Advances in Chemistry

Volume 2014 (2014), Article ID 838162, 6 pages

http://dx.doi.org/10.1155/2014/838162

Research Article

GC Analyses of Salvia Seeds as Valuable Essential Oil Source

Mouna Ben Taârit,¹ Kamel Msaada,¹ Karim Hosni,² and Brahim Marzouk¹

¹Laboratoire des Substances Bioactives, Centre de Biotechnologie, Technopôle de Borj-Cédria, BP 901, 2050 Hammam-Lif, Tunisia

²Laboratoire des Substances Naturelles, Institut National de Recherche et d’Analyse Physico-Chimique (INRAP), Sidi Thabet, 2020 Ariana, Tunisia

Received 23 April 2014; Accepted 19 August 2014; Published 27 August 2014

Academic Editor: Constantinos Pistos

Copyright © 2014 Mouna Ben Taârit et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract.

The essential oils of seeds of Salvia verbenaca, Salvia officinalis, and Salvia sclarea were obtained by hydrodistillation and analyzed by gas chromatography (GC) and GC-mass spectrometry. The oil yields (w/w) were 0.050, 0.047, and 0.045% in S. verbenaca, S. sclarea, and S. officinalis, respectively. Seventy-five compounds were identified. The essential oil composition of S. verbenaca seeds showed that over 57% of the detected compounds were oxygenated monoterpenes followed by sesquiterpenes (24.04%) and labdane type diterpenes (5.61%). The main essential oil constituents were camphor (38.94%), caryophyllene oxide (7.28%), and 13-epi-manool (5.61%), while those of essential oil of S. officinalis were α-thujone (14.77%), camphor (13.08%), and 1,8-cineole (6.66%). In samples of S. sclarea, essential oil consists mainly of linalool (24.25%), α-thujene (7.48%), linalyl acetate (6.90%), germacrene-D (5.88%), bicyclogermacrene (4.29%), and α-copaene (4.08%). This variability leads to a large range of naturally occurring volatile compounds with valuable industrial and pharmaceutical outlets.

1. Introduction

The genus Salvia (Lamiaceae) comprises nearly 900 species widely spread throughout the world, which display marked morphological and genetic variations according to their geographical origin [1]. Several Salvia species, namely, Salvia officinalis, Salvia sclarea, and Salvia verbenaca, are widely used in folk medicine [2]. Potential therapeutic activities of these Salvia species are due to their essential oils [3], since these species are known to possess antioxidant, antimicrobial, antifungal, and aromatic properties [4]. Chemical composition of essential oils reveals differences among these Salvia species [5–7]. Numerous investigations on Salvia officinalis show that 1,8-cineole, α-thujone, β-thujone, and camphor are the main compounds of the essential oil [8–10]. Linalool, linalyl acetate, and germacrene-D characterize S. sclarea plants [11]. Salvia species also display great intraspecific essential oil variations according to geographical origin, since sabinene, cadinene, terpinen-4-ol, and pinene are shown to be typical compounds of S. verbenaca essential oil originated from Saudi Arabia [4], while β-phellandrene and ()-caryophyllene prevail in essential oil from Greece [7]. In Tunisia, S. verbenaca essential oil shows variations of composition according to the region origin [12, 13] and in respect to the studied plant part [14].

These numerous studies are focused on aerial parts of these species, while works interested in seeds are scanty in spite of their interest. In fact, Salvia seeds provide dietary and healthy oil rich in essential fatty acids (linolenic and linoleic acids) [12, 15, 16] that promote decrease in coronary heart diseases [17]. Besides, the seeds of Salvia species often produce mucilage on wetting [18]. This mucilage is used for lacquerware [19]. In eastern countries, the mucilage is used for the treatment of eye diseases [20].

In our continuing research on essential oil with pharmacological potential and food industry applications, we report in this paper chemical analysis of S. verbenaca, S. sclarea, and S. officinalis seeds as a new valuable essential oil source.

2. Materials and Methods

2.1. Plant Material

The seeds of Salvia verbenaca (accession PI 420430) were originated from Spain; Salvia officinalis (accession W6 20659) and Salvia sclarea (accession W6 20660) which are from Italy were kindly supplied by the US National Plant Germplasm System (NPGS).

