Int. J. Pharm. Sci. Rev. Res., 48(1), January - February 2018; Article No.

14, Pages: 58-61 ISSN 0976 – 044X

Research Article

Phytochemical Screening of the Essential Oil of Syzygium aromaticum and Antibacterial

Activity against Nosocomial Infections in Neonatal Intensive Care
Amal Taroq*1, Fatima El Kamari1, Bouchra Oumokhtar2, Imane Aouam1, Yassine El Atki1, Badiaa Lyoussi1, Abdelfattah Abdellaoui1
1
Laboratory of Physiology Pharmacology and Environmental Health, Department of Biology, Faculty of Sciences DharMahraz, University
Sidi Mohamed Ben Abdellah, B.P1796, Atlas, Fez, Morocco.
2
Laboratory of Microbiology and Molecular Biology, Faculty of Medicine and Pharmacy, University Sidi Mohamed Ben Abdellah, 200
Route de Sidi Harazem, Fez, Morocco.
*Corresponding author’s E-mail: taroq.amal@gmail.com

Received: 06-12-2017; Revised: 28-12-2017; Accepted: 14-01-2018.

ABSTRACT
Nosocomial infection in pediatric intensive care, which is dominated by bacterial strains, represents a real public health problem.
Indeed, most pathogens showed a low resistance to antibiotics which induced diseases. This is the reason why new alternatives
were established in order to overcome the incidence of nosocomial infections. Hence, secondary metabolites of plants are known
as an efficient source of new natural products especially to tackle these kinds of infections. Throughout this study, a phytochemical
screening of clove (Syzygium aromaticum) essential oil was carried out by using chromatography-flame ionization detector and gas
chromatography-mass spectrometer analysis. Thus, the antibacterial activity of the extracted oil was evaluated against four bacteria
(Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus aureus and Proteus mirabilis) that cause nosocomial infections in
neonatal intensive care by using disc diffusion method. Indeed, the analysis showed that two major compounds known as Eugenol
(87.03%) and Eugenol Acetate (11.25%) are present in Syzygium aromaticum essential oil. Moreover, a strong antibacterial activity
of the extracted oil was identified and could be exploited as a natural antibacterial drugs bacterial strains resistance.
Keywords: Syzygiyum aromaticum; Essential oils; Antibacterial activity; Neonates, nosocomial infections.

INTRODUCTION rates in intensive care units especially in neonatal

service.10 Nosocomial infection in pediatric intensive care,

S yzygium aromaticum (synonym:

Cariophylata) commonly known as clove, from the
Mirtaceae family native to islands in east
Eugenia

Indonesia.1 Clove oil is used as a first aid remedy for

which is dominated by bacterial strains, represents a real
public health problem.
A relevant low resistance of numerous pathogens to
mucosal inflammations of the mouth and toothache. The antibiotics was identified. Therefore, new alternatives
ethanol extract of Syzygium aromaticum flower bud were developed to decrease the prevalence of these
demonstrated anti-nociceptive and anti-inflammatory infections. Clove contains a variety of bioactive
effects in mice and Wistar rats2, and the hydro-alcoholic compounds such as triterpenoids and sesquiterpenes. 11
3
extract showed an efficient anti-stress activity. It was Extracts and essential oils of Syzygium aromaticum
12, 13, 14
reported that Clove extract may act as insulin substance demonstrated strong antioxidant properties.
inside hepatocytes and hepatoma cells by inducing According to recent studies the essential oils may be an
15
glucose 6-phosphate(G6Pase) gene expression and by alternative remedy against pathogenic microorganisms.
reducing phosphoenolpyruvatecarboxykinase (PEPCK).4 In addition, among these new approaches, these natural
Besides to this, it was reported that clove oil could inhibit extracted elements could be useful in terms of treating
hepatic cell proliferation and decrease oxidative stress and preventing antibacterial infections.
which is behind the mechanism of protection against liver
The previous work related to natural oils inhibitors of
cirrhosis.5 A recent review revealed that lower dose of
nosocomial infections are promoting new investigations
Syzygium aromaticum flower bud extract is androgenic in
in this area of research.16, 17 however, research based
nature and may have a viable future as an indigenous
clove essential oil still limited in terms of preventing
sexual rejuvenator.6 Furthermore, Cloves are natural
antibacterial activity against bacteria especially in
products with an excellent cytotoxicity toward MCF-7
neonatal intensive care.18, 10
cells; thus, they are promising sources for the
development of anticancer agents 7 ,and its essential oil The major aim of the current study was to emphasize the
exerts the eradication of dermatophagoidespteronyssius antibacterial effects of Syzygium aromaticum essential oil
8
house dust mites. Moreover, solvent extracts of Syzygium against some causing nosocomial infections in neonate
aromaticum showed a strong antimicrobial activity.9, 15 reanimation service, at the University Hospital Centre in
Fez-Morocco, and to determine the chemical constituents
Nosocomial infections acquired in hospitals are known as
of bud clove essential oil used in this study.
healthcare associated with infections. An average of 5 à
10% of patients has nosocomial infection with the highest

