Volatile Constituents of the Aerial Parts
Gary R. Takeoka,*
Western Regional Research Center, Agricultural Research Service, US Department of Agriculture, 800 Buchanan Street,
Albany, CA 94710
Department of Integrative Biology, University of California, Berkeley, California 94720-2465
Institute of Ecological Phytochemistry, Hankyong National University, Ansung-City, Kyonggi-Do 456-749, Korea
Volatile constituents of the aerial parts of fresh white sage (Salvia apiana
Jepson) were isolated by extraction
with diethyl ether followed by high vacuum distillation with a solvent assisted flavor evaporation (SAFE) apparatus.
The isolated volatiles were analyzed by GC and CC/MS. A total of 84 constituents were identified (constituting
95.1% of the total area), 11 of which were tentatively identified. The volatiles were characterized by a high content
of hydrocarbon and oxygenated monoterpenes. The major constituents identified were 1,8-cineole (34.5%), camphor
(21.7%), 3-pinene (7.4%), a-pinene (6.4%), -3-carene (6.4%), camphene (3.9%), limonene (3.5%), myrcene (3.2%),
and terpinolene (1.3%).
Key Word Index
Salvia apiana, white sage, Lamiaceae, essential oil composition, 1 ,8-cineole, camphor, 1 ,3,5-undecatriene isomers,
Jepson is one of approximately 900 world-
wide species of
found in the Lamiaceae family (1). It
generally grows below 1500
Baja California, in the South
Coast (SCo), Transverse Ranges (TR), and Peninsular Ranges
(PR) sub-regions of southwestern California as well as in the
western edge of the Desert Province (DR) in the southeastern
portion of California (1). The plant has been used to treat chest
colds, coughs, sore throats, systemic poison oak rashes and
acute candidal vaginitis, and has been widely used by natives
(2,3) and in traditional Churnash healing (4). The leaves were
eaten, smoked and used in sweat baths by Cahuilla Indians to
treat upper respiratory infections (3). Four new
6,7-didehydroferruginol; 6,7-didehydrosempervirol; 16-hy-
abeo-abieta-1(10),6,8,11,13-pentaene, two new diterpene quino-
nes, 16-hydroxyroyleanone and 6-deoxo-5,6-didehydrolanugon
Q as well as the known compounds, ferruginol, miltiodiol,
cryptanshinone, lanugon Q and salvicanol have been isolated
and characterized from the roots of S. apaina (5).
a-Amyrin, oleanolic acid and ursolic acid were identi-
fied in the dried aerial parts of S. apiana (6).
Hoffmann (7) identified two abietane acids, 16-hydroxy-
carnosic acid and carnosic acid in the leaves of
Luis and co-workers (8,9) identified the new C terpenoids,
(22,6)-olide along with the known diterpenes, 16-hycir
carnosic acid, 16-hydroxycarnosol, 16-hydroxyrosmanol,
16-hydroxy-7-methoxyrosmanol, rosmanol, 7-epirosmanol
and salvicanol in the aerial parts of
constituents from S.
Jepson leaves have been shown
to inhibit the root growth of
seedlings (10). It was postulated the volatiles may be deposited
when dew condenses on the seedlings in the field though the
active constituent(s) was not characterized.
While the composition of volatiles from the essential oils
species have been reported (11-18) there
is only limited knowledge of the volatile constituents in white
sage (19-21). The aim of this study was to provide a more
*Address for correspondence
Received: February 2008
Revised: April 2008
1041 -2905/10/0003-0241$14.00/O—© 2010 Allured Business Media
Accepted: August 2008
Vol. 22, May/June 2010
Journal of Essential Oil Research/241
trans-linalool oxide A furanoid
1 (E E) 3 5undecatriene
Table I. Volatile constituents
of theaerial parts of
Peak area percentage of total FID area (assuming all response factors of 1)
represents a % area <0 1
peak area of this constituent and the following constituent
weré cIclated 6n the basis of the GC/MS total ion chromatàgrrn;
identifications (in parentheses) assigned based on mass spectra and
reported in Adams
(2007); 'Tentative identifications (in parentheses) assigned based on mass spectràreported in Wiley Registry of Mass Spectral Data,
Edition; Tentative identifications (in
parentheses) assigned based on mass spectra and
reported in MassFinder 3 (Dr. Hochmuth Scientific Consulting Hamburg Germany)
This constituent and the next
eluting constituent were resolved by GC/MS but were not separated by GC FID correct isomer not identified
comprehensive;knowledge of the volatiles in
stored in the dark after addition of 1-2 ppm of antioxidant 330
of S. apiana Jepson.
