Industrial Crops and Products 76 (2015) 809816

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Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

Chemical composition and yield of essential oils from Bakhtiari savory

(Satureja bachtiarica Bunge.) under different extraction methods
Sayadeh Mansoreh Memarzadeh a , Abdollah Ghasemi Pirbalouti b, ,
Mohammad AdibNejad a
a

Flavarjan Branch, Islamic Azad University, Department of Essential Oil Chemistry, Isfahan, Iran
Shahrekord Branch, Islamic Azad University, Research Center for Medicinal Plants & EthnoVeterinary, Department of Medicinal Plants, PO Box: 166,
Shahrekord, Iran
b

a r t i c l e

i n f o

Article history:
Received 1 April 2015
Received in revised form 26 July 2015
Accepted 28 July 2015
Keywords:
Essential oil
Chemical composition
Microwave-assisted hydro-diffusion
Innovative technique steam distillation
Microwave-assisted steam hydro-diffusion
Satureja bachtiarica

a b s t r a c t
Bakhtiari savory is a perennial aromatic herb distributed in Zagros mountain range, southwestern Iran. To
determine the effect of extraction methods on qualitative and quantitative characteristics of the essential
oil of Bakhtiari savory was extracted by different methods. Conventional methods [hydrodistillation by
two Clevenger-type apparatus: British Pharmacopeia (HDBP ) and Research Institute of Forests and Rangelands, Iran (HDRIFR ), and traditional steam and water distillation (TSWD)], an innovative technique steam
distillation (SDinnov ), microwave-assisted steam hydro-diffusion (MSHD 400 and 800 W), and microwaveassisted hydro-diffusion (MHD 400 and 800 W) were used to extract essential oil from the aerial parts of
savory and their results were compared. The essential oils of all samples were analyzed using GCFID,
GC/MS, and for comparison, a sample was analyzed using head space solid-phase microextraction (HSSPME). The highest essential oil yields were obtained from two methods, including HDBP , and MSHD
400 W. Signicant differences occurred among the major constituents in essential oils from extraction
methods, including thymol, carvacrol, -cymene, and -terpinene. The highest amounts of thymol and
carvacrol were obtained from MHD (800 W) method in comparison other methods. The highest content of
monoterpene hydrocarbons was obtained from HS-SPME, whereas the highest percentage of oxygenated
monoterpenes was achieved from MHD (800 W) method. In conclusion, extraction of the essential oil from
Bakhtiari savory with microwave-assisted hydro-diffusion (MHD) was better in terms of energy saving,
extraction time, oxygenated fraction (thymol and carvacrol), and product quality.
2015 Elsevier B.V. All rights reserved.

1. Introduction
The genus Satureja L. (savory) belonging to the family Lamiaceae contains 200 species of herbs and shrubs, often aromatic,
widely distributed in the Mediterranean area (Cronquist, 1988).
Satureja L. is represented by 14 species commonly observed among
the mountains in the southwestern part of Iran (Mozaffarian,
2008). A total of nine Satureja species have been reported in
ora of Iran (Mozaffarian, 2008), two of which, S. bachtiarica
Burge and S. kallarica Jamzad are distributed in southwestern Iran
(Ghasemi-Pirbalouti et al., 2014a). S. bachtiarica (Persian name:
Marzeh-e-Koohi) is well-known aromatic plant which is frequently used as spices and as traditional medicinal herbs in Iran. S.

Corresponding author. Fax: +98 381 336 1012.

E-mail addresses: ghasemi@iaushk.ac.ir, aghasemipir@psis.umass.edu
(A. Ghasemi Pirbalouti).
http://dx.doi.org/10.1016/j.indcrop.2015.07.068
0926-6690/ 2015 Elsevier B.V. All rights reserved.

bachtiarica is a perennial herb, which it distributed in Zagros mountain range, southwestern Iran (Ghasemi-Pirbalouti et al., 2013a).
Infusions and decoctions of aerial parts of S. bachtiarica are used for
the treatment of colds, and also are used as analgesic and antiseptic by the indigenous people of Bakhtiar va Chaharmahal province,
southwestern Iran (Ghasemi-Pirbalouti et al., 2014b). S. bachtiarica contains various biologically active components, especially
volatile compounds and monoterpenes (Sedkon and Jamzad,
2000). The essential oil isolated from the aerial parts of S. bachtiarica
has been shown to have biological and pharmacological activities, including antibacterial (Ghasemi-Pirbalouti and Dadfar, 2013;
Ghasemi-Pirbalouti et al., 2014b), antifungal (Ghasemi-Pirbalouti
et al., 2011a,b), antioxidant (Ghasemi-Pirbalouti et al., 2014c), and
immunomodulatory effects (Ghasemi-Pirbalouti et al., 2011c).
The conventional methods for the essential oil extraction of
herb materials, including hydrodistillation and steam distillation
have some disadvantages. Losses of some volatile constitutes, low
extraction efciency, degradation of unsaturated or ester com-

