Key words: Hyptis suaveolens (L.) poit phytochemical analysis Antimicrobial activity, Antifungal, Antibacterial.
Plants have been used to treat or prevent illness since before recorded history. The sacred Vedas dating back
between 3500 B.C and 800 B.C give many references of medicinal plants .Plants and plant based medicaments
are the basis of many of the modern Pharmaceuticals we used today for our various ailments . However, the
knowledge of medicinal plant is rapidly dwindling due to the influence of Western lifestyle, reducing in number of
generations to carry on the use of plant species in traditional medicine which has increased the interact throughout
the world .
Chemical constituents in the plant are responsible for their medicinal as well as their toxic properties . They are
source of medicine since the earliest time. The most important of these bioactive constituents of plants are steroids,
terpenoids, carotenoids, flavanoids, alkaloids, tannins, and glycosides. Plants in all facet of life have served a
The traditional medicinal methods, especially the use of medicinal plants, still play a vital role to cover the basic
countries in the last decade.
The present study was carried out to evaluate the antibacterial and antifungal activity of aqueous, ethanol, methanol,
The plant, Hyptis suaveolens (L) Poit commonly known as Wilayati tulsi belongs to the family Lamiaceae and is an
ethno botanically important medicinal plant. The plant has been considered as an obnoxious weed, distributed
throughout the tropics and subtropics. Almost all parts of this plant are being used in traditional medicine to treat
various diseases. The leaves of H. suaveolens have been utilized as a stimulant, carminative, sudorific, galactogogue
and as a cure for parasitic cutaneous diseases . Crude leaf extract is also used as a relief to colic and stomach-
ache. Leaves and twigs are considered to be antispasmodic and used in antirheumatic and antisuporific baths ,
anti-inflammatory, antifertility agents , and also applied as an antiseptic in burns, wounds, and various skin
complaints. The decoction of the roots is highly valued as appetizer and is reported to contain urosolic acid, a natural
HIV-integrase inhibitor .
Collection of plant materials
Fresh leaves of Hyptis suaveolens (L.) Poit were collected from Kannukudi, Thanjavur (DT), Tamilnadu, India.
They were identified by Dr.S.John britto, The Director, The Rapinet herbarium St.Joseph’s college, Thiruchirapalli
and a voucher specimens were deposited in the Rapinet herbarium of St.Joseph’s College, Thiruchirapalli (Voucher
number K G 001/ 2013).
Preparation of power
The leaves of plants were dried under shade. The dried materials were mechanically powdered sieved using 80
meshes and stored in an air tight container. The powdered material was used for further photochemical, and
Extraction of plant material
Aqueous, Methanol, Chloroform and ethanol extracts were prepared according to the methodology of Indian
pharmacopoeia. The coarse powder material was subjected to soxhlet extraction separately and successively with
ethanol and distilled water, .methanol, chloroform, these extracts were concentrated to dryness in flash evaporator
under reduced pressure and controlled temperature (40
c to 50
c) the aqueous and ethanol extracts put in air tight
container stored in a refrigerator
Qualitative phytochemical analyses were done by using the procedures of kokate et al. (1995). Alkaloids, Tannins,
Phenols, Flavonoids, Steroids, Terpenoids, Glycosides Carbohydrates, Protein, Quinones, and Saponins were
Totally eight human pathogens were selected for the present investigation four microbial strains namely E.coli,
Thayaar Educational Trust, Mannargudi.
Bacterial inoculam preparation
The young microbial inoculam was prepared and used in the entire research period. The nutrient broth were
prepared and poured into tubes and sterilized. The culture was inoculated in the tube using inoculation needles or
loops. The tubes were incubated at 37°c for 24 hours for bacteria.
Fungal inoculum preparation
The invitro method proposed by National committee for clinical Laboratory standard for testing molds  was
followed for the present study. The fungal stock inoculam suspension was prepared from seven days old culture
grown on PDA medium. The fungal colonies were covered with 10 ml of sterile distilled water and suspension well
made by gently probing the surface with the tip of the Pasteur pipette and transferred to sterile tube. Heavy particles
were allowed to settle for 5 – 20 minutes, and the homogenous suspension were collected and mixed with a vortex
mixture. The density of this suspension was adjusted with a spectrophotometer at a wavelength of 530 nm for 80 –
85 % to obtain the standard inoculum.
