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Journal of Experimental Sciences Vol. 2, Issue 3, Pages 04-06 [2011]

www.jexpsciences.com

Regular Article

Antimicrobial and Phytochemical Screening of Plumbago zeylanica Linn.

(Plumbaginaceae) Leaf
D.A. Dhale1* and S.K. Markandeya2
1
Post-Graduate Department of Botany, SSVP Santha’s L.K. Dr. P. R. Ghogrey Science
College, Dhule-424005 (India); 2Government Institute of Science, Caves Road,
Aurangabad-431004 (India)
al., 2001), antimicrobial (Ahmad et al., 2000), antifungal (Mehmood
ABSTRACT: The antimicrobial effect of Plumbago zeylanica Linn. et al., 1999), anti-inflammatory (Oyedapo, 1996), antihyperglycemic
(Plumbaginaceae) leaf extract was evaluated on microbial strains (Olagunju et al., 1999), hypolipidaemic and antiatherosclerotic
like gram positive species Staphylococcus aureus, and Bacillus activities (Sharma et al., 1991).
subtilis and gram negative species Escherichia coli and
Pseudomonas aeruginosa. The solvent used for extraction of plant Materials and Methods
were petroleum ether, chloroform and alcohol. The alcoholic extract Sample collection and Authentication
of leaves of Plumbago zeylanica shows maximum antimicrobial The fresh, mature healthy leaves of Plumbago zeylanica Linn
activity. The in vitro antimicrobial evaluation was carried out by agar (Plumbaginaceae) were collected from were collected from the
disc diffusion method. The significant antibacterial activity of active Government institute of Science, Aurangabad (M.S.) campus. The
extract was compared with standard antibiotic Amphicillin. The plant materials were identified using the Flora of Marathwada (Naik,
samples of leaves were further used for the phytochemical studies. 1998a) at Post graduate Department of Botany, Government
Results of the phytochemical analysis indicated the presence of institute of Science, Caves Road, Aurangabad (M.S.) India.
alkaloids, glycoside, reducing sugars, simple phenolics, tannins,
Lignin, saponins and flavonoids. The antibacterial activities of the Sample preparation
leaves were due to the presence of various secondary metabolites. Fully grown leaves of P. zeylanica were weighed (1kg). The plant
samples were shade dried ground and sieved with 2mm copper
Key words: Antimicrobial, Physico-chemical, Phytochemical, sieve to form uniform powder and stored in airtight bottles.
Plumbago zeylanica
Preparation of extract
Introduction The dried plant material was pulverized into fine powder using a
In fact, plants produce a diverse range of bioactive molecules grinder (mixer). About 50 g of powdered material was extracted in
making them a rich source of different types of medicines. Higher soxhlet extraction apparatus with 250 ml of each of the following
plants, as sources of medicinal compounds, have continued to play a solvents; petroleum ether, chloroform, and alcohol (Vogel, 1988).
dominant role in the maintenance of human health since ancient The extracts obtained with each solvent were filtered through
times (Farombi, 2003). Over 50% of all modern clinical drugs are of Whatman filter paper No. 1 and the respected solvents were
natural product origin (Stuffness and Douros, 1982). Natural evaporated (at 40ºC) with the help of heating mantle. The sticky
products play on important role in drug development programmes in greenish-brown substances were obtained and stored in refrigerator
the pharmaceutical industry (Baker et al., 1995). There are a few for prior to use (Beyer and Walter, 1997). Some of the extracts of
reports on the use of plants in traditional healing by either tribal each solvent were used for the qualitative phytochemical screening
people or indigenous community (Sandhy et al., 2006; Ayyanar and for the identification of the various classes of active chemical
Ignacimuthu, 2005; Rajan et al., 2002; Natarajan et al., 1999 and constituents, using standard prescribed methods (Harborne, 1984;
Ignacimuthu et al., 1998). The antimicrobial activity have been Trease and Evans, 1987; Ajaiyeoba, 2000; Edeoga et al., 2005). The
screened because of their great medicinal relevance with the recent positive tests were noted as present (+) and absent (-).
years, infections have increased to a great extent and resistant
against antibiotics, becomes an ever increasing therapeutic problem Preparation of microorganism
(Austin et al., 1999). Natural products of higher plants may give a Isolation of bacterial species of Gram positive (Staphylococcus
new source of antimicrobial agents. There are many research groups aureus and Bacillus subtilis ) and Gram negative (Echerichia coli and
that are now engaged in medicinal plants research (Samy et al., Pseudomonas aeruginosa) takes place. The cultures of these
1998; Hamil et al., 2003; Motsei et al., 2003). Silver and Bostian bacteria were checked for purity by doing gram staining and
(1993) have documented the use of natural products as new biochemical test and they were grown in nutrient broth at 37˚C and
antibacterial drugs. There is an urgent need to identify novel maintained in nutrient agar slants at 2-8˚C. Nutrient agar medium
substances active towards highly resistant pathogens (Recio, 1989; was used as bacterial culture medium in the antibacterial assays.
Cragg et al., 1997). In an effort to discover new compounds, many
research groups screen plant extracts to detect secondary Selection of Reference antibiotic
metabolites with the relevant biological activities. In this regard, Reference antibiotic Amphicillin was obtained from authorized
several simple bioassays have been developed for screening medical shop Aurangabad (M.S.). The purity of the antibiotic is
purposes (Hostettmann, 1991). 99.8%
The present study was carried out on the phytochemical and
antibacterial activity of leaf of Plumbago zeylanica Linn. Dilutions and Inoculum preparations
(Plumbaginaceae) is a tropical shrub. It grows wild as a garden plant The dried plant extracts of P. zeylanica and antibiotic Amphicillin
in eastern, northern and southern India and Ceylon. The leaves of P. were weighed and dissolved in sterile distilled water to prepare
zeylanica are widely used medicinally in India and China. appropriate dilution to get required concentration of 50, 100mg/ml.
Traditionally, P. zeylanica is believed to kill intestinal parasites, and The inoculums of Staphylococcus aureus, Bacillus subtilis,
it is used clinically to treat rheumatism, intestinal parasites, anemia Escherichia coli and Pseudomonas aeruginosa were prepared in
due to "stagnant blood”, external and internal trauma, toxic swelling nutrient broth medium and kept incubation at 37˚C for 8 hours.
and malignant furunculous scabies (Jiangsu, 1979). In India it is After growth was observed, the cultures are stored in the
usually used to treat fever or malaria. Pharmacological studies have refrigerator at 2-8˚C for analysis.
indicated that P. zeylanica extract has antiplasmodial (Simonsen et

