Journal of Essential Oil Research

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Quali-quantitative variation of essential oil from

Iranian rosemary (Rosmarinus officinalis L.)
accessions according to environmental factors

Iman Bajalan, Razieh Rouzbahani, Abdollah Ghasemi Pirbalouti & Filippo

Maggi

To cite this article: Iman Bajalan, Razieh Rouzbahani, Abdollah Ghasemi Pirbalouti & Filippo
Maggi (2017): Quali-quantitative variation of essential oil from Iranian rosemary (Rosmarinus
officinalis L.) accessions according to environmental factors, Journal of Essential Oil Research,
DOI: 10.1080/10412905.2017.1380542

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Published online: 23 Oct 2017.

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 Journal of Essential Oil Research, 2017
https://doi.org/10.1080/10412905.2017.1380542

Quali-quantitative variation of essential oil from Iranian rosemary (Rosmarinus

officinalis L.) accessions according to environmental factors
Iman Bajalana, Razieh Rouzbahania, Abdollah Ghasemi Pirbaloutib,c and Filippo Maggid
a
Young Researchers and Elite Club, Borujerd Branch, Islamic Azad University, Borujerd, Iran; bMedicinal Plants Department, Shahrekord Branch,
Islamic Azad University, Shahrekord, Iran; cMedicinal Plants Program, College of Natural Sciences, Massachusetts University, Amherst, MA, USA;
d
School of Pharmacy, University of Camerino, Camerino, Italy

ABSTRACT ARTICLE HISTORY

The aim of this study was to assess environmental factors effects on quality and quantity of Iranian Received 23 February 2017
Downloaded by [The Library at Queen's University] at 23:55 24 October 2017

rosemary oil. Chemical components were identified by GC-FID and GC/MS. Results indicated that Accepted 10 September 2017
essential oil yields exhibited significant variation among accessions. The highest contents were KEYWORDS
recorded for the Bo2 and Bo3 accessions (2.6 and 2.3 ml/100 g, respectively). The major constituents Rosmarinus officinalis L.;
were 1,8-cineole (5.32–28.29%), camphor (1.58–25.32%) and α-pinene (14.19–21.43%). The Iranian environmental factors;
accessions also exhibited chemical variability for other major compounds such as borneol, piperitone, 1,8-cineole; correspondence
camphene, bornyl acetate, verbenone, and trans-pinocamphone. According to multivariate analysis, analyses; principal
including CA, HCA, DCA, and PCA the rosemary accessions belonged to two main chemotypes component analysis;
(I camphor type and II α-pinene/1,8-cineole/camphor type). Results of CA and DCA analyzes were correlation
similar and both of them were in agreement with the results of PCA analyzes. Furthermore, there
were positive and negative correlations between the major constituents and environmental factors.

Introduction essential oil possesses a wide range of biological activi-

Natural products derived from aromatic and medicinal
plants have been widely used in the food, pharmaceuti-
cal, cosmetic and perfumery industries. They have been
largely employed for their antimicrobial, fungicidal, anti-
viral, insecticidal and antioxidant activities (1).
Rosemary (Rosmarinus officinalis L.), belonging to
the Lamiaceae family, is a popular aromatic and medici-
nal plant (2). Some authors consider it as the most used
plant worldwide because of its essential oil and phenolic
compounds which are endowed with important biolog-
ical properties (3). Rosemary essential oil is obtained by
steam or hydrodistillation of twigs, leaves and inflores-
cences. According to the literature, its content may vary
depending on the harvesting time and extraction tech-
niques (4).
Studies on the chemical compositions and biologi-
cal activity of rosemary extracts using the aerial parts
collected in different stages of the phenological cycle
have been performed. In these studies, the quality of
the essential oils was significantly influenced by the geo-
graphic origin, environmental conditions and develop-
mental stage of collected plant materials (5,6). Rosemary
ties (7,8). According to our knowledge, there are limited
reports on the variation of content and chemical compo-
sition in the essential oil from different populations of
Iranian rosemary. Thus, the present study was designed
to quantify the variation in chemical constituents and oil
yield of rosemary accessions from various geographical
regions of Western Iran and to assess their relationships
with the environmental factors characterizing the dif-
ferent ecosystems.
Materials and methods
Plant material and site description
The leaves of 21 rosemary accessions, belonging to
seven rosemary populations, were harvested accord-
ing to nested method (each sample was the leaves of
three individual plants in its location) at flowering
stage from the Zagros Mountains rang, western prov-
inces of Iran in April 2015 (Table 1). A voucher speci-
men, representative for each population, was deposited
in the herbarium of I. A. U. (Borujerd Branch) from
Iran. All the plants growing in the regions exhibited

CONTACT Iman Bajalan I.bajalan@iaub.ac.ir

© 2017 Informa UK Limited, trading as Taylor & Francis Group
 2  I. BAJALAN ET AL.

