Phytochemical and Bioactivity of Commercially Available Eucalyptus Oil against Human Pathogens

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South Asian Journal of Life Sciences 2 (1): 8 – 11
http://dx.doi.org/10.14737/journal.sajls/2014/2.1.8.11
Neelam et al (2014). Commercially available
Eucalyptus
Oil
8
ISSN: 2311–0589
Research Article
Asia Neelam1, Omm–e–Hany1*, Sikander Khan Sherwani2, Suraiya Jabeen1, Hasnain Nangyal3
1Institute of Environmental Studies, University of Karachi, Karachi, Pakist an, 75300; 2Department of Microbiology, Federal u niversity of Arts, sc ience and
Technology, Karachi, Pakistan, 75300; 3Departm ent of Botany, Hazara University, Mansehra, KPK, Pakistan
*Corresponding author: hany786@yahoo.com
ARTICLE HISTORY
ABSTRACT
Received:
Revised:
Accepted:
2014–04–16
2014–05–30
2014–05–31
Many species of Eucalyptus are used as a folk medicine. Traditionally, they are used in the
treatment of many respiratory tract infections like bronchitis, sinusitis and pharyngitis as it
is rich with the antimicrobial inhibitory agent. The present study aimed to determine the
phytochemical and bioactivity of commercially available Eucalyptus oil against human
pathogens which cause many infections in human population. The “AXE Brand Eucalyptus
oil” which are used for the relief of colic and stomachache show significant bioactivity against
Staphylococcus aureus, Escherichia coli, Salmonella paratyphi A, Salmonella paratyphi B, Shigella
dysenteriae, Bacillus licheniformis and Pseudomonas aeruginos, which is seen as a significant pathogen.
However, Highest bioactivity observed against Bacillus licheniformis whereas no activity or
zeroes zone of inhibition shows against Bacillus subtilis and Salmonella typhi. Phytochemical tests
to know about the secondary metabolites show the presence of phytosterols, phenols,
tannins, flavanoids, protein and amino acid. On the bases of this result, the oil of eucalyptus
can be effectively utilized in the treatment of infectious disease as well as in Sanitizers and
disinfectant as an innate and environmental friendly product.
All copyrights reserved to Nexus® academic publishers
Key Words: pathogens,
antibacterial, eucalyptus oil,
phytochemical
ARTICLE CITATION: Neelam A, Omm–e–Hany, Sherwani SK, Jabeen S, Nangyal H (2014). Phytochemical and bioactivity of
commercially available eucalyptus oil against human pathogens. S. Asian J. Life Sci. 2 (1): 8 – 11.
INTRODUCTION
In today’s world large numbers of medicinal plants and their
compound have shown beneficial therapeutic activity (Aqil
et al., 2006). These medicinal plants are used for the
mitigation of various diseases and characterize as a rich
source of antimicrobial agent. In different countries plants
are used medicinally for the extraction of potential active
and powerful drugs (Srivestava et al., 1996).
The Eucalyptus belongs to the Myrtaceae family and
includes about 900–700 species over the entire world
(Broker and klerining 2006, 2004). It is the native genus of
Australia, but few numbers of these plants are also the
native of New Guinea, Indonesia and the Phillipines. The
importance of eucalyptus is due to their wood, proteins,
tannins, gum, and dyes, but the most important product of
eucalyptus is their oil. Approximately 300 species of this
genus have volatile oils in their leaves. Less than 20, within
these species, have a high content of 1,8– (more than 70%
and up to 90%), which is commercially applied for the
synthesis of essential oils in cosmetic and pharmaceutical
industries (Pino et al., 2002). There are many studies that
support the antimicrobial activity of eucalyptus oil. One of
the best study showed that the Eucalyptus oil is effective
against 22 kinds of bacteria and 11 kinds of fungus (Pattnaik
et al., 1995). According to one of the researcher three
different species of Eucalyptus – Eucalyptus globulus,
Eucalyptus tereticornis and Eucalyptus robusta showed excellent
activity against Escherichia coli, S almonella typhi, Streptococcus
Lactobacillus, Pseudomonas aeruginosa and Staphylococcus aureus
(Bachheti et al., 2011). For instance, E. globules contain 1,8–
channel which showed toxicity against Pediculus humanus
capitis (human head lice) (Yang et al., 2004), it has also
strong antibacterial activity against Es, Streptococcus pyogenes,
Candida albicans Acinetobacter baumannii, and Klebsiella
pneumoniae (Biljana et al., 2011).
