Biochemical Systematics and Ecology 31 (2003) 249–259

www.elsevier.com/locate/biochemsyseco

Chemical composition of the essential oil of

different varieties of thyme (Thymus
pulegioides) growing wild in Lithuania
Kristina Ložienė a, Jonė Vaičiūnienė a, Petras R. Venskutonis b,∗
a
Institute of Botany, Žaliu
ju
ežeru
47, LT-2021, Vilnius, Lithuania
b
Department of Food Technology, Kaunas University of Technology, Radvilėnu
pl. 19, Kaunas, LT-
3028, Lithuania

Received 23 February 2001; accepted 15 April 2002

Abstract

Twenty-five samples of Thymus pulegioides L. representing two varieties of subsp. silvestris

and four varieties of subsp. pulegioides were collected from 11 localities in Lithuania and the
composition of their essential oil was analysed by capillary gas chromatography (GC) and
mass spectrometry (MS). Five chemotypes have been defined for the species using hierarchical
cluster analysis: (1) linalool (L)—one sample; (2) geranial/geraniol/neral (G/G/N)—11
samples; (3) thymol (T)—two samples; (4) carvacrol/γ-terpinene/p-cymene (C/γT/pC)—seven
samples; (5) thymol/carvacrol/p-cymene/γ-terpinene (T/C/pC/γT)—four samples. It was con-
cluded that there is no clear relation in essential oil composition between the studied taxa and
the chemical types detected. Four different chemotypes were defined within the subsp. sil-
vestris var. conglomeratus, three chemotypes within the subsp. pulegioides var. pulegioides
and var. ovatus. The G/G/N-chemotype is the most frequently found, followed by the phenolic
types, the linalool chemotype being recorded only once. Both infra-specific and intra-popu-
lation chemical polymorphisms were recorded.
 2003 Elsevier Science Ltd. All rights reserved.

Keywords: Thymus pulegioides L.; Variety; Chemotype; Thymol; Carvacrol; Linalool; Geranial; Geraniol;
Neral; p-Cymene; γ-Terpinene

∗
Corresponding author. Tel.: +370-7-356426; fax: +370-7-456647.
E-mail address: rimas.venskutonis@ctf.ktu.lt (P.R. Venskutonis).

0305-1978/03/$ - see front matter  2003 Elsevier Science Ltd. All rights reserved
doi:10.1016/S0305-1978(02)00142-4
250 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

1. Introduction

Chemical polymorphism is characteristic for the plants belonging to the genus

Thymus. Two Thymus species grow naturally in Lithuania, T. pulegioides and T.
serpyllum (Gudžinskas, 1999). It was reported that the pharmacognostically valuable
components thymol and carvacrol are absent in T. serpyllum growing wild in Lithu-
ania (Ložiene et al., 1998).
T. pulegioides L. growing wild in Lithuania can be characterised as a plant with
a high morphological and chemical polymorphism (Ložiene, 1997; Ložiene and Vaič-
iūniene, 1999). Lekavičius and Jaskonis (1968, 1969) described three T. pulegioides
subspecies and 11 of their varieties exist in Lithuania according to the Lyka (1927)
and Jalas (1947) system. The same authors analysed the content of the essential oil
in T. pulegioides varieties from Lithuania. They found that different varieties
accumulate different amounts of essential oil, e.g. var. pulegioides—0.43–1.21%;
var. adscendens Wimm. et Grab.—0.41–0.49%; var. minutus Lyka—0.80%; var.
conglomeratus Lyka—0.21–0.73% (Lekavičius and Jaskonis, 1968). However, the
samples of selected varieties for these studies were collected in different growing
sites with different environmental conditions. The environmental conditions such as
light, soil acidity and chemical composition can be of a greater importance to the
essential oil content than the taxonomical adscription of the plant to any particular
variety.
Chemical polymorphism is characteristic to T. pulegioides as well as to many
other Thymus species. For instance, five chemotypes were described in Slovakia,
namely fenchone, linalool, citral/geraniol, thymol and carvacrol (Mártonfi, 1992).
Stahl-Biskup (1986) reported the presence of thymol and carvacrol chemotypes in
Norwegian populations of T. pulegioides by analysing individuals from different
populations. Carvacrol was dominating in three T. pulegioides samples from Sweden,
while thymol in those samples was absent (Lundgren and Stenhagen, 1982). In Italy
T. pulegioides essential oils were characterised by a high content of γ-terpinene, p-
cymene and thymol (up to 39.1%) (Senatore, 1996). The essential oils of T. pule-
gioides from Yugoslavia were rich in p-cymene (6–15%), thymol (1–22%) and carva-
crol (10–30%), while one of the samples analysed contained 50% of linalool (Kustrak
et al., 1990). Linalool was the major component of T. pulegioides from Poland (22.3–
45.0%) followed by carvacrol (4.9–31.0%); thymol in this case constituted only 1.0–
4.0% (