The results are expressed in Table 1. Following the
antibacterial assay, all the extracts were tested for their
activity against the following bacterial culture.
Test microorganisms:
The bacteria used for antibacterial study were
Staphylococcus aureus (MTCC 087), Escherichia coli
(MTCC 729), Klebsiella pneumoniae (MTCC 432),
Pseudomonas aeruginosa (MTCC 1688) and Proteus
mirablis (MTCC 425).
CONTROL:
Chemicals:
Chloramphenicol (1mg/ml) – (MERCK)
RESULTS AND DISCUSSION:
Plants have been a source of medicinal compounds since
pre-historic time. It is well established that all parts of plants
were used in Unani systems of medicine for centuries.
However, the discovery and use of synthetic drugs led to a
dramatic decline in the popularity of herbal products used in
the therapy. Nevertheless the realization of harmful toxic
effect of a large number of synthetic drug led to for
alternative sources which would be safe and effective in
various ailments.
Antibacterial Activity of Leaves Extract:
The screening results of various leaves extract of Lawsonia
inermis (Table 1) exhibited activity against the gram
positive and gram negative organisms, whereas n-hexane
and chloroform extracts were found active against
Staphylococcus auereus, Klebsiella pneumoniae, Proteus
mirablis and Pseudomonas aeruginosa (zone of inhibition
range 16mm-24mm) and inactive against Escherichia coli.

Table-1: Evaluation of Antibacterial Activity of Lawsonia inermis

CRUDE Zone of inhibition(mm)
EXTRACTS Antibacterial Activity
Staphylococcus aureus Klebsiella pneumonia E.coli Proteus mirablis Pseudomonas aeruginosa
n-Hexane (5mg/ml) 14 22 0 18 21
Choloroform 18 16 0 20 18
(5mg/ml)
Methanol (5mg/ml) 22 24 21 22 20
Chloramphenicol 24 28 26 28 24
(1mg/ml)

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Research J. Pharmacognosy and Phytochemistry 2010; 2(3): 103-108 P Arun et.al.

Methanol extracts have shown maximum activity against

Staphylococcus auereus, Klebsiella pneumoniae, Proteus
mirablis, Escherichia coli and Pseudomonos aeruginosa
(zone inhibition 21mm-24mm). Consequently it can be
suggested that the activity of Methanol extract is much
higher as compared to the n-Hexane and Choloroform
extract of Lawsonia inermis. As such the different
chemical constituents elaborated by different spp. of
Lawsonia inermis displayed are coumarin, Alkaloids,
Flavonoids, Quinones and Tannins and the antibacterial
activity of Lawsonia inermis may emencipate due to either
of these constituents. In conclusion, the summary of the
biological activity evaluation of the leaves extracts of
Lawsonia inermis can be delineated as follows: The
antibacterial activity of leaves extract were promising
against Staphylococcus aureus, Klebsiella pneumoniae,
E.coli, Proteus mirablis and Pseudomonas aeruginosa.
Crude methanol extract posses maximum activity against
both gram positive and gram negative bacteria.

ACKNOWLEDGEMENTS:
We are grateful to our Prof. Dr. R.Vasantha kumari for
her constant support, encouragement and guidance
throughout the study.

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