International Journal of Science Research Volume 01, Issue 02, September 2012, pp.

83-86
Phytochemical Analysis, Antibacterial and Antioxidant Activity of Leaf Extract of Datura
Stramonium
S. Sreenivasa1*, K.Vinay2, N. R. Mohan1
1
2

Department of Studies and Research in Chemistry, Tumkur University, Tumkur 572103, India
Department of Chemistry, Channabasaveshwara Institute of Technology, Gubbi, Tumkur - 572216, India

*Author for Correspondence: drsreenivasa@yahoo.co.in

Abstract: The present study presents the phytochemical analysis, antibacterial activity and antioxidant activity of leaf extract of
Datura stramonium. The extraction of the active phytochemicals from Datura stramonium leaves was achieved by soxhlet assembly
successively with petroleum ether, benzene, solvent ether, chloroform, acetone, ethanol and methanol. The qualitative analysis of
the extracts for various phytochemicals revealed the presence of alkaloids, glycosides, terpenoids, steroids, flavonoids, tannins and
saponins in extracts of leaf. Further D. stramonium phytochemicals were investigated for their in vitro activity against bacterial
pathogens by disk diffusion method. D. stramonium leaf extracts exhibited a considerable antibacterial activity even at low
concentrations. Of various fractions obtained from leaf, methanol extracts showed maximum inhibitory effect. D. stramonium leaf
extracts also exhibited potent antioxidant property in in vitro studies.
Key Words: Datura stramonium, phytochemicals, antibacterial activity, antioxidant activity, disk diffusion

1.

INTRODUCTION

Even though pharmacological industries have produced a number of new antibiotics in the last three decades,
resistance to these drugs by microorganisms has increased. In general, bacteria have the genetic ability to transmit and
acquire resistance to drugs which are utilized as therapeutic agents. Such a fact is cause for concern, because of
increase in the number of patients in hospitals with suppressed immunity and also multi-resistant new bacterial strains.
Consequently, it results in new infections in hospitals leading to high mortality. Medicinal plants are used for treating
the ailments due to several microbial and non-microbial diseases due to their valuable effects in health care. The
affordability, reliability, availability and low toxicity of medicinal plants in therapeutic use has made them popular and
acceptable by all religions for implementation in medical health care all over the world. Plants are indeed the first
material used in alternative medicine type of remedy against many diseases. Several plants have therapeutic and
pharmaceutical effects, for antimicrobial, antioxidant, anti-infectious and anti-tumour activities [1, 2]. Herbal medicine
has been widely used as an integral part of primary health care in many countries [3]. In view of the above facts and
reported antibacterial activities of plant extracts, present study describes the phytochemical analysis, antibacterial
activity and antioxidant activity of leaf extract of Datura stramonium plant species.
2. MATERIAL AND METHODS
2.1 Plant materials:
2.1.1. Datura stramonium

Datura stramonium also called datura is an erect annual plant, on average 30 to 150 cm (1-5 feet) tall with erect,
forking and purple stems. The leaves are large 7 to 20 cm (3-8 in) long and have irregular teeth similar to those of
oakleaves. The flowers are one of the most distinctive characteristics of D. stramonium. They are trumpet-shaped,
white to purple and 5-12.5 cm (2-7 in) long. The flowers open and close at irregular intervals during the evening,
earning plant the nickname Moonflower. The fruit are walnut-sized, egg-shaped and covered in prickles. They split