2.2. Essential Oil Isolation

The seeds of each species were ground in a mortar before essential oil isolation [12] and were subjected to conventional hydrodistillation for 90 min followed by a liquid-liquid extraction using diethyl ether and -pentane mixture (v/v) as solvent. The concentration step was carried out at 35°C using a Vigreux column and the essential oils obtained were dried over anhydrous sodium sulphate and stored in amber vials at −18°C until they were analyzed.

3. Chromatographic Analysis

3.1. Gas Chromatography (GC-FID)

The essential oils were analysed by gas chromatography using a Hewlett-Packard 6890 gas chromatograph (Palo Alto, CA, USA) equipped with a flame ionization detector (FID) and an electronic pressure control (EPC) injector. A polar HP Innowax (PEG) column and an apolar HP-5 column (30 m × 0.25 mm, 0.25 μm film thicknesses) were used. The carrier gas was N₂ with a flow rate of 1.6 mL/min; split ratio was 60 : 1. The analysis was performed using the following temperature program: oven temps isotherm at 35°C for 10 min, from 35 to 205°C, at the rate of 3°C/min, and isotherm at 205°C during 10 min. Injector and detector temperatures were held, respectively, at 250 and 300°C. The volume injected was 1 μL.

3.2. Gas Chromatography-Mass Spectrometry (GC-MS)

GC-MS analysis was performed on a gas chromatograph HP 5890 (II) interfaced with a HP 5972 mass spectrometer with electron impact ionization (70 eV). A HP-5MS capillary column (30 m × 0.25 mm, 0.25 μm film thickness) was used. The column temperature was programmed from 50°C to rise to 240°C at a rate of 5°C/min. The carrier gas was helium with a flow rate of 1.2 mL/min; split ratio was 60 : 1. Scan time and mass range were 1 s and 40–300 , respectively.

3.3. Compounds Identification

The identification of the essential oil constituents was based on the comparison of their retention indexes relative to (C₈–C₂₂) -alkanes with those of literature or with those of authentic compounds available in our laboratory. Further identification was made by matching their recorded mass spectra with those stored in the Wiley/NBS mass spectral library of the GC-MS data system and other published mass spectra [21].

3.4. Statistical Analyses

Data were subjected to statistical analysis using “Statistica” statistical program package [22]. The percentages of volatile compounds are means of three experiments; the one-way analysis of variance (ANOVA) followed by Duncan multiple range test was employed and the differences between individual means were deemed to be significant at .

4. Results

Hydrodistillation of full ripened seeds of S. verbenaca, S. sclarea, and S. officinalis offered essential oils with average yields of 0.050, 0.047, and 0.045% (w/w on the dry weight basis), respectively.

Essential oil constituents of Salvia seeds were presented in Table 1. The results of analysis of essential oil of S. verbenaca seeds by GC and GC-MS techniques revealed the occurrence of thirty-two compounds. The essential oil composition showed that over 57% of the detected compounds were oxygenated monoterpenes followed by sesquiterpenes (24.04%) and labdane type diterpenes (5.61%). Apart from camphor, the main essential oil constituents of this sample were caryophyllene oxide (7.28%), 13-epi-manool (5.61%), δ-elemene (3.97%), and β-eudesmol (3.76%).

Table 1: Essential oil composition (%w/w) of Salvia verbenaca (Sv), Salvia officinalis (So), and Salvia sclarea (Ss) seeds.