The antimicrobial activities of the aqueous, ethanol, methanol, chloroform, extracts were tested using spread plate
method. The disc was prepared by using whatmann no 1 filter paper, then the filter paper disc of 5 mm in diameter
were sterilized and soaked in the different extract of plant material. The culture media is inoculated when the
medium maintains a temperature of 45°c so that the cells can be distributed thoroughly , then the medium containing
the culture is poured in the sterilized petriplates, allow to solidify and then incubated colonies appear on the agar
surface. The filter paper disc soaked in aqueous, ethanol, methanol, and chloroform, extract of. Hyptis suaveolens
(L) poit. Were placed on the surface of the microbial seeded nutrient plate and then the plate was incubated at 37°c.
Preparation of Antibiotic disc
Commercially available ciprofloxacin antibiotic was used for the study and the antibiotic (500 mg) was prepared for
disc by dissolving antibiotic powder in 100 ml of distilled water.
Antibiotic sensitivity method
The isolated microorganisms were tested with antibiotic for the susceptibility by disc diffusion method. Antibiotic
disc were used to detect antibiotic sensitivity of the microbial suspension from nutrient broth. The antibiotic disc
were placed on the inoculated plates and incubated at 38°c for 24 hours antibiotic sensitivity was assayed from the
diameter of zones.
RESULTS AND DISCUSSION
Plants are rich in a wide variety of secondary metabolites such as tannins, terpenoids, alkaloids, and flavonoids
which have been found invitro to antimicrobial properties. The biologically active compounds are screened by
dissolving the crude powder in various solvent. Phytochemical studies of various extract of Hyptis suaveolens (L.)
result showed that aqueous extract contains alkaloids, glycosides, carbohydrates, proteins, steroids, flavonoids,
phenols, terpenoids and devoid of quinines, saponins. In case of ethanolic extract alkaloids, glycosides,
carbohydrates, proteins, steroids, flavonoids, phenols, terpenoids, Quinones were present. Methanolic extract
contains alkaloids, glycosides, carbohydrates, proteins, steroids, flavonoids, phenols. Chloroform extract gives
positive result to the test for alkaloids, glycosides, carbohydrates, proteins, steroids, flavonoids, phenols.
(+) Indicates Presence, (-) Indicates Absence
The antibacterial activity was observed by measuring the width of the inhibitory zones. The result obtained in this
present study is summarized below
Aqueous extract of Hyptis suaveolens (L.) Poit showed 6mm in diameter of inhibitory zone against both
10mm against Escherichia coli, Staphylococcus aureus respectively. Methanolic extract also showed significant
inhibitory action (14mm) against Escherichia coli and 10mm against staphylococcus aureus. Chloroform extract
of Hyptis suaveolens (L.) poit Showed 15mm in diameter of inhibitory zone against Escherichia coli and the
Aqueous extract showed no inhibitory action against both Pseudomonas aeruginosa, Proteus vulgaris. Ethanolic
against Proteus vulgaris was found to be 15mm in diameter. The inhibitory zone of Methanolic extract against
exhibits 12mm of inhibitory zone against Pseudomonas aeruginosa, 13mm against Proteus vulgaris
Aqueous extract showed no inhibitory activity against both .Aspergillus Niger, Aspergillus flavus. Methanolic
extract showed highest activity of 14mm inhibitory zone against Aspergillus Niger followed by 13mm against
Aspergillus flavus. Ethanolic extract showed 12mm inhibitory zone against Aspergillus Niger where as chloroform
extract showed 10mm inhibitory zones against Aspergillus Niger. Ethanolic extract showed 14mm of inhibitory
zones against Aspergillus flavus where as chloroform extract showed 15mm against Aspergillus flavus.
Ethanolic extract showed 12mm inhibitory zones against Fusarium and 10mm against Rhizopus, inhibitory zone of
significant inhibitory activity (11mm, 14mm) against both Fusarium, and Rhizopus.
The extent of antimicrobial activity is varied depending upon solvent that has been used. Finally it can be concluded
that the active chemical compounds present in Hyptis suaveolens (L.) poit showed certainly fine place in treatment
of bacterial and fungal infections. The results from the present study is very encouraging and it indicates that the
mechanism of activity of antimicrobial ability should be studied more extensively to explore its potential in the
treatment of infectious disease
Authors acknowledge the valuable help rendered by Dr. V. Dhivakaran, Managing Trustee, STET Women’s
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