Composition of the essential oil of Salvia officinalis L. from various European countries

Abstract

Variations in the essential oil composition of Salvia officinalis L. growing in Estonia and in other European countries were determined. The oils were obtained in yields of 2.2-24.8 mL kg(-1). In three samples, the content of essential oil did not conform to the EP standard (10 mL kg(-1)). Variations in the essential oil composition of sage were studied using capillary gas chromatographic methods. A total of 40 components were identified. The principal components in the sage oils were 1,8-cineole, camphor, alpha-thujone, beta-thujone, borneol, and viridiflorol. The chemotypes of sage were not determined in investigated samples. The concentration of the main compounds in the drugs cultivated in Estonia varied in about the same range as the concentrations of these compounds in the oils of drugs obtained from other countries. The comparatively high concentration of toxic thujones seem to be characteristic to sage leaves cultivated in Estonia.

Full text

Naturcl
Protlut't
Researt'lt,
Vol.2l, No. 5, I
May 2007,
406 411
Taylor
&
Francis
Taylor &F.ancis Crolrp
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Composition
of the
essential
oil of Salvia
officinalis
L.
from Yarious
European
countries
AIN RAAL*t,
ANNE
ORAVI
and ELMAR
ARAI(f
tlnstitute
of
Pharmacy, University
of
Tartu,
Noontse 1,5041 l, Tartu
(Estclnia)
{lnstitute
ol Cherristry,
Tallinn
University of
Technology,
Akadeemia tee
15, 12618,
Tallinn
(Estonia)
,
(
Received 22
November
2005:
in.finul
.lbrtn
14
December
2005\
Variations
in the
essential oil
composition
of
Sulvkr ofJicinulis
L.
growirtg
in
Estonia ancl in
other
European
countries
were determined.
The oils
rvere
obtained
in
yields
ol
2.2-24,8mLkg-r.
In three
samples,
the content
of
essentiirl oil did
not conform to the
EP
standarcl
(l0mLkg-r).
Varialions in the
essential
oil composition
of serge
were
studied
using
capillary
gas
chromatographic
methods.
A total ol
40 comportents were
identified. The
principal colnponents
in
the sage oils
rvere
1,8-cineole,
ciunphor, a-thujone,
p-thu.ione.
borneol,
and
yiridiflorol.
The
chenotypes
ol sage
were
not determined
in investigated
samples. The
concentration
of the
rnain compounds
in the
drugs
cultivated in
Estonia varied in
about the
same
ralnge
as the
concentrations
ol'these
compounds
in the oils of drugs obtained
fronr other
countries.
The comparatively
high concentration
oltoxic
thujones seenl
to be chirtacteristic
to
sage
leaves
cultivated
in
Estoniir.
Keytvord.u;
satvitt
oJJit:inulls
L.; European
countriesl
Essential oil;
Terpenes; C-hentotypes;
European
PharmacoPocia
1. Introduction
Many
,snlvia
species
ancl
varieties
are
known,
They belong to
the
mint
family
(Labiatae).
The
most
important
of
these
are
Dalmatian
(common)
sage
(Salvitr
officinali,s
L.), Greek
sage
(.S.
tiloba
L.) and
Spanish
sage
(S.
lat'andulaqfttlia
(L.)
Vahl.).
S.
o/ficinalis
L. is an
odorous
small
perennial shrub
native to the
Mediterranean
region
(Dalmatia,
Albania,
Turkey,
Italy)
and
is cultivated
in many
parts
of the
world.
The
sage
leaves
and
their
essential
oils
are
used
commonly
in the
food, drug
and
perfitmery
industries
[1].
Sage
leaves
and
essential
oils
are stated
to
possess
carminative,
antispasmodic,
antiseptic,
astringent
and
antihidrotic
properties
[2].
External
indica-
tions of
the
drug,
essenti?tl
oil
and their
preparations
are
inflammations
and infections
of
the
mucous
membranes
of
throat
and
mouth
such as
stomatitis,
gingivitis and
pharyngitis,
interrral
indications
are
dyspeptic
symptoms
and
excessive
perspiration
[Z+1.
fn.
essential
oil
is
reported
to
be a
moderate
skin
irritant
[2].
*Corresponcling
author.
Tel.:
+3727375281.
Fax: +3727375289.
Email:
ain
raal(li)ut'ee
Nalural
Producl
Rcsecu'clt
ISSN
1478-6419
prinr/ISSN 1029-2349
online
@
2007 Taylor
& Frnncis
httP://www.tandl.co.uk/journals
DOI:
I 0.
1080/14786410500528478

