Analgesic effects of rosemary essential oil and its interactions with codeine and paracetamol in mice

Abstract

The use of herbal medicinal products in the management of pain has been increasing steadily in recent years, often in combination with conventional analgesics, which can induce significant interactions. In traditional medicine, rosemary was used as mild analgesic, for relieving renal colic pain and dysmenorrhea. The aim of our study was to examine analgesic effects of rosemary essential oil and its pharmacodynamic interactions with codeine and paracetamol in mice.
The identification and quantification of chemical constituents of the essential oil isolated from air-dried aerial parts of rosemary were carried out by GC/FID and GC/MS. The hot plate test was performed on NMRI mice by placing them individually on hot plate and assessing their response to the thermal stimulus.
In this research, we identified 29 chemical compounds of the studied rosemary essential oil, and the main constituents were 1,8-cineole, camphor, and α-pinene. Administration of investigated essential oil increased significantly the latency time of animal response to heat-induced pain between 20th and 50th minute of the test, when compared to saline-treated group. Rosemary essential oil in the dose of 20 mg/kg was shown to be more efficient than in the dose of 10 mg/kg, in combinations with both codeine and paracetamol.
Our findings support the use of rosemary in the management of pain and indicate a therapeutic potential of rosemary essential oil in combination with analgesic drugs. The mechanisms involved in analgesic effects of rosemary essential oil and the potential influence on cytochromes and drug metabolism should be more in-depth investigated.

Full text

Abstract. – OBJECTIVE : The use of herbal
medicinal products in the management of pain
has been increasing steadily in recent years, often
in combination with conventional analgesics,
which can induce significant interactions. In tradi-
tional medicine, rosemary was used as mild anal-
gesic, for relieving renal colic pain and dysmenor-
rhea. The aim of our study was to examine anal-
gesic effects of rosemary essential oil and its
pharmacodynamic interactions with codeine and
paracetamol in mice.
MATERIALS AND METHODS: The identifica-
tion and quantification of chemical constituents
of the essential oil isolated from air-dried aerial
parts of rosemary were carried out by GC/FID
and GC/MS. The hot plate test was performed on
NMRI mice by placing them individually on hot
plate and assessing their response to the ther-
mal stimulus.
RES ULTS: In this research, we identified 29
chemical compounds of the studied rosemary
essential oil, and the main constituents were 1,8-
cineole, camphor, and αα-pinene. Administration
of investigated essential oil increased signifi-
cantly the latency time of animal response to
heat-induced pain between 20th and 50th minute
of the test, when compared to saline-treated
group. Rosemary essential oil in the dose of 20
mg/kg was shown to be more efficient than in
the dose of 10 mg/kg, in combinations with both
codeine and paracetamol.
CONCLUSIONS: Our findings support the use
of rosemary in the management of pain and indi-
cate a therapeutic potential of rosemary essen-
tial oil in combination with analgesic drugs. The
mechanisms involved in analgesic effects of
rosemary essential oil and the potential influ-
ence on cytochromes and drug metabolism
should be more in-depth investigated.
Key Words:
Rosmarinus officinalis, Essential oil, Antinociceptive,
Hot plate, Interaction.
European Review for Medical and Pharmacological Sciences
Analgesic effects of rosemary essential oil
and its interactions with codeine and
paracetamol in mice
A. RASKOVIC, I. MILANOVIC1, N. PAVLOVIC, B. MILIJASEVIC,
M. UBAVIC2, M. MIKOV
Department of Pharmacology, Toxicology and Clinical Pharmacology, Faculty of Medicine,
University of Novi Sad, Novi Sad, Serbia
1High Medical School of Professional Skills, Zemun, Serbia
2Health Care Institution for Laboratory Diagnostics ’Medlab’, Novi Sad, Serbia
Corresponding Author: Nebojsa Pavlovic, MPharm; e-mail: nebojsa.pavlovic@gmail.com 165
Introduction
The use of herbal medicinal products to re-
lieve pain has been increasing steadily in recent
years because they are often perceived as being
natural and therefore harmless1,2. Herbal medi-
cines are often taken in combination with con-
ventio nal dr ug the rapie s and some of thei r
pharmacologically active ingredients might in-
teract with synthetic drugs3. Drugs that are sub-
strates for metabolism mediated by cytochrome
P450 (CYP) enzymes are particularly subject to
herb-drug interactions, which can be attributed
to a large extent to CYP polymorphism and
ability of many herbal compounds to induce or
inhibit these enzymes4.