Eucalyptus contains many chemical complexes
that play significant roles in the plant. These chemicals
consist of the many chemical compounds that are providing
safety against UV radiation, vertebrate, insects and cold
stress. The main phytochemical present in Eucalyptus oil
are steroids, alkaloids, tannins, saponins, phenolics,
flavonoids, terpenoids glycosides (Saxena et al., 2010,
Pathmanathan et al., 2010, Jain et al., 2010). Among of these
chemicals the better–known chemicals constituted are
the terpenoids, which giving eucalyptus foliage, its natural
smell and synthesis most of the essential oil (Lassak and
MacCarthy 1992). In many countries Eucalyptus species are
traditionally used as folk medicine. The Aborigines (native
Australians) have used eucalyptus leaves to cure wounds
and fungal infections (Takahashi 2004). While in Tunisian
essential oil of eucalyptus species has been used for the
treatment of respiratory tract disorders such as sinusitis,
pharyngitis, and bronchitis (Boukef 1986).
Today these essential oils have great value in the
market (Ajaj 1995) since they are employ for the treatment
of chest pain, or cough, influenza, and skin rashes while
Phytochemical and Bioactivity of Commercially Available
Eucalyptus
Oil
against Human Pathogens

South Asian Journal of Life Sciences 2 (1): 8 – 11
http://nexusacademicpublishers.com/journal/25
Neelam et al (2014). Commercially available
Eucalyptus
Oil
9
ISSN: 2311–0589
their vapour is inhaled to cure the inflammation (Musyimi
and Ogur 2008).There application are also in the stringent,
anesthetic, anodyne, deodorant, diaphoretic, expectorant,
febrifuge , disinfectant, fumigant, hemostat,
rubefacient,cancer, diabetes, diarrhea, diphtheria
encephalitis, enteritis, laryngitis,erysipelas,leprosy, malaria,
mastitis, miasma, rhinitis, laryngalgia, sores, sore throat, and
trachalgia.((Elliot and Jones 1980).To the best of our
information there are no such studies on phytochemical
and bioactivity of market available eucalyptus oil, therefore
the main objective of this study was conducted it and
evaluate its activity.
MATERIALS AND METHOD
Product and Culture
The synthesis oil of eucalyptus of “AXE brand universal oil
“was purchased from the local market of Saudia Arabia. The
microbial culture used for the antibacterial test was Bacillus
subtilis,Staphylococcus aureus,Escherichia coli,Salmonella
typhi,Salmonella paratyphi A,Salmonella paratyphi B,Shige lla
dysenteriae,
Bacillus licheniformis and Pseudomonas aeruginosa
which was obtained from the Institute of Environmental
studies and Department of Microbiology, University of
Karachi.
Antibacterial Assay
The antibacterial sensitivity of eucalyptus oil was
performed by using two methods the disc diffusion method
(Anonymous 1993) and by agar well diffusion method
(Irobiet et al., 1994). For control Cefixime antibiotic disc
was applied. The plate was incubated at 37 ◦C for 24 hours.
The antibacterial activity was measured through the
diameter of the zone of inhibition, which is expressed in
millimeter (mm). For more accuracy the test was conducted
in triplicate.
Phytochemical Screening
To investigate the different compound, the oil of eucalyptus
was tested by follow the following procedures.
Detection of Alkaloids
Wagner`s Test: Few drops of oil were dissolved in 1 ml of
hydrochloric acid than filter it. In Filtrated with a few drops
of Wagner`s reagent added (Iodine in potassium Iodide).
Brown/reddish color precipitate indicates the presence of
alkaloids (Tiwari et al., 2011)
Detection of Saponins
Foam Test: 2ml of oil was treated with 6ml of water into a
test tube. The mixture was trembling vigorously and
observed in the formation of persistent foam that verifies the
presence of saponins (Ugochukwu et al., 2011).
Detection of Phytosterols
Salkowski’s Test: 1 ml of oil was treated with chloroform
and later filtered. The filtrates were treated with a few
drops of Concentrated Sulfuric acid, shaken for some times
and allowed to stand. The presences of golden
yellow color indicate the oil contain triterpenes (Tiwari et
al., 2011).