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Contposition
o/
the essential oil o./
S. officinalis
L.
40l,
The
content of essential
oil in
the dried savage leaves
is usually
up
to 2.5(2.8)% (1,j)
or between
1.5-3.5%
t4l.European
Pharmacopoeia (EP)
standard
for
whole
s. offic:inal.is.folium is
not less
than l5mLkg-r
(-1j%)
and
for
cut
drug minimum
l0mLkg-r
(-1.0%)
t5l.
The essential
oil composition
of different
Sailin species
has
been
previously
stuclied
[6-
20]. Main
oil components
of S.
lavqnduhefolia were
camphor,
borneol
and 1,8-cineol
[13].
ln
the oil from
S. ,stenoJulla,
a-phellandrene
and
cv-bisabolene
were
found
in high
quantities
[12].
a-copaene
and
germacrene
D dominated
in
the
oil of
S. aethiopr,r
[ll],
but sclareol, linalool
and linalyl
oxide in the
oil of S. scalare[7,111.
The biological
properties
of essential oil of
S. ofJicinalis
are attributed
mainly
to
cv- and
B-thujone,
camphor
and
1,8-cineole
[2-4,6,10,15,17,r9].
Because
of the
thujone-
rich
essential oil. the drug
is antimicrobial
and antiviral
(11).
cy- and
B-thujone
are
the
major components
of the essential
oil
(35-50%,
mainly
cu-thujone)
(2).
The
rnaximum
contents of o- and
B-thujone
are up to
63 and
l3%o
respectively.
a-thujone,
which
is
present
as
a
higher
proportion
in
the essential oil, is rnore
toxic
than
p-thujone.
It is
a
codvulsant, and
the
mechanism
ofa-thujone neurotoxicity
has recently
been
elucrdated.
The
oral
LD5e
of both thujones in rats
was found to
be 192mgkg-r
[3].
Thujones-rich
essential oil of sage is known
to be abortifacient
and
emmenagogic.
Expectorant
and antibacterial activities
have been reported
for 1,S-cineole
[2].
The
antioxidant
activity
of sage
polyphenols
has also been evaluated
[9,14,20].
In Estonia, S.
fficinalis
L.
plants
are cultivated. The
composition
of
Estonian
sage
oil
has not
been
previously
investigated using
capillary
gas
chromatographic
methods.
ln tlris work, the chemical
composition of essential oil
of S.
fficinalis
L.
cultivated in
Estonia was compared with samples from other European
countries
and with
EP
standards.
2. Results and discussio
The
essential oil
isolated fi'om
the
leaves
of S.
fficinali:;
L.
by hydrodistillation
was
obtained
in
a
yield
of
2.2-24,8mLkg-r
(table
l). The
content of
essential oilin samirles
from Scotland
(4.2mLkg-r),
Moldavia
(2.2mLkg-t)
ancl Estonia
(5.1mLkg-r;
dicl
not conform
to the EP standard
(l0ml.kg-r)
for cut
drug of sage folium
[5].
The
highest content
(24.tlmLkg-t)
of essential oil was determined
in sample from
France.
Also drugs from Greece and Ukraine are
rather
essential oil rich
(21.8
and 2l.l niLkg-r
respectively).
Thus, differences between
minimum and
maximum contents are
more
than l0
times.
Forty constitnents,
representing
89.7-98.4%
of the
totai oil,
have been
identified
in
the l2 studied
samples
(table
l).
Also identified tentatively
were
(E)-anethole
and
isobornyl acetate
(totally
0-0.4%), cr-bergarnotene
(0-0.6%),
viridiflorenc
(0-0.6%),
ledol
(0-0.8%),
B-bisabolol
(0-1.7%),
farnesol, and u-santalol
(0-0.a%)
ancl manool
(0.3-8.2%).
With mentioned terpenes,
all analysed constituents represent 96.3-99.8o/o
of the
total oil.
Oxygenated
nronoterpenes
(54.9*74.5o/o)
arc the major fraction
of the sage oil, fi'om
which
1,S-cineole,
camphor, o-thujone,
p-thujone,
borneol and
bornyl acetate
were
the
main components
(table
I). The monoterpene
fraction ranged liom
0.7
to 23.0o/o,
a-pinene,
camphene,
p-pinene
and
myrcene being its main constituents. In the