Rosemary (Rosmarinus officinalis L., Lami-
aceae) is widely cultivated all over the world as
an ornamental and aromatic plant, and has been
commonly used for flavoring food, but also for
diff eren t medicinal purpose s. In traditi onal
medicine, rosemary was used as mild analgesic,
for relieving renal colic pain, dysmenorrhea,
respiratory disorders, due to its antispasmodic
prop erties5, 6. Rece ntly, essen tial o il isolated
from rosemary and monoterpenes as its main
active compounds have been of great interest
due to their various health benefits and thera-
peutic effects. According to the recommenda-
tion of European Medicines Agency (EMA)
from 2010, rosemary essential oil (REO) can be
used for treating dyspepsia and mild spasmodic
disorders of the gastrointestinal tract, as well as
an adjuvant in the relief of minor muscular and
articular pain and in minor peripheral circulato-
ry disorders7. Besides, the experiments con-
ducted with REO have demonstrated its several
notable pharmacological effects, such as an-
2015; 19: 165-172

A. Raskovic, I. Milanovic, N. Pavlovic, B. Milijasevic, M. Ubavic, M. Mikov
and CYP2A6 subfamilies of cytochromes P450,
resulting in the formation of the highly reactive
and hepatotoxic N-acetyl-p-benzoquinoneimine
(NAPQI), which is quickly conjugated with glu-
tathione to form non-toxic cysteine and mercap-
turic acid conjugates19. Hepatic enzymes induction
may increase paracetamol hepatotoxicity, and
therefore the potential for interactions of paraceta-
mol with herbal medicinal products should be
carefully considered.
Based on the above mentioned facts, the aim
of our study was to examine analgesic effects of
REO and its pharmacodynamic interactions with
codeine and paracetamol in mice.
Materials and Methods
Plant Material and Chemicals
Aerial parts of cultivated plants of rosemary
were obtained from the Institute for Studies on
Medicinal Plants, Dr Josif Pancic, Belgrade, in
2010. A voucher specimen of the plant (Rosmari-
nus officinalis L. 1753 subsp. officinalis No 2-
1746, det.: Goran Anackov) was confirmed and
deposited in the Herbarium of the Department of
Biology and Ecology, Faculty of Natural Sci-
ences, University of Novi Sad. The essential oil,
used in our experiments, was isolated from the
obtained plant material.
Parace tamol was pur chase d fro m Si gma-
Aldrich (St Louis , M O, USA). C odeine hy-
drochloride was obtained from Fampharm
(Kruševac, Serbia).
Isolation and Analysis of Essential Oil
The essential oil was isolated from air-dried
aerial parts of rosemary by hydrodistillation, ac-
cording to the procedure of the European Phar-
macopoeia 420. N-hexane was used as a collect-
ing solvent, which was afterward removed under
vacuum from the obatined essential oil.
The identification and quantification of chemi-
cal constituents of the essential oil were carried
out by gas chromatography coupled with flame
ionization detection (GC/FID) and mass spectro-
metric detection (GC/MS). GC/FID analysis was
performed using a Hewlett-Packard HP 5890 se-
ries II chromatograph equipped with an autosam-
pler and a split/splitless injection system. The
capillary column used in this study was HP-5 (25
m × 0.32 mm; film thickness of 0.52 µm), cou-
pled to the flame ionization detector (FID). The
injector and detector temperatures were set at
tioxidant and antimicrobial8, anti-inflammatory
and antinociceptive9, antidepressant10, cogni-
tion-enhancing11, among others.
On the other hand, there are several well-estab-
lished interactions of rosemary preparations with
different drugs, such as antibiotics, anxiolytics and
anticoagulant medicines, in terms of potentiation
of their activity3,12. Given that REO was found to
exert analgesic effects in different experimental
models of nociception, its potential interactions
with analgesics may be assumed. Besides, it was
demonstrated that REO and its main components
induce catalytic activities of microsomal enzymes,
particularly CYP2B, but also slightly the activity
of UDP-glucuronosyltransferase (UGT) and,
therefore, may interact with drugs metabolized by
cytochromes or UGT13,14.
