Bioresource Technology 101 (2010) 25102514

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Composition and biological activity of the essential oil from leaves of Plinia
cerrocampanensis, a new source of a-bisabolol
Roser Vila a, Ana Isabel Santana b, Renato Prez-Ross a, Anayansi Valderrama c, M. Victoria Castelli d,
Sergio Mendonca e, Susana Zacchino d, Mahabir P. Gupta b, Salvador Caigueral a,*
a

Unitat de Farmacologia i Farmacognsia, Facultat de Farmcia, Universitat de Barcelona, Av. Diagonal 643, 08028 Barcelona, Spain
Centro de Investigaciones Farmacognsticas de la Flora Panamea (CIFLORPAN), Facultad de Farmacia, Universidad de Panama, Panama, Republic of Panama
Instituto Conmemorativo Gorgas en Estudio de Salud, Panama, Republic of Panama
d
Laboratorio de Farmacognosia, Facultad de Ciencias Bioqumicas y Farmacuticas, Universidad Nacional de Rosario, Rosario, Argentina
e
Laboratorio de Microbiologa, Mestrado Prossional em Farmcia, Universidade Bandeirante de Sao Paulo, Brazil
b
c

a r t i c l e

i n f o

Article history:
Received 28 July 2009
Received in revised form 28 October 2009
Accepted 5 November 2009
Available online 16 December 2009
Keywords:
Plinia cerrocampanensis
Essential oil
a-Bisabolol
Antimicrobial activity
Aedes aegypti

a b s t r a c t
The essential oil from fresh leaves of Plinia cerrocampanensis Barrie (Myrtaceae), obtained by hydrodistillation, was analysed by GCFID and GCMS. Forty components, representing more than 91% of the oil,
were identied. Oxygenated sesquiterpenes represented the main fraction with a-bisabolol (42.8%) as
the major constituent, making this plant a new and good source of this substance.
Biological activity of the essential oil was evaluated against several bacterial and fungal strains as well
as larvae from Aedes aegypti. The highest activity was found against Staphylococcus aureus, Pseudomonas
aeruginosa, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum with MIC values
from 32 to 125 lg/ml. The essential oil also showed potent inhibitory and bactericidal activities against
three H. pylori strains, with MIC and MBC values of 62.5 lg/ml, and caused 100% mortality of A. aegypti
larvae at a concentration of 500 lg/ml.
2009 Elsevier Ltd. All rights reserved.

1. Introduction
Plants provide a multitude of avours and fragrances which
have found their way into everyday life. According to different
authors, approximately 3000 plant species contain essential oils,
from which only 300 are commercially important. Essential oils
and some of their constituents are used not only in pharmaceutical
products for their therapeutic activities but also in agriculture, as
food preservers and additives for human or animal use, in cosmetics and perfumes, and other industrial elds. In many cases, they
serve as plant defence mechanisms against predation by microorganisms, insects, and herbivores (Bakkali et al., 2008).
The complex composition of the essential oils and the variety of
chemical structures of their constituents are responsible of a wide
range of biological activities many of which are of increasing interest in the elds of human and animal health. Particularly, many
essential oils and their constituents have traditionally been used
for their antimicrobial activity which has long been recognized.
In addition, some of them may be useful in the control of mosquito
larvae that are responsible of the transmission of several diseases

* Corresponding author. Tel.: +34 93 4024531; fax: +34 93 4035982.

E-mail address: s.canigueral@ub.edu (S. Caigueral).
0960-8524/$ - see front matter 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2009.11.021