Number
Compounds*
RIa
RIb
Sv
So
Ss
Identification

1
Hexanal
800
1093
0.42
—
—
GC-MS, Co-GC

2
(E)-2-Hexenal
852
1232
—
—
0.03
GC-MS, Co-GC

3
1-Hexanol
878
1360
—
1.29
—
GC-MS, Co-GC

4
Tricyclene
927
1014
0.96a
0.23b
0.08c
GC-MS

5
-Thujene
930
1035
—
3.08b
7.48a
GC-MS

6
-Pinene
939
1032
0.44b
1.26a
0.27c
GC-MS

7
-3-Carene
1012
1159
0.50a
0.03b
—
GC-MS

8
p-Cymene
1022
1280
0.37b
1.52a
0.12c
GC-MS

9
Limonene
1030
1203
—
—
0.17
GC-MS, Co-GC

10
-Phellandrene
1030
1218
—
—
0.03
GC-MS

11
1,8-Cineole
1033
1213
—
6.66a
0.12b
GC-MS, Co-GC

12
cis-Sabinene hydrate
1068
1556
—
0.19
—
GC-MS

13
cis-Linalool oxide
1072
1478
—
0.13
—
GC-MS

14
-Fenchone
1087
1406
0.26
—
—
GC-MS

15
trans-Linalool oxide
1087
1450
1.05
—
—
GC-MS

16
Linalool
1098
1553
0.84b
0.68b
24.25a
GC-MS, Co-GC

17
n-Undecane
1100
1100
2.65a
0.48c
0.75b
GC-MS, Co-GC

18
-Thujone
1102
1429
0.52b
14.77a
—
GC-MS

19
cis-allo-Ocimene
1113
1382
1.55a
0.14b
1.46b
GC-MS

20
-Thujone
1114
1451
—
4.30
—
GC-MS

21
Geijerene
1140
1338
—
1.01
—
GC-MS

22
Camphor
1143
1532
38.94a
13.08b
—
GC-MS

23
Borneol
1165
1719
—
3.54
—
GC-MS

24
Terpinen-4-ol
1177
1611
0.88a
0.09c
0.37b
GC-MS

25
p-Cymen-8-ol
1185
1864
—
0.17
—
GC-MS

26
-Terpineol
1187
1682
0.49b
2.42a
0.20c
GC-MS

27
-Terpineol
1189
1706
2.03a
0.91b
0.20c
GC-MS

28
Myrtenal
1190
1648
—
0.55
—
GC-MS

29
Myrtenol
1194
1804
—
0.28
—
GC-MS

30
cis-Sabinol
1210
1800
—
0.18
—
GC-MS

31
Nerol
1228
1797
—
—
0.98
GC-MS, Co-GC

32
Linalyl acetate
1239
1565
2.53b
0.08c
6.90a
GC-MS

33
Geraniol
1254
1857
—
0.33b
2.79a
GC-MS, Co-GC

34
Bornyl acetate
1285
1597
0.79a
0.16b
—
GC-MS

35
Thymol
1293
2198
—
0.37a
0.10b
GC-MS

36
Carvacrol
1296
nd
—
0.83b
1.93a
GC-MS

37
-Elemene
1337
1479
3.97a
0.07c
0.85b
GC-MS

38
-Cubebene
1348
1456
—
—
2.86
GC-MS

39
-Terpinyl acetate
1353
1709
4.77a
1.81b
0.29c
GC-MS

40
Eugenol
1357
2192
—
0.83
—
GC-MS, Co-GC

41
Neryl acetate
1366
1733
2.40a
—
0.36b
GC-MS, Co-GC

42
-Ylangene
1372
1493
—
0.04b
0.24a
GC-MS

43
-Copaene
1376
1497
—
0.01b
4.08a
GC-MS

44
-Damascenone
1381
1838
—
0.06
—
GC-MS

45
Geranyl acetate
1383
1765
—
—
1.94
GC-MS, Co-GC

46
-Bourbonene
1384
1533
1.73a
—
0.35b
GC-MS

47
-Elemene
1389
1600
—
0.16
—
GC-MS

48
Methyl eugenol
1402
2030
—
0.18
—
GC-MS, Co-GC

49
-Caryophyllene
1413
1612
0.27a
0.19b
0.24a
GC-MS

50
-Cubebene
1419
1549
1.50a
—
0.72b
GC-MS

51
Aromadendrene
1439
1628
0.66a
0.18b
—
GC-MS

52
(Z)--Farnesene
1441
1668
1.76
—
—
GC-MS

53
-Humulene
1454
1687
0.67b
3.71a
0.08c
GC-MS, Co-GC

54
allo-Aromadendrene
1460
1661
1.29b
1.43a
0.06c
GC-MS

55
-Amorphene
1474
1680
—
0.47
—
GC-MS

56
-Muurolene
1477
1704
0.05
—
—
GC-MS

57
Germacrene-D
1479
1726
—
1.18b
5.88a
GC-MS

58
epi-Cubebol
1491
1900
—
0.34a
0.24b
GC-MS

59
Bicyclogermacrene
1494
1755
—
1.29b
4.29a
GC-MS

60
(E,E)--Farnesene
1506
1758
—
0.24
—
GC-MS

61
-Bisabolene
1508
1741
1.10a
0.72b
—
GC-MS

62