Since first isolated from Papaver somniferum
L. in 1832, codeine has been used as an anal-
gesic, antitussive and antidiarrhoeal drug. For its
analgesic effect, codeine is regarded as a prodrug
that is metabolized through O-demethylation to
morphine by CYP2D6 and N-demethylation to
norcodeine by CYP3A4. Codeine, morphine and
norcodeine can be further glucuronidated. The
majority of the analgesic effect of codeine is at-
tibu t e d to the potent µ-receptor ag o n i s t
morphine15. Genetic polymorphism of CYP2D6
is common and can affect therapeutic response of
codeine. Poor CYP2D6 metabolizers are at risk
of r educed or abolished analgesic effects of
codeine, while people who are ultra-rapid metab-
olizers are at greater risk of opiate related side-
effects when given codeine at commonly used
therapeutic doses16.
Paracetamol (acetaminophen) is one of the most
widely used drugs for the treatment of both acute
and chronic pain. It has a unique position among
analgesic drugs, having a spectrum of action simi-
lar to that of NSAIDs, but with negligible anti-in-
flammatory and antirheumatic activities. The
mode of action of paracetamol has still not been
fully elucidated, but there are some evidences sup-
porting a central analgesic effect. It is now gener-
ally accepted that it inhibits COX-1 and particular-
ly COX-2 through metabolism by the peroxidase
function of these isoenzymes17,18. Recently, it was
discovered that paracetamol may act as a prodrug
by triggering the CB1receptor - mediated effects
of the cannabinoid system. At therapeutic doses,
majority of paracetamol is conjugated with glu-
curonic acid and, to a lesser extent, with sulphate
or cysteine. A fraction usually ranging from 5 to
15% is oxidized by CYP2E1, CYP1A2, CYP3A4,
166

•Par: saline solution p.o. for 7 days, where the
last dose on the 7th day was applied 30 minutes
before i.p. administration of paracetamol
•Cod R10: REO (10 mg/kg) p.o. for 7 days,
where the last dose on the 7th day was applied 30
minutes before i.p. administration of codeine
•Cod R20: REO (20 mg/kg) p.o. for 7 days,
where the last dose on the 7th day was applied 30
minutes before i.p. administration of codeine
•Par R1 0: REO (10 mg/kg) p.o. for 7 days,
where the last dose on the 7th day was applied
30 minutes before i.p. administration of parac-
etamol
•Par R2 0: REO (20 mg/kg) p.o. for 7 days,
where the last dose on the 7th day was applied
30 minutes before i.p. administration of parac-
etamol
Applied daily doses of REO for mice were 10
mg/kg and 20 mg/kg. Recommended human dai-
ly dose of REO of 40 mg/day for a male of ap-
proximately 70 kg weight7,21 was adapted for the
experimentation on mice. Each animal received
appropriate dose of REO in the volume of 10 ml
of emulsion per kg of body weight, by per os
gavage. The te ste d c ompounds, codeine hy-
dr ochlo ride (30 mg/ kg) or par acet amol (60
mg/kg), were administered intraperitoneally (i.p.)
30 min after the last REO intake. The control
group of animals received an equivalent volume
of saline solution. All experiments were carried
out during the daytime.
Hot Plate Test
The hot plate test was performed by placing
mice individually on hot plate and assessing their
response to the thermal stimulus. The tempera-
ture of the metal plate enclosed by plexiglas
walls was maintained at 52.5°C. The response
time was measured in seconds at which the ani-
mal licked or flinched one of the hind paws, or
jumped off the plate. To prevent tissue damage, a
cut-off time was used as a double value of laten-
cies measured before drug application. Response
latencies were first determined two times before
the application of the tested compound in order
to determine a pre-treatment response for each
mouse, and then 5, 10, 15, 20, 30, 40, 50, 60
minutes following the drug administration. After
responding or reaching the cut-off time, mice
were removed from the plate. Analgesic effect
determined in seconds was expressed as percent-
age of prolongation of measured reaction time
compared to control reaction time22.
167
Analgesic effects of rosemary essential oil
250°C and 300°C, respectively, and the column
temperature was programmed from 40 to 260°C
at a rate of 4°C/min. The flow rate of hydrogen
as a carrier gas was 1 ml/min. A sample of 1%
solution of the oil in ethanol (1 µl) was injected
in split mode (split ratio, 1:30). GC/MS analysis
was carried out using a Hewlett-Packard HP
G1800C series II GCD system under the same
analytical conditions as in GC/FID. The column
HP-5MS (30 m × 0.25 mm; film thickness 0.25
µm) and helium as a carrier gas were used in this
analysis. The system was operated in electron
ionization (EI) mode at 70 eV, in the mass (m/z)
range 40-450 Da.
Identification of essential oil constituents was
performed by comparison of obtained mass spec-
tra and retention indexes with those of reference
compounds or those from mass spectra libraries
and literature data. The quantitative analysis pro-
vided the percentage composition of the essential
oil components, calculated by FID peak area nor-
malization method.
