FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fra`r[ J[ 1999^ 04] 004Ð007

Essential oil composition of Nepeta argolica Bory et Chaub.

subsp. argolica
O. Tzakou,1 C. Harvala,1 E. M. Galati2 and R. Sanogo2
1
Department of Pharmacognosy, School of Pharmacy, University of Athens, Panepistimioupolis Zografou, 157 71 Athens, Greece
2
Pharmaco-Biological Department, School of Pharmacy, University of Messina, SS. Annunziata, 98168 Messina, Italy

Received 12 April 0888

Revised 12 November 0888
Accepted 15 November 0888

ABSTRACT] The chemical composition of the essential oils obtained from the herbal parts and di}erent plant
parts of Nepeta ar`olica Bory et Chaub[ subsp[ ar`olica was examined by GC and GCÐMS[ Sixty!two con!
stituents were identi_ed[ The major constituents were found to be stereoisomers of nepetalactones "83[4Ð57[9)#[
3aa\6a\6ab!nepetalactone was isolated and identi_ed by 0H! and 02C!NMR[ Copyright Þ 1999 John Wiley +
Sons\ Ltd[

KEY WORDS] Nepeta ar`olica Bory et Chaub[ subsp[ ar`olica^ Lamiaceae^ essential oil^ nepetalactones

Introduction Experimental
In the frame of our research of Greek aromatic plants\ Plant Material
we found a natural population of Nepeta ar`olica Bory
Aerial parts of N[ ar`olica subsp[ ar`olica were collected
et Chaub[ subsp[ ar`olica "syn[ N[ sibthorpii Benth[
from wild!growing plants from Mt[ Parnes "Attica#\ at
subsp[ sibthorpii# grown in Mt[ Parnes[ N[ ar`olica
an altitude of 0199 m during the ~owering stage from
"Lamiaceae#\ an aromatic perennial herb\ is an endemic
0883 to 0887[ Voucher specimens are deposited in the
species of the south Balkan peninsula[ It is usually
Laboratory of Pharmacognosy\ University of Athens
found in dry stony places and scree[ It is a variable
"Ref[ Nos OT!8\ OT!8A\ OT!8B\ OT!8C\ OT!8D#[
species\ which has been further divided into four sub!
species[0
The genus Nepeta comprises about 149 species dis! Isolation of Oils
tributed in the central and southern parts of Europe\
Air!dried\ above!ground parts of the plant material
Asia and Middle East[ Many reports on phytochemical
were subjected to hydrodistillation for 2 h using a
analysis of this species!rich genus\ including essential
Clevenger!type apparatus[ Analyses for the years 0883Ð
oil analyses\ are found in the literature[1Ð01
0887 were performed on the herbal parts[ In addition\
Terpenes\ including terpenoid lactones of the nep!
the sample of 0885 was divided into leaves\ ~owering
etalactone!type\ are found to be the main constituents
tops and stems with branches[ The di}erent organs were
in most of the oils[ However\ some di}erences in the
analysed separately[ The essential oils were light yellow
essential oil composition were detected in several Nep!
in colour\ heavier than water and with a strong pleasant
eta oils\02Ð06 as well as di}erences in the oil composition
odour[ The oil yields were 9[75Ð9[50) "herbal parts\
of the same species from di}erent localities[4\07
0883Ð0887#\ 0[22) "leaves\ 0885#\ 9[41) "~owering
N[ cataria L[ commonly known at catnip\ is the most
tops\ 0885# and 9[29) "stems with branches\ 0885#[ The
intensively studied species[8\08\19 Some species of the
yield was estimated on the basis of dry weight of the
genus are used in folk medicine as diuretic\ diaphoretic\
samples[ The distillates were saturated with NaCl and
antitussive\ antispasmodic\ anti!asthmatic\ febrifuge\
the oils were extracted successively with hexane[ The
emmenagogue and sedative agents[1\3\10\11
organic phases were combined\ dried over anhydrous
sodium sulphate and kept at 3>C until analysed[ The
physicochemical characteristics of the oil from the
 Correspondence to] O[ Tzakou\ Department of Pharmacognosy\ School of
Pharmacy\ University of Athens\ Panepistimioupolis Zografou\ 046 60 Athens\
herbal parts of 0885 sample were] d14  0[05\
Greece^ e!mail] tzakouÝpharm[uoa[gr ðnŁ14 19
D  0[3763\ ðaŁD  −04[30> "CHCl2\ c 1[22#[