-Cadinene
1513
1776
—
0.08
—
GC-MS

63
-Cadinene
1517
1773
—
0.53a
0.24b
GC-MS

64
-Calacorene
1542
1942
—
—
0.10
GC-MS

65
Germacrene-B
1558
1854
—
0.08b
1.29a
GC-MS

66
(E)-Nerolidol
1563
2050
—
1.41
—
GC-MS

67
Spathulenol
1575
2144
—
0.08a
0.03b
GC-MS

68
Caryophyllene oxide
1579
2008
7.28a
0.16c
3.18b
GC-MS

69
Viridiflorol
1592
2104
—
2.66
—
GC-MS

70
Humulene epoxide I
1596
2045
—
—
1.55
GC-MS

71
Humulene epoxide II
1606
2071
—
0.25
—
GC-MS

72
T-Cadinol
1642
2187
—
0.15b
0.52a
GC-MS

73
-Cadinol
1643
nd
—
0.41b
0.88a
GC-MS

74
-Eudesmol
1649
nd
3.76a
0.47c
1.72b
GC-MS

75
13-epi-Manool
Nd
nd
5.61a
2.22b
0.59c
GC-MS

Compound classes

Alcohols
2.46a
0.42b
0.03c

Aliphatic hydrocarbons
Alkanes
—
1.29
—

Aldehydes
0.84a
0.48c
0.75b

Monoterpene hydrocarbons
3.82c
6.45b
9.60a

Oxygenated monoterpenes
57.30a
50.14b
38.38c

Sesquiterpene hydrocarbons
13.00b
11.39b
21.27a

Norisoprenoids with 13 carbons
—
0.06
—

Phenylpropanes
—
1.01
—

Phenols
—
1.10b
2.03a

Oxygenated sesquiterpenes
11.04a
5.93c
8.12b

Labdane type diterpenes
5.61a
2.22b
0.59c

Total

*Components are listed according to their elution on apolar column (HP-5). RI: retention indices relative to C₈–C₂₂  n-alkanes on the aHP-5 and bHP-Innowax columns; nd: not detected; GC/MS: identification based on comparison of mass spectra; Co-GC: identification based on retention time comparison to authentic compounds. Values (means of three replicates) in the same lines with different letters (a–c) are significantly different at .

The essential oil of S. officinalis seeds showed a higher percentage of monoterpenes (56.59%) than sesquiterpenes (17.32%). Oxygenated derivatives were major among the monoterpenes (50.14%), while they represented only 5.93% of sesquiterpenes. Seeds essential oils were characterised by the predominance of α-thujone (14.77%), camphor (13.08%), and 1,8-cineole (6.66%). Viridiflorol (2.66%) and α-humulene (3.71%) were also detected in seeds essential oil of sage. Furthermore, other minor compounds were found, especially the labdane type diterpene 13-epi-manool.

Essential oil compounds of S. sclarea seeds were representing 80.79% of total essential oil components. The essential oil is predominated by monoterpenes accounting for 47.98%; their oxygenated derivatives (38.38%) prevailed on hydrocarbon ones (9.60%). The sesquiterpenes pool is less numerous (29.39%). In addition, one diterpene compound, 13-epi-manool, is detected at a level of 0.59% and some phenols (2.03%) such as thymol and carvacrol were produced in small amounts (0.1% and 1.93%, resp.). The oxygenated monoterpenes were characterised by linalool (24.25%), geraniol (2.79%), and their ester derivates (linalyl acetate (6.90%) and geranyl acetate (1.94%)). The sesquiterpenes characteristics of seeds were germacrene-D (5.88%), bicyclogermacrene (4.29%), and α-copaene (4.08%).