Table 1. Collection site, geographical and soil characteristics of different Rosmarinus officinalis accessions.
Latitude and
Longitude Altitude Nitrogen phosphorus potassium E. C.b
No. A. n. a
Region (UTM) (m a.s.l) CaO (%) (%) (mg/kg) (mg/kg) pH (dS m−1) Soil texture
1 Do1 Dorud, Lorestan 322029 E 1455 16.6 0.11 10.1 389 7.6 0.73 Clay loam
Province, Iran 3707478 N
2 Do2 Dorud, Lorestan 322029 E 1455 16.6 0.11 10.1 389 7.6 0.73 Clay loam
Province, Iran 3707478 N
3 Do3 Dorud, Lorestan 322029 E 1455 16.6 0.11 10.1 389 7.6 0.73 Clay loam
Province, Iran 3707478 N
4 Kh1 Khoramabad, 261185 E 1368 17.4 0.12 5.8 645 7.7 1.35 Clay loam
Lorestan 3707356 N
Province, Iran
5 Kh2 Khoramabad, 261185 E 1368 17.4 0.12 5.8 645 7.7 1.35 Clay loam
Lorestan 3707356 N
Province, Iran
6 Kh3 Khoramabad, 261185 E 1368 17.4 0.12 5.8 645 7.7 1.35 Clay loam
Lorestan 3707356 N
Province, Iran
7 Bo1 Borujerd, 290028 E 1581 15.2 0.19 12.8 447 7.4 0.96 Clay loam
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Lorestan 3753829 N
Province, Iran
8 Bo2 Borujerd, 290028 E 1581 15.2 0.19 12.8 447 7.4 0.96 Clay loam
Lorestan 3753829 N
Province, Iran
9 Bo3 Borujerd, 290028 E 1581 15.2 0.19 12.8 447 7.4 0.96 Clay loam
Lorestan 3753829 N
Province, Iran
10 Ma1 Malayer, 302088 E 1840 14.6 0.14 11.3 352 7.7 0.72 Silty clay
Hamedan 3793470 N
Province, Iran
11 Ma2 Malayer, 302088 E 1840 14.6 0.14 11.3 352 7.7 0.72 Silty clay
Hamedan 3793470 N
Province, Iran
12 Ma3 Malayer, 302088 E 1840 14.6 0.14 11.3 352 7.7 0.72 Silty clay
Hamedan 3793470 N
Province, Iran
13 Jo1 Jowkar, Hamed- 287114 E 1705 15.7 0.16 10.2 378 7.5 0.68 Sandy loam
an Province, 3811412 N
Iran
14 Jo2 Jowkar, Hamed- 287114 E 1705 15.7 0.16 10.2 378 7.5 0.68 Sandy loam
an Province, 3811412 N
Iran
15 Jo3 Jowkar, Hamed- 287114 E 1705 15.7 0.16 10.2 378 7.5 0.68 Sandy loam
an Province, 3811412 N
Iran
16 Bi1 Bisotun, 724468 E 1312 10.48 0.12 9.54 329 7.3 0.54 Clay loam
Kermanshah 3808439 N
Province, Iran
17 Bi2 Bisotun, 724468 E 1312 10.48 0.12 9.54 329 7.3 0.54 Clay loam
Kermanshah 3808439 N
Province, Iran
18 Bi3 Bisotun, 724468 E 1312 10.48 0.12 9.54 329 7.3 0.54 Clay loam
Kermanshah 3808439 N
Province, Iran
19 Ke1 Kermanshah, 693621 E 1343 22.23 0.11 15.12 227 7.7 0.47 Clay
Kermanshah 3807294 N
Province, Iran
20 Ke2 Kermanshah, 693621 E 1343 22.23 0.11 15.12 227 7.7 0.47 Clay
Kermanshah 3807294 N
Province, Iran
21 Ke3 Kermanshah, 693621 E 1343 22.23 0.11 15.12 227 7.7 0.47 Clay
Kermanshah 3807294 N
Province, Iran
a
Association name.
b
Electrical conductivity.

the same phenotype. For each population, to be more Garmin) receiver has been used in order to record the
exact about sampling, three leaf samples were collected location data.
from one shrub (individual) and then were mixed into Soil samples were taken from each region. In each
one sample. A Global Positioning System (GPS, Vista area, six samples were taken from depths of 0–40 cm and
 JOURNAL OF ESSENTIAL OIL RESEARCH  3

mixed. The soil properties were analyzed using standard Compound identification
laboratory procedures. Soil pH was determined using
The components of the oils were identified by compari-
an automatic probe. Nitrogen (N) was analyzed accord-
son of their spectra with those from available MS (NIST
ing to Kjeldahl method. Phosphorus (P) and Potassium
08 and Wiley MS) libraries and by comparison of their
(K) were determined by Olsen and flame photometer
KI (Kovats index) relative to C5-C24 n-alkanes obtained
method, resp. Electrical conductivity (EC) was also
on an apolar HP-5MS column with those reported in the
determined by an electrical conductivity meter (EC
literature (12). The percentage composition (average of
meter) (9).
three independent analyses) was computed from the GC
peak areas without using any correction factors.
Essential oil isolation
Fresh leaves were dried for 10 days at room tempera- Statistical analysis
ture. One hundred grams of dried material were submit-
Data were analyzed using the SPSS ver. 21 statistical soft-
ted to hydrodistillation for 3 h, using a Clevenger-type
ware. A probability value at p ≤ 0.05 was considered sta-
apparatus, according to the method recommended in
tistically significant using Duncan’s multiple range test.
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the European Pharmacopoeia (10). The essential oil was

