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Antimicrobial activity and
phytochemical analysis of crude
extracts and essential oils from
medicinal plants
N.C.C. Silva
a
, L. Barbosa
b
, L.N. Seito
c
& A. Fernandes Junior
a
a
Department of Microbiology and Immunology, Institute of
Biosciences, So Paulo State University, UNESP, Botucatu, So
Paulo, Brazil
b
Department of Biostatistics, Institute of Biosciences, So Paulo
State University, UNESP, Botucatu, So Paulo, Brazil
c
Department of Pharmacology, Institute of Biosciences, So Paulo
State University, UNESP, Botucatu, So Paulo, Brazil
Available online: 19 Oct 2011
To cite this article: N.C.C. Silva, L. Barbosa, L.N. Seito & A. Fernandes Junior (2011): Antimicrobial
activity and phytochemical analysis of crude extracts and essential oils from medicinal plants,
Natural Product Research: Formerly Natural Product Letters, DOI:10.1080/14786419.2011.564582
To link to this article: http://dx.doi.org/10.1080/14786419.2011.564582
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Natural Product Research
2011, 15, iFirst
SHORT COMMUNICATION
Antimicrobial activity and phytochemical analysis of crude extracts
and essential oils from medicinal plants
N.C.C. Silva
a
, L. Barbosa
b
, L.N. Seito
c
and A. Fernandes Junior
a
*
a
Department of Microbiology and Immunology, Institute of Biosciences, Sao Paulo State
University, UNESP, Botucatu, Sao Paulo, Brazil;
b
Department of Biostatistics,
Institute of Biosciences, Sao Paulo State University, UNESP, Botucatu, Sao Paulo, Brazil;
c
Department of Pharmacology, Institute of Biosciences, Sao Paulo State University, UNESP,
Botucatu, Sao Paulo, Brazil
(Received 1 July 2010; final version received 16 February 2011)
We aimed to establish a phytochemical analysis of the crude extracts and
performed GC-MS of the essential oils (EOs) of Eugenia uniflora L.
(Myrtaceae) and Asteraceae species Baccharis dracunculifolia DC,
Matricaria chamomilla L. and Vernonia polyanthes Less, as well as
determining their antimicrobial activity. Establishment of the minimal
inhibitory concentrations of the crude extracts and EOs against 16
Staphylococcus aureus and 16 Escherichia coli strains from human
specimens was carried out using the dilution method in MuellerHinton
agar. Some phenolic compounds with antimicrobial properties were
established, and all EOs had a higher antimicrobial activity than the
extracts. Matricaria chamomilla extract and E. uniflora EO were efficient
against S. aureus strains, while E. uniflora and V. polyanthes extracts and
V. polyanthes EO showed the best antimicrobial activity against E. coli
strains. Staphylococcus aureus strains were more susceptible to the tested
plant products than E. coli, but all natural products promoted antimicro-
bial growth inhibition.
Keywords: plant crude extracts; essential oils; medicinal plants; minimal
inhibitory concentration; antibacterial activity; phytochemical analysis
1. Introduction
Eugenia uniflora L. (Myrtaceae) is used as anti-diarrhoeal, diuretic, anti-rheumatic,
anti-febrile and anti-diabetic agents in Brazilian folk medicine and its leaf extracts
possess anti-inflammatory (Schapoval, Silveira, Miranda, Alice, & Henriques, 1994)
and antimicrobial activities (Oliveira, Lopes, Cabral, & Eberlin, 2006).
Terpenoids and flavonoids are the chemical groups most commonly found in the
genus Baccharis (Asteraceae) (Verdi, Brighente, & Pizzolatti, 2005) and Baccharis
dracunculifolia DC also has phenolic compounds (Funari, Ferro, & Mathor, 2007)
with antimicrobial activity (Duarte, Figueira, Pereira, Magalha es, & Delarmelina,
2004; Ferronato, Marchesan, Pezenti, Bednarski, & Onofre, 2007).