such as malaria, dengue fever or yellow fever, that are among the
greatest health problems in the world (Cheng et al., 2003, 2009).
The need of new anti-infective agents due to the emergence of
multiple antibiotic resistances has lead to the search of new
sources of potential antimicrobials (Carson and Riley, 2003).
Among them, the plant kingdom offers a wide range of biodiversity
of great value for the pharmaceutical industry.
Within the framework of our ongoing research on aromatic
ora from Panama, in view of potentiate the use of its natural resources, the present work deals with the study of the essential
oil from fresh leaves of Plinia cerrocampanensis Barrie (Myrtaceae),
in particular its chemical composition and the evaluation of its biological activity against several bacterial and fungal strains, as well
as against larvae from Aedes aegypti.
P. cerrocampanensis, which has recently been described as a new
species of the genus Plinia, is a tree that grows between 800 and
1000 m of altitude in the surroundings of Cerro Campana (Republic
of Panama), reaching a height of about 8 m (Barrie, 2004). Until
now no data on its chemical constituents or its biological activity
are available in the scientic literature, although the composition
of the essential oils (Apel et al., 2006; Pino et al., 2002, 2003) and
the xanthine oxidase inhibitory activity of hydro-alcoholic extracts
(Theoduloz et al., 1988) of other Plinia sp. have been reported
previously.

R. Vila et al. / Bioresource Technology 101 (2010) 25102514

2. Methods
2.1. Plant material
Fresh leaves of P. cerrocampanensis Barrie were collected in the
National Park Altos de Campana (N08410 09.800 ; W079560 04.400 ),
Province of Panama (Republic of Panama) in February 2005. A voucher specimen No. FLORPAN 6623 is deposited in the Herbarium of
the University of Panama (PMA).
2.2. Isolation and analysis of the essential oil
The essential oil was obtained from 100 g of fresh leaves by
hydrodistillation, using the standard apparatus described in the
European Pharmacopeia (Council of Europe, 2005).
Analysis of the oil was carried out by GCFID and GCMS using
two fused silica capillary columns (30 m  0.25 mm i.d.; 0.25 lm
lm thickness) of different stationary phases: Supelcowax 10
and methylsilicone SE-30. GCFID analyses were performed on a
HewlettPackard 6890 instrument, equipped with a HP ChemStation data processor software, using the following analytical conditions: carrier gas, Helium; ow rate, 1 ml/min; oven temperature
programmed from 60220 C at 4 C/min, 220 C (10 min); injector
temperature, 250 C; detector temperature, 270 C; split ratio 1:80.
The essential oil was injected undiluted (0.1 ll). Mass spectra were
obtained with a computerized system constituted by a GC HewlettPackard 6890 coupled to a mass selective detector Hewlett
Packard 5973N, using the same analytical conditions as above.
Mass spectra were taken over m/z 35400, using an ionizing voltage of 70 eV.
Identication of components was achieved by means of their GC
retention indices in two stationary phases, determined in relation
to a homologous series of fatty acid methyl esters, and by comparison of fragmentation patterns in the mass spectra with those
stored in our own library, in the GCMS database and with literature data (Adams, 1995; McLafferty, 1993). Quantication of each
compound was performed on the basis of their GC peak areas on
the two columns, using the normalisation procedure without corrections for response factor.

2511

and bacterial density adjusted to 6  108 CFU. Viability control

was done by Gram staining and colony count.
MIC and MBC determinations: broth microdilution was performed in brucella broth supplemented with 2% fetal calf serum
(Cultilab, Brasil) and 0.2% DMSO. Two fold dilutions of essential
oil ranging from 1000 to 7.8 lg/ml were used. The standardized
inoculum was diluted to achieve a nal inoculum concentration
of approximately 6  106 CFU per well. The microplates were incubated at 37 C under microaerophilic conditions. All assays were
performed in duplicate using amoxicillin (ranging from 16 to
0.125 lg/ml) as positive internal control. The microplates were
aseptically examined for the presence of turbidity after 72 h of
incubation, and MIC were the lowest concentration of essential
oil that inhibited detectable bacterial growth. After that, 2 ll of
each sample were spread onto brucella-sheep blood agar plates
in order to determine the MBC. These plates were monitored for
the presence of bacterial growth after 4872 h of incubation, and
MBC were the lowest concentration that killed at least 99.9% of
the original inoculum. Amoxicillin was used as a positive control.
2.5. Antifungal activity