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pounds through thermal or hydrolytic effects and toxic solvent

residue in the extract may be encountered using these extraction methods (Tigrine-Kordjani et al., 2006; Prino-Issartier et al.,
2013). In addition, these methods have distinct drawbacks such
as time-consuming and labor intensive operations, handling of
large volumes of hazardous solvents (Khajeh, 2011). The development of new extraction techniques for the food and herbal
drugs industries has received a lot of attention lately due to the
environmental restrictions, reducing waste water, and the need
for minimizing the energy costs. In addition, there is a constant
demand to improve the quality of essential oils because consumers
demand this quality in their food, pharmaceutical or perfumery
products (Tigrine-Kordjani et al., 2006). However, in order to reduce
the extraction time, and the operation costs and possibly improve
the oil yield and the quality of the essential oil, new approaches
such as microwave-assisted extraction, supercritical uid extraction, and ultrasound-assisted extraction have also been sought
(Kaufmann and Christen, 2002). Driven by these goals, advances in
microwave extraction have resulted in a number of techniques such
as microwave steam distillation (Farhat et al., 2011), microwave
hydrodiffusion and gravity (Vian et al., 2008; Bousbia et al., 2009),
microwave steam diffusion (Farhat et al., 2009), microwave dry
distillation or microwave accelerated distillation (Tigrine-Kordjani
et al., 2006) and solvent free microwave extraction (Lucchesi et al.,
2007) being used. Microwave-mediated processes bring a number
of advantages to essential oil extraction thanks to their reduced
equipment size, ease-of-use, speed, ability to control a process
via mild increments in heating and low solvent consumption, all
of which contribute to reducing environmental impact and costs
(Prino-Issartier et al., 2013). Over the years procedures based on
microwave extraction have replaced some of the conventional processes and other thermal extraction techniques that have been
used in chemical laboratories for decades. Some recently studies
(Stashenko et al., 2004; Tigrine-Kordjani et al., 2006; Golmakani
and Rezaei, 2008; Prino-Issartier et al., 2013) have successfully
utilized a microwave oven for the extraction of active components from medicinal plants/herbs. However, to the best of authors
knowledge no work has been published on the microwave-assisted
of savory species. In this study, we present a comparative study of
the ability of a number of different methods to extract the essential
oils from Bakhtiari savory (Satureja bachtiarica Bunge.) in order to
nd the most advantageous in term of extraction kinetics, essential
oil quality, and quantity.
2. Material and methods
2.1. Plant material
The aerial parts (up to 5 cm, 100 g) of Bakhtiari savory (Satureja
bachtiarica Bunge.) were collected from Chaharmahal va Bakhtiari,
southwestern Iran (32 17 N and 50 33 E) about 2200 m above sea
level in July 2012. Plant identities were conrmed by Dr. Shrimardi
and voucher specimen (no. 1999) has been placed in the Herbarium of Research Center for Agricultural and Natural Resources of
Chaharmahal va Bakhtiari, Iran. The fresh aerial of S. bachtiarica
were dried inside for ve days at room temperature (30 5 C) in
dark conditions, and the ground to ne a powder using Endecott
food processor and passed through a 16 mesh sieve to remove large
pieces of debris.
2.2. Chemicals
Homologous series of C5 C24 n-alkanes (SigmaAldrich, Steineheim, Germany) were used for identication of all constituents by
calculation of the retention indexes. Anhydrous sodium sulphate

