Afsharypuor, S., Rahiminezhad, M., Ghaemmaghami, L., Soleimani, M., Khanmohammadic, M., Afsharipour, N. (2007). Essential Oil Constituents of Leaves of the Male and Female Shrubs of Juniperus chinensis L. from Isfahan. Iranian Journal of Pharmaceutical Sciences, 3(3), 177-180.
Suleiman Afsharypuor; Mohammadreza Rahiminezhad; Leili Ghaemmaghami; Majid Soleimani Soleimani; Mohammadreza Khanmohammadic; Neda Afsharipour. "Essential Oil Constituents of Leaves of the Male and Female Shrubs of Juniperus chinensis L. from Isfahan". Iranian Journal of Pharmaceutical Sciences, 3, 3, 2007, 177-180.
Afsharypuor, S., Rahiminezhad, M., Ghaemmaghami, L., Soleimani, M., Khanmohammadic, M., Afsharipour, N. (2007). 'Essential Oil Constituents of Leaves of the Male and Female Shrubs of Juniperus chinensis L. from Isfahan', Iranian Journal of Pharmaceutical Sciences, 3(3), pp. 177-180.
Afsharypuor, S., Rahiminezhad, M., Ghaemmaghami, L., Soleimani, M., Khanmohammadic, M., Afsharipour, N. Essential Oil Constituents of Leaves of the Male and Female Shrubs of Juniperus chinensis L. from Isfahan. Iranian Journal of Pharmaceutical Sciences, 2007; 3(3): 177-180.
Essential Oil Constituents of Leaves of the Male and Female Shrubs of Juniperus chinensis L. from Isfahan
1Faculty of Pharmacy and Pharmaceutical Sciences, Isfahan University
of Medical Sciences, Isfahan, Iran
2Faculty of Sciences, Department of Botany, University of Isfahan, Isfahan, Iran
3Faculty of Sciences, Department of Chemistry, Imam Khomeini International University, Ghazvin, Iran
Abstract
The composition of essential oil of leaves of the male and female shrubs of Juniperus chinensis L. (growing in Isfahan) was analyzed by gas choromatography(GC) and GC-mass spectrometery (MS). The main constituents of leaf oil of the male shrub were: bornyl acetate (26.1%), sabinene (18.3%), trans-sabinyl acetate (11.1%), myrcene (9.2%), limonene (6.1%), elemol (5.5%), α-cadinol (2.3%), δ-cadinene (2.2%), trans-thujone (1.6%), α-pinene (1.3%) and α-muurolol (1.1%); while the major constituents of leaf oil of the female shrub were: sabinene (18.0%), bornyl acetate (12.3%), carotol (11.4%), elemol (9.9%), myrcene (8.8%), α-pinene (6.4%), δ-cadinol (4.8%), limonene (3.9%), δ-cadinene (3.4%) and α-muurolol (2.7%).
Juniperus chinensis L., commonly knownas Chinese juniper (Fam. Cupressaceae) is a dioecious or rarely monoecious shrub or tree growing in different parts of China, Japan, Korea, Myanmar and eastern parts of Russia [1]. The ornamental dioecious shrubs of this plant are cultivated widely in different parts of Iran [2]. Leaves of the plant claimed to have antitumour promoting and antitumor activities [3].
As the chemical composition of the essential oils of leaves of both the male and female cultivated shrubs of Chinese juniper in Iran has not been investigated previously, and as a part of our long term research programs is to identify new sources of valuable terpenoids in our country, therefore, in the present study the above-mentioned oils were isolated separately, and their active constituents were determined using gas choromatography (GC) and GC-mass spectrometery (MS) methods.
2. Materials and methods
2.1. Plant material
Leaves of the male and female shrubs were collected in May 2004 from populations growing in Isfahan (Iran). The shrubs were identified by the Botany Department of the Faculty of Sciences at the University of Isfahan (Iran). Voucher specimens of the leaves (no.1773) were deposited in the Herbarium of the Pharmacognosy Department, Faculty of Pharmacy and Pharmaceutical Sciences at the Isfahan University of Medical Sciences (Iran).
2.2. Isolation of the essential oil
Powdered air-dried leaves of the male and female shrubs were separately hydrodistilled for 4 h. Pale yellow oils (1.29% and 0.66% v/w) were obtained having the refractive indices of 1.4475 and 1.4476 measured at 25 °C.
2.3. GC analysis
Gas chromatographic determinations were run on a Perkin Elmer 8500 instrument using a BP1 capillary column (30 m×0.25 mm; film thickness: 0.25 μm). The carrier gas was nitrogen with a flow rate of 2 ml/min. The oven temperature was programmed from 60-275 °C at 4 °C/min. Injector and detector temperatures were 275 °C and 280 °C, respectively.
2.4. GC/MS analysis
Analysis of the oils was performed on a Hewlett Packard 6890 GC/MS instrument under the following conditions: Injection of 0.1 μl samples, HP-5 MS capillary column (30 m×0.25 mm ; film thickness 0.25 μm); carrier gas He, flow rate 2 ml/min., injector temperature 250 °C, temperature program: 60-275 °C at 4 °C/min.; mass spectra: electronic impact, ionization potential 70 eV, ion source temperature 250 °C, ionization current 1000 μA, resolution 1000, and mass range 30-300.
Identification of the constituents was based on computer matching against the library spectra (Library Database Wiley 275), their retention indices with reference to an n-alkane series in a temperature programmed run, interpreting their fragmentation pattern and comparison of the mass spectra with the literature data [4].