International Journal of Science Research Volume 01, Issue 02, September 2012, pp. 83-86
into four chambers each with a few kidney-shaped seeds. All parts of the plant emit a foul odour when crushed or
bruised (Willard & Meritt, 1986)[5].
The plant is strongly narcotic, but has a peculiar action on the human frame which renders it very valuable as a
medicine. The whole plant is poisonous, but the seeds are the most active neither drying nor boiling destroys the
poisonous properties. The usual consequences of the poison when taken in sufficient quantity are dimness of sight,
dilation of the pupil, giddiness, delirium and sometimes amounting to mania, but its action varies greatly on different
persons (Yoshikawa K et al., 1989) [9].
D. stramonium is known to have several benefits to human health care as a wonderful therapeutic agent. The D.
stramonium were collected from the surrounding areas of Urdigere, situated in the regions of Devarayana durga hills
of Tumkur district which has a rich ecological resource.
2.2 Extraction of Phytochemicals from Datura Stramonium
Leaves of plant were collected during the rainy season in the August month of 2007 and were dried for about 15 days.
Alcohol was sprayed in order to prevent microorganism growth. The dried plant material in powder form was
subjected to phytochemical extraction with Soxhlet assembly successively with petroleum ether, benzene, solvent
ether, chloroform, acetone, ethanol and methanol as solvents depending upon its solubility. Each time before
extracting with the next solvent, the powdered material is dried in hot-air oven below 50oC. Here the solvent selection
was made by trial and error method.
A known quantity of dried and powdered plant material was packed in a Whatman paper. Thimble was then placed in a
Soxhlet extractor assembly fitted with a round bottom flask containing th volume of a suitable solvent. The
temperature in the thermostat was adjusted nearly to the boiling point of the solvent. Finally the extract was
concentrated by distilling off the solvent using rotavapour on water-bath. The extract obtained was air-dried and
weighed to calculate yield.
2.3 Determination of the composition of Phytochemicals in Datura stramonium extracts
In order to know the phytochemical composition, the leaf extract powder was defatted by methanol and subjected to
successive continuous extraction in soxhlet apparatus with different solvents with increase in polarity, viz., acetone,
chloroform and finally with water. The extracts were filtered in each step, concentrated and the solvent was removed
by vacuum distillation. The extracts were dried in the vacuum desiccators and the residues were weighed. The
presence or absence of phytoconstituents such as alkaloids, glycosides, terpenoids, steroids, flavonoids, tannins and
saponins leaf extracts were assessed by standard phytochemical assay methods.
2.4 Determination of the Antibacterial activity of phytochemicals from Datura stramonium
2.4.1 Test organisms
The Microorganisms used in this study were clinical isolates of pathogenic bacteria such as Staphylococcus aurers
ATCC25923, Methicillin-resistant S. aureus, Enterococcus sp., Escherichia colii ATCC25922, Enteroinvasive
Escherichia coli and Pseudomonas aeruginosa.
2.4.2 Bacterial Culture media and preparation of inoculums
The quantity of Muller Hinton Agar (MHA) medium as per manufacturers manual was dissolved in distilled water and
autoclaved (20min at 15 psi). The MHA plates were prepared by pouring 15 ml of autoclaved media into sterile
petriplates and allowed to solidify. The agar surface of the plates was inoculated with 0.1% of inoculum suspension by
swabbing uniformly and allowed to dry.
2.4.3 Determination of Anti Bacterial Action of Datura stramonium extracts
The disk diffusion method was used to screen the antibacterial activity of the plant extracts. Sterile 6-mm diameter
paper disks (Schleicher and Schuell, germany) were impregnated with 10mg (10l) of each fraction dissolved in 95%

International Journal of Science Research Volume 01, Issue 02, September 2012, pp. 83-86
ethanol. Air dried disks were placed on the bacteria inoculated MHA surface. Commercially available antibiotic disks
of Vanomycin (30 g), Tetracycline (30 g) and Gentamicin(10 g) were used as standard antibiotics and disks
impregnated with 10 l of 95% ethanol as negative controls. Plates were incubated at 370 C for 18 hours and the
inhibition zone was measured in mm. The tests were performed in duplicate.
2.7 Assay of Anti oxidant ability of Datura stramonium extracts
The antioxidant activity of the plant extracts was assessed on the basis of the free radical scavenging effect on the
stable 1, 1-diphenyl-2-picrylhydrazyl (DPPH) by modified method (Reference?). The diluted working solutions of the
test extracts were prepared in methanol. Ascorbic acid was used as standard in 1-100 g/ml solution. 0.002% of DPPH
was prepared in methanol and 1 ml of this solution was mixed with 1 ml of sample solution and standard solution
separately. These solution mixtures were kept in dark for 30 min and optical density was measured at 517 nm using
Cecil-Elect Spectrophotometer. Methanol (1 ml) with DPPH solution (0.002%, 1 ml) was used as blank. The optical
density was recorded and % inhibition was calculated using the formula given below: Percent (%) inhibition of DPPH
activity = A B 100
A
Where A = optical density of the blank; B = optical density of the sample
3.

RESULTS AND DISCUSSION

The preliminary phytochemical investigation of the plant extracts of Datura stramonium revealed the presence of
secondary metabolites such as alkaloids, flavonoids, terpenes, tannins, saponins, iridoids, gycosides and sterols (Table
1).
Table 1: Phytochemicals detected from the Datura stramonium plant leaf extracts
Tests
Ethanol Methanol Water Chloroform Benzene
(Phytochemical)
Hagers Test
++
(Alkaloids)
Trim Hill Test
(Iridoids)
Ferric Chloride Test
+
(Flavonoids)
Salkowanski Test
(Terpenoids)
Foam Test
+
(Saponins)
Libermann-Burchard Test
+
(Sterols)
Ferric Chloride Test
+
(Tannins)
Bromine-water Test
(Glycosides)
Lead acetate Test
(Flavonols)
(-) Absence; (+) Low concentration; (++) High concentration; (+++) Very high concentration

a.

Pet. ether
+++

++

Antibacterial property of Datura stramonium leaf extracts

The antibacterial activity of the datura stramonium extract was studied by using the following microorganisms:
Bacteria Escherichia coli ATCC25922 and Staphylococcus aureus ATCC25923. Commercially available antibiotic
disks of Gentamicin and Vanomycin of same concentrations were used as standards. The disk diffusion method was
used to screen the antimicrobial activity of the Datura stramonium plant extracts. After incubation of the organisms
along with plant extracts the inhibition zone was measured (Table 2).

International Journal of Science Research Volume 01, Issue 02, September 2012, pp. 83-86

Table 2: The antibacterial activities of the Datura stramonium leaf extracts

Datura stramonium fraction
Disk A
Test
MIC/
Microorganisms IZ
MBC
(mm)
(mg/ml)
Escherichia
9.8
10/>10
coli ATCC25922
Staphylococcus
aureus
6.8
10/>10
ATCC25923

Control drug

Disk C

Disk B
Disk D
MIC/
MIC/
IZ
IZ
MBC
MBC
(mm)
(mm)
(g/ml)
(g/ml)

IZ
(mm)

MIC/
Drug
MBC
(mg/ml)

0.5

1/>1

Gentamicin 20.1

1/1

19.4

1/1

0.2

1/>1

Vanomycin 16.5

1/1

16.3

1/1

Note: IZ = Inhibition Zone, MIC = Minimum Inhibitory Concentration, MBC = Minimum Bactericidal
Concentration
The results revealed marked that the fraction Datura stramonium disk C exhibited low inhibitory activity against the
same fraction Datura stramonium disk A with the Bacteria Escherichia coli ATCC25922 measuring the inhibition
zones of 0.5 9.8 mm and the MIC values of 1 10 mg/ml. However, the fraction Datura stramonium disk C
exhibited low inhibitory activity against the same fraction Datura stramonium disk A with the Bacteria
Staphylococcus aureus ATCC25923 measuring the inhibition zones of 0.2 6.8 mm and the MIC values of 1 10
mg/ml. The extracts of Datura stramonium fraction have been found to possess an antibacterial activity, although a
very lack of antibacterial activity has also been reported at very lower concentrations.
Table 3: Percentage Inhibition of DPPH activity
Sl No

Solution to be
examined

Absorbance/
% inhibition of DPPH
Optical
activity
density

1.

Blank solution

0.492

2.

Datura stramonium
Plant extract

0.376

23.5

From the above observations, the absorbance value for standard compound ascorbic acid was found to be 0.684.
Datura stramonium plant extracts shows lower optical density compared to blank solution. Hence, we can predict that
the constituents present in the extracted crude plant samples of Datura stramonium exhibits anti oxidant property.
Among the plant extract and standard tested for the in vitro antioxidant activity using the DPPH method, the crude
methanolic extracts of Datura stramonium showed antioxidant activity with IC50 values of 6.7 0.1 (Table 4).
Table 4: In vitro anti oxidant activity of methanolic extracts of species
Sl
No.

Test Compound

IC50 (g/ml)
(Mean + SD)

Weight of extract
(g % ,w/w)

Datura Stramonium
Extracts

6.7 + 0.1

22.4

Ascorbic acid

8.9 + 0.1

Plant extracts have great potential as antimicrobial compounds against microorganisms and other medicinal
benefits. Thus they can be used in the treatment of infectious diseases caused by resistant microbes and other ailments.

International Journal of Science Research Volume 01, Issue 02, September 2012, pp. 83-86
The synergistic effect from the association of antibiotic with plant extracts against resistant bacteria leads to new
choices for the treatment of infectious diseases. This effect enables the use of the respective antibiotic when it is no
longer effective by itself during therapeutic treatment.
The antibacterial activities of the studied plant extracts were comparable to the reference antibiotic used.
Therefore, this study offers a scientific basis for the use of the plant extracts for the treatment of infections that could
be caused by the strains of the test bacterial organisms. The phytochemical tests indicated the presence of alkaloids,
glycosides and flavonoids in the crude methanolic extract. Majority of the phytochemical compound identified in the
ethanol extract have been reported to be highly of therapeutic importance. The IC50 value for standard compounds
ascorbic acid was 8.9 0.1 g/ml. The free radical scavenging activity of methanolic extracts was also noticed in the
present investigation. However, the chemical constituents like alkaloids, glycosides, tannins and flavonoids present in
the extract may be responsible for such activity. Several of such compounds are known to possess potent antioxidant
activity. Some of these constituents have already been isolated from this plant. Hence, the observed antioxidant
activity may be due to the presence of any of these constituents. These properties may be due to its antioxidant
activity. The compounds such as flavonoids, which contain hydroxyls, are responsible for the radical scavenging effect
in plants. According to our study, the highest radical scavenging activity was showed by Datura stramonium with IC50
value 6.7+ 0.1 g/ml. The promising result obtained has subjected this plant extract to further analysis for its toxicity
and side effects for possible therapeutic importance.
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