* Corresponding Author, Email: datta.dhale@yahoomail.com JES

ISSN: 2218-1768
J Exp Sci Vol. 2, Issue 3, Pages 04-06 [2011]

Procedure for performing the Disc Diffusion test (Bayer et zeylanica Linn. leaves were tested against various Gram-negative
al., 1997) and Gram-positive bacteria (Table.1). Among the extracts assayed,
The required amount of Petri plates is prepared and autoclaved at the alcohol leaf extracts of P. zeylanica exhibited good activity
121˚C for 15 minutes. And they were allowed to cool under Laminar against P. aeruginosa at 100mg/ml for example, 17 mm was
air flow. Aseptically transfer about 20 ml of media into each sterile recorded as diameter zone of inhibition. This was followed by 16
Petri dishes and allowed to solidify. 1 ml inoculum suspension was mm E. coli, 11 mm B. subtilis and Staphylococcus aureus 10 mm
spread uniformly over the agar medium using sterile glass rod to get respectively. The least activity 3 mm against E. coli, 4 mm and
uniform distribution of bacteria. The readily prepared sterile discs B.subtilis at 50mg/ml was recorded by petroleum ether extracts.
were loaded with different concentrations of about 50, 100mg/ml of Activities of the various extracts were comparable to those of
plant extract of P. zeylanica and antibiotic amphicillin into each standard antibacterial agent Ampicillin.
separate disc of about 100μl. The paper diffuse discs were placed on In the present antimicrobial activity of both plant extract towards
the medium suitably apart and the plate were incubated at 5˚C for 1 drug resistant or clinically significant microbes are reported and it
hour to permit good diffusion and then transferred to an incubator was observed that active constituent of plant material seep out in
at 37˚C for 24 hours. The antibacterial activity was recorded by organic solvent to display biological activity. Further phytochemical
measuring the width of the clear inhibition zone around the disc studies for identification and elucidation of active constituent in plant
using zone reader (mm). material tested in expected to serve as lead in the development of
novel bioactive antimicrobial compound.
Results and Discussion Phytochemical evaluation
Indian systems of medicine such as Ayurveda and Siddha uses The results of qualitative screening of phytochemical components
majority of the crude drugs that are of plant origin. It is necessary in leaves of P. zeylanica revealed the presence of alkaloids,
that standards have to be laid down to control and check the glycoside, reducing sugars, simple phenolics, tannins, Lignin,
identity of the plant an ascertain its quality before use. A detailed saponins and flavonoids presented in Table 2.
pharmacognostic evaluation therefore is highly essential prerequisite
(Ramana, 2007). Methanol, chloroform and alcoholic extracts of P.

Table 1. Antibacterial efficacy of different solvent extracts of Plumbago zeylanica leaves

Strain Zone of inhibition (mm)

Sr. +/- Concentration
Microorganism Petroleum Amphicillin
No. (mg/ml) Chloroform Alcohol
ether (40 μg/ml)
-ve 50 03 06 08
1. Escherichia coli 19
100 05 09 16
Pseudomonas -ve 50 06 07 12
2. 23
aeruginosa 100 12 06 17
+ve 50 05 08 08
3. Staphylococcus aureus 21
100 06 12 10
+ve 50 04 09 12
4. Bacillus subtilis 100 06 14 11 20

Table 2: Phytochemical components of different solvent leaves extracts of Plumbago zeylanica leaves

Phytochemical constituents Petroleum ether Chloroform Alcohol

Alkaloids - - +
Coumarins - + -
Glycoside - - +
Reducing sugars + - +
Simple phenolics - - +
Tannins - - +
Lignin - - +
Saponins - - +
Flavonoids - - +
Terpenoid - - -

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