A.
Raal et
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410
A.
Raal et al.
sesquiterpene
fraction
(3.4-15.0%),
(E)-B-caryophyllene
and
cy-humulene
were
founcl
in
the
highest
quantities.
The oxygenated
sesquiterpenes (3.8-275%)
were
clominated
by
viridiflorol
and manool
(0.3
8.2%,
tentatively
identified),
Comparison
between
the
sage
oil compositions
from
different
European
countries
showed
some differences
in
the
quantities
of the main
constituents
(table
1). The
highest
content
of camphor
(29.8%),
borneol
(11.8%)
and bornyl
acetate (7.8%)
was founcl
in
the
oil from
Scotland
origin.
In contrast
to other
samples,
Scottish
sage
contained
terpinen-4-ol,
a-terpineol,
thyrnol,
carvacrol, caryophyllene
oxide
anci humulene
epoxide
from 2.1
to 4.3oh
(other
samples contained
below
1.8%
of
these
compounds).
The sarnple from
Greece was
rich in 1,8-cineole
(45.3%)
and
monoterpencs
cr-pinele,
camphetre,
B-pinene
and myrcetre
(total23.\oh).
The drug
from
Moldavia
containecl
less
monoterpenes
(0.7%)
and
oxygenated monoterpenes (54.9%)
than
the
other
samples
studied,
but was rich
in
(E)-B-caryophyllene
(7.5%),
humulene
(7.5%)
an<l viridiflorol
(15.7%).
The chemotypes
of
sage were not
determined
in
the investigated
samples.
The
concetttrations
of the
main compounds
of
three
sage
samples
fi'om
Estqnia
(Sainples
9*11) cultivated
in 2000*2002were quite
similar
to
the samples
from
France,
Hungary, Belgium, Russia
and
ukraine. In the
sample
12.
cultivated
in 2003,
the
fraction
of monoterpenes
wits less
(2.8%)
and
the fractions
of sesquiterpenes (13,3%)
and
oxygenated sesquiterpenes
(13.2%)
was higher
than
that in
the other
Estonian
samples. The
total content
of toxic thujones
(a-
and
B-thujone)
in
the
samples
9*12
from Estonia was
28.1, 29.3,30.2
and 36.9oh respectively,
in
the samples
from
other
courrtries
their
concentration
was comparatively low:
(3.4-
14.2o/o
in
the samples
fron-t
Ukraine,
Greece, Scotland and
Belgium) or medium
(in
samplcs
f}om
Russia
(23.3%)
ancl Hungary
(25.2%)).
The
concentration of main
compounds in the sage leaves
cultivated
in Estonia
varied
about in
the
same range as the
concentrations
of
these
compounds
in
the oils
of drugs
obtained
from
other European
countries.
3.
Experimental
3.1. Plont nrcterials
Plant materials
(commercial
S.
fficinalis Jblium
)
were obtained
from
retail
pharmacies
of different European countries
in
2000
(Estonia,
France),
in 2001
(Hungary,
Belgium).
in2002
(Russia,
Estonia,
Greece, Ukraine) and in
2003
(Scotland,
Moldavia, Estonia).
The sage seeds from
British
origin
(Cornpany
CN
sceds.
Sample
l0) were cultivated
in the Experimental Garden of the University of
Tartu,
in the summer
2001. Voucher
specimens have been deposited at the
Institute of Pharmacy,
lJniversity of Tartu,
Estonia.
3.2, Isolution
of tlrc
essential oil
Essential oil was
isolated
from the dried sage herb by tho distillation method described in
the
EP
[5]
using
20
g
of cut drug, a 500
mL round-bottomed flask and
250
mL
distilled
water
as
the distillation
liquid. Also, 0.5 mL of
xylene in the
graduated
tube
was added
to take
up the essential
oil. The distillation
time
was 2h
at
a rate
of
2.'3nrl.min-1.

Conrposition
o/ the essentiul
oil
o.f S. officinalis
l.
3.3, Capillary
gqs
clrromatography
The essential
oil extracts
were analyzed using
a Chlom-S
chromatograph
with
FID
on
two
fused silica
capillary columns
(50mx0.20mm)
with
two stationary
phases:
nonpolar
poly(dimethylsiloxane)
(NB-30,
Nordion, Finland) and
polar poly(ethylene
glycol)
(NB-20M,
Nordion,
Finland). The
film
thickness
of both stationary
phases
was 0.25pm.
The carrier
gas
was
helium
with the split ratio
of
l:150, and a flow
rate of
2H25cms-'*as
appliecl. The
temperature
program
was from 50 to 250'C
at
2'Cmin-1,
and the
iniector temperature
was 200"C.
The
identification
of the oil componcnts
was acconrplished
by
comparing their:
retention
indices
(RI)
on
two colnmns
with the RI
values
of
reference
standards,
the
authors
RI
data bank,
and litelature
data.
The
percentage
composition
of
the oils was
calculated in
peak
areas
(nonpolar
column)
using
a normalization
method
without
using
correction
factors.
The relative
standard
deviation
of
percentages of oil components of
three repeated
GC analyses
of
a
single oil
did
not exceed
57n.
Acknowledgenrent
Financial
support
for the
work
reported
here
was
provided
by the Estonian Science
Foundation
(grant
No.
4332).
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