Copyright Þ 1999 John Wiley + Sons\ Ltd[

005 O[ TZAKOU ET AL[

Table 1. Percentage composition of the essential oils of Nepeta argolica subsp. argolica
Component KIa Year of harvest
b b
0883 0884 0885b 0886b 0887b
"H# "H# "H# "B¦S# "L# "F# "H# "H#

0\2!Cyclopentadiene\ trimethyl tr tr
2!Methyl!cyclopentanone 9[5 tr 9[2 tr
2!Methyl!butanoic acid\ ethyl ester tr
a!Thujene 829 tr tr
a!Pinene 825 tr 9[0 4[4 2[4 9[4 7[7 9[6
Camphene 841 tr tr tr
Thuja!1\3"09#!diene 845 tr tr tr
Benzaldehyde 850 tr tr tr
Sabinene 864 tr tr tr tr tr tr
b!Pinene 868 9[0 9[8 0[9 9[4 0[0 9[2
Myrcene 889 tr tr tr tr tr
2!Octanol 881 tr 9[7 tr tr
p!Cymene 0914 tr 9[4 tr tr tr tr
0\7!Cineole 0918 9[3 1[6 0[0 3[3 3[9 1[5 1[3 0[2
Benzene acetaldehyde 0934 tr tr tr
"E#!b!Ocimene 0940 9[0
g!Terpinene 0952 tr
Linalol 0988 9[0 tr tr tr tr
n!Nonanal 0097 tr tr tr
endo!Fenchol 0006 tr tr
a!Campholenal 0029 tr 0[6 0[1 tr tr
trans!Pinocarveol 0031 9[7 9[3
trans!Verbenol 0035 9[6
trans!Pinocamphone 0053 tr tr
trans!Dihydro!a!terpineol 0054 9[0 ! tr
Borneol 0058 tr tr
p!Mentha!0\4!dien!7!ol 0069 tr tr tr
cis!Pinocamphone 0068 tr tr
Terpin!3!ol 0070 tr tr tr tr tr
Napthalene 0074 tr !
p!Cymen!7!ol 0078 tr tr tr tr tr
a!Terpineol 0084 9[6 tr 9[2 9[7 9[7 9[3 tr tr
Methyl salicylate 0087 tr tr
Myrtenol 0190 tr tr
Verbenone 0103 tr tr tr tr tr
1!Ethyl!1\4!dimethylcyclopent!1!one 0010 0[6 2[2 1[6 0[9
trans!Carveol 0112 9[5 9[2 tr
Carvone 0137 tr tr
Geranial 0169 tr
Benzene propanoic acid\ methyl ester 0179 9[2 9[1 9[8 9[6 tr tr
Benzene propanoic acid\ ethyl ester 0244 9[0 tr tr tr tr
3aa\6a\6aa!Nepetalactone 0260 2[7 2[1 09[4 2[4 6[5 3[3 5[3 09[3
a!Copaene 0270 tr tr 9[4 2[6 1[8 0[1 0[9
b!Bourbonene 0289 tr tr tr tr 9[6 tr tr tr
3aa\6a\6ab!Nepetalactone 0394 89[4 80[2 70[9 53[4 55[7 71[4 65[6 71[3
Nepetalactone 0305 tr tr 9[1
Longifolene 0307 tr
trans!Caryophyllene 0313 tr tr tr
Germacrene D 0379 9[5
cis!Calamenene 0415 tr tr 0[3 tr tr
d!Cadinene 0429 tr tr 9[4 tr 0[7 1[8 0[2 9[2
a!Calacorene 0449 tr tr tr 9[5 9[3 tr tr
0!nor!Bourbonanone 0457 tr
b!Calacorene 0469 tr
Caryophyllene oxide 0470 tr tr 9[0 tr tr 9[6 tr
epi!a!Cadinol 0537 tr
epi!a!Muurolol 0549 9[4 tr
a!Cadinol 0551 tr tr tr tr tr
Cadalene 0571 tr tr 9[5 9[5 tr tr
Khusinol 0563 tr 9[5
cis!03!Muurol!4!en!3!one 0584 tr 0[9 9[5 tr tr
Hexadecanoic acid 0864 0[7
0
Retention indices relative to C8 ÐC13 n!alkanes on the HP 4MS column[
b
Relative percentage obtained from peak area[
 Identi_cation based on mass spectrum^ the correct isomer was not assigned[
tr  trace "³ 9[94)#^ H  herbal parts^ B¦S  branches¦stems^ L  leaves^ F  ~owering tops[