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Int. J. Pharm. Sci. Rev. Res., 48(1), January - February 2018; Article No. 14, Pages: 58-61 ISSN 0976 – 044X

MATERIALS AND METHODS Antimicrobial activity assessment

Isolation of clove essential oil The tested microorganisms included Pseudomonas
aeruginosa (P. aeruginosa), Klebsiella pneumonia (K.
The buds of Syzygium aromaticum were purchased from a
pneumonia), Staphylococcus aureus (S. aureus), and
local supermarket in Fez province (Morocco).
Proteus mirabilis (P. mirabilis). These bacterial strains
Identification was confirmed by Professor Amina Bari,
were isolated in the hospital’s environment from patients
botanist (Department of Biological Sciences, Faculty of
in the neonate intensive care unit (CHU Fez Morocco).
Science, Sidi Mohammed Ben Abdellah University, and
Fez, Morocco). A total of 100 g clove were subjected to The agar-disc diffusion was used for bacterial
hydro-distillation for 3 h with 600 ml distilled water using susceptibility screening test as previous studies. 20, 16 Each
a Clevenger-type modified apparatus: the hydrosol was microorganism stock was suspended in Mueller-Hinton
collected in a separator funnel (1 liter) so that the heavy (MH) broth and then incubated at 37°C for 18–24 h. The
oil was decanted to the bottom of the flask , while the overnight cultures were diluted and adjusted in order to
hydrosol water was recycled into the flask containing the obtain the density test 100 CFU/ml (0.5 McFarland
boiling plant material. The essential oil was collected and turbidity standards). Microorganisms were flood-
dried over anhydrous sodium sulphate and stored at 4°C inoculated on to the surface of MH agar. Sterile 6 mm
prior to analysis. The distillation yield was calculated diameter filter discs (Whatman paper No. 3) were
based on the dried weight of the sample. impregnated with 10µg/disc of the essential oil and were
put onto the surface of the inoculated agar (MH). The
Gas chromatography analysis - FID
plates were incubated for 18 h at 37°C. Antimicrobial
The isolated oil was diluted with hexane (dilution ratio activity was evaluated by measuring the inhibition zones
10:100), and 1 µl was sampled to perform gas on the tested bacterial strains. The antibiogram discs of
chromatographic analysis. Trace gas chromatograph (GC) Imipenem (IMP), Cefotaxime (CTX), Kanamycine (K),
(ULTRA S/N 20062969, Thermo Fischer) and gas Pristinamycine (PT) were used asstandard drugs for
chromatograph equipped with HP 5MS non polar fused comparison. The tests were carried out in triplicates.
silica capillary column (60 m × 0.32 mm, film thickness Results were interpreted in terms of a diameter of
0.25 mm) were used. Operating conditions: oven inhibition zone: resistant (D < 6 mm), intermediaries (6
temperature program from 50 °C (2 min) to 280 °C at 5 mm < D < 13 mm) and sensitives (D > 13 mm). An average
°C/min and the final temperature kept for 10 min; 2 “split zone of inhibition was calculated for three replicates.
mode” ratio 1:20; carrier gas Nitrogen (N), flow rate 1 ml/ ANOVA test was used to determine whether there are
min; temperature of injector and detector (flame any significant statistical differences between all
ionization detector) were fixed at 250 °C and 280 °C, inhibition tests.
respectively.
RESULTS AND DISCUSSION
Gas chromatography-mass spectrometry (GC-MS)
Table 1: Chemical composition of the essential oil from
The analysis of the volatile constituents was run on a buds of Syzygium aromaticum.
Thermo Fischer capillary gas chromatograph directly
coupled to the mass spectrometer system (model GC Kovats Area Chemical
Compounds
index (%) formula
ULTRA S/N 20062969; Polaris QS/N 210729), using an HP-
5MS non polar fused silica capillary column (60 m × 0.32 Eugenol 1353 87.03 C10H12O2
mm, 0.25 mm film thickness). The operating condition of
β-Caryophyllene 1428 <1 C15H24
GC were: oven temperature was maintained as initial
temperature 40 °C for 2 min, programmed rate 2 °C/min Acetate d’eugenol 1538 11.25 C12H14O3
up to a final temperature of 260 °C with isotherm for 10
min; injector temperature 250 °C. The carrier gas was Caryophylleneoxide 1689 <1 C15H24O
helium, flow rate 1 ml/ min. Samples were run in hexane The essential oils obtained from the buds of Syzygium
with a dilution ratio of 10:100. 1µl of the diluted oil aromaticum appeared with white colour and isolated
specimen was injected using the split less injection with a yield of 12%.
technique with ionization energy of 70 eV in the
electronic ionization mode. The ion source temperature The chemical composition of buds clove essential oils was
was 200 °C; the scan mass range was 40–650 m/z and the determined by GC-MS and FID analysis. Four compounds
interface line temperature was 300°C.Components accounting for more than 99.28% of the total essential oil
identification was made by determination of their were identified (table1). Several studies on the chemical
retention indices (KI) relative to those of a homologous composition of the essential oil of Clove were reported
series of n-alkanes (C8–C20) (Fluka, Buchs/sg, Switzerland) and showed the similar compounds: Eugenol(87.03%) and
and by matching their recorded mass spectra with those Eugenol Acetate(11.25%).21, 22Approximately, 89% of the
stored in the spectrometer database (NIST MS Library v. clove essential oil represents eugenol substance and 5%
2.0) and the bibliography.19 to 15% belongs eugenol acetate and β-caryophyllene.23