(Na2 SO4 ) for drying extracted oil was purchased from Merck Co.
(Darmstadt, Germany).
2.3. Extraction procedures
The extraction of the essential oils from S. bachtiarica was
performed using different methods, including three conventional
methods, three innovative techniques, and head space solid-phase
microextraction method. All the results are reported in grams of
essential oils per 100 g of dried savory aerial parts.
2.3.1. Hydrodistillation using two Clevenger apparatus
A 100 g of S. bachtiarica was submitted to hydrodistillation using
two Clevenger-type apparatus, including British Pharmacopeia
(HDBP ) (Fig. 1a) and Research Institute of Forests and Rangelands,
Iran (HDRIFR ) (Fig. 1b) and extracted with 1 L of water for 240 min
(until no more essential oil was obtained). The essential oil was
dried by anhydrous sulphate and stored at 4 C until analysis. Each
extraction was performed at least three times.
2.3.2. Traditional steam and water distillation procedure
The aerial parts of S. bachtiarica (100 g) were submitted to traditional steam and water distillation (TSWD). The vapor produced
by the steam generator passed through the essential oil rich plant
material before being condensed into a receiving Clevenger-type
apparatus (British Pharmacopeia approach) with 1 L of water for
240 min. The essential oil was obtained, and then dried under anhydrous sulphate. Each extraction was performed at least three times.
2.3.3. Steam distillation apparatus and procedure
The essential oil from the aerial parts of S. bachtiarica was
extracted by an innovative technique steam distillation (SDinnov )
that designed by Ghasemi Pirbalouti (Fig. 1c). The vapor produced
by the steam generator passed through the essential oil rich plant
material before being condensed into a receiving Clevenger-type
apparatus with 1 L of water for 240 min. The collected essential
oil was dried under anhydrous sulphate and stored at 4 C until
analysis. Each extraction was performed at least three times.
2.3.4. Microwave-assisted steam hydro-diffusion (MSHD)
apparatus and procedure
A domestic microwave oven (MG-4012 WM/00, LG, Korea) was
modied for MHD and MSHD operation. 100 g of S. bachtiarica that
had been soaked in 1L of distilled water for 30 min were placed in
a 1 L ask containing deionized water. The ask was setup within
the microwave oven cavity and a condenser was used on the top
(outside the oven) to collect the extracted essential oils. The vapor
produced by the microwave passed through the essential oil rich
plant material before being condensed into a receiving Clevengertype apparatus (Fig. 1d). The microwave oven was operated at 400
and 800 W power level for a period of 30 min. This period was sufcient to extract all the essential oils from the sample. During 30 min,
the collected essential oils were decanted from the condensate in
10 min intervals. To remove water, the extracted essential oils were
then dried over anhydrous sodium sulfate, weighed and stored in
amber vials at 4 C until they were used for analysis. Each extraction
was performed at least three times.
2.3.5. Microwave-assisted hydro-diffusion (MHD)
A 100 g of S. bachtiarica that had been soaked in 1 L of distilled
water for 30 min were placed in a 1 L ask containing deionized
water. The ask was setup within the microwave oven cavity and
a condenser was used on the top (outside the oven) to collect the
extracted essential oils (Fig. 1d). The microwave oven was operated
at 400 and 800 W power level for a period of 30 min. This period was
sufcient to extract all the essential oils from the sample. During

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

Fig. 1. (a) HDBP , (b) HDRIFR , (c) SDinnov , (d) MSHD and MHD.

811

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S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

