Original Article

Natural Product Communications

Effects of Inhalation of Geranium Essential Volume 14(10): 1–4

© The Author(s) 2019

Oil on Blood Pressure and Heart Rate

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DOI: 10.1177/1934578X19881534
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Rhuichiro Masubuchi1, Saori Watanabe1, and Tadaaki Satou1

Abstract
For a better scientific understanding of the basics of aromatherapy, our research group attempted to clarify the effectiveness of the
essential oil from Pelargonium graveolens (EOPG, geranium essential oil), which is used to try and balance the mind-body connection.
In order to eliminate any possible placebo effect, we used animal experiments that are considered to be insensitive to the placebo
effect. Measurements of blood pressure and heart rate in the mouse tail artery were used as a reflection of the mind-body balance.
Thirty minutes after inhalation of EOPG (5 µL/L air) for 90 minutes, blood pressure and heart rate of the mice were measured.
EOPG significantly reduced blood pressure and heart rate. To further clarify the factors responsible for these effects, gas chroma-
tography analysis was performed in order to determine the components transferred into the brain after the EOPG inhalation.
Linalool, citronellol, and geraniol were detected at concentrations around 0.1 nL/L tissue from the brain after 10 µL/L air inhala-
tion of EOPG. However, these were not detected after a 5 µL/L air inhalation of EOPG, as the levels were below the detection
limit. These results suggest EOPG inhalation might lower blood pressure and heart rate, with the expressed effects associated with
the transfer of components such as linalool into the brain.

Keywords
monoterpene, volatile compounds, autonomic nerve, blood brain barrier, noninvasive blood pressure monitor

Received: July 25th, 2019; Accepted: August 27th, 2019.

Although aromatherapy has been the subject of research for a
long time, scientific elucidation of the mechanisms involved
has yet to be definitively established. Our research group has
recently been conducting basic studies for the purpose of
determining a scientific method for elucidating mechanisms of
aromatherapy.1,2 One of the factors that makes aromatherapy
research difficult is the placebo effect caused by the aroma. To
eliminate this placebo effect, we performed experiments using
an animal model that made it possible to evaluate the scientific
mechanism of aromatherapy.
Aromatherapy consists of both inhalation and treatment.
There are 2 mechanisms of action involved with aromatherapy
inhalation. These include olfactory stimulation (neurotransmis-
sion) pathways and pathways that specifically enter the body,
such as the brain (pharmacological transmission). Our research
group has been focusing on the pharmacological transmission,
as this is thought to be both persistent and potent. Our current
study additionally focused on the route of pharmacological
transmission.
Studies of essential oil from Pelargonium graveolens (EOPG,
geranium essential oil) have suggested that EOPG can be used
to balance the mind-body connection. In the current study,
assessment of the mind-body balance was performed using
measurements of the blood pressure and heart rate in mice.
EOPG has been reported to have antiviral, antibacterial,
and antifungal activities. Inhibitory effects of EOPG and
essential oil from Cymbopogon citratus have been reported for the
Ross River virus3; antioxidant and antibacterial activities have
been shown for the essential oil from Pelargonium asperum and
Ormenis mixta4; antifungal activity against pathogenic Candida
strains has been shown for the essential oil from Cinnamomum
verum, Thymus capitatus, Syzygium aromaticum, and Pelargonium gra-
veolens5; and protective antifungal activity has been reported for
the essential oils from Buddleja perfoliata and Pelargonium graveo-
lens,6 among others. However, there have yet to be any basic
1
Department of Pharmaceutical Sciences, International University of Health
and Welfare, Kitakanemaru, Otawara City, Tochigi, Japan
Corresponding Author:
Tadaaki Satou, Department of Pharmaceutical Sciences, International
University of Health and Welfare, 2600-1 Kitakanemaru, Otawara City,
Tochigi 324-8501, Japan.
Email: ​tsatou@​iuhw.​ac.​jp

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2 Natural Product Communications

Figure 1.  Blood pressure (a) and heart rate (b) after EOPG
inhalation (5 µL/L air). Values are mean ± SE, n = 8. *P < 0.05.
EOPG, essentialoil from Pelargonium graveolens.