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Int. J. Pharm. Sci. Rev. Res., 48(1), January - February 2018; Article No. 14, Pages: 58-61 ISSN 0976 – 044X

Overall that the main components of buds essential oil of synergistic effect between the diverse components of the
clove varies by grinding, maturity, geographical oil is possible.
conditions, climate, harvest season, period of distillation
CONCLUSION
and extraction method used. Eugenol (4-allyl-2-
methoxyphenol), the active substance, makes up 90–95% The results reported herein indicated that essential oil of
24,25
of the clove oil . These results are in good agreement Syzygium aromaticum is a potential source of bioactive
with several reported studies.21, 22, 23, 24 Moreover, it was compounds against some bacteria inducing nosocomial
reported that some volatile minor compounds could be infections in neonate intensive care units. Therefore,
present at lower concentrations in clove essential oil, clove essential oil could be used for further therapeutic
such as: β-pinene, limonene, farnesol, benzaldehyde,2- use against P.Aeruginosa, which had acquired a resistance
heptanone and ethyl hexanoate.1 to several conventional antibiotics. On one hand, clove
essential oil may be used as natural antibacterial drugs for
Clove represents one of the major plant sources of
nosocomial infections treatment. On the other hand, it
phenolic compounds as flavonoids, hidroxibenzoic acids,
could be used for sterilizing air, surfaces and equipment
hidroxicinamic acids and hidroxiphenylpropens.
in neonate intensive care units. Our findings could be
Aromatic plants and their EOs have traditionally been exploited for further investigations in terms of purifying
used since antiquity for their biological properties certain compounds to produce new and efficient
(fungicidal, virucidal, bactericidal, insecticidal and antibacterial drugs.
26, 27
antiparasitical). The antibacterial activity of clove was
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Source of Support: Nil, Conflict of Interest: None.

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