The antibacterial activity was assayed against Escherichia coli

(ATCC 9637), Klebsiella pneumoniae (ATCC 10031), Mycobacterium
smegmatis (ATCC 607), Pseudomonas aeruginosa (ATCC 27853), Salmonella gallinarum (ATCC 9184) and Staphylococcus aureus (ATCC
6538), following the method described by Mitscher et al. (1971),
using streptomycin sulphate as positive control. The essential oil
was rst dissolved in DMSO (1000 lg/ml) and the bacteriostatic
activity was determined by measuring the minimum inhibitory
concentration (MIC) from diluted aqueous samples of 500, 250,
125, 62.5, 31.25, 15.6 and 7.8 lg/ml. All the experiments were performed in triplicate and the results are expressed as mean values.

Antifungal activity was assayed against several yeasts and lamentous fungi strains from the American Type Culture Collection
(ATCC, Rockville, MD, USA) and Centro de Referencia en Micologa
CEREMIC (C, Facultad de Ciencias Bioqumicas y Farmacuticas,
Rosario, Argentina): Candida albicans (ATCC 1023), C. tropicalis (C
131), Saccharomyces cerevisiae (ATCC 9763), Cryptococcus neoformans (ATCC 32264), Aspergillus avus (ATCC 9170), A. fumigatus
(ATCC 26934), A. niger (ATCC 9029), Trichophyton rubrum (C 110),
Trichophyton mentagrophytes (ATCC 9972) and Microsporum gypseum (C 115).
MIC values were determined using broth dilution techniques as
described by the Clinical and Laboratory Standards Institute (CLSI,
formerly National Committee for Clinical and Laboratory Standards) for yeasts (M27-A2) (NCCLS, 2002a) as well for lamentous
fungi (M38-A) (NCCLS, 2002b) in microtiters of 96 wells. RPMI1640 (Sigma, St. Louis, Mo, USA) buffered to a pH 7.0 with MOPS
was used. The starting inocula were approximately 1  103 to
5  103 CFU/ml. Microtiter trays were incubated at 35 C for yeasts
and hyalohyphomycetes and at 2830 C for dermatophytes in a
moist, dark chamber. MIC values were recorded at 48 h for yeasts
and at a time according to the control fungus growth, for the other
fungi. The susceptibility of the standard drugs ketoconazole, terbinane and amphotericin B was dened as the lowest concentration
of drug which resulted in total inhibition of fungal growth.
For the assay, the essential oil of P. cerrocampanensis was twofold diluted with RPMI from 1000 to 0.98 lg/ml (nal volume = 100 ll) and a nal DMSO concentration 61%. A volume of
100 ll of inoculum suspension was added to each well with the
exception of the sterility control where sterile water was added
to the well instead. The MIC was dened as the minimum inhibitory concentration of the essential oil which resulted in total inhibition of the fungal growth.

2.4. Anti-Helicobacter pylori activity

2.6. Larvicidal activity

Stock cultures of H. pylori 26695 (ATCC 700392), J99 (ATCC

700824) and SS1 (Sidney Strain 1) were reactivated on Columbia
agar plates (CA) (Merck, Germany) supplemented with 10% debrinated sheep blood (BBV, Brazil), 10 mg/L vancomycin, 20 mg/L
nalidixic acid, 2 mg/L amphotericin B and 40 mg/L 2,3,5-triphenyltetrazolium chloride (Sigma, Germany), and cultured at 37 C in a
humidied 12% CO2 incubator (Revco, USA). The identity of the colonies was conrmed by Gram staining and oxidase, catalase and
urease production. The colonies were suspended in PBS (pH 7.2)

For this purpose, larvae of A. aegypti (Culicidade: Diptera) of III

and IV Stage obtained after 68 days post oviposition were used.
The insectarium was kept at temperature of 22.525 C, relative
humidity of 80% and with a photoperiod of 12:12 h. The larvae
were fed with granulated yeast suspended in water (1:4).
The essential oils were dissolved in ethanol and were tested at a
nal concentration of 1, 100 and 500 ppm in triplicate. Water and
alcohol were used as control. For each replicate, 20 individuals of
mosquito species were placed in a foam container. After 24 h, per