30 min, the collected essential oils were decanted from the condensate in 10 min intervals. The essential oil was obtained, dried
under anhydrous sodium sulphate and stored at 4 C until analysis.
The extractions were performed at least three times, and the mean
values were reported.
2.3.6. Head space solid-phase microextraction (HS-SPME)
The samples were stored in sealed plastic bags and placed
in a cooler on ice until analysis by HS-SPME no more than ve
days after collection. The SPME device coated with divinyl benzene/carboxen/poly dimethyl siloxane (30 m and 20 mm length,
cross linked, bipolar) was used for extraction of volatile compounds. The ber was extended through the needle and exposed
to the head space above (2 g) of sample contained in a (30 mL glass
vial) for extraction 60 min at 40 C. The ber was then retracted and
inserted into the injector of the gas chromatograph coupled to mass
spectrometer equipped with a quadrupole mass analyzer, where
the metabolites were thermally desorbed in the splitless mode at
250 C for 6 min, and transferred directly to the analytical capillary column (HP-5MS 5% phenyl methyl silicone, 30 m 0.25 mm,
0.25 m lm thicknesses).
2.4. Identication of the oil constituents
Chemical composition of the essential oils was determined by
gas chromatography (GC) and mass spectrophotometry (GC/MS).
The GC analysis was done on an Agilent Technologies 7890 GC
(Agilent Technologies, Santa Clara, CA) equipped with a single injector and a ame ionization detector (FID). An apolar HP-5 capillary
column (30 m 0.25 mm, 0.25 m lm thicknesses) coated with 5%
phenyl, 95% methyl polysiloxane were used. The ow of the carrier
gas (N2) was 0.8 mL/min. Initial column temperature was 60 C and
programmed to increase at 4 C/min to 280 C. The injector temperature was set at 280 and 300 C. Split injection was conducted with
a ratio split of 1:40. Samples of the essential oil of 0.1 L were injected
neat.
GCMS analyses of the samples of the essential oil were
performed on an Agilent Technologies 7890 gas chromatograph
coupled to Agilent 5975C mass selective detector (MSD) and
quadrupole EI mass analyzer (Agilent Technologies, Palo Alto,
CA, USA). A HP5MS 5% column (coated with methyl silicone)
(30 m 0.25 mm, 0.25 m lm thicknesses) was used as the stationary phase. Helium was used as the carrier gas at 0.8 mL/min
ow rate. The temperature was programmed from 60 to 280 C at
4 C/min ramp rate. The injector and the GCMS interface temperatures were maintained at 290 C and 300 C, respectively. Mass
spectra were recorded at 70 eV. Mass range was from m/z 50550.
The ion source and the detector temperatures were maintained at
250 and 150 C, respectively.
Chemical compositions of the essential oils were identied
based on their retention indices (determined with reference to
homologous series of C5C24 n-alkanes), by comparison of their
mass spectra with those reported in the literature (Adams, 2007)
and stored in NIST 08 (National Institute of Standards and Technology) and Willey (ChemStation data system) libraries. The peak area
percentages were computed from HP-5 column without the use of
FID response factors.
2.5. Statistical analyses
The data was statistically analyzed using one-way ANOVA by
SPSS (19.0), and comparison of the means of the main constituents
and extraction yields for the essential oils was evaluated by Duncans multiple range test (p 0.05).

3. Results and discussion

3.1. Effect of extraction methods on the essential oil yield
All essential oils extracted from the aerial parts of Bakhtiari
savory (S. bachtiarica) by different extraction methods produced
a clear, yellow liquid. Results indicated that the extraction method
had a signicant effect on essential oil yield of Bakhtiari savory
(p 0.01) with the highest essential oil yields (v/w on dry weight
basis) obtained by HDBP and MSHD 400 W. The lowest essential oil
yields occurred by HDRIFR and MHD 400 W (Fig. 2). HDBP and MSHD
show similar oil yield: they reach maximal yields of 1.4 0.01
and 1.37 0.06 mL/100 g dry plant, in 4 h and 30 min extraction
times, respectively. HD is an approved method that is used as reference for the quantication of essential oils (Golmakani and Rezaei,
2008). Extraction with MSHD started at much earlier time than
that with HD. This is due to the more efcient heat ow involved
with microwaves. During microwave assisted extraction of secondary metabolites, such as essential oil from plants, microwave
rapidly delivers energy to extractant and plant matrix (Zhang
et al., 2011). Results of a study by Golmakani and Rezaei (2008)
indicated that microwave-assisted hydrodistillation destroyed the
glands of garden thyme (Thynus vulgaris L.) leaves in 30 min, but the
extraction by conventional hydrodistillation destroyed the glands
in about 75 min. Unlike the classical conductive heating methods,
microwaves can heat the entire sample almost simultaneously and
at a higher rate (Kaufmann and Christen, 2002). These results conrm results from the literature, which indicate that the use of
microwaves allows extractions to be accelerated (Golmakani and
Rezaei, 2008; Prino-Issartier et al., 2013). In addition, a result this
study indicated other microwave-assisted methods, does not have a
positive effect on gives a maximum yield (Fig. 2), whereas has a positive effect on extraction kinetics. These results mean a substantial
saving in time.
3.2. Effect of extraction methods on the essential oil composition
The identities of the extracted essential oils from different
extraction methods are shown in Table 1 . In total, the 49 components shown in Table 1 consist of about 96100% of total GC
peak area. The main components of the essential oils from different extraction methods were -cymene, -terpinene, linalool,
thymol, carvacrol, and -caryophyllene (Table 1). All essential oils
were found to be rich in the active monoterpene phenols (thymol
and carvacrol) and their corresponding monoterpene hydrocarbon precursors (-cymene and -terpinene). Thymol and carvacrol
were the most abundant components in S. bachtiarica essential
oils extracted using extraction methods expect head space solidphase microextraction (HS-SPME) method. This is in agreement
with the values previously reported for the same species, but
using conventional extraction method (Sedkon and Jamzad, 2000;
Ghasemi-Pirbalouti et al., 2013a; Ghasemi-Pirbalouti and Dadfar,
2013). A comparison of our results with the previous report by
researchers suggests few differences in the volatile composition
of the plant material could be attributed to the geographic origin
of the plant and to the methods of extraction (Mirhosseini et al.,
2015).
In this study, results of analysis of variance indicated that signicant differences occurred among the major constituents in the
essential oils from extraction methods. The highest percentages of
-cymene (32.7 0.49%) and -terpinene (23.54 1.09%) obtained
from and head space solid-phase microextraction (HS-SPME)
method, whereas the lowest percentages of thymol (6.69 0.31%)
and carvacrol (4.80 0.08%) obtained from this method. In addition,
results indicated that the percent of thymol (34.53 1.48%) and
carvacrol (31.66 0.61%) in the MHD (800 W) method were higher