Freslileaves and flowering tops of S. apiana
Jesdnwee collected in the UC Davis Bbtanical Gardens in
June 2007. The samples were prepared the same day that they
were picked. A voucher specimen was deposited in the Jepson
Herbariñrn, University of California, Berkeley, CA.
Diethyl ether was freshly distilled through
60 cth long Pyrex column packed With glassheliées and
benzene; Ethyl Corporation, Richmond, VA).
crushed with a mortar and pestle under liquid nitrogen. The
material was divided into equal portion and added to two 250
mL Pyrex glass bottles with Teflon lined screw caps. Approxi-
mately 125 mL of ether was added to each bottle. The bottles
were covered with aluminum foil and were sonicated in an
ultrasonic bath for 15
The bottles were shaken throughout
the clay every 2 h and allowed to stand overnight. The dark
242/Journal of Essential Oil Research
Vol. 22, May/June 2010
Takeoka et al.
green extract was filtered through pre-rinsed (ether) filter
paper (ED fluted filter paper, grade 513, size 24 cm, Eaton-
Dikeman, Mount Holly Springs, PA). The extract was dried
overnight over anhydrous sodium sulfate (previously heated to
150°C for several hours to remove volatiles). The extract was
subjected to high vacuum distillation using a solvent assisted
flavor evaporation (SAFE; 22) apparatus. The SAFE apparatus
was heated to 40°C with a circulating water bath and the ex-
tract was added to the dropping funnel of the apparatus. The
distillation flask (500 mL) was heated to 40°C in a water bath.
The receiving flask for the distillate and the safety-cooling
trap of the SAFE apparatus were cooled with liquid nitrogen.
The SAFE apparatus was connected to a high vacuum pump
(<0.01 Pa) and then the mixture in the dropping funnel was
added in small aliquots into the distillation flask over 20 mm.
The distillate was concentrated using a Vigreux column (15 x
1 cm) and water bath at 40°C. The extract (0.6637
for CC and CC/MS analyses.
A Hewlett-Packard (Avondale,
PA) 6890 gas chromatograph equipped with a flame ionization
detector (FID) was used. A 60 m X 0.32 mm DB-1 (d
gm; J&W Scientific, Folsom, CA) fused silica capillary column
was employed. The oven temperature was programed from
30°C (4 min isothermal) to 200°C at 2°C/mm (final hold was
25 mm). Split injections (1:20) were used. Helium was used as
the carrier gas at a linear velocity of 38.3 cm/s (30°C).
Gas chromatography/mass spectrometry (GC/MS):
The GC/MS system consisted of an Agilent Technologies 6890
gas chromatograph coupled to an Agilent Technologies 5973
Network MSD (Agilent Technologies, Palo Alto, CA). A 60 m
X 0.25 mm DB-1 MS fused silica capillary column was used
= 0.25 i.tm). The CC oven was programed from 30°C (4
min isothermal) to 200°C at 2°C/mm (final hold was 35 mm).
Helium was used as the carrier gas at a headpressure of 22
psi. The injector, transfer line, ion source and quadrupole tem-
peratures were 180°C, 200°C, 170°C and 130°C, respectively.
The mass spectrometer was operated in the electron impact
mode with an ionization voltage of 70 eV. A scan range of
35-320 at 4.94 scans/s was employed.
Identification of volatiles:
Volatile constituents were
identified by comparing the component's mass spectrum and
experimental retention index (I) with that of an authentic
reference standard. The retention system proposed by Kováts
(23) was utilized. When standards were not available, tentative
identifications were assigned based on mass spectra and reten-
tion indices reported in Wiley Registry of Mass Spectral Data,
Edition (John Wiley & Sons, Inc., Hoboken, NJ), NIST/
EPAJNIH Mass Spectral Library 2005 (U.S. Department of
Commerce), MassFinder3 (Dr. Hochmuth Scientific Consult-
ing, Hamburg, Germany) and Identification of Essential Oil
Components by Gas Chromatography/Mass Spectrometry, 4"
Results and Discussion
Aerial portions of S.