2.3. Antibacterial activity

2512

R. Vila et al. / Bioresource Technology 101 (2010) 25102514

cent mortality was calculated. Tetramethrin with a LC100 value of

0.25 0.01 lg/ml was used as a positive control.
3. Results and discussion
The fresh leaves of P. cerrocampanensis gave by hydrodistillation
an essential oil yield of 0.64% (v/w). Qualitative and quantitative
analysis of the oil by GC and GCMS allowed the identication of
forty components, representing more than 91% of the total sample.
The oil was very rich in oxygenated sesquiterpenes (65.9%), especially a-bisabolol (42.8%), bisabolol oxide B (10.3%) and trans-nerolidol (9.4%). Among monoterpenes, linalool (10.3%) was found in
the highest percentage. Further details of the essential oil composition are shown in Table 1.
Table 1
Composition of the essential oil from leaves of Plinia cerrocampanensis.
Constituentsa

RICWb

RISEc

%d

a-Pinene

112
204
220
225
237
271
323
337
378
380
399
434
439
447
457
462
474
483
489
510
521
544
615
623
656
662
676
700
710
763
824

209
251
255
266
246
224
270
278
287
296
323

475
329
487
492
494
477

518

562
568

579
555
593
640
208
243
260
281
428
445
458
489
508

0.2e,f,g
0.4e,f,g
0.1e,f,g
0.5e,f,g
1.1e,f,g
0.2e,f,g
0.6e,f,g
0.7e,f,g
10.3e,f,g
0.1e,f,g
1.1e,f,g
0.3e,f
0.1e,f,g
1.2e,f,g
0.1e,f,g
4.8e,f,g
0.6e,f,g
0.8e,f,g
0.1e,f
0.3e,f
0.7e,f
0.1e,f
9.4e,f,g
te,f
10.3e,f,g
1.4e,f,g
0.5e,f
42.8e,f,g
0.6e,f,g
0.5e,f,g
0.4e,f,g
0.1e,g
te,g
0.1e,g
0.1e,g
0.1e,g
0.2e,g
0.1e,g
0.3e,g
0.3e,g

Limonene
cis-Ocimene
c-Terpinene
p-Cymene
6-Methyl-5-hepten-2-one
trans-Linalool oxide
cis-Linalool oxide
Linalool
4-Acetyl-1-methyl-1-cyclohexene
Terpinen-4-ol
Estragole
a-Amorphene
a-Terpineol
a-Muurolene
b-Bisabolene
d-Cadinene
a-Curcumene
Nerol
Calameneneh
Geraniol
a-Calacorene
trans-Nerolidol
Cubenol
a-Bisabolol oxide B
b-Bisabolol
10-epi-Cadinol
a-Bisabolol
b-Eudesmol
E,E-Farnesol
a-Bisabolol oxide A
Benzaldehyde
d-3-Carene
trans-b-Ocimene
Terpinolene
a-Copaene
a-Cedrene
trans-a-Bergamotene
cis-a-Bisabolene
trans-a-Bisabolene