Hierarchical cluster analysis (HCA) was performed using
separated from water, dried over anhydrous sodium sul-
Minitab ver. 16. based on Ward variance method. Analysis
phate and stored in dark glass bottle (sealed brown vials)
related to correspondence was, respectively, performed
at 4 °C until chemical analysis (11).
by Correspondence Analyses (CA), Principal Component
Analysis (PCA) and Detrended Correspondence Analyses
GC-FID/MS analysis (DCA) using PC-ORD software.
Compositions of the essential oil were determined by
GC-FID and GC-MS analyses. They were achieved on Results and discussion
an Agilent Technologies 7890 GC equipped with FID
Essential oil yield
and mass spectrometer detectors using a HP-5MS 5%
capillary column (30.00 m × 0.25 mm, 0.25 μm film The essential oil yields obtained from the leaves of rose-
thicknesses; J & W Scientific, Folsom). The carrier gas mary showed a significant variability (p ≤ 0.01) among the
was helium at a flow of 0.8 ml/min. Initial column tem- Iranian populations (Figure 1). The highest content was
perature was 60 °C and programmed to increase up to obtained from the plants collected in the Bo2 population
280 °C at 4 °C/min. The split ratio was 40:1. The injector with 2.6 ml/100 g air dry matter. The soil of this region was
temperature was set at 300 °C. The acquisition range was rich in nitrogen (Table 1). The lowest values were obtained
50–550 m/z in electron impact (EI) mode using an ion- from the Jo2, Jo1 and Jo3 populations (0.6 ml/100 g air dry
ization voltage of 70 eV. The essential oils were diluted matter) (Figure 1). Similar to our study, Singh et al. (13)
1:100 in n-hexane, then 0.1 μL were injected into the reported that the yield of rosemary normally responds
GC systems. to the application of N fertilizer. Boutekedjiret et al. (14)

3
2.6 a
2.43 b
2.35 bc

2.16 d
2.3 c

2.5
Essential oil yield (ml/100 g)

1.78 e
1.69 ef

2
1.64 f
1.61 f

1.51 g
1.4 h

1.4 h

1.28 i
1.22 i

1.5
1.11 j
0.95 k
0.9 kl

0.84 l

0.61 m

1
0.6 m
0.53 m

0.5

Figure 1. Essential oil yield of rosemary accessions. Means with same letters are not significant (p ≤ 0.05), according to Duncan’s multiple
range test.
 4  I. BAJALAN ET AL.

reported that the essential oil yield of R. officinalis L. from Multivariate analysis
Algeria, obtained by steam and hydrodistillation methods
The chemical composition of Iranian rosemary acces-
were 1.2 and 0.44%, respectively. In addition, in another
sions was analyzed using Minitab and PC-ORD programs
investigation by Jamshidi et al. (15) the essential oil yields
for grouping of population. The HCA was based on the
of rosemary from central Iran (Lalehzar and Kerman)
major essential oil compounds. As a result, the samples
were 2.1–2.6%. Khorshidi et al. (16) reported that the
were classified into two main groups (Figures 2 and 3).
essential oil yield of rosemary from Karaj province (Iran)
The first group consisted of six accessions (Bo1, Bo2,
was 1.8%. Overall, the essential oil yields of rosemary from
Bo3, Ke1, Ke2 and Ke3 populations). They were richer
different countries ranged from 0.7 to 2.1% (17–20).
in camphor (21.41–25.32%) and poorer in 1,8-cineole
(5.32–8.38%). The second group was classified in two
Essential oil composition subgroups. The main component of first subgroup (Kh1,
Kh2, Kh3, Ma1, Ma2 and Ma3 populations) was 1,8-cine-
The essential oil constituents of 21 Iranian rosemary
ole (22.85–28.29%), whereas camphor occurred in low
accessions are reported in Table 2. A total of 34 constitu-
amounts (1.58–4.33%). On the other hand, the main
ents were identified by GC-FID and GC/MS, representing
components of second subgroup (Bi1, Bi2, Bi3, Do11,
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90.33–98.96% of the total compositions. The major com-

Do2, Do3, Jo1, Jo2 and Jo3 populations) were α-pinene
pounds were 1,8-cineole (3.32% in the Bo3 population
(14.19–18.87%), 1,8-cineole (12.75–16.80%) and camphor
to 28.29% in the Ma1), camphor (from 1.6% in the Ma2
(9.13–11.63%) (Figures 2 and 3). Results of HCA and CA
population to 25.3% in the Bo3 population), α-pinene
were in agreement with the results of principal component
(14.19% in the Do2 population to 21.43% in the Kh1 pop-
analyses (PCA) (Figure 4). Furthermore, DCA were used
ulation). Generally, among major components, 1,8-cineole
to determine the most effective environmental factors in
showed high variation. Other components occurring in
the separation of groups and relationships between veg-
noteworthy levels were borneol (0.16–8.65%), piperitone
etation and environmental factors. The results of the CA,
(2.24–7.71%), camphene (3.25–7.58%), bornyl acetate
PCA and DCA ordination are presented in Table 3. The
(4.14–6.95%), verbenone (0.42–6.86%) and trans-pino-
eigenvalues for the data-set indicate that the first DCA is
camphone (0.26–6.62%) (Table 2).
by far the most important for representing the eigenvalue,
1,8-Cineole, also known as eucalyptol, has typical
variation and coefficient of correlation of the environment
fresh and camphoraceous fragrance and pungent taste
species of the groups. As shown in Table 4, the first axis
and hence used extensively in food, pharmaceutical and
(eigenvalue = 0.191) accounted for 89% of the variation
cosmetic industries. It showed many biological activities
in environmental factors (Figure 4).
(21,22).
PCA and DCA are the most popular statistical methods
Similar results were reported by Lakušić et al. (23) who
in plant ecology at the present time. They have the advan-
found 1,8-cineole and camphor are the major compounds
tage of analyzing simultaneously all the environmental
of the essential oil of rosemary leaves from the Balkan
variables potentially determining species habitat selection.
Peninsula. There are many reports regarding the variation
The distribution of the vegetation groups could compre-
of rosemary essential oil according to geographic origin.
hensively reflect the influence of environmental factors
Gurbuz et al. (19) reported that camphor (14.3–17.5%),
(28,29). PCA statistical analyses indicated the existence of
1,8-cineole (11.0–21.6%) and α-pinene (10.2–12.5%) were
two main clusters according to the chemical differences
the major essential oil components in populations grow-
in essential oil composition. This variability may result
ing in Turkey. The rosemary essential oil from Morocco
from genetic diversity and differences in environmental
was rich in 1,8-cineole (47.44%), α-pinene (12.51%) and
conditions and their interactions (28) (Figure 5).
camphor (7.9%), while α-pinene was the major compo-
According to literature data, rosemary can be classified
nent of rosemary from Spain (24.7%) and France (35.80%)
into three chemotypes such as cineoliferum (high content
(24). Alike, Boutekedjiret et al. (14) reported 1,8-cine-
in 1,8-cineol), camphoriferum (camphor > 20%) and ver-
ole (52.4%), camphor (12.6%) and β-pinene (5.7%) for
benoniferum (verbenone > 15%) (30). The results of our
rosemary populations growing in Algeria. Pino et al. (25)
study confirmed those previously obtained by Angioni
reported higher camphor (34.80%) content than borneol
et al. (31), Pintore et al. (32) and Cioni et al. (33) for
(11.6%), 1,8-cineole (11%) and α-pinene (8.17%) for rose-
samples from Sardinia (Italy) and Corsica (France); and
mary from Cuba.
were in agreement with those by Napoli et al. (34), who
The variability of essential oil contents can be mainly
classified rosemary populations from Sicily (Italy) into
attributed to environmental conditions as well as agro-
cineoliferum and camphoriferum chemotypes. Limonene
nomic management including irrigation, plant density
was often reported as a major essential oil component, for
and soil tillage (26,27).
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Table 2. Chemical composition of the essential oils from 21 rosemary populations.