Jepson were extracted with
diethyl ether and the volatiles were isolated by high vacuum
distillation using a SAFE apparatus. CC analysis of the extract
(0.6637 g) revealed that volatiles constituted
while ether made up 42.5% (0.2821
The yield of volatiles
from the sample was 0.4%. A total of 84 constituents were
identified (constituting 95.1% of the total area), 11 of which
were tentatively identified. The major constituents identified
were 1,8-cineole(34.5%), camphor (21.7%), -pinene(7.4%), a-
pinene (6.4%), -3-carene(6.4%), camphene (3.9%),limonene
(3.5%), myrcene (3.2%), and terpinolene (1.3%). Emboden
and Lewis (20) reported similar percentages of 1,8-cineole
(39.5-46.6%), combined camphor and borneol(30.6-40.1%),
pinene(6.7-7.6%), a-pinene (5.5--6.2%), camphene(3.9-4.9%)
and limonene (3.3-5.1%) in S.
more recent study on S.
(21) found similar levels
of 3-pinene (9.1%), a-pinene (9.0%), and limonene (2.0%)
but lower levels of 8-3-carene (1.3%), camphene (0.4%) and
camphor (2.1%). 1,8-Cineole was also reported as the main
constituent though its percentage (71.6%) was higher in the
previous investigation (21). 1,8-Cineole has been reported to
be useful for the treatment of brochial asthma, cough, and
liver failure induced by endotoxemic shock (25-27). This
monoterpene oxide has been shown to possess gastroprotective
activity, an effect related to both its antioxidant activity and its
lipoxygenase inhibitory effects (28). Juergens and co-workers
(29) demonstrated that 1,8-cineole was a strong inhibitor
of TNF-a and IL-1P production in stimulated lymphocytes
and monocytes. They also showed that 1,8-cineole at known
therapeutic blood concentrations had inhibitory effects on
the chemotactic cytokmne of IL-8 and IL-5. The reduction of
cytokine production suggested an anti-inflammatory mode of
action and consequently inhibition of cytokine induced airway
mucus hypersecretion rather than simple secretolytic activity.
Isolated monoterpenes such as 1,8-cineole may offer a new
opportunity for initial and long-term treatment of asthma and
chronic obstructive pulmonary disease (COPD).
Salviafructicosa oil and its major compounds, thujone and
1,8-cineole, showed relatively low antimicrobial activity against
eight bacterial strains, Escherichia coli (NCIMB 8879 and
(NCIMB 9518 and NCIMB 8625),
3610), while camphor was exhibited almost no activity against
the bacteria tested (30).
camphor and thujone exhibited cytotoxic activity against
African Green Monkey kidney (Vero) cells and high levels of
virucidal activity against herpes simplex virus 1 (30). Pitarokili
and co-workers(15) tested the antifungal activity of 1,8-cineole
and camphor (the main constituents identified in S.
oil) against five phytopathogenic fungi,
f. sp. dianthi, Fusarium proliferatum, Fusarium solani f. sp.
cucurbitae, Rhizoctonia solani
Camphor showed moderate activity against S.
but displayed lower activity against the three
species. 1,8-Cineole exhibited only slight activity
against the five fungal species. The oil of
activity than camphorwhich led the research-
ers to conclude that other components exert direct activity or
possibly a synergistic effect with camphor. Three previously
constituents, cymene, oc-pinene oxide and
3-caryophyllene oxide, were not detected in this study (21).
Vol.-22, May/June 2010
Journal of Essential Oil Research/243
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L. oil (12,14,16). To
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been reported in
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1-(E,ZZ)-3,,8-undecatetraene has been described by Berger
et aL (31). The configuration of the double bond in the C-5
position is crucial as the corresponding isomers, 1-(E,E)-3,5-
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times higher, respectively (31). 1-(E,Z)-
3,5-Undecatriene has a balsamic, spicy, pinewood odor while
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odor (31). .Sesquiterpene hydrocarbons were identified for the
first time in S. apiana,
though they have been reported in other
species (13,17,18). The most abundant sesquiterpenes
were 3-caiyophyllene(1.0%), 8-cadinene(0.3%), germacrene B
(0.2%); guaia-6,9-diene(0.2%), 13-bisabolene(0.2%),y-cadinene
(0:2%), a-copaène (0.1%), a-gurjunene (0.1%), a-humulene
(0.1%), y-muurolene (0.1%), bicyclogermacrene (0.1%), and
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Vol. 22, May/June 2010