Table 1
Effect of different extraction methods on chemical compositions of essential oils from the aerial parts of Bakhtiari savory (Satureja bachtiarica Bunge.).
a

Components

RI-cal

RI-lit HDBP

TSWD

SDinnov

HDRIFR

MHD400 W

MHD800 W

MSHD400 W

MSHD800 W

HS-SPME

ANOVA

927
934
949
979
991
1006
1012
1018
1027
1029
1036
1047
1059
1089
1131

931
936
953
980
991
1005
1011
1018
1026
1031
1040
1050
1062
1088
1129

0.286 0.038b
1.073 0.231
0.870 0.130
0.133 0.023
0.690 0.103
0.177 0.029
0.043 0.006
1.857 0.22b
14.043 1.75b
0.503 0.136
0.133 0.055
0.183 0.251
12.653 1.18b
0.217 0.021

32.861 4.17b

0.250 0.046
1.110 0.227
0.850 0.142
0.130 0.020
0.700 0.085
0.187 0.050
0.043 0.006
1.757 0.24b
14.570 1.81b
0.507 0.075
0.113 0.030
0.493 0.690
12.257 1.52b
0.220 0.020

33.187 4.97b

0.187 0.051
0.930 0.199
0.813 0.167
0.107 0.023
0.587 0.110
0.130 0.026
0.037 0.006
1.470 0.27bc
13.880 2.06b
0.413 0.072
0.077 0.015
0.057 0.015
10.163 1.8c
0.183 0.021

29.034 4.83bc

0.223 0.011
0.853 0.085
0.763 0.085
0.120 0.010
0.597 0.060
0.133 0.015
0.037 0.006
1.523 0.16bc
12.533 1.29b
0.427 0.040
0.077 0.015
0.070 0.010
10.573 0.69bc
0.170 0.030

28.099 2.49bc

0.170 0.014
0.750 0.042
0.705 0.021
0.110 0.000
0.575 0.021
0.120 0.000
0.035 0.007
1.425 0.0bc
13.975 0.11b
0.445 0.007
0.070 0.000
0.065 0.007
9.970 0.04c
0.180 0.000

28.595 0.27bc

0.095 0.007
0.420 0.042
0.400 0.056
0.060 0.000
0.340 0.000
0.080 0.000

0.920 0.0d
9.120 0.78c
0.270 0.000
0.045 0.007
0.040 0.000
7.210 0.38d
0.115 0.007

19.115 1.278d

0.143 0.030
0.610 0.121
0.623 0.105
0.090 0.010
0.503 0.023
0.113 0.006
0.030 0.000
1.290 0.043cd
12.806 0.38bc
0.380 0.026
0.070 0.010
0.053 0.006
9.350 0.23c
0.170 0.010

26.231 1cd

0.130 0.036
0.540 0.144
0.597 0.145
0.093 0.021
0.510 0.132
0.113 0.029
0.033 0.011
1.290 0.27 cd
12.887 2.61bc
0.387 0.051
0.080 0.017
0.063 0.011
9.747 1.41c
0.177 0.006