%d
a b c
Components RI RI lit. BI1 BI2 BI3 Bo1 Bo2 Bo3 Do1 Do2 Do3 Jo1 Jo2 Jo3 Ka1 Ka2 Ka3 Kh1 Kh2 Kh3 Ma1 Ma2 Ma3
Tricyclene 926 921 0.23 0.36 0.32 0.52 0.23 0.41 0.43 0.51 0.29 0.21 0.35 0.28 0.18 0.41 0.30 0.35 0.15 0.67 0.19 0.31 0.11
α-Pinene 935 932 17.74 16.80 18.12 16.84 16.23 17.63 14.78 14.19 15.11 18.87 18.49 17.35 17.28 17.42 18.90 21.43 20.38 20.63 17.45 17.28 17.85
Camphene 950 946 6.44 5.21 6.42 5.11 6.32 6.21 5.92 6.65 4.47 5.35 6.71 5.40 5.71 5.52 5.39 6.12 7.58 6.30 3.99 3.25 3.64
Verbenene 955 961 0.84 0.41 0.75 0.73 0.48 0.67 0.73 0.81 0.35 0.56 0.54 0.71 0.48 0.52 0.23 0.61 0.55 0.32 0.94
β-Pinene 978 974 1.26 2.11 2.26 1.94 1.39 1.75 2.58 1.86 2.29 2.11 2.66 2.26 2.31 1.77 2.10 1.52 1.95 2.49 2.22 2.39 2.67
3-Octanone 987 979 0.42 0.53 0.41 2.13 1.20 2.23 1.74 1.45 1.92 1.96 2.25 2.17 2.11 2.46 2.25 2.21 1.85 2.07 0.20 0.33 0.11
β-Myrcene 991 988 2.74 2.23 2.94 3.09 3.41 3.85 2.93 1.85 2.60 3.11 2.63 2.99 2.97 2.72 2.81 2.71 1.96 3.22 1.40 1.56 1.71
α-Phellandr- 1008 1002 0.01 0.67 0.45 0.12 0.47 0.33 0.26 0.41 0.29 0.34 0.20 0.47 0.38
ene
α-Terpinene 1018 1014 0.45 0.39 0.37 0.11 0.17 0.49 0.52 0.68 0.93 0.29 0.42 0.23 0.11 0.31 0.69 0.31 0.62 0.44
1,8-Cineole 1032 1026 16.80 16.46 15.93 6.12 5.41 5.32 14.17 15.21 14.57 14.25 13.60 12.75 8.18 8.46 7.95 24.12 22.85 23.34 28.29 25.79 26.58
γ-Terpinene 1061 1054 0.51 0.63 0.72 0.59 0.16 0.38 0.82 0.62 0.93 0.39 0.19 0.31 0.52 0.63 0.15 0.21 0.77 0.68 0.47 0.35 0.72
cis-Sabinene 1069 1065 0.11 0.22 0.15 0.14 0.26
hydrate
α-Terpinolene 1089 1086 0.11 0.82 0.49 0.81 0.76 0.11 0.86 0.38 0.11 0.41 0.39 0.14 0.59 0.40 0.72 0.60 0.31 0.97 0.29 0.63 0.70
Linalool 1100 1095 1.02 1.70 1.82 3.37 2.51 2.09 2.70 3.12 2.39 1.52 2.29 2.11 1.98 2.31 2.13 1.33 2.27 2.06 2.19 2.11 2.63
Chrysanthe- 1123 1124 0.32 0.15 0.85 0.63 1.17 1.63 1.91 1.23 1.76 1.15 1.44 1.64 0.87 0.87 0.87 1.10 1.14 0.19
none
Camphor 1145 1141 10.54 11.21 9.69 25.18 23.95 25.32 11.30 10.17 11.63 9.23 9.13 9.96 21.41 21.41 21.41 4.33 4.19 3.55 1.79 1.58 1.61
trans-Pino- 1160 1158 1.81 1.23 0.89 0.35 0.26 0.63 0.43 0.24 0.19 0.59 0.42 0.31 0.82 0.82 0.82 0.28 0.84 0.37 5.99 6.21 6.62
camphone
Pinocarvone 1162 1160 0.24 0.17 0.43 0.30 0.11 0.12 0.45 0.17 0.29 0.11 0.37 0.58 0.18
Iso-Borneol 1156 1160 0.87 1.69 1.43 1.47 1.58 2.22 1.88 1.54 1.76 0.55 1.64 1.22 6.87 5.96 6.29
Borneol 1167 1165 7.06 8.21 9.42 8.21 8.65 6.43 7.97 8.41 7.55 8.23 8.12 8.41 8.04 8.04 8.04 6.31 7.49 6.12 0.49 0.31 0.16
Terpinen-4-ol 1177 1174 1.42 1.11 0.19 1.47 1.11 1.29 1.43 1.32 1.64 0.65 1.11 1.25 1.38 1.38 1.38 1.23 1.65 1.74 1.23 1.57 1.29
α-Terpineol 1189 1186 2.75 2.09 2.25 0.85 1.15 1.62 2.24 2.15 2.53 2.47 2.89 2.25 2.21 2.43 1.08 0.85 1.53 1.65 1.33 1.76 1.44
Myrtenol 1198 1195 0.56 0.21 0.34 0.27 0.16 0.37 0.50 0.21 0.74 0.37 0.23 0.36 0.74 0.25 3.13 3.40 3.88
Verbenone 1204 1204 4.32 4.15 3.37 5.22 5.31 6.32 4.51 4.51 4.51 5.22 4.64 4.90 6.86 6.22 6.39 5.32 5.11 4.59 0.73 0.66 0.42
Piperitone 1246 1249 4.81 5.97 5.18 2.24 3.76 2.46 4.02 4.35 3.96 7.71 6.65 7.23 4.73 4.54 4.23 4.21 3.35 4.69 5.11 5.88 5.29
Bornyl acetate 1286 1287 5.03 5.84 5.17 5.43 6.25 6.20 6.60 6.82 6.95 5.39 5.22 5.52 6.35 6.11 6.50 6.19 5.73 6.54 4.48 5.63 4.14
Neryl acetate 1365 1359 0.17 0.20 0.11 0.19 0.35 0.27 0.13 0.19 0.33 0.54
Methyl euge- 1401 1403 0.21 0.52 0.36 0.36 0.36 0.28 0.14 0.28 0.37 0.12 0.14 0.22
nol
β-Caryophyl- 1418 1417 0.45 0.74 0.93 1.35 1.46 1.18 1.42 1.97 1.11 1.32 1.54 1.12 0.66 0.51 0.94 1.15 1.33 1.41 0.81 0.72 0.24
lene
α-Humulene 1456 1452 0.11 0.21 0.19 0.82 0.27 0.21 0.33 0.49 0.65 0.39 0.20 0.14 0.24 0.18 0.62 0.53 0.14 0.13 0.86 0.39
Caryophyllene 1587 1583 1.15 1.09 1.13 1.46 1.70 1.82 1.18 1.77 1.31 1.52 1.19 1.78 0.44 0.98 1.13 0.91 0.42 0.73 0.78 0.39 0.98
oxide
Widdrol 1597 1599 0.35 0.25 0.25 0.25 0.28 0.53 0.70 0.65 0.41 0.23 0.41 0.20 0.31 0.25 0.13 0.17 0.18 0.09 0.31
α-Bisabolol 1683 1685 0.64 0.79 0.26 0.39 0.46 0.23 0.50 0.45 0.19 0.17 0.40 0.63 0.12 0.19 0.35 0.41 0.30 0.26 0.29 0.19 0.43
Total (%) 90.96 93.03 91.89 97.09 97.12 98.08 96.60 96.28 95.24 96.20 96.12 94.00 97.86 98.42 99.54 96.41 98.87 98.19 91.09 90.33 91.33
a
Compounds are listed in order of their elution from a HP-5MS column.
b
Linear retention index on HP-5MS column, experimentally determined using homologous series of C8-C30 alkanes.
c
Linear retention index taken from Adams (12).
d
Relative percentage values are means of three determinations.
JOURNAL OF ESSENTIAL OIL RESEARCH 
5
 6  I. BAJALAN ET AL.