Figure 2.  Brain transferability of major components in inhaled

EOPG (10 µL/L air). Values are mean ± SE, n = 3. EOPG,
research studies that have examined the effect of EOPG on essentialoil from Pelargonium graveolens.
the mind-body balance.
Presently, there has been no definitive overall mechanism
established for the basis of aromatherapy. However, mecha-
nisms of action have been reported for essential oils adminis-
tered by inhalation, for the neurotransmission pathways
associated with olfactory stimulation,7 for the pharmacological
transduction pathways associated with the periphery,8,9 and for
the pharmacological transduction pathways associated with the
central nervous system.1,2 Our current report presents infor-
mation on a potential mechanism of action for essential oils in
aromatherapy.
Figure 3.  Blood pressure (a) and heart rate (b) after inhalation of
linalool (5 µL/L air). Values are mean ± SE, n = 8.
Experiment 1
The EOPG components tested included citronellol (21.9%),
geraniol (13.3%), citronellyl formate (8.6%), and linalool
tissue), and geraniol (0.097 ± 0.029 nL/g tissue) were detected
(6.3%). Mice were lightly stressed by being individually housed
in the brain after inhalation of 10 µL/L air EOPG (Figure 2).
for 1 to 3 days. Individual housing is carried out so that individ-
ual differences do not occur during the housing. After mice
inhaled EOPG (5 µL/L air) for 90 minutes, the blood pressure Experiment 3
and heart rate were measured using the caudal artery at 30 min-
The results of Experiment 2 clearly indicated that linalool, cit-
utes after the administration of the anesthesia. Purified water
ronellol, and geraniol were transferred into the brain after inha-
was used as a negative control. Thus, EOPG inhalation resulted
lation of EOPG. Since the results for linalool were the most
in significant reductions in the blood pressure and heart rate in
stably detected from the brain, we examined the effects of lin-
mice as compared to the inhalation of purified water (Figure 1).
alool inhalation on the blood pressure and heart rate. Under
the same conditions as used in Experiment 1, we examined the
influence of inhalation of linalool (5 µL/L air) on the blood
Experiment 2 pressure and heart rate of mice. Although the inhalation of
The results of Experiment 1 were considered to indicate the linalool (5 µL/L air) exhibited no significant differences as
intracerebral transfer of the component, thereby making it compared to the negative control, purified water, it tended to
possible to examine the intracerebral transferability of the decrease both blood pressure and heart rate (Figure 3).
component. Similar to Experiment 1, EOPG was also inhaled Current experimental results show that EOPG lowered
for 90 minutes, immediately thereafter the brain was removed blood pressure and heart rate in mice. In addition, results sug-
and analyzed by gas chromatography (GC) for its hexane gested that the effect occurred due to the transfer of compo-
extract. Although none of the components could be detected nents such as linalool into the brain. Thus, EOPG is thought to
in the brain after inhalation of 5 µL/L air EOPG, linalool regulate the autonomic nervous system, as EOPG inhalation
(0.095 ± 0.002 nL/g tissue), citronellol (0.071 ± 0.001 nL/g lowers both the blood pressure and heart rate. These results
Masubuchi et al. 3

can be considered to be scientific evidence documenting the
effect of EOPG on the mind-body balance.
Prior to this study, there has been little research on the
effects of EOPG on the blood pressure and heart rate.
However, there have been several reports on linalool. For
example, clinical studies on the inhalation of linalool have doc-
umented antioxidant activity and reductions in the blood pres-
sure and heart rate.10 Furthermore, it has also been reported
that olfactory stimulation affects the anxiolytic-like activity of
linalool after inhalation in mice.7 Linalool and essential oil from
Citrus bergamia have been reported to induce vasorelaxation in
mice.8,9 Other previous studies have suggested that the inhala-
tion effect of essential oils is mainly dependent on the transfer
of individual components into the brain.1,2 Anxiolytic-like
effects of intraperitoneal administration of lavender essential
oil and linalool have also been reported.11
Although we can conclude that transfer of the components
to the brain was responsible for observed changes in our
experimental study, we cannot completely rule out the poten-
tial effects of olfactory stimulation and changes in the periph-
eral blood vessels. A further detailed study will need to be
performed in the future. Our current and expected future find-
ings suggest that EOPG may be able to be clinically applied for
therapeutic treatments in patients.
Experimental
Analysis Using GC
Component analysis was performed using GC-MS-QP2010
(Shimadzu Corporation, Kyoto, Japan) and GC-2010
(Shimadzu Corporation, Kyoto, Japan). Samples were prepared
by diluting EOPG with n-hexane. The capillary column for
analysis is a DB-5ms capillary column (30 m x 0.25 mm ID,
0.25 µm, non-polar column; Agilent Technologies Inc., Tokyo,
Japan). Helium (99.99995%, 1.82 mL/min) was used as the
carrier gas. Inlet line temperature and source temperature are
250°C. The temperature of column oven is 40°C for 2 minutes,
40°C to 200°C at 5°C/min, and then 200°C for 2 minutes. The
voltage of electron impact (electron ionization) was 70 eV.
Components were detected by comparing standard com-
pounds or the GC-MS NIST 02 spectral library (National
Institute of Standards and Technology, Gaithersburg, MD,
USA) and retention indices.12 GC-FID was analyzed under the
same conditions as GC-MS.
Essential Oil and Components
EOPG extracted from the leaves of Pelargonium graveolens
(Geraniaceae) by steam distillation was purchased from Green
Flask Co., Ltd. (Tokyo, Japan). The components of EOPG pri-
marily include citronellol (21.9%), geraniol (13.3%), citronellyl
formate (8.6%), and linalool (6.3%) (Table 1).
Animals
Male ICR mice (Kumagai-Shigeyasu Co., Ltd., Miyagi, Japan)
that were 5 weeks of age at the start of each experiment were
used in the study. All mice were individually housed in cages
for 1 to 3 days. The cages were placed in a room that was arti-
ficially illuminated by fluorescent lamps on a 12 hours light and
dark schedule (light period from 8:00 to 20:00) and maintained
at 24°C ± 5°C. The mice had free access to food (Labo MR