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 004Ð007

Identification of Components
The identi_cation of the chemical constituents was
based on comparisons of the their Kovats indices "KI#\
retention times "RT# and mass spectra with those
obtained from authentic samples and:or the NIST:
NBS\ Wiley libraries spectra and the literature[12
Isolation of 4aaa ,7aa ,4ab
b -nepetalactone
An aliquot part "9[1 ml# of the essential oil "herbal
parts\ collection 0885# was applied to Merck silica gel
59 F143 plates "19×19 cm\ 0 mm layer thickness# and
chromatographed using n!hexane!ethyl acetate "84]4#
as mobile phase[ The major band was scrapped and
extracted with dichloromethane[ The spectral analyses
of the concentrated extracts showed that this band con!
tained a 84]4 mixture of 3aa\6a\3ab!nepetalactone and
3aa\6a\3aa!nepetalactone "Table 0#[ The 0H! and 02C!
NMR data of this mixture contain in the appropriate
ratio all the resonances of the two isomers[4\13 The iso!
meric ratio\ even though determined from the 0H!NMR
integration\ was further veri_ed through the integration
of the inverse gated 02C!NMR spectrum "Brucker
AC299#[
Gas Chromatography
GC analyses were carried out in two gas chromato!
graphs] "a# Varian 2299 equipped with a J+W DB!4
column "29 m×9[21 mm i[d[#[ Oven temperature was
performed as follows] 49>C for 4 min and then up to
149>C at 2>C:min^ injector temperature\ 149>C^ detec!
tor temperature\ 179>C^ carrier gas\ He "1 ml:min#^ split
ratio\ 0]04[ "b# Carlo!Erba HRGC 4299 Series Mega
Apparatus equipped with a Supelcowax 09 column "59
m×9[14 mm i[d[#[ Oven temperature was programmed
as follows] 64Ð129>C at 3>C:min^ injector temperature\
129>C^ detector temperature\ 159>C^ carrier gas\ He "0[5
ml:min#^ split ratio\ 0]099[
Gas Chromatography–Mass Spectrometry
The GCÐMS analyses were carried out using a Hewlett!
Packard 4862!5789 GCÐMS system operating on EI
mode at 69 eV\ equipped with a HP 4MS capillary
column "29 m×9[14 mm^ _lm thickness\ 9[14 mm#[ The
initial temperature of the column was 59>C and was
heated to 179>C at 2>C:min[ Carrier gas\ He^ ~ow rate\
0 ml:min[
Results and Discussion
The qualitative and quantitative composition of the oils
analysed are presented in Table 0\ where the com!
Copyright Þ 1999 John Wiley + Sons\ Ltd[
ESSENTIAL OIL OF NEPETA ARGOLICA 006
ponents are listed in the order of elution from the HP
4MS column[ Sixty!two components were identi_ed\
constituting 84[6Ð87[8) of the total oil[ The major
component in all collections was found to be con!
sistently 3aa\6a\6ab!nepetalactone "72[0Ð83[4)#[ Low
concentrations of normal terpenoids were found\ prob!
ably due to the high nepetalactone content[ Apart from
nepetalactones\ the main components of Nepeta oils
were found] 0\7!cineole "9[3Ð1[3)#\ germacrene D\
trans!caryophyllene\ caryophyllene oxide and cyclo!
pentanoid idiridodial derivatives[1Ð04
The essential oils of the di}erent plant organs "col!
lection 0885# are also characterized by a high content
of nepetalactones "57[9Ð75[8)#\ especially in the oil of
~owering tops "75[8)#[ These oils contain mono!
terpenes "0[8Ð04[1)# with 0\7!cineole "1[5Ð3[3)# and
a!pinene "9[4Ð4[4)# as principal constituents[ The ses!
quiterpene fraction of these oils amounted to 0[2Ð8[1)[
Since nepetalactones are structurally similar to vale!
potriates\ it is suspected that they might possess similar
sedative properties[14 In that case\ N[ ar`olica subsp[
ar`olica\ with a constantly high content in nep!
etalactones could be shown to be a pharmaceutically
interesting plant[
Acknowled`ements * We thank Dr Th[ Constandinidis "Institute of
Botany\ University of Patras# for the identi_cation of the plant
material[
References
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Flavour Fra`r[ J[ 1999^ 04] 004Ð007
007 O[ TZAKOU ET AL[
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Flavour Fra`r[ J[ 1999^ 04] 004Ð007