*Corresponding author. Email: ary@ibb.unesp.br
ISSN 14786419 print/ISSN 14786427 online
2011 Taylor & Francis
http://dx.doi.org/10.1080/14786419.2011.564582
http://www.tandfonline.com
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Vernonia polyanthes Less (Asteraceae) are used in folk medicine in Brazil against
flu, colds, coughs, bronchitis, bruises, haemorrhoids and infections of the uterus
(Corre a, Bertolucci, Pinto, Reis, & Alves, 2004), showing antibacterial activity
against mycobacterial strains (Oliveira et al., 2007).
Matricaria chamomilla L (Asteraceae) is used as tea against gastrointestinal and
nervous diseases (Fragoso, Esparza, Burchiel, Ruiz, & Torres, 2008) as well as
antimicrobial activities (Romero et al., 2005).
We aimed to establish the antimicrobial activity and phytochemical analysis of
crude methanolic extracts and essential oils (EOs) from E. uniflora,
B. dracunculifolia, V. polyanthes and M. chamomilla samples against
Staphylococcus aureus and Escherichia coli strains isolated from human specimens.
2. Results and discussion
Extracts dry weight, EO density and yield, phytochemical analysis of extracts and
EO are presented in Table 1. Phenols, flavonoids and fixed strong acids were found
in all extracts, which were previously reported for their antimicrobial activity
(Burt, 2004). Most compounds identified in the oils were monoterpenes or
sesquiterpenes, which may be involved in the EO antimicrobial effect (Cowan,
1999), although other compounds may also have increased their antimicrobial
effects, in a synergistic way. The crude extracts showed around 10% of those
obtained in the EO samples and were used for processing the minimum inhibitory
concentration (MIC) values (mg mL
1
) in the antibacterial assays.
MIC
90%
values are presented in Table 2, and found that S. aureus strains were
susceptible to both extracts and EOs from all studied plants. The crude extracts from
M. chamomilla, followed by V. polyanthes and B. dracunculifolia, were those with the
highest activities against S. aureus strains, while in contrast, the EO from E. uniflora
showed the highest inhibitory activity against this bacterium, with a value of MIC
90%
2.2 mg mL
1
. On the other hand, E. coli strains were less susceptible to plant
products, except to E. uniflora extract, which MIC
90%
(2.2 mg. ml
1
) showed the best
inhibitory effect, althought all EOs showed a similar result. Therefore, all plants
showed an antimicrobial activity, either as extracts or oils, revealing their potential
use as antimicrobial agents.
Gonc alves, Alves Filho, and Menezes (2005) verified the antimicrobial activity of
hydroalcoholic extract of E. uniflora against E. coli, S. aureus, Staphylococcus
coagulase-negative, Streptococcus pyogenes, Proteus mirabilis, Providencia spp and
Shigella sonnei. These data corroborate our results since E. uniflora had a potential
as an antimicrobial agent among the assayed plants. Auricchio and Bacchi (2003)
demonstrated the presence of flavonoids and tannins in E. uniflora crude extract,
allowing us to conclude that these compounds are characteristic of this plant species,
although further studies should characterise flavonoid and tannin groups.
Terpenoids and flavonoids (Verdi et al., 2005) and phenolic compounds (Funari
et al., 2007) were found in the genus Baccharis. Ferronato et al. (2007) found that
B. dracunculifolia EO showed antimicrobial effect against E. coli, S. aureus and
Pseudomonas aeruginosa, while our results showed higher MIC values assaying
S. aureus and E. coli strains. Duarte et al. (2004) reported that the MIC for
B. dracunculifolia extract against Enterococcus faecium strains was 10-fold higher for
2 N.C.C. Silva et al.
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the Gram-positive bacteria, although different bacterial strains were studied. The
differences between the MICs may be due to the chemical composition and
edaphoclimatic conditions (harvest season, time, location, among others), changing
secondary compounds (Gobbo-Neto and Lopes, 2007). Souza, Sena, Maranho,
Oliveira, and Guimara es (2008) found compounds such as fixed acids, alkaloids,
coumarins, flavonic and saponin glycosides and the presence of alkaloids, coumarins
Table 1. Physico-chemical characterisation of plant (crude extracts and EOs) antimicrobials.
