FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fragr. J. 2000; 15: 96±99

Essential oil analysis of Nepeta argolica Bory & Chaub.

subsp. argolica (Lamiaceae) growing wild in Greece
Helen D. Skaltsa,1* Diamanto M. Lazari,1 Anargyros E. Loukis1 and Theophanis Constantinidis2
1
Division of Pharmacognosy, Department of Pharmacy, University of Athens, Panepistimiopolis, Zografou, GR-157 71,
Athens, Greece
2Botanical Institute, Department of Biology, University of Patras, GR-26 500 Patras, Greece

Received 10 February 1999

Revised 2 November 1999
Accepted 4 November 1999

ABSTRACT: The chemical composition of the essential oils obtained from the aerial parts of two di€erent
populations of Nepeta argolica Bory & Chaub. ssp. argolica (Lamiaceae) was examined by GC and GC±MS.
Their components were mainly represented by oxygenated sesquiterpenes. Copyright # 2000 John Wiley & Sons,
Ltd.

KEY WORDS: Nepeta argolica subsp. argolica; Lamiaceae; essential oils; GC±MS; 1,8-cineole; nepetal-
actones; Greece

Introduction taxon distributed in northern Peloponissos and south-

The genus Nepeta (Lamiaceae) comprises some 250
species, most of which are spread out over the larger
part of central and southern Europe, the Near East and
central and southern Asia. Nepeta species are used in
folk medicine.1,2 Antibacterial, fungicidal2,3 and anti-
viral3 activities have been attributed to nepetalactones.
These types of iridoids are known to elicit a unique
sequence of responses in Felidae.4 It is also reported
that they are powerful insect repellents5 and that they
are also found in the defensive secretions of some
insects.6,7
Although sectional classi®cation of the genus Nepeta
as a whole appears to be problematic, Nepeta argolica
(syn. Nepeta sibthorpii Benth.), examined here, should
be referred to Nepeta sect. Stenostegiae, according to
recent data.8 The group to which Nepeta argolica
belongs has been recently revised taxonomically9,10 and
four infraspeci®c taxa have been proposed at subspe-
ci®c rank.
Certain species of Nepeta are characterized by a high
content of one or more diastereomers of nepetalac-
tones,1,11±21 most frequently of the (S)-con®guration, at
C-7; while others have a much higher concentration of
di€erent components,22±24 mainly of 1,8-cineole.25±28
This is the ®rst report on the essential oil composi-
tion of Nepeta argolica ssp. argolica, a Greek endemic
* Correspondence to: H. Skaltsa, University of Athens, School of Pharmacy,
Department of Pharmacognosy, Panepistimiopolis±Zografou, Greece 157 71,
Athens, Greece; e-mail: skaltsa@pharm.uoa.gr
eastern Sterea Ellas (east central Greece), and found in
dry stony places, rocky habitats and scree at an altitude
of almost sea level to ca. 1700 m.
Experimental
Collection of Plant Material
Two di€erent populations of Nepeta argolica subsp.
argolica, both collected during ¯owering time, were
examined. The ®rst population (No. 1) was collected in
August 1994 on Mt. Erimanthos (Peloponnisos). The
second population (No. 2) was found on the slopes of
Mt. Pateras (Attiki, Sterea Ellas), and was collected
in July 1998. Voucher specimens have been kept in the
Herbarium of the Laboratory of Pharmacognosy of the
University of Athens and in the Herbarium of Patras
University (UPA), under accession numbers Lazari &
Skaltsa 103 (No. 1) and Constantinidis 2561 (No. 2),
respectively.
Isolation of Essential Oils
Air-dried plant material was cut in small pieces and the
essential oil was obtained by steam distillation for 3 h.29
The oil was dried over anhydrous sodium sulphate and
kept at ÿ208C, in order to avoid conversion of nepe-
talactones into nepetalic acid and dehydronepetalac-
tone.11 The composition of the volatile constituents was

Copyright # 2000 John Wiley & Sons, Ltd.