Figure 2. Hierarchical cluster analysis (HCA), based on the main compositions of the essential oils using Ward’s minimum variance.
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Table 3. Components of CA, DCA and PCA according to the re-

1.0

-Pinene gions and main constituents of the essential oils.

Axes 1 2 3 Total inertia
Correspondence analyses (CA)
Eigenvalues 0.191 0.022 0.002 0.215
Cumulative percentage vari- 89.0 99.0 99.8
ance of species data
Sum of all eigenvalues 0.215
Kh1 Jo2 Jo1 Ke3 Detrended correspondence analyses (DCA)
BI3 Jo3
Kh2 Kh3 Ke2
Ma3 BI1 Do1 Bo2 Eigenvalues 0.191 0.001 0.000 0.215
BI2 Bo3 Bo1
Ma2 Ke1 Lengths of gradient 1.183 0.359 0.000
Ma1 Do2 Do3
Cumulative percentage vari- 89.0 89.4 0.000
ance of species data
Sum of all eigenvalues 0.215
-0.6

Camphor
Principal component analyses (PCA)
1,8-Cineole
Eigenvalues 0.755 0.227 0.017 1.000
-1.0 1.0 Cumulative percentage vari- 75.3 98.1 99.8
ance of species data
Sum of all eigenvalues 1.000
Figure 3. Correspondence analyses of the oil constituents
obtained from rosemary essential oils.
instance in samples from Poland (35), Balkan Penninsula
(23) and Serbia (36). This constituent was missing in the
Iranian rosemary.
1.0