5. Discussion

As for S. verbenaca, the oil yield was higher than that offered by seeds originated from Tunisia [12]. Compared with leaves of S. verbenaca from Tunisia [14], seeds herein studied appeared as moderately rich in volatile oil. Recovered essential oil from S. sclarea seeds appeared to be near to literature data which showed that inflorescences oil yield of a cultivated strain developed in India [23]. In S. officinalis, the fruits of sage cultivated in Tunisia are distinguished by oil yields of 0.39% [24]. Thereby seeds appeared as oil-moderate organs contrary to the different other parts of the species owing to the genus Salvia its aromatic reputation amongst Lamiaceae family.

As regards essential oil composition of S. verbenaca, it is worthy to note that tricyclene and camphor are also common to the seeds sample from Tunisia [12] so we may suggest that these two compounds are significant markers of essential oil compounds of S. verbenaca seeds whatever the sample origin is. Interestingly, the essential oil of the studied seeds could be employed as antimicrobial agent since their high percentage of camphor is associated with an efficient antimicrobial activity according to Magiatis et al. [25] and Bougatsos et al. [26].

In S. officinalis, the predominance of α-thujone, camphor, and 1,8-cineole endowed to the essential oil an antimicrobial activity [27]. Viridiflorol and α-humulene identified in several S. officinalis essential oils showed antiacetylcholinesterase activity used in the treatment of Alzheimer’s disease [28], antifungal property [29], and cytotoxic activity against some tumor cell lines [30]. Furthermore, the labdane type diterpene 13-epi-manool displays in vitro a cytotoxic activity against human leukemic cell lines [31]. We noted that seeds had a similar qualitative composition to aerial parts with the predominance of α-thujone, β-thujone, camphor, and 1,8-cineole. These monoterpenes are taken as significant parameter to differentiate S. officinalis from other species [32]. Their amount is lower than that in leaves but matches the ranges of the standard ISO 9909 for official sage oil except for the toxic ketone α-thujone which had a lower amount (14.77%). These findings promote the use of the seeds essential oil in food industry.

Similar to the studied S. sclarea essential oil, the sesquiterpenes were mainly composed of germacrene-D, α-copaene, and bicyclogermacrene in plant inflorescence according to Carrubba et al. [11] and Lorenzo et al. [33]. In our sample, the essential oil of S. sclarea flowering shoots raised in experimental plots in India was characterised by linalool (36.6–41.9%) and linalyl acetate (13.2–19.2%) as main compounds [23]. The wild-growing S. sclarea collected at flowering stage from central Greece [7] and Spain [6] showed a close similar composition regarding linalool and linalyl acetate (30.43%, 32.97% and 19.75%, 16.85%, resp.). Generally, S. sclarea essential oil is extracted from flowering shoots which are found to be rich in linalool and linalyl acetate. According to Carrubba et al. [11] high amounts of linalool and linalyl acetate are typical of good quality oil suitable for flavouring purposes. Furthermore, linalool plays a major role as anti-inflammatory suggesting that linalool-producing species are potentially anti-inflammatory agents [34]. Moreover, linalool has an ecological role since it constitutes one of the common components of floral scent that can attract a large variety of insects that convey pollen [35]. The present study showed that essential oil derived from seeds had a similar composition to the flowering parts. Thus, seeds seemed to display the same enzymatic patterns of the essential oil biosynthesis as the flowers, while the essential oil composition of S. sclarea vegetative organs displayed different qualitative trends from reproductive parts.

6. Conclusions

Overall, it emerges that tricyclene and camphor were biochemical markers of the essential oil of S. verbenaca seeds. Being rich in camphor, seeds could be used as antimicrobial agent. Another point that should be highlighted is that S. officinalis seeds had the same α-thujone chemotype as leaves, whereas these two organs showed some quantitative differences leading to the safe use of seeds essential oil in food industry. From a qualitative standpoint, seeds of S. sclarea seemed to have the same enzymatic trend as flowers characterized by the prevalence of linalool. It is noteworthy to mention that linalool-producing seeds as S. sclarea were suitable for flavouring purposes and constitute potential anti-inflammatory agents.

Conflict of Interests

The authors declare that there is no conflict of interests regarding the publication of this paper.

References

I. C. Hedge, A Global Survey of the Biogeography of Labiatae, Royal Botanical Gardens, Kew, UK, 1992.

G. Penso, Index Plantarum Medicinalium Totius Mundi Eorumque Synonymorum, OEMF, Milano, Italy, 1983.

A. Y. Leung and S. Foster, Encyclopedia of Common Natural Ingredients Used in Food, Drugs and Cosmetics, John Wiley & Sons, New York, NY, USA, 2nd edition, 1996.