26.647 4.89cd

7.19 0.13
5.04 0.29
0.85 0.03
1.80 0.05
0.50 0.00

2.99 0.65a
32.7 3.98a

0.17 0.00
23.54 5.56a
0.54 0.00
0.41 0.03
75.73 9.45a

p > 0.05
p < 0.01
p < 0.01
p > 0.05
p < 0.01
p > 0.05
p > 0.05
p < 0.01
p < 0.05
p > 0.05
p > 0.05
p < 0.05
p < 0.01
p < 0.01
p < 0.01
p < 0.01

Oxygenated monoterpenes
Alcohols
16
Octan-3-ol
17
trans-Sabinene hydrate
18
Linalool
19
Borneol
Terpinene-4-ol
20
-Cymene-8-ol
21
-Terpineol
22
cis-Piperitol
23
24
Geraniol

996
1068
1101
1167
1179
1187
1192
1181
1257

993
1068
1098
1165
1177
1183
1189
1193
1255

0.067 0.021
2.717 0.185b
0.560 0.138
0.560 0.020
0.053 0.006
0.130 0.017

0.127 0.032

0.237 0.038
3.05 0.151ab
3.600 0.466
0.447 0.049

0.127 0.025

0.080 0.026

0.247 0.060
3.040 0.451ab
3.900 0.404
0.420 0.046
0.053 0.006
0.140 0.010

0.127 0.025

0.203 0.075
3.080 0.170ab
4.060 0.400
0.540 0.132

0.147 0.023

0.223 0.118

0.250 0.000
2.820 0.014b
3.370 0.014
0.365 0.035
0.050 0.014
0.150 0.042

0.095 0.021

0.160 0.000
1.855 0.162c
2.785 0.289
0.275 0.007

0.100 0.000

0.085 0.007

0.037 0.006
0.303 0.045
3.377 0.319a
3.950 0.23
0.430 0.026
0.050 0.010
0.127 0.006

0.113 0.025

0.033 0.006
0.310 0.010
3.357 0.262a
3.737 0.274
0.427 0.061
0.043 0.006
0.113 0.032

0.140 0.053

1.16 0.43
5.49 1.11
3.71 0.78

0.35 0.06

p < 0.05
p < 0.01
p < 0.05
p < 0.05
p < 0.05
p > 0.05
p > 0.05
p < 0.01
p > 0.05

Phenol
25
26

Thymol
Carvacrol

1296
1306

1290
1298

28.610 2.99bc 26.806 2.07c

24.980 1.07b 24.203 0.61b

28.510 2.38bc 28.063 1.52bc 30.285 0.13b

24.827 2.75b 26.087 2.55b 26.885 0.95b

34.525 1.48a
31.660 0.61a

31.190 0.53b
27.300 0.52b

30.990 1.88b
26.993 1.93b

6.69 1.11d
4.80 0.45c

p < 0.01
p < 0.01

Aldehydes
27

trans-Chrysanthemal

1151

0.060 0.000

0.063 0.006

p < 0.05

0.060 0.000

0.067 0.006

0.050 0.000

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

Monoterpene hydrocarbons
1
-Thujene
2
-Pinene
Camphene
3
-Pinene
4
Myrcene
5
6
-Phellandrene
7
3-Carene
8
-Terpinene
9
-Cymene
10
Limonene
cis--Ocimene
11
trans--Ocimene
12
-Terpinene
13
-Terpinolene
14
E,Z-Alloocimene
15