Animals and Treatment
Experiments were carried out on adult, sexual-
ly mature NMRI mice of both sexes, weighing
25-35 g, which were obtained from the Veteri-
nary Institute Novi Sad, Serbia. Animals were
housed in standard laboratory cages at a con-
trolled temperature (23 ± 1°C) and humidity (55
± 1. 5 % ) under standard ci r c a d i a l r hythm
(day/night), with free access to pelleted food and
water. Animal care and experimental procedures
were conducted in accordance with the Guide for
the Care and Use of Laboratory Animals edited
by Commission of Life Sciences, National Re-
search Council (USA). The experimental proce-
dures were approved by Ethical Committee for
Animal Use in Experiments of the University of
Novi Sad (No. 01-153/6-2).
All animals were divided into 8 experimental
groups, each containing 6 individuals, and treat-
ed as follows:
•ConS: control group, saline solution p.o. for 7
days, where the last dose on the 7th day was
applied 30 minutes before i.p. administration
of saline solution
•REO: REO (20 mg/kg) p.o. for 7 days, where
the last dose on the 7th day was applied 30
minutes before i.p. administration of REO
•Cod: saline solution p.o. for 7 days, where the
last dose on the 7th day was applied 30 minutes
before i.p. administration of codeine

168
Statistical Analysis
The level of significance between the groups
was assessed with the Student’s t-test for small
independent samples using MedCalc 9.2.0.1 soft-
ware. All data are expressed as a mean ± stan-
dard deviation (SD). A value of p< 0.05 was
considered to be statistically significant.
Results
The essential oil obtained from rosemary had a
pale yellow color and a strong odor, and the ob-
tained yield of the essential oil was 1.03% (v/w
in dry matter). The total number of identified
ch emica l co nstit uents was 29, represe nting
99.87% of the total oil content. As presented in
Table I, the isolated essential oil contains a com-
plex mixture of 95.10% of monoterpenes and
4.77% of sesquiterpenes. It was found to be com-
po s ed ma i nly o f oxy g enated monot e rpenes
(63.88%), followed by monoterpene hydrocar-
bons (31.22%) and sesquiterpene hydrocarbons
(4.77%). The major compounds that were identi-
fied and quantitated by GC/FID and GC/MS
were 1,8-cineole (43.77%), camphor (12.53%),
α-pinene (11.51%), β-pinene (8.16%), camphene
(4.55%), and β-caryophyllene (3.93%).
In our study, REO (20 mg/kg) increased sig-
nificantly the latency time of animal response to
heat-induced pain between 20thand 50thminute
of the test, when compared to saline-treated
group (Table II). Codeine and paracetamol were
used as antinociceptive reference drugs. We
A. Raskovic, I. Milanovic, N. Pavlovic, B. Milijasevic, M. Ubavic, M. Mikov
Compounds RT-FIDaRT-MSbRRTcPercentage content (% m/m)
Monoterpene hydrocarbons 31.22
Tricyclene 12.640 6.82 0.579 0.23
α-Thujene 12.777 7.00 0.585 0.13
α-Pinene 13.100 7.19 0.600 11.51
Camphene 13.737 7.62 0.629 4.55
Sabinene 14.697 8.47 0.673 0.05
β-Pinene 14.887 8.50 0.681 8.16
β-Myrcene 15.292 9.04 0.700 0.99
α-Phellandrene 15.955 9.43 0.730 0.19
δ3-Carene 16.212 9.62 0.742 0.13
α-Terpinene 16.456 9.85 0.753 0.14
p-Cymene 16.782 10.13 0.768 1.23
Limonene 16.970 10.25 0.777 2.80
γ-Terpinene 18.178 11.30 0.832 0.92
α-Terpinolene 19.416 12.31 0.889 0.19
Oxygenated monoterpenes 63.88
1,8-cineole 17.124 10.43 0.784 43.77
Linalool 19.746 12.82 0.904 0.46
Camphor 21.845 14.30 1.000 12.53
Isoborneol 22.318 14.71 1.022 0.53
Borneol 22.655 15.05 1.037 2.97
Terpinen-4-ol 23.059 15.44 1.056 0.56
α-Terpineol 23.548 15.93 1.078 1.53
γ-Terpineol 23.802 16.17 1.090 0.40
Bornyl acetate 27.185 19.14 1.244 1.13
Sesquiterpene hydrocarbons 4.77
α-Copaene 30.598 22.03 1.401 0.12
Longifolene 31.866 22.93 1.459 0.18
β-Caryophyllene 32.240 23.41 1.476 3.93
α-Humulene 33.406 24.45 1.529 0.36
Germacrene D 34.029 25.18 1.558 0.08
δ-Cadinene 35.528 26.59 1.626 0.10
Total identified 99.87
Number of compunds identified 29
Table I. Chemical composition of the rosemary essential oil.
aRT-FID - retention time in GC/FID system; bRT-MS - retention time in GC/MS system; cRRT - relative retention time with re-
spect to camphor.