T. A. Al-Howiriny, “Chemical composition and antimicrobial activity of essential oil of Salvia verbenaca,” Biotechnology, vol. 1, pp. 45–48, 2002.

F. Chialva and F. Monguzzi, “Composition of the essential oils of five Salvia species,” Journal of Essential Oil Research, vol. 4, pp. 447–455, 1992.

M. E. Torres, A. Velasco-Negueruela, M. J. Pérez-Alonso, and M. G. Pinilla, “Volatile constituents of two Salvia species grown wild in Spain,” Journal of Essential Oil Research, vol. 9, no. 1, pp. 27–33, 1997.

D. Pitarokili, O. Tzakou, and A. Loukis, “Essential oil composition of Salvia verticillata, S. verbenaca, S. glutinosa and S. candidissima growing wild in Greece,” Flavour and Fragrance Journal, vol. 21, no. 4, pp. 670–673, 2006.

V. Radulescu, S. Chiliment, and E. Oprea, “Capillary gas chromatography-mass spectrometry of volatile and semi-volatile compounds of Salvia officinalis,” Journal of Chromatography A, vol. 1027, no. 1-2, pp. 121–126, 2004.

P. Avato, I. M. Fortunato, C. Ruta, and R. D’Elia, “Glandular hairs and essential oils in micropropagated plants of Salvia officinalis L.,” Plant Science, vol. 169, no. 1, pp. 29–36, 2005.

S. Marie, M. Maksimovic, and M. Milos, “The impact of the locality altitudes and stages of development on the volatile constituents of Salvia officinalis L. from Bosnia and Herzegovina,” Journal of Essential Oil Research, vol. 18, no. 2, pp. 178–180, 2006.

A. Carrubba, R. La Torre, R. Piccaglia, and M. Marotti, “Characterization of an Italian biotype of clary sage (Salvia sclarea L.) grown in a semi-arid Mediterranean environment,” Flavour and Fragrance Journal, vol. 17, no. 3, pp. 191–194, 2002.

M. B. Taarit, K. Msaada, and B. Marzouk, “Chemical composition of fatty acids and essential oils of Salvia verbenaca L. Seeds from Tunisia,” Agrochimica, vol. 54, no. 3, pp. 129–141, 2010.

M. B. Taarit, K. Msaada, K. Hosni, T. Chahed, and B. Marzouk, “Essential oil composition of Salvia verbenaca L. growing wild in Tunisia,” Journal of Food Biochemistry, vol. 34, no. 1, pp. 142–151, 2010.

M. Ben Taarit, K. Msaada, K. Hosni, N. Ben Amor, B. Marzouk, and M. E. Kchouk, “Chemical composition of the essential oils obtained from the leaves, fruits and stems of Salvia verbenaca L. from the northeast region of Tunisia,” Journal of Essential Oil Research, vol. 22, no. 5, pp. 449–453, 2010.

R. Ayerza, W. Coates, and M. Lauria, “Chia seed (Salvia hispanica L.) as an ω-3 fatty acid source for broilers: influence on fatty acid composition, cholesterol and fat content of white and dark meats, growth performance, and sensory characteristics,” Poultry Science, vol. 81, no. 6, pp. 826–837, 2002.

B. Heuer, Z. Yaniv, and I. Ravina, “Effect of late salinization of chia (Salvia hispanica), stock (Matthiola tricuspidata) and evening primrose (Oenothera biennis) on their oil content and quality,” Industrial Crops and Products, vol. 15, no. 2, pp. 163–167, 2002.

J. E. Kinsella, K. S. Broughton, and J. W. Whelan, “Dietary unsaturated fatty acids: interactions and possible needs in relation to eicosanoid synthesis,” Journal of Nutritional Biochemistry, vol. 1, no. 3, pp. 123–141, 1990.

I. C. Hedge and L. Salvia, Flora of Turkey and the East Aegean Islands, Edinburgh University Press, Edinburgh, UK, 1982.

A. Estilai, A. Hashemi, and K. Truman, “Chromosome number and meiotic behavior of cultivated chia, Salvia hispanica (Lamiaceae),” Hortscience, vol. 25, pp. 1646–1647, 1990.