This composition pattern is quite unusual among essential oils

from leaves of other Plinia sp. previously reported. Although most
of them are also characterized by the predominance of oxygenated
sesquiterpenes (Apel et al., 2006; Pino et al., 2003), only the one
from P. cordifolia showed remarkable percentages of compounds
with a bisabolane nucleus, such as a-bisabolol oxide A (28.0%),
a-bisabolol oxide B (7.0%) and a-bisabolol (5.8%) (Apel et al., 2006).
Results on the antibacterial activity of the essential oil of P. cerrocampanensis are summarized in Table 2. It exhibited the strongest activity against P. aeruginosa and S. aureus with MIC values
of 62.5 and 125 lg/ml, respectively. This essential oil also showed
potent inhibitory and bactericidal activities against three H. pylori
strains (including SS1, traditionally used for in vivo assays), with
MIC and MBC values of 62.5 lg/ml. H. pylori, a gastric pathogen
whose infection is associated with chronic supercial gastritis,
peptic ulceration and gastric cancer, chronically infects more than
half of the worlds population. To be effective, therapies require the
use of more than one antimicrobial in combination. Unfortunately,
increased primary resistance to recommended antibiotics modies
the therapy effectiveness and negatively affects its eradication,
requiring the search for new strategies (Ortiz Godoy et al., 2003).
Therefore, the oil of P. cerrocampanensis stands out as a new contribution in this search.
MIC values towards several yeasts and fungi strains are shown
in Table 3. Dermatophytes were the most sensitive ones, particularly T. mentagrophytes, T. rubrum and M. gypseum with MIC values
of 32, 62.5 and 125 lg/ml, respectively.
All these activities can be related to the major constituents,
mainly a-bisabolol (Kedzia, 1991; Szalontai et al., 1976), linalool
(Pattnaik et al., 1997; Sonboli et al., 2006) and nerolidol (Kubo
et al., 1992) whose antimicrobial properties have been previously
reported. Particularly, nerolidol has been recently found to inhibit
the hyphal growth of T. mentagrophytes causing destruction and
disorganization of organelles in the fungal cytoplasm (Park et al.,
2009).
A. aegypti is the major vector of dengue and yellow fever which
have experimented a recrudescence due to the increasing resistance of mosquitoes to current commercial insecticides. Although
yellow fever has been reasonably brought under control with its
vaccine, no vaccine is available for dengue. Several secondary
metabolite plant products have been successfully assayed for their
larvicidal activity against A. aegypti, particularly essential oils and
their constituents. (Barreira Cavalcanti et al., 2004; Urano Carvalho
et al., 2003). In the present work, the essential oil of P. cerrocampanensis showed, at concentrations of 100 and 500 lg/ml, 53%
and 100% mortality of A. aegypti larvae, respectively, constituting
a potential alternative to the conventional chemical control.
Table 2
Antibacterial activity of the essential oil of Plinia cerrocampanensis.
Bacteria

EOPCa

Streptomycin
sulphate
MIC

Amoxicillin

>1000
>1000
>1000

6.25
3.12
12.50

62.5
>1000
125
62.5/62.5
62.5/62.5
62.5/62.5

1.56
3.12
12.50

0.125/0.5
0.125/0.5
0.125/0.5

Monoterpene hydrocarbons
Oxygenated monoterpenes
Sesquiterpene hydrocarbons
Oxygenated sesquiterpenes
Others
Total identied

MIC /
MBCc

2.5
14.7
7.8
65.9
0.7
91.6

Escherichia coli
Klebsiella pneumoniae
Mycobacterium
smegmatis
Pseudomonas aeruginosa
Salmonella gallinarum
Staphylococcus aureus
Helicobacter pylori 26695
Helicobacter pylori J99
Helicobacter pylori SS1

Components are listed in increasing order according to their retention indices in

Supelcowax 10 except the last nine constituents, which were only detected in
methylsilicone.
b
RICW: retention indices in Supelcowax10 column.
c
RISE: retention indices in methylsilicone (SE-30) column.
d
t: traces (60.05).
e
Identication method: GCMS.
f
Identication method: retention index in Supelcowax10.
g
Identication method: retention index in methylsilicone.
h
Isomer not assigned.

MIC/MBC

EOPC: essential oil of P. cerrocampanensis.