813

814

Table 1 (Continued)
a

TSWD

SDinnov

HDRIFR

MHD400 W

MHD800 W

MSHD400 W

MSHD800 W

HS-SPME

ANOVA

Geranial

1276

1270

0.037 0.006

0.060 0.017

p < 0.05

Ketone
29
30

Camphor
Carvone

1147
1246

1143
1242

0.047 0.006

0.130 0.028

0.043 0.006

p < 0.05
p < 0.05

Esters
31
32

Thymyl acetate
Carvacryl acetate

1355
1374

1355
1371

0.293 0.089
0.187 0.580

0.367 0.095
0.257 0.070

0.510 0.050
0.377 0.032

0.583 0.055
0.447 0.049

0.185 0.205
0.255 0.035

0.505 0.063
0.410 0.028

0.370 0.020
0.267 0.021

0.413 0.107
0.310 0.087

p > 0.05
p > 0.05

Ether
33
34

Thymyl methyl ether

1,8-Cineole

1244
1033

1235
1033

0.08 0.030

0.073 0.023

0.060 0.010

0.160 0.084

0.050 0.017
0.067 0.006

0.037 0.015
0.063 0.011

p > 0.05
p < 0.05

Epoxides
35

cis-Linalool oxide

1073

1074

58.363 5.17b

59.307 3.63b

62.271 6.23b

63.547 5.1b

65.050 1.58b

72.360 2.64a

67.737 1.8c

0.033 0.006
67.059 4.75c

22.2 3.11d

p < 0.05
p < 0.01

1422
1442
1464
1456
1510
1544
1387
1412
1484
1526

1418
1439
1461
1452
1509
1536
1384
1409
1480
1524

2.713 0.37b
0.097 0.015

0.160 0.036

3.870 0.51a
0.147 0.025

0.243 0.071
0.253 0.060

4.390 0.23a
0.167 0.006
0.077 0.011
0.340 0.026
0.310 0.035

3.023 0.14 b

0.087 0.032
0.203 0.011
0.200 0.026

2.355 0.19b
0.075 0.007

0.155 0.007
0.130 0.014

3.060 0.81b
0.105 0.035

0.230 0.042
0.210 0.070

2.250 0.02b
0.073 0.006

0.150 0.000
0.120 0.000

2.617 0.85b
0.093 0.032

0.220 0.091
0.157 0.073

2.08 0.22b

p < 0.01
p > 0.05
p > 0.05
p > 0.05
p > 0.05

0.043 0.006

0.050 0.010
3.063 0.44abc

0.073 0.011

0.073 0.011
4.659 0.68a

0.080 0.000

0.080 0.010
5.444 0.32a

0.047 0.006

0.053 0.006
3.613 0.22ab

0.065 0.049
0.040 0.000
0.065 0.049
0.075 0.035
2.960 0.35d

0.050 0.014

0.080 0.042
3.735 1.01bc

0.030 0.000

0.037 0.006
2.66 0.03cd

0.043 0.015

0.027 0.011
3.157 1.07cd

2.08 0.41d

p > 0.05
p > 0.05
p < 0.05
p > 0.05
p < 0.01

Sesquiterpene hydrocarbons
-Caryophyllene
36
Aromadendrene
37
Alloaromadendrene
38
-Humulene
39
-Bisabolene
40
41
cis--Bisabolene
-Bourbonene
42
-Gurjunene
43
44
Germacrene-d
45
-Cadinene

RI-cal

RI-lit

Oxygenated sesquiterpenes
Alcohol
46
(+) Spathulenol

1581

1576

0.083 0.035

0.083 0.038

0.160 0.030

0.223 0.144

0.090 0.000

0.290 0.113

0.107 0.035

0.120 0.070

Epoxide
47

Caryophyllene oxide

1587

1581

0.467 0.150
0.550 0.19c

0.590 0.219
0.673 0.26c

1.090 0.166
1.250 0.19abc

1.283 0.710
1.506 0.85 ab

0.595 0.007
0.685 0.01c

1.595 0.445
1.885 0.56a

0.713 0.175
0.820 0.21bc

0.810 0.410
0.93 0.48bc

p < 0.01
p < 0.05

Tricyclene
Leden

923
1498

926
1490

0.033 0.006

94.87

0.033 0.006
0.137 0.011
98

0.037 0.006
0.143 0.006
98.18

0.030 0.000
0.127 0.055
96.92

0.030 0.00
0.095 0.035
97.41

0.105 0.035
97.2

0.023 0.006
0.063 0.006
97.53

0.080 0.036
97.97

99.99

p > 0.05
p < 0.05

Other
48
49
Total
a
b

RI (cal): retention indices determined on HP-5MS capillary column and RI (lit): retention indices of literature values.
Mean with different letter in a row are statistically signicant at 5% level probability), according to Duncans multiple range test. Values of major compounds are given as means SD.

p < 0.05

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

HDBP

28

Components

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

815

Fig. 2. Comparison of essential oil yield of savory (v/w on dry weight basis) by the extraction methods.
Means with common letters are not signicant (p 0.05), according to Duncans multiple range test.