3. Results and discussion
Forty-one constituents in the male shrub leaf oil (representing 96.2% of the total oil) and thirty nine components in the female shrub leaf oil (representing 95.4% of the total oil) were identified (Table 1). Bornyl acetate (26.1%), sabinene (18.3%), trans-sabinyl acetate (11.1%), myrcene (9.2%), limonene (6.1%), elemol (5.5%), α-cadinol (2.3%), δ-cadinene (2.2%), α-thujone (1.6%), α-pinene (1.3%) and α-muurolol (1.1%) were the main constituents of the leaf oil of male shrub; while the major constituents of leaf oil of the female shrub were sabinene (18.0%), bornyl acetate (12.3%), carotol (11.4%), elemol (9.9%), myrcene (8.8%), α-pinene (6.4%), α-cadinol (4.8%), limonene (3.9%), δ-cadinene (3.4%) and α-muurolol (2.7%). In a previous work which has been done by other workers, ethyl acetate was reported to be one of the major constituents of leaf oils of Juniperus chinensis growing in China (three samples: 20.65%, 12.55% and 19.6%), but it was absent in the leaf oil of Juniperuschinensis var. kaizuca [5]. The lattermentioned compound could not be detected in our analyzed leaf oils as well. As in case of of J. chinensis cv. pyramidalis and J. chinensis var. kaizuca leaf oils, our examined leaf oils did not contain the cedrene/cedrol components [5].
The amount of essential oil obtained from the leaf of the male shrub (1.29% v/w) was about twice the amount of that produced by the leaf of the female shrub (0.66%). As it is obvious from Table 1, the relative percentage production of sabinene and myrcene in both leaf oils is about the same, while the production of bornyl acetate by the leaf of the male shrub (26.1%) is higher than that produced by the leaf of the female shrub (12.3%). On the other hand, the leaf of the male shrub produces 11.1% trans-sabinyl acetate, while the leaf of the female shrub doesn't produce this monoterpenoid.
Table 1. Percentage composition of leaf essential oils of the male and female shrubs of Juniperus chinensis from Isfahan.
No.
Components
Retention indices
Male leaf
(%)
Female leaf (%)
1
tricyclene
927
0.8
0.8
2
α-thujene
931
0.3
1.2
3
α-pinene
940
1.3
6.4
4
camphene
954
0.6
1.1
5
sabinene
976
18.3
18.0
6
β-pinene
981
t
t
7
myrcene
991
9.2
8.8
8
α-phellandrene
1004
t
0.8
9
α-terpinene
1018
0.1
0.6
10
limonene
1031
6.1
3.9
11
(E)-β-ocimene
1053
0.1
0.3
12
γ-terpinene
1062
0.4
0.8
13
cis-sabinene hydrate
1069
0.2
0.5
14
terpinolene
1089
0.1
0.9
15
trans-thujone
1117
1.6
0.1
16
trans-sabinol
1142
0.3
-
17
camphor
1145
0.2
0.3
18
camphene hydrate
1150
0.1
0.1
19
borneol
1168
0.1
0.3
20
terpinen-4-ol
1177
0.8
1.3
21
bornyl acetate
1287
26.1
12.3
22
trans-sabinyl acetate
1293
11.1
-
23
δ-elemene
1341
t
0.1
24
α-copaene
1379
t
t
25
β-elemene
1392
t
0.2
26
germacrene-D
1482
0.4
1.7
27
α-muurolene
1500
0.2
0.4
28
7-epi-α-selinene
1520
0.3
0.6
29
δ-cadinene
1525
2.2
3.4
30
cadina-1,4-diene
1533
t
t
31
α-cadinene
1539
t
0.1
32
elemol
1551
5.5
9.9
33
carotol
1591
5.0
11.4
34
β-oplopenone
1609
0.4
0.4
35
γ-eudesmol
1632
0.3
0.3
36
α-muurolol
1643
1.1
2.7
37
β-eudesmol
1651
0.2
t
38
α-cadinol
1654
2.3
4.8
39
caryophyllene acetate
1698
t
0.2
40
epi-13-manool
1962
t
0.1
41
manool
2058
0.5
0.6
Grouped constituents:
Hydrocarbon monoterpenes
Oxygenated monoterpenes
Hydrocarbon sesquiterpenes
Oxygenated sesquiterpenes
Oxygenated diterpenes
Total identified
37.2
40.6
3.1
14.8
0.5
96.2
42.7
15.8
6.5
29.7
0.7
95.
t = trace (less than 0.1%)
Acknowledgement
We are grateful to Mr. Akbar Akoochekian for his help to prepare the voucher specimens of the plant materials and Mrs. Armita Jamshidi for her help in analyzing the oil samples by GC and GC/MS methods.
References
[1] Brickell C. Gardener's encyclopedia of plants and flowers. 1sted. London:Dorling Kindersley, 1994; p. 525.
[2] Mozaffarian V. A dictionary of iranianplant names. Tehran: Farhang Moasser,1996; p. 279.
[3] Ali AM, Mackeen MM, Intan-safinar I, Hamid M, Lajis NH, el-Sharkawy SH, Murakoshi M. Antitumor promoting and antitumor activities of the crude extract from the leaves of Juniperus chinensis. J.Ethnopharmacol 1996; 53:165-9.
[4] Adams RP. Identification of essential oilcomponents by gas chromatography /quadrupole mass spectroscopy. CarolStream: Allured Publishing, 2004.
[5] Adams RP, Ge-lin C, Shao-Zhen Z. Composition of the volatile oils of Juniperus chinensis L., J. chinensis var. kaizuca Hort. and cv. pyramidalis fromChina. J Essent Oil Res 1994; 6:149-54.