MIC: minimum inhibitory concentration (lg/ml).
c
MBC: minimum bactericidal concentration (lg/ml). MBC was only determined
against Helicobacter pylori.
b

2513

R. Vila et al. / Bioresource Technology 101 (2010) 25102514

Table 3
Antifungal activity of the essential oil of Plinia cerrocampanensis.
EOPCa
MICb

Amphotericin B
MIC

Ketoconazole
MIC

Terbinane
MIC

Yeasts
Candida albicans
Candida tropicalis
Cryptococcus neoformans
Saccharomyces cerevisiae

>250
>250
>250
>250

0.78
1.56
0.78
0.78

6.25
6.25
1.56
3.12

Filamentous fungi
Aspergillus avus
Aspergillus fumigatus
Aspergillus niger

>250
>250
>250

0.78
3.12
0.78

6.25
12.5
6.25

Dermatophytes
Microsporum gypseum
Trichophyton mentagrophytes
Trichophyton rubrum

125
32
62.5

0.25
0.75
0.75

0.50
0.25
0.25

0.04
0.02
0.01

Fungi

a
b

EOPC: essential oil of P. cerrocampanensis.

MIC: minimum inhibitory concentration (lg/ml).

The essential oil of P. cerrocampanensis contains a 42% of abisabolol, its major constituent, being a potential industrial source
of this interesting compound. a-Bisabolol, a well known monocyclic unsaturated sesquiterpene alcohol which is one of the main active principles of chamomile (Chamomilla recutita), is widely used
in cosmetic preparations due to its anti-inammatory activity
and low toxicity (Habersang et al., 1979; Hempel and Hirschelmann, 1998; Isaac, 1979; Jellinek, 1984; Madhavan, 1999; Yakovlev and von Schlichtegroll, 1969). Furthermore, both a-bisabolol
and nerolidol, also one of the main constituents of the essential
oil (9.4%), may increase dermal absorption of other substances by
more than 20-fold, being useful vehicles for other drugs (Cornwell
and Barry, 1994). These two sesquiterpenes are also able to enhance bacterial permeability and susceptibility to clinically important antibiotic compounds (Brehm-Stecher and Johnson, 2003).
Moreover, a-bisabolol has been found to be a promising inducer
of apoptosis in highly malignant glioma cells (Cavalieri et al.,
2004). Finally, it is interesting to highlight that linalool, the major
monoterpene of the oil of P. cerrocampanensis (10.3%), could also
enhance the activities of a-bisabolol in the oil, since it has shown
anti-inammatory, anti-hyperalgesic and anti-nociceptive effects
in several animal models, which have been ascribed to different
mechanisms of action (Peana et al., 2002, 2004, 2006a,b; Re
et al., 2000).
4. Conclusions
In conclusion, the essential oil from leaves of P. cerrocampanensis from Panama is an outstanding new source of a-bisabolol, a
compound highly appreciated in the pharmaceutical and cosmetic
industry. In addition, it shows interesting antimicrobial and larvicidal activities which make this essential oil a potential industrial
resource of new products.
Acknowledgements
Authors are grateful to National Secretariat for Science, Technology, and Innovation (SENACYT) of Panama, Project No. 112004 and the Organization of American States for nancial support
to CIFLORPAN. Also, thanks are due to Cristina Minguillon and
Antoni Riera (University of Barcelona) for helping to conrm the
identication of a-bisabolol by means chiral chromatography. R.
Prez-Ross was supported by the Generalitat de Catalunya (Education and Universities Department) and the European Social Fund.
S. Zacchino is grateful to ANPCyT (PICT R 260). M.V. Castelli
acknowledges CONICET.