showed that analgesic effect of REO was slightly
higher than that of paracetamol, but significantly
lower than that of codeine, especially from 5th to
20th minute of the experiment.
As shown in Figure 1, the administration of
codeine in combination with REO in the dose of
20 mg/kg significantly supressed the termal pain
response of animals, and induced more pro-
nounced analgesic effect when compared to mice
treated only with codeine. On the other hand,
REO in the dose of 10 mg/kg slightly lowered
codeine analgesic effect.
We demonstrated that analgesic effect of REO
was comparable to those of paracetamol, alone
and in combinations with REO (Figure 2). The
administration of REO in the dose of 20 mg/kg
with paracetamol significantly prolonged the re-
action time of animals provoked by heat stimuli,
when compared to both saline- and paracetamol-
treated group, with a maximal response observed
between 30th and 50th minute of the test.
Discussion
In this research, we determined chemical com-
position of the studied REO, its analgesic activity,
and its potential to interact with opiod analgesic
codeine and non-opioid analgesic paracetamol.
Two main chemotypes of essential oils isolated
from rosemary have been reported considering the
chemical composition. The main component of
169
Analgesic effects of rosemary essential oil
Time [min]
Group 0510 15 20 30 40 50 60
ConS 10.9 ± 2.8 11.1 ± 2.5 9.9 ± 2.2 12.5 ± 3.4 10.1 ± 1.8 10.9 ± 2.8 10.3 ± 1.7 11.6 ± 1.6 11.9 ± 2.5
REO 10.9 ± 2.8 12.9 ± 3.3 13.8 ± 7.7 15.2 ± 3.0 14.2 ± 3.2* 14.7 ± 5.2 14.2 ± 6.5 15.6 ± 3.1* 14.4 ± 3.9
Cod 10.9 ± 2.8 20.1 ± 8.7* 20.8 ± 12.8* 21.1 ± 14.2 15.9 ± 5.1* 13.7 ± 4.8 15.4 ± 5.8* 15.7 ± 3.0* 14.5 ± 3.9
Par 10.9 ± 2.8 10.3 ± 1.1 12.4 ± 3.9 11.7 ± 2.3#11.1 ± 3.8 11.6 ± 2.5 12.4 ± 1.8* 13.3 ± 2.3 12.1 ± 5.1
Table II. Response latency times to the termal stimulus in seconds.
All values are expressed as mean ± standard deviation. *Significantly different from ConS group; #Significantly different from
REO group; p< 0.05.
Figure 1. Interactions of REO with codeine. Analgesic
effect is expressed as percentage of prolongation of mea-
sured reaction time compared to control reaction time.
Figure 2. Interactions of REO with paracetamol. Anal-
gesic effect is expressed as percentage of prolongation of
measured reaction time compared to control reaction time.

170
the Tunisian, Turkish, Moroccan and Italian oils is
1,8-cineole with usually over 40%, whereas most
Spanish, French and Greek oils have 1,8-cineole,
α-pinene and camphor with approximately equal
ratios (20-30%)8. The main constituents in the
rosemary essential oil investigated in our study
were 1,8-cineole (43.77%), camphor (12.53%),
and α-pinene (11.51%), and therefore it can be
categorized in the Morocco/Tunisian type. It
should be noted that there are several factors that
contribute to significant variations in the chemical
composition of rosemary essential oils, including
the geographic origin, part of the plant, season of
harvesting, hence the phenological stage of the
plant, and also the essential oil isolation method23.
All effects of REO should be, therefore, carefully
examined, considering the chemical composition
of the investigated oil.