Detection of Tannins
Gelatin Test: 1% gelatin solution containing sodium
chloride was added in a few drops of oil. White precipitate
in tube confirms the presence of tannins (Tiwari et al., 2011).
Detection of Flavonoids
Ferric chloride Test: Ferric chloride solution treated with
drops of oil in the test tube that result in the formation of
blackish red color point out the presence of flavonoids
(Bhandary et al., 2012).
Detection of Proteins and Amino Acids
Xanthoproteic Test: 5–6 drops were treated with a few
drops of conc. Nitric acid. Appearance of yellowish
color verifies the presence of proteins and amino acid
(Tiwari et al., 2011).
Detection of Quinones
Few drops of oil were treated with concentrated
Hydrochloric acid and observed for the appearance of
yellow precipitation (or coloration) (Ugochukwu et al.,
2011).
Detection of Terpenoids
1 ml of chloroform was treated with 2ml of oil, then added a
few drops of Concentrated Sulfuric acid. A reddish brown
precipitation generates immediately indicated the presence
of terpenoids (Ugochukwu et al., 2011).
Detection of Phenol
Ferric Chloride Test: 0.5 ml of ferric chloride solution was
added in a test tube containing oil. The appearance of bluish
black color indicates the presence of phenols (Tiwari et al.,
2011).
Data Analysis
All the date and graphs which is given, analysis on
Microsoft Office Excel–2007 and all the zones of inhibitions
were expressed as mean ± standard deviation (SD).
Table 1: Zone of inhibition of eucalyptus oil from Disc
Diffusion method
Bacteria Spp.
Gram reaction
Zone of inhibition
in mm (0.025 mg)
Bacillus subtilis
Gram positive
NE
SalmonellaTyphi
Gram negative
NE
Staphylococcus
aureus
Gram positive
7.2±0.4
Salmonella
Paratyphi A
Gram positive
5.8±0.4
Salmonella
Paratyphi B
Gram positive
3.9±0.1
Shigella
dysenteriaes
Gram negative
4±0.2
Bacillus
lichniformis
Gram negative
10.5±0.2
Pseudomonas
aeruginosa
Gram negative
1.1±0.2
Escherichia coli
Gram negative
7.9±0.2
Proteus spp
Gram negative
3.9±0.2
*For control Cefixime antibiotic used; *NE = No zone of inhibition

South Asian Journal of Life Sciences 2 (1): 8 – 11
http://nexusacademicpublishers.com/journal/25
Neelam et al (2014). Commercially available
Eucalyptus
Oil
10
ISSN: 2311–0589
RESULT AND DISCUSION:
Plants are important sources for the investigation of many
drugs against different diseases (Jean et al., 2005). Essential
oils have many volatile constitute natures, which have
strong antibacterial activities (Cowan 1999). Eucalyptus
plant also has a volatile compound in their oil, therefor its
shows significant antimicrobial activity. The results in
Table 1 and 2 revealed that the commercially available
eucalyptus oil showed antibacterial activity with different
magnitudes, depending on the size of inoculums and the
concentration of oil. Diameter of inhibition zone oil varied
from 1 to 7mm.In disc diffusion method Bacillus lichniformis
shows highest susceptibility with eucalyptus oil and in agar
well diffusion method highest susceptibility shows by
Salmonella paratyphi B. whereas there is no effect of
Eucalyptus oil is measure against Bacillus.subtilis and
Salmonella typhi by both the techniques i.e. agar well diffusion
and disc diffusion method.Eucluptus oil also shows
significant restrain activity against Escherichia coli and
Staphylococcus aureus that are consider as opportunistic
pathogens that cause life–threatening and severe infections
in immunocompromised patients (Lestari, 2004).
Table 2: Zone of inhibition of Eucluptus oil from agar well
diffusion method
Bacteria Spp.