References
Adams, R.P., 1995. Identication of Essential Oils by Gas Chromatography/Mass
Spectroscopy. Allured Carol Stream, IL.
Apel, M.A., Sobral, M., Zuanazzi, J.A., Henriques, A., 2006. Essential oil composition
of four Plinia species (Myrtaceae). Flavour Fragr. J. 21, 565567.
Bakkali, F., Averbeck, S., Averbeck, D., Idaomar, M., 2008. Biological effects of
essential oils. Rev. Food Chem. Toxicol. 46, 446475.
Barreira Cavalcanti, E.S., Maia de Morais, S., A.Lima, M.A, Pinho Santana, E.W, 2004.
Larvicidal activity of essential oils from Brazilian plants against Aedes aegypti L.
Mem. Inst. Oswaldo Cruz Rio de Janeiro 99, 541544.
Barrie, F.R., 2004. Synopsis of Plinia (Myrtaceae) in Mesoamerica. Novon 14, 380400.
Brehm-Stecher, B.F., Johnson, E.A., 2003. Sensitization of Staphylococcus aureus and
Escherichia coli to antibiotics by the sesquiterpenoids nerolidol, farnesol,
bisabolol, and apritone. Antimicrobial. Agents Chemother. 47, 33573360.
Carson, C.F., Riley, T.V., 2003. Non-antibiotic therapies for infectious diseases.
Commun. Dis. Intell. 27 (Suppl.), S143S146.
Cavalieri, E., Mariotto, S., Fabrizi, C., Carcereri de Prati, A., Gottardo, R., Leone, S.,
Berra, L.V., Lauro, G.M., Ciampa, A.R., Suzuki, H., 2004. a-Bisabolol, a nontoxic
natural compound, strongly induces apoptosis in glioma cells. Biochem.
Biophys. Res. Commun. 315, 589594.
Cheng, S.S., Chang, H.T., Chang, S.T., Tsai, K.H., Chen, W.J., 2003. Bioactivity of
selected plant essential oils against the yellow fever mosquito Aedes aegypti
larvae. Bioresour. Technol. 89, 99102.
Cheng, S.S., Huang, C.G., Chen, Y.J., Yu, J.J., Chen, W.J., Chang, S.T., 2009. Chemical
compositions and larvicidal activities of essential oils from two eucalyptus
species. Bioresour. Technol. 100, 452456.
Cornwell, P.A., Barry, B.W., 1994. Sesquiterpene components of volatile oils as skin
penetration enhancers for the hydrophilic permeant 5-uorouracil. J. Pharm.
Pharmacol. 46, 261269.
Council of Europe, 2005. European Pharmacopeia, vol. 1, fth ed. Directorate for the
Quality of Medicines of the Council of Europe, Strasbourg, pp. 217218.
Habersang, S., Leuschner, F., Isaac, O., Thiemer, K., 1979. Pharmacological studies of
chamomile constituents IV. Studies on the toxicity of ()-a-bisabolol. Planta
Med. 37, 115123.
Hempel, B., Hirschelmann, R., 1998. Chamomilla inammation inhibiting effects of
the contents and formulations in vivo. Deutsch. Apoth. Zeit. 138, 42374242.
Isaac, O., 1979. Pharmacological investigations with compounds of chamomile I.
The pharmacology of ()-a-bisabolol and bisabolol oxides. Planta Med. 35,
118124.
Jellinek, S., 1984. a-Bisabolol: an anti-inammatory agent for cosmetic products.
Parfums Cosmetiques Aromes 57, 5557.
Kedzia, B., 1991. Antimicrobial activity of chamomile oil and its components. Herba
Pol. 37, 2938.
Kubo, I., Muroi, H., Himejima, M., 1992. Antimicrobial activity of green tea avor
components and their combination effects. J. Agr. Food Chem. 40, 245248.
Madhavan, B.N., 1999. Final report on the safety assessment of bisabolol. Int. J.
Toxicol. 18, 3340.
McLafferty, F.W., 1993. Registry of Mass Spectral Data. John Wiley & Sons, New York.
Mitscher, L.A., Leu, R.P., Bathala, M.S., Wu, W.N., Beal, J.L., 1971. Antibiotic agents
from higher plants I: introduction, rationale and methodology. J. Nat. Prod. 35,
157166.
NCCLS National Committee for Clinical and Laboratory Standards, 2002a.
Reference method for broth dilution antifungal susceptibility testing of
yeasts; approved standard, second ed. NCCLS Document M 27-A2, vol. 22, No.
15. NCCLS, Wayne, PA, pp. 129.
NCCLS National Committee for Clinical and Laboratory Standards, 2002b.
Reference method for broth dilution antifungal susceptibility testing of
lamentous fungi; approved standard, second ed., NCCLS Document M 38-A,
vol. 22, No. 16. NCCLS, Wayne, PA, pp. 129.