In hot plate assay 7-day treatment with REO
(20 mg/kg) induced significant analgesic effects
in mice, which is in accordance with results of
previous studies. Analgesic activity of REO was
confirmed in different nociceptive experimental
models. The treatment with REO inhibited paw
edema induced by carrageenan in rats in a dose-
dependent manner, significantly reduced the
number of writhing movements induced by the
i.p. administration of acetic acid, and produced
an inhibition during both early (neurogenic pain)
and the late phase (inflammatory pain) of the for-
malin test, suggesting antinociceptive and anti-
inflammatory activity of REO2 4. Besides, REO
has also produced a dose-dependent antinocicep-
tive effect in the pain-induced functional impair-
ment model in the rat (PIFIR model)25. In addi-
tion to REO, analgesic effect was confirmed for
different monoterpenes that constitute more than
90% o f th e es sential oils26 , but als o fo r the
ethanol extract of rosemary27 and its major con-
stituent carnosol28, and triterpenes fractionated
from the rosemary extract29.
The observed analgesic effect of REO in hot
plate test in our study indicates its central mecha-
nism of analgesia, since both paw licking and
jumping are supraspinally integrated responses30.
Serotonergic and opioid endogenous systems were
suggested to be involved in the mechanism of ac-
tion as an antinociceptive of the essential oil isolat-
ed from rosemary25. Furthermore, the GABAergic
system may be another possible route involved in
the pharmacological activity of rosemary27.
Several studies using different nociceptive in
vivo models demonstrated analgesic activity of
ma ny monoter pene s31 . Al l th ree major con-
stituents of REO in our study (1,8-cineole, cam-
phor and α-pinene) were shown to exert anal-
gesic effects in acetic acid-induced writhing test
in mice32. The mechanisms of antinociceptive ef-
fects of monoterpenes are still to be elucidated,
but are suggested to involve different members of
the TRP channel family, as confirmed for cam-
phor33 . It is g enera lly assum ed t hat acyc lic
monoterpenes modulate the opioid system, while
monocyclic and bicyclic monoterpenes produce
analgesic effects mostly by peripheral
pathways26. The potent analgesic and anti-inflam-
matory activity of 1,8-cineole, as a dominant
component of REO, was shown to be mediated
through the inhibition of COX enzymes and sup-
pression of cytokines (TNF-αand IL-1β) pro-
duction as well34.
Although it possesses analgesic activity, REO
can also influence catalyti c activities of cy-
tochromes and, thus, efficacy of other analgesic
drugs. REO rich in 1,8-cineole was found to in-
duce catalytic activities of microsomal enzymes,
particularly CYP2B, but also slightly the activity
of UDP-glucuronosyltransferase (UGT), and
therefore may interact with codeine and paraceta-
mol14. The main REO component 1,8-cineole al-
so increased the levels of cytochromes CYP2B1
and 3A213. It was demonstrated that water ex-
tracts of rosemary significantly increased activity
of CYP2E1, 1A2 and 3A, which are involved in
formation of hepatotoxic metabolite of paraceta-
mol35,36. As shown in Figures 1 and 2, the admin-
istration of REO in combination with codeine
and paracetamol exerted distinct effects when ap-
plied in different doses. REO in the dose of 20
mg/kg was shown to be more efficient than in the
dose of 10 mg/kg, in combinations with both
codeine and paracetamol, which suggests that
dose determines whether REO will predominant-
ly induce cytochromes or act as analgesic and
have additive effect with administered antinoci-
ceptive drugs. This is in agreement with previous
findings that many monoterpenes do not exhibit
dose-dependent effects and that it is necessary to
find the most appropriate dose range that shows
effectiveness37.
Conclusions
REO possess es centrally actin g analgesi c
properties, as determined in hot plate assay, al-
though a more in-depth evaluation of the mecha-
nisms involved should be investigated. Our find-
A. Raskovic, I. Milanovic, N. Pavlovic, B. Milijasevic, M. Ubavic, M. Mikov

ings support the use of rosemary in the manage-
ment of pain, but also indicate a therapeutic po-
tential of REO in combination with analgesic
drugs. Considering nonlinear dose-response rela-
tionship of most monoterpenes, the appropriate
dose of REO has to be determined in order to ob-
tain an improved therapeutic effect without ad-
verse reactions due to interactions.
––––––––––––––––––––
Acknowledgements
This study was funded by Ministry of Education, Science
and Technological Development, Republic of Serbia (grant
No 41012) and by Provincial Secretariat for Science and
Technological Development, Autonomous Province of Vo-
jvodina (grant No 114-451-3551/2013-02). The authors are
grateful to Mr. Goran Anačkov, Department of Biology and
Ecology, Faculty of Sciences, University of Novi Sad, for
determination of plant material.
–––––––––––––––––-––––
Conflict of Interest
The Authors declare that there are no conflicts of interest.
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