 ESSENTIAL OIL OF NEPETA ARGOLICA SUBSP. ARGOLICA 97

Table 1. Chemical composition of Nepeta argolica ssp. argolica essential oils

Compound RIa Peak area (%)b

Chemotype

No. 1 No. 2
a-Pinene 940 0.25 0.24
Sabinene 974 0.55 ±
b-Pinene 978 1.17 0.26
Myrcene 991 0.28 0.04
p-Cymene 1023 0.70 0.05
1,8-Cineole 1030 39.79 0.59
cis-Ocimene 1035 ± 0.03
trans-Ocimene 1048 0.96 ±
Benzenacetaldehyde 1040 ± 0.04
a-Pinene oxide 1090 ± 0.04
Linalol 1098 0.26 0.23
Nonanal 1101 0.49 0.11
a-Campholenal 1122 ± 0.40
Pinocarveol 1134 ± 0.11
p-Mentha-1(7),2-dien-8-ol 1163 ± 0.12
Terpinen-4-ol 1174 0.24 0.03
p-Cymen-8-ol 1182 ± 0.12
a-Terpineol 1189 0.62 0.25
Myrtenal 1191 ± 0.10
Verbenone 1205 ± 0.10
trans-Carveol 1215 ± 0.09
Pulegone 1235 0.26 ±
Benzyl propanate 1255 0.24 ±
Methyl ester of 1a,2a,5a-nepetonic acidc 1303 1.21 ±
2-Phenylethylpropionate 1349 0.24 ±
4ab,7a,7aa-Nepetalactonec 1360 1.93 29.38
4aa,7a,7aa,Nepetalactonec 1375 ± 26.52
a-Copaene 1376 4.06 ±
b-Bourbonene 1384 0.48 ±
4aa,7a,7ab-Nepetalactonec 1390 ± 12.93
b-Elemene 1391 0.51 ±
4ab,7a,7ab-Nepetalactonec 1398 0.38 14.45
3,4a-Dihydro,4aa,7a,7aa-nepetalactonec 1414 ± 0.25
b-Caryophyllene 1418 0.29 ±
b-Gurjunene 1435 0.34 0.07
epi-Bicyclosesquiphellandrene 1478 1.46 ±
g-Muurolene 1479 0.92 0.58
Germacrene D 1480 0.88 ±
b-Selinene 1484 1.39 tr
Butylated hydroxytoluene 1513 0.79 ±
g-Cadinene 1514 1.23 0.08
cis-Calamenene 1517 ± 0.53
d-Cadinene 1524 8.67 ±
3-Hydroxy-4a,4aa,7a,7aa-dihydronepetalactonec 1535 4.73 ±
trans-Calamenene 1536 ± 0.14
11-nor-Bourbonan-1-one 1554 ± 0.06
b-Calacorene 1563 1.99 ±
Caryophyllene oxide 1581 0.78 ±
1-Hexadecene 1593 0.38 ±
1,10-di-epi-Cubenol 1616 0.78 ±
g-Eudesmol 1630 1.12 ±
Torreyol 1642 0.68 0.05
a-Cadinol 1655 1.84 0.48
Cadalene 1676 1.26 0.08
14-nor-Cadin-5-en-4-one, isomer B 1687 2.44 0.72
Palmitic acid 1962 0.78 ±
Abietatriene 2054 ± 0.13
Total 87.37 89.45
Monoterpene hydrocarbons 3.91 0.62
Oxygenated monoterpenes 48.45 85.76
Sesquiterpene hydrocarbons 23.48 1.48
Oxygenated sesquiterpenes 5.99 0.53
a
RI ˆ KovaÂts index on HP-5 column.
b Percentage of total FID area obtained on HP-5 column; tr ˆ trace (50.03%).
c Tentatively identi®ed according to their mass spectra, their retention indices and to bibliography.

Copyright # 2000 John Wiley & Sons, Ltd. Flavour Fragr. J. 2000; 15: 96±99
98 H. D. SKALTSA ET AL.
carried out by GC and GC±MS analyses. The yield of
the oils was ca. 0.27% for population No. 1 and 0.66%
for population No. 2, based on dry weight of samples.
Gas Chromatography
Analysis was carried out on a Perkin-Elmer 8500 gas
chromatograph with FID, ®tted with a Supelcowax-10
fused silica capillary column (30 m  0.32 mm i.d.; ®lm
thickness, 0.25 mm). The column temperature was
programmed from 758C to 2008C at a rate of 2.58C/
min. The injector and detector temperatures were
programmed at 2308C and 3008C, respectively.
Gas Chromatography±Mass Spectrometry
Analysis was carried out on a Hewlett-Packard 6890
gas chromatograph ®tted with a fused silica HP-5 MS
capillary column (30 m  0.25 mm; ®lm thickness;
0.25 mm). The column temperature was programmed
from 508C (5 min) to 2808C at a rate of 48C/min.
Helium was used as carrier gas at a ¯ow rate of 0.6 ml/
min. The chromatograph was coupled to a MS 5973
mass selective detector at 70 eV.
Retention indices for all compounds on HP 5MS
capillary column were determined according to the Van
der Dool approach30 using n-alkanes as standards. The
identi®cation of the components was based on com-
parison of their mass spectra with those of Wiley and
NBS Libraries31 and literature references.11,12,32
Results and Discussion
Nepetalactones are a group of cyclopentanoid mono-
terpenoids, which are found in varying amounts
exclusively as components of di€erent Nepeta oils. As
can be seen from Table 1 the chemical composition of
the studied samples varies largely in the two collected
populations.
Chemotype No. 1 produces an essential oil of
herbaceous, mildly menthole-like odour, which may
be attributed to the high amount of 1,8-cineole
(39.79%), while the nepetalactone content is much
lower (8.25%), compared to that of chemotype No. 2
(83.53%). The speci®c chemical composition of the
latter chemotype gives a di€erent olfactory pro®le to
the essential oil, which is characterized by a high
oxygenated monoterpene content (85.76%) consisting
mainly of nepetalactones, and by a low portion of
monoterpene (0.62%) and sesquiterpene hydrocarbons
(1.48%). The same oil is also characterized by a very
low amount of 1,8-cineole (0.59%).
Copyright # 2000 John Wiley & Sons, Ltd.
The essential oil composition of population No. 1,
when compared to other Nepeta species, shows many
similarities with N. parnassica Heldr. & Sart. ex Boiss,25
a Greek endemic taxon, N. sulfuri¯ora P. H. Davis,27
N. viscida Boiss.26 and N. discolor Benth.28 However,
most oils of Nepeta species contain nepetalactones as
their main components, such as the essential oils of
N. cataria L.,12,15,16 N. grandi¯ora M. B.,15 N. nuda L.,15
N. racemosa Lam.14 (syn. N. mussini Spreng, ex
Henckel,18,19 N. teydea Webb. & Berth.,11 N. elliptica
Royle ex Benth.,20 N. nepetella,1,12,21 but also di€erent
compositions have been detected (e.g. N. cataria L. cv.
citriodora,17 N. nuda L. subsp. nuda,22 N. govaniana
Benth.,23 N. tuberosa L. subsp. tuberosa24).
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Copyright # 2000 John Wiley & Sons, Ltd. Flavour Fragr. J. 2000; 15: 96±99