2514

R. Vila et al. / Bioresource Technology 101 (2010) 25102514

Ortiz Godoy, A.P., Lima Ribeiro, M., Borges Benbengo, Y.H., Vitiello, L., Bueno
Miranda, M.C., Mendona, S., Pdrazzoli, J., 2003. Analysis of antimicrobial
susceptibility and virulence factors in Helicobacter pylori clinical isolates. BMC
Gastroenterol. 3, 20.
Park, M.J., Gwak, K.S., Yang, I., Kim, K.W., Jeung, E.B., Chang, J.W., Choi, I.G., 2009.
Effect of citral, eugenol, nerolidol and a-terpineol on the ultrastructural changes
of Trichophyton mentagrophytes. Fitoterapia 80, 290296.
Pattnaik, S., Subramanyam, V.R., Bapaji, M., Kole, C.R., 1997. Antibacterial and
antifungal activity of aromatic constituents of essential oils. Microbios 89, 39
46.
Peana, A.T., DAquila, P.S., Panin, F., Serra, G., Pippia, P., Moretti, M.D.L., 2002. Antiinammatory activity of linalool and linalyl acetate constituents of essential
oils. Phytomedicine 9, 721726.
Peana, A.T., De Montis, M.G., Sechi, S., Sircana, G., DAquila, P.S., Pippia, P., 2004.
Effects of ()-linalool in the acute hyperalgesis induced by carrageenan, Lglutamate and prostaglandin E2. Europe. J. Pharmacol. 497, 279284.
Peana, A.T., Marzocco, S., Popolo, A., Pinto, A., 2006a. ()-Linalool inhibits in vitro
NO formation: Probable involvement in the antinociceptive activity of this
monoterpene compound. Life Sci. 78, 719723.
Peana, A.T., Rubattu, P., Piga, G.G., Fumagalli, S., Boatto, G., Pippia, P., De Montis,
M.G., 2006b. Involvement of adenosine A1 and A2A receptors in ()-linaloolinduced antinociception. Life Sci. 78, 24712474.

Pino, J.A., Bello, A., Urquiola, A., Correa, T., Rosado, A., 2002. Essential oil of Plinia
rubrinervis Urb from Cuba. J. Essent. Oil Res. 14, 372.
Pino, J.A., Bello, A., Urquiola, A., Aguero, J., 2003. Chemical composition of the leaf oil
of Plinia dermatodes. J. Essent. Oil Res. 15, 2324.
Re, L., Baroci, S., Mencarelli, A., Vivani, C., Paolucci, G., Scarpantonio, A., Rinaldi, L.,
Mosca, E., 2000. Linalool modies the nicotinic receptor-ion channel kinetics at
the mouse neuromuscular junction. Pharmacol. Res. 42, 177181.
Sonboli, A., Babakhani, B., Mehrabian, A.R., 2006. Antimicrobial activity of six
constituents of essential oil from Salvia. Z. Naturforsch C J. Biosci. 61, 160
164.
Szalontai, M., Verzar-Petri, G., Florian, E., 1976. Data on the antifungal effect of the
biologically active components of Matricaria chamomilla L. Acta Farm. Hung. 46,
232247.
Theoduloz, C., Franco, L., Ferro, E., Schmeda Hirschmann, G., 1988. Xanthine oxidase
inhibitory activity of Paraguayan Myrtaceae. J. Ethnopharmacol. 24, 179183.
Urano Carvalho, A.F., Maciel Melo, V.M., Arago Craveiro, A., Lacerda Machado, M.I.,
Braga Bantim, M., Fontenele Rabelo, E., 2003. Larvicidal activity of the essential
oil from Lippia sidoides Cham. against Aedes aegypti Linn. Mem. Inst. Oswaldo
Cruz Rio de Janeiro 98, 569571.
Yakovlev, V., von Schlichtegroll, A., 1969. Anti-inammatory activity of ()-abisabolol, an essential oil component of chamomile oil. ArzneimittelForsch. 19,
615616.