Jo1 Jo3
Jo2 α-Pinene
Do1 BI3
BI1BI2
Ke3Ke1
Correlation between compounds and soil
Kh3 Do3 Ke2 Bo3
characteristics
Kh2
Kh1 Bo2
Do2 Bo1
Ma3 Analyses of correlations revealed that there were sig-
Ma2
Ma1 nificant correlations between camphene and borneol
Camphor
(0.714), camphene and verbenone (0.638), 1,8-cineole
1,8-Cineole and trans-pinocamphone (0.651), camphor and borneol
(0.559), camphor and verbenone (0.714), camphor and
-1.0

phosphorous (0.711), trans-pinocamphone and alti-

-1.5
Figure 4. Principal component analysis of the oil constituents
obtained from rosemary essential oils.
1.5
tude (0.613), borneol and verbenone (0.823), bornyl
acetate and verbenone (0.633) at 1% probability level,
and between bornyl acetate and borneol (0.517), bor-
nyl acetate and CaO (0.498) at 5% probability level. The
 JOURNAL OF ESSENTIAL OIL RESEARCH  7

1.5
X16

1
-Pinene

−0.355
X15

ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns

1
0.977
−0.227
Kh2
X14

BI2 Jo2 Kh3

ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns

1
*

Ma3
Ke3 BI1 Kh1 Ma2
Jo3 BI3
Ke2 Do1 Ma1
Bo3 Do3 Do2

−0.673
−0.177
−0.801
Bo2 Jo1
Ke1
X13

Bo1

ns
ns
ns
ns
ns
ns
ns
ns
ns

**
**
*

1
1,8-Cineole

0.210
−0.235
0.164
0.195
X12

ns
ns
ns
ns
ns
ns
ns
ns
ns
ns
ns

-1.0
Camphor

0.021
−0.694
−0.115
0.398
−0.265

-1.0 2.0
X11
Downloaded by [The Library at Queen's University] at 23:55 24 October 2017

ns
ns
ns
ns
ns
ns
ns
ns

*
*

Figure 5. Detrended correspondence analyses (DCA) of the oil

−0.015
−0.446
−0.051
0.135
0.584
−0.179

constituents obtained from rosemary essential oils.

X10

ns
ns
ns
ns

**
**
**
**

*
1

highest negative correlations were between camphene and

0.167
−0.261
0.140
0.030
−0.306
0.498
−0.547

trans-pinocamphone (−0.740), 1,8-cineole and camphor

ns
ns
ns
ns
X9
ns

**
**
*

(−0.952), 1,8-cineole and borneol (−0.711), verbenone

(−0.767) and phosphorous (−0.637), trans-pinocam-
−0.401
0.164
−0.292
−0.118
−0.069
−0.200
0.353
−0.376

phone and borneol (−0.943), verbenone (−0.880) and

ns
ns
ns
ns
ns
X8
ns

bornyl acetate (−0.654), altitude and camphene (−0.598),

borneol (−0.651) and verbenone (−0.618) at 1% prob-
0.058
−0.116
0.032
0.204
0.010
0.513
−0.618
0.633
−0.298

ability level, and between α-pinene and phosphorous

X7
ns

**
**
**
**
**

(−0.442), camphor and trans-pinocamphone (−0.517),

camphor and piperitone (−0.479), bornyl acetate and alti-
−0.088
0.219
−0.020
0.025
−0.042
0.131
−0.651
0.517
−0.100
0.823

Notes: No significant; *Significant at the 5% probability levels; **Significant at the 1% probability levels.
Table 4. Correlation of major components of rosemary and some environmental factors.
X6
ns

**
**
**
**

tude (−0.547) at 5% probability level (Table 4). Probably,

1

these compounds are double bond isomers or are both

−0.163
−0.023
−0.214
0.105
0.008
−0.235
0.631
−0.654
0.211
−0.880
−0.943

produced from the same biosynthetic pathway (37).

X5
ns

**
**

*
1

Studies conducted by Tigrine-Kordjani et al. (38) in the

north of Algeria indicated a strong correlation between
−0.256
−0.073
−0.351
0.711
0.341
0.336
−0.300
0.393
−0.479
0.714
0.559
−0.517

the main essential oil components and the sample geo-

ns
X4
ns

**
1

graphic origin. Results of this study are also in agreement

with the results by Zaouali et al. (7), who reported that
0.320
0.053
0.369
−0.637
−0.338
−0.293
0.286
−0.426
0.290
−0.767
−0.711
0.651
−0.952

the organ sources influence the essential oil composition

ns
X3
ns

in Tunisian rosemary.
0.331
0.161
0.391
−0.342
−0.058
0.054
−0.598
0.333
−0.239
0.638
0.714
−0.740
0.218
−0.316
X2
ns
1

Conclusion
This study highlighted a wide variability in the essential
0.509
−0.022
0.499
−0.442
−0.014
0.128
−0.124
−0.234
0.112
0.096
−0.109
−0.109
−0.001
−0.308
0.235

oil content and composition of different Iranian rose-

X1
1

mary populations according to environmental condi-

tions. Indeed, significant correlations were observed
trans-Pinocamphone

between essential oil content and composition and soil

conditions in the growing sites. The significant differences
Bornyl acetate
1,8-Cineole

Verbenone

observed could provide useful information for breeding

Camphene

Piperitone
Camphor
α-Pinene

Altitude
Borneol

programmes and to develop improved cultivars. It can

CaO

pH
E. C.

be recommended also to commercial companies, which

N

K
P

seek higher amounts of each component, to cultivate

ns

this plant in the most suitable soil condition with higher

X16
X15
X14
X13
X12
X11
X10
No.