T. Baytop, Türkiye’de bitkilerle tedavi (geçmiste ve bugün), Baskı, Nobel Tıp Kitapevleri, Çapa-Ýstanbul, Konak-Ýzmir, Sıhhıye-Ankara, Turkey, 1999.

R. P. Adams, Identification of Essential Oil Components by Gas Chromatography/Quadrupole Mass Spectroscopy, Allured, Carol Stream, Ill, USA, 2001.

Statsoft, STATISTICA for Windows (Computer Program Electronic Manual), StatSoft, Tulsa, Okla, USA, 1998.

S. K. Lattoo, R. S. Dhar, A. K. Dhar, P. R. Sharma, and S. G. Agarwal, “Dynamics of essential oil biosynthesis in relation to inflorescence and glandular ontogeny in Salvia sclarea,” Flavour and Fragrance Journal, vol. 21, no. 5, pp. 817–821, 2006.

M. Ben Taarit, K. Msaada, K. Hosni, M. Hammami, M. E. Kchouk, and B. Marzouk, “Plant growth, essential oil yield and composition of sage (Salvia officinalis L.) fruits cultivated under salt stress conditions,” Industrial Crops and Products, vol. 30, no. 3, pp. 333–337, 2009.

P. Magiatis, A. L. Skaltsounis, I. Chinou, and S. Haroutounian, “Chemical composition and in vitro antimicrobial activity of the essential oils of three Greek Achillea species,” Zeitschrift für Naturforschung, vol. 57, pp. 287–290, 2002.

C. Bougatsos, O. Ngassapa, D. K. B. Runyoro, and I. B. Chinou, “Chemical composition and in vitro antimicrobial activity of the essential oils of two Helichrysum species from Tanzania,” Zeitschrift fur Naturforschung, vol. 59, no. 5-6, pp. 368–372, 2004.

V. Jalsenjak, S. Peljnjak, and D. Kuštrak, “Microcapsules of sage oil: essential oils content and antimicrobial activity,” Pharmazie, vol. 42, no. 6, pp. 419–420, 1987.

M. Miyazawa, H. Watanabe, K. Umemoto, and H. Kameoka, “Inhibition of acetylcholinesterase activity by essential oils of Mentha species,” Journal of Agricultural and Food Chemistry, vol. 46, no. 9, pp. 3431–3434, 1998.

H. J. M. Gijsen, J. B. P. A. Wijnberg, G. A. Stork, A. de Groot, M. A. De Waard, and J. G. M. Van Nistelrooy, “The synthesis of mono- and dihydroxy aromadendrane sesquiterpenes, starting from natural (+)-aromadendrene-III,” Tetrahedron, vol. 48, no. 12, pp. 2465–2476, 1992.

S. L. da Silva, P. M. Figueiredo, and T. Yano, “Cytotoxic evaluation of essential oil from Zanthoxylum rhoifolium Lam. leaves,” Acta Amazonica, vol. 37, no. 2, pp. 281–286, 2007.

K. Dimas, C. Demetzos, M. Marsellos, R. Sotiriadou, M. Malamas, and D. Kokkinopoulos, “Cytotoxic activity of labdane type diterpenes against human leukemic cell lines in vitro,” Planta Medica, vol. 64, no. 3, pp. 208–211, 1998.

J. Bruneton, Pharmacognosy, Phytochemistry, Medicinal Plants, Tech. & Doc. Lavoisier, Paris, France, 1999.

D. Lorenzo, D. Paz, P. Davies et al., “Characterization and enantiomeric distribution of some terpenes in the essential oil of a Uruguayan biotype of Salvia sclarea L,” Flavour and Fragrance Journal, vol. 19, no. 4, pp. 303–307, 2004.

A. T. Peana, M. D. L. Moretti, and C. Juliano, “Chemical composition and antimicrobial action of the essential oils of Salvia desoleana and S. sclarea,” Planta Medica, vol. 65, no. 8, pp. 752–754, 1999.

E. Pichersky, J. P. Noel, and N. Dudareva, “Biosynthesis of plant volatiles: nature’s diversity and ingenuity,” Science, vol. 311, no. 5762, pp. 808–811, 2006.