10
9

SDinnove

8
Energy (Kwh)

HDBP

SWDBP

240

240

HDRIFR

7
6
5
4
3
2
1

MSHD400

MHD400

40

40

MSHD800

MHD800

40

40

0
240

240

Different extraction methods-time (min)

Fig. 3. Energy consumption in different extraction methods.

than other methods (Table 1). The MHD is more selective than conventional hydrodistillation. Furthermore, the MHD is less tedious
and minimizes the risk of compound degradation due to heat. The
MHD also presents some advantages over conventional hydrodistillation such as extraction rate. Therefore, microwave does not
involve in any deterioration of the extracted components and it
can be introduced as a safe method for the extraction of essential
oils.
Our results demonstrated that different extraction methods
could signicantly change (p < 0.01) the chemical proles of essential oils from the aerial parts of S. bachtiarica. These variations occur
from losses or increase in oil constituents due to the formation
of new compounds by oxidation, glycoside hydrolysis, esterication, and/or other processer (Ghasemi-Pirbalouti et al., 2013b). The
highest content of monoterpene hydrocarbons was obtained from
HS-SPME method (75.73 9.45%), whereas the lowest percentages
of monoterpene hydrocarbons were achieved from microwaveassisted methods (Table 1). This results show that the monoterpene
hydrocarbons had more susceptible than other chemical groups to
high temperature during the extraction process. In Table 1, it can
be seen that the essential oils obtained from HDBP , HDRIFR , TSWD,
SDinnov , and MHD (400 W), MSHD (400 W), and MSHD (800 W)
methods share similar compositions, except for the MHD (800 W)

and HS-SPME methods. In MHD (800 W) method, the amounts of

-cymene, -terpinene, and linalool were lower than other methods. These differences are probably due to the degradation of these
compounds of monoterpene hydrocarbons into phenol components, including thymol and carvacrol (Table 1). This result was
in agreement with many other studies (Bousbia et al., 2009; Okoh
et al., 2010) reported on other plant species, which the amounts
of oxygenated compounds in the essential oils of the aromatic
plants (Rosmarinus ofcinalis L.) isolated by microwave-assisted
were higher in comparison with conventional hydrodistillation.
Compounds with high and low dipolar moments could be extracted
in various proportions by microwave extraction. Organic compounds like oxygenated monoterpenes that have a high dipolar
moment will interact more vigorously with microwaves and can be
extracted more easily in contrast with aromatic compounds which
have low dipolar moments, such as monoterpene hydrocarbons
(Bousbia et al., 2009).
3.3. Cost and energy
The reduced cost of extraction is clearly advantageous for the
proposed MHD and MSHD methods in terms of time and energy.
Hydrodistillation and steam distillation using HDBP , HDRIFR , TSWD,

816

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

and SDinnov required an extraction time of 60 min for heating 1 L of

water and 100 g of savory to the extraction temperature, followed
by evaporation of water and essential oil for 180 min. The MAD and
MSHD methods required heating for 5 min only and evaporation for
25 min of the in situ water and essential oil of savory. The energy
required to perform the different extraction methods shown in
Fig. 3. The power consumption was determined with a Wattmeter
at the microwave generator entrance and the electrical heater
power supply. In addition, microwave-assisted hydro-diffusion is
a very clean method, which avoids residue generation and the use
of a large quantity of water and voluminous extraction vessels.
4. Conclusion
Extraction is one of the crucial steps for research and development of plant secondary metabolites, such as avonoids, quinones,
phenylpropanoids, terpenoids and alkaloids. Microwave assisted
extraction is one of the important techniques for extracting valuable compounds from herbs, and it is quite adaptable on a small
or large scale (i.e., on a laboratory or industry scale). The method
which gave the best results was the microwave hydro-diffusion
(MHD) method which gave reduced extraction time (30 min against
240 min for conventional hydro-distillation and steam distillation methods) and gave no differences in essential oil yield. The
essential oils from savory have been extracted using seven different methods and a comparison of the different conventional
and innovative techniques has been carried out. The characteristics analyzed were extraction time, essential oil composition,
power consumption and yield. Results identied the optimal
extraction technique as being microwave hydro-diffusion (MHD).
Indeed, it gave the maximum yield (1.4% v/w) in only 30 min
(240 min for conventional hydro-distillation and steam distillation methods) and consumed 0.30.5 kWh (against 6.68.3 kWh for
conventional hydro-distillation and steam distillation methods).
Generally, extraction of the essential oil from S. bachtiarica with
microwave-assisted hydro-diffusion (MHD) was better in terms of
energy saving, extraction time, oxygenated fraction (thymol and
carvacrol), and product quality.
Acknowledgments
The authors would like to acknowledge Mr. Behzad Hamedi for
his technical assistance in essential oil extraction. This work was
supported by Deputy of Research & Technology, Islamic Azad University, Iran.
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