X9
X8
X7
X6
X5
X4
X3
X2
X1
 8  I. BAJALAN ET AL.
acidity. According to our results, the main volatile com-
ponents of Iranian rosemary were 1,8-cineole, camphor
and α-pinene. Applications of CA, DCA and PCA revealed
differences in the chemical composition depending on the
geographical origin and determined the membership of
the Iranian rosemary accessions to two main chemotypes.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This work was supported by Young Researchers and Elite Club
of I.A.U. (Borujerd Branch, Iran) [grant number 93532].
Downloaded by [The Library at Queen's University] at 23:55 24 October 2017
References
1. P.A. Ghasemi, P. Jahanbazi, S. Enteshari, F. Malekpoor and
B. Hamedi, Antimicrobial activity of some of the Iranian
medicinal plants. Archives of Biological Sciences, 62, 633–
641 (2010).
2. J. Tai, S. Cheung, M. Wu and D. Hasman, Antiproliferation
effect of rosemary (Rosmarinus officinalis L.) on human
ovarian cancer cells in vitro. Phytomedicine, 19, 436–443
(2012).
3. T. Rozman and B. Jersek, Antimicrobial activity of rosemary
extracts (Rosmarinus officinalis L.) against different species
of Listeria. Acta Agriculturae Slovenica, 93, 51–58 (2009).
4. M. Singh and N. Guleria, Influence of harvesting stage
and inorganic and organic fertilizers on yield and oil
composition of rosemary (Rosmarinus officinalis L.) in a
semi-arid tropical climate. Industrial Crops and Products,
42, 37–40 (2013).
5. M.J. del Baño, J. Lorente, J. Castillo, O. Benavente-
García, J.A. del Río, A. Ortuño, K.W. Quirin and
D. Gerard, Phenolic diterpenes, flavones, and rosmarinic
acid distribution during the development of leaves,
flowers, stems, and roots of Rosmarinus officinalis L. and
antioxidant activity. Journal of Agricultural and Food
Chemistry, 51, 4247–4253 (2003).
6. Z. Yosr, C. Hnia, T. Rim and B. Mohamed, Changes
in essential oil composition and phenolic fraction in
Rosmarinus officinalis L. var. typicus Batt. organs during
growth and incidence on the antioxidant activity. Industrial
Crops and Products, 43, 412–419 (2013).
7. Y. Zaouali, T. Bouzaine and M. Boussaid, Essential oils
composition in two Rosmarinus officinalis L. varieties and
incidence for antimicrobial and antioxidant activities. Food
and Chemical Toxicology, 48, 3144–3152 (2010).
8. M.M. Özcan and D. Arslan, Antioxidant effect of essential
oils of rosemary, clove and cinnamon on hazelnut and poppy
oils. Food Chemistry, 129, 171–174 (2011).
9. I. Bajalan, R. Rouzbahani, A. Ghasemi Pirbalouti and F.
Maggi, Variation in chemical composition and antibacterial
activity of the essential oil of wild populations of Phlomis
olivieri. Chemistry & Biodiversity, 14, e1600444 (2017).
10. European Pharmacopoeia, 6th ed., Council of Europe,
Strasbourg (2008).
11. I. Bajalan and A. Ghasemi Pirbalouti, Variation in chemical
composition of essential oil of populations of Lavandula ×
intermedia collected from Western Iran. Industrial Crops
and Products, 69, 344–347 (2015).
12. R.P. Adams, Identification of Essential Oil Components by
Gas Chromatography/Mass Spectrometry, 4th ed., Allured
Publ. Corp, Carol Stream, IL (2007).
13. M. Singh, R. Ganesha Rao and S. Ramesh, Effects of N and
K on growth, herbage, oil yield and nutrient uptake pattern
of rosemary (Rosmarinus officinalis L.) under semi-arid
tropical conditions. Journal of Horticultural Science and
Biotechnology, 82, 414–419 (2007).
14. C. Boutekedjiret, F. Bentahar, R. Belabbes and J. Bessiere,
The essential oil from Rosmarinus officinalis L. Journal of
Essential Oil Research, 10, 680–682 (1998).
15. R. Jamshidi, Z. Afzali and D. Afzali, Chemical composition of
hydrodistillation essential oil of rosemary in different origins
in Iran and comparison with other countries. American-
Eurasian Journal of Agricultural and Environmental
Science, 5, 78–81 (2009).
16. J. Khorshidi, R. Mohammadi, T. Fakhr and H. Nourbakhsh,
Influence of drying methods, extraction time, and organ type
on essential oil content of rosemary (Rosmarinus officinalis
L.). Natural Science, 7, 42–44 (2009).
17. G. Fournier, J. Habib, A. Reguigui, F. Safta, S. Guetari and
R. Chemli, Study of different samples of the essential oil of
Tunisian rosemary. Plantes Medicinales et Phytotherapie,
23, 180–185 (1989).
18. R.S. Verma, L.U. Rahman, S. Mishra, R.K. Verma,
A. Chauhan and A. Singh, Changes in essential oil content
and composition of leaf and leaf powder of Rosmarinus
officinalis cv. CIM-Hariyali during storage. Maejo Int.
Journal of Science and Technology, 5, 181–190 (2011).
19. B. Gurbuz, R.B. Bagdat, M. Uyanik and K.A.P. Rezaeieh,
Rosemary (Rosmarinus officinalis L.) cultivation studies
under Ankara ecological conditions. Industrial Crops and
Products, 88, 12–16 (2016).
20. H. Bendif, M. Boudjeniba, M.D. Miara, L. Biqiku, M.
Bramucci, G. Caprioli, G. Lupidi, L. Quassinti, G. Sagratini
and L.A. Vitali, Rosmarinus eriocalyx: an alternative
to Rosmarinus officinalis L. as a source of antioxidant
compounds. Food Chemistry, 218, 78–88 (2017).
21. U. Juergens, U. Dethlefsen, G. Steinkamp, A. Gillissen,
R. Repges and H. Vetter, Anti-inflammatory activity of
1,8-cineole (eucalyptol) in bronchial asthma: adouble-blind
placebo-controlled trial. Respiratory Medicine, 97, 250–
256 (2003).
22. R.C. Padalia, R.S. Verma, A. Chauhan, P. Goswami,
S.K. Verma and M.P. Darokar, Chemical composition of
Melaleuca linarrifolia Sm. from India: a potential source of
1, 8-cineole. Industrial Crops and Products, 63, 264–268
(2015).
23. D.V. Lakušić, M.S. Ristić, V.N. Slavkovska, J.B. Šinžar-
Sekulić and B.S. Lakušić, Environment-related variations of
the composition of the essential oils of rosemary (Rosmarinus
officinalis L.) in the Balkan Penninsula. Chemistry &
Biodiversity, 9, 1286–1302 (2012).
24. J.C. Chalchat, R.P. Garry, A. Michet, B. Benjilali and J.
Chabart, Essential oils of rosemary (Rosmarinus officinalis
L.). The chemical composition of oils of various origins
(Morocco, Spain, France). Journal of Essential Oil Research,
5, 613–618 (1993).