Vegetal source
Density
(mg mL
1
)
(EOs)
Dry weight
(mg mL
1
)
(crude
extracts)
Yield (%)
(EOs)
Phytochemical
analysis of crude
extracts GC-MS of EOs
E. uniflora 924.0 145.0 0.19 Phenols, tannins,
chalcones,
aurones, fla-
vones, cate-
chins, flavo-
noids, saponins,
fixed strong
acids, quater-
nary com-
pounds, free
steroids and
quinones
Selina 1,3,7(11)
trien-8-one
(30.1%), Selina
1,3,7(11) trien-
8-one-epoxide
(21.89%),
cariofilene
(6.51%)
V. polyanthes 856.0 62.5 0.15 Phenols, tannins,
chalcones, aur-
ones, flavo-
noids, fixed
strong acids,
saponins, free
steroids, qui-
nones and
flavananois
Germacrene D
(27.79%),
"-Cariofilene
(16.2%),
Germacrene B
(15.01%)
B. dracunculifolia 857.0 76.0 0.20 Phenols, tannins,
flavones, cate-
chins, flavo-
noids, saponins,
fixed strong
acids, quater-
nary bases and
xanthones
Nerolidol
(18.77%),
Germacrene D
(10.45%), limo-
neme (8.75%)
M. chamomilla 940.0 100.0 0.17 Phenols, flavones,
flavonoids,
fixed strong
acids, quater-
nary com-
pounds, qui-
nones, xan-
thones, free
triterpenes
Chamazulene
(31.48%)
-bisabolol and
Bisabolone
oxide (15.71%)
Note: GC-MS Gas chromatography-mass spectrometry.
Natural Product Research 3
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and flavonoids in V. polyanthes samples. Oliveira et al. (2007) reported the
anti-mycobacterial activity with V. polyanthes, crude extracts as well as viable count
of inhibited cells after 3 h of contact in the time kill curve assay. Despite different
methodologies performed, our results were also satisfactory with V. polyanthes crude
extract.
Matricaria chamomilla showed antibacterial activity against S. aureus and
phenolic compounds were responsible for its antimicrobial properties (Romero et al.,
2005). Nogueira, Diniz, and Lima (2008) found no inhibition against P. aeruginosa
growth with chamomile EO, but 4% EO inhibited S. aureus and Candida albicans
strains, with inhibition zones ranging between 10 and 12 mm. Thus, M. chamomilla
extract and EO showed important inhibitory action against S. aureus strains, and
phenolic compounds were found in the phytochemical analysis. However, we believe
that other compounds are involved with the antimicrobial property together with the
phenols.
3. Conclusion
The results showed the potential use of these plants as antimicrobial agents and
S. aureus strains were more susceptible to plant products than E. coli and all natural
products promoted antimicrobial growth inhibition. Further studies should inves-
tigate the toxicity and chemical characterisation, aiming its use in the treatment of
infectious diseases, either alone or as an adjuvant with conventional antimicrobial
drugs.
Supplementary material
Experimental details relating to this article are available online.
Acknowledgements
The authors thank Dr Margarida Saeki and Dr Julio Doyama for the chromatographic
analysis of essential oils, and Dr Jose Mauricio Sforcin for the critical review of the
manuscript.
Table 2. Minimal inhibitory concentration (mg mL
1
) values for 90% (MIC
90%
) of S. aureus
and E. coli strains according to the crude extracts and EOs.
S. aureus E. coli
Vegetal source Crude extracts Essential oils Crude extracts Essential oils
B. dracunculifolia 5.4
a
3.7
ac
32.4
ab
25.8
a
V. polyanthes 3.3
ab
2.8
a
26.9
ac
24.1
a
M. chamomilla 1.2
b
2.9
bc
43.4
b
28.2
b
E. uniflora 24.1
c
2.2
b
15.9
c
27.6
c
Note: Different letters in columns represent significant differences in antimicrobial activity
between products when p 0.05.
4 N.C.C. Silva et al.
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