Table 1.  Concentration of the Components of EOPG.

LRI LRI Concentration
(measurement) (reference) Name (mL/L)
1 986 988 β-Myrcene 3.6
2 1024 1024 Limonene 10.1
3 1102 1095 Linalool 63.0
4 1111 1107 Phenyl ethyl alcohol 2.9
5 1152 1148 Menthone 7.5
6 1194 1186 α-Terpineol 2.3
7 1233 1223 Citronellol 219.0
8 1256 1249 Geraniol 133.0
9 1273 1271 Citronellyl formate 86.0
10 1297 1298 Geranyl formate 71.1
11 1376 1379 Geranyl acetate 26.7
12 1445 1452 Neryl propionate 12.5
13 1467 1476 Geranyl propionate 21.3
14 1522 1530 Citronellyl butanoate 3.3
15 1618 1624 Citronellyl pentanoate 9.3
16 1660 1670 Bulnesol 12.7
EOPG, essential oil from Pelargonium graveolens; LRI, Linear retention index.
(-)-Linalool was purchased from Tokyo Chemical Industry (Tokyo, Japan).
4 Natural Product Communications
Stock, Nosan Corporation, Kanagawa, Japan) and water. One
mouse is used only for 1 experiment. The number of mice
used in this experiment is 41. For this experiment, it was
assumed that mice that were independently housed experi-
enced more stress as compared to mice kept in group cages. All
experiments were conducted in accordance with the guidelines
regarding the care of experimental animals as approved by the
Animal Research Committee at the International University of
Health and Welfare.
Procedure
A mouse was placed in a stainless-steel container (10 L, φ240 ×
240 mm, As One Corporation, Osaka, Japan). A filter paper
(GE Healthcare Japan, Tokyo, Japan) soaked in either EOPG
(5, 10 µL/L air), linalool (5 µL/L air), or water (5 µL/L air) was
placed on the upper side of the stainless-steel container. Mice
inhaled EOPG, linalool, or water for 90 minutes. The tempera-
ture in the room was 24°C ± 5°C.
Three types of mixed anesthesia (IP) were used. The tem-
perature of the mouse was maintained at 37°C, and the blood
pressure and heart rate were measured after 30 minutes using a
noninvasive blood pressure monitor (MK-2000, Muromachi
Kikai, Tokyo, Japan) (n = 8). The 3 types of anesthesia used in
the mixture were medetomidine hydrochloride (0.3 mg/kg
mouse, Nippon Zenyaku Kogyo Co., Ltd., Fukushima, Japan),
butorphanol tartrate (5 mg/kg mouse, Meiji Seika Pharma Co.,
Ltd., Tokyo, Japan), and midazolam (4 mg/kg mouse, Astellas
Pharma Inc., Tokyo, Japan).13
Quantitative Analysis of the EOPG Components in the
Brain
As linalool, citronellol, and geraniol were detected in the brain
after inhalation of EOPG, a measurement index was con-
structed based on linalool. Immediately after inhalation, the
mice were decapitated, and whole brains were removed (n = 3).
Each brain was homogenized in 1 mL n-hexane with an ultra-
sonic crusher (THU-80, As One Corporation). This GC analy-
sis was similar to the analysis performed for the EOPG
components. The obtained results are quantitatively calculated
for wet brain using the standard compound linalool.
Statistical Analysis
Results are expressed as the means ± SE (n = 8). Statistical
differences were determined by a 2-sided Student’s t-test using
a bell curve in Excel (Social Survey Research Information Co.,
Ltd., Tokyo, Japan). Differences with P < 0.05 were considered
significant.
Acknowledgments
We are grateful to Mr Shinichiro Hayashi of Green Flask Laboratory
for providing the EOPG.
Declaration of Conflicting Interests
The author(s) declared no potential conflicts of interest with respect
to the research, authorship, and/or publication of this article.
Funding
The author(s) received no financial support for the research,
authorship, and/or publication of this article.
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