25. J.A. Pino, M. Estarrón and V. Fuentes, Essential oil of
rosemary (Rosemary officinalis L.) from Cuba. Journal of
Essential Oil Research, 10, 111–112 (1998).
26. A. Lubbe and R. Verpoorte, Cultivation of medicinal and
aromatic plants for specialty industrial materials. Industrial
Crops and Products, 34, 785–801 (2011).
27. A. Ghasemi Pirbalouti, M. Hashemi and F.T. Ghahfarokhi,
Essential oil and chemical compositions of wild and
cultivated Thymus daenensis Celak and Thymus vulgaris L.
Industrial Crops and Products, 48, 43–48 (2013).
28. M. Lange, M. Türke, E. Pašalić, S. Boch, D. Hessenmöller,
J. Müller, D. Prati, S.A. Socher, M. Fischer and W.W.
Weisser, Effects of forest management on ground-dwelling
beetles (Coleoptera; Carabidae, Staphylinidae) in Central
Europe are mainly mediated by changes in forest structure.
Forest Ecology and Management, 329, 166–176 (2014).
29. N. Enright, B. Miller and R. Akhter, Desert vegetation and
vegetation–environment relationships in Kirthar National
Downloaded by [The Library at Queen's University] at 23:55 24 October 2017
Park, Sindh. Journal of Arid Environments, 61, 397–418
(2005).
30. M.J. Jordán, V. Lax, M.C. Rota, S. Lorán and J.A. Sotomayor,
Effect of bioclimatic area on the essential oil composition
and antibacterial activity of Rosmarinus officinalis L. Food
Control, 30, 463–468 (2013).
31. A. Angioni, A. Barra, E. Cereti, D. Barile, J.D. Coïsson,
M. Arlorio, S. Dessi, V. Coroneo and P. Cabras, Chemical
composition, plant genetic differences, antimicrobial and
antifungal activity investigation of the essential oil of
Rosmarinus officinalis L. Journal of Agricultural and Food
Chemistry, 52, 3530–3535 (2004).
32. G. Pintore, M. Usai, P. Bradesi, C. Juliano, G. Boatto,
F. Tomi, M. Chessa, R. Cerri and J. Casanova, Chemical
JOURNAL OF ESSENTIAL OIL RESEARCH  9
composition and antimicrobial activity of Rosmarinus
officinalis L. oils from Sardinia and Corsica. Flavour and
Fragrance Journal, 17, 15–19 (2002).
33. P. Cioni, G. Flamini, C. Buti Castellini, L. Ceccarini and
M. Macchia, Composition and yield of the essential oils from
whole plant, leaves and branches of Rosmarinus officinalis L.
growing in minor islands of ‘Parco Nazionale dell’ Arcipelago
Toscano’. Acta Horticulturae, 723, 255–260 (2006).
34. E.M. Napoli, G. Curcuruto and G. Ruberto, Screening
of the essential oil composition of wild Sicilian rosemary.
Biochemical Systematics and Ecology, 38, 659–670
(2010).
35. A. Szumny, A. Figiel, A. Gutiérrez-Ortíz and Á.A.
Carbonell-Barrachina, Composition of rosemary essential
oil (Rosmarinus officinalis L.) as affected by drying method.
Journal of Food Engineering, 97, 253–260 (2010).
36. B. Bozin, N. Mimica-Dukic, I. Samojlik and E. Jovin,
Antimicrobial and antioxidant properties of rosemary and
sage (Rosmarinus officinalis L. and Salvia officinalis L.,
Lamiaceae) essential oils. Journal of Agricultural and Food
Chemistry, 55, 7879–7885 (2007).
37. I. Bajalan and A. Ghasemi Pirbalouti, Variation in
antibacterial activity and chemical compositions of essential
oil from different populations of myrtle. Industrial Crops
and Products, 61, 303–307 (2014).
38. N. Tigrine-Kordjani, F. Chemat, B. Meklati, L. Tuduri,
J. Giraudel and M. Montury, Relative characterization
of rosemary samples according to their geographical
origins using microwave-accelerated distillation, solid-
phase microextraction and Kohonen self-organizing maps.
Analytical and Bioanalytical Chemistry, 389, 631–641
(2007).