Gram reaction
Zone of inhibition
in mm (0.1 ml)
Bacillus. subtilis
Gram positive
NE
SalmonellaTyphi
Gram negative
NE
Staphylococcus
aureus
Gram positive
4.1±0.6
Salmonella
Paratyphi A
Gram positive
2.9±0.2
Salmonella
Paratyphi B
Gram positive
6.9±0.2
Shigella
dysenteriaes
Gram negative
4.6±0.2
Bacillus
lichniformis
Gram negative
3.5±0.5
Pseudomonas
aeruginosa
Gram negative
1.9±0.2
Escherichia coli
Gram negative
3±0.5
Proteus spp
Gram negative
3.3±0.3
*For control Cefixime antibiotic used ; *NE = No zone of inhibition
This result is more conformed with the previous research on
eucalyptus oil which result showed the significant
inhibition effect on Escherichia coli and Staphylococcus aureus
(Bachir and Benali 2012) however, Farah et al., 10, Babayi et al.,.
11, Gamal and Sabrin12 and Nair et al.,4 who also reported the
inhibitory activity of eucalyptus essential oil against S. aureus and E.
coli. In both the test inhibition rate of gram negative
bacteria and gram positive bacteria are almost similar
however, according to the reports of some authors gram–
negative bacteria are slightly more sensitive to essential oils
of eucalyptus species when compared to gram–positive
(Bachir and Benali 2012). Although the amount of inoculum
and the concentration of oil may also affect the growth of
the organisms. This antibacterial activity of Eucalyptus
extracts has been due to the presence of many chemical
components like citronella, 1,8–cineole, Citronellol, p–
cymene, alloocimene, eucamalol, limonene, citronellyl
acetate, linalool, β – pinene, γ –terpinene, α – terpinol, and
aroma dendrene (Nezhad et al., 2009).
Table 3: Phytochemical investigation of Eucalyptus oil
Phytochemical
Eucalyptus oil
Alkaloids
–
Phytosterols
+
Phenols
+
Tannins
+
Flavanoids
+
Proteins and amino acid
+
Saponins
–
Quinones
–
Terpenoids
+
Key: Present = +, Absent =–
Phytochemical Analysis
Preliminary phytochemical testing for the presence of
various compounds in eucalyptus oil reveals the positive
presence of Phytosterols, Phenols, Tannins, Flavanoids,
Terpenoids, Proteins and amino acid and negative presence
of Saponins, Quinones and Alkaloids. Phytochemicals are
aromatic compounds synthesized by plants which have
been used as protective agents to microbial infection, insect
and herbivore killer. The macromolecules produced by
plants have been divided into primary and secondary
metabolites (Rauha et al., 2005). The secondary metabolites
act as, repellent, antibacterial agent or even toxic agents to
pests and herbivores (Dewick, 1997). Our finding is
somehow verifying with the result of Pathmanathan et al.,
the presence of tannins in eucalyptus specie was also
reported in one of the previous study (Babayi et al.,
2004).The eucalyptus oil, which is under tested used against
many diseases. The plants which have terpenoids and
tannins are used in anti inflammatory activities (Okwu and
C. Jasiah 2006). Tannins are also responsible for
antimicrobial characteristics in different plants (Chung
1998). Phenol is also present in eucalyptus oil. The presence
of phenolic compounds in plant is responsible for the
protection of carcinogen, inflammation, atherosclerosis and
cardiovascular. Flavanoides also positive in eucalyptus oil,
this is synthesized by plants in response to microbial
infection (Nyarko and add 1990). Hence, the inhibitory
mechanism of these oils on the microorganisms may
therefore be due to the presence of the above phytochemical
components.
The commercially available eucalyptus oil showed
varying degrees of antibacterial activity against a reference
strain of bacteria. The presence of important phytochemical
in this oil also reveals its medical and herbal importance.
This information provides a scientific ground for the
application of this oil in the drug formulation for the
prevention and treatment of bacterial infections caused by
various pathogenic bacteria.
CONCLUSION
Eucalyptus oil has been shown a wide spectrum
antimicrobial activity and has important phytochemical
compound. After few modifications it is effectively used

South Asian Journal of Life Sciences 2 (1): 8 – 11
http://nexusacademicpublishers.com/journal/25
Neelam et al (2014). Commercially available
Eucalyptus
Oil
11
ISSN: 2311–0589
against human pathogens and this oil will use in
manufacturing of different disinfectant and Sanitizers.
ACKNOWLEDGEMENTS
All authors are thankful to the IES for providing the fully
indentified bacterial cultures and relevant necessary
facilities for conducting this research.
CONFLICT OF INTEREST
There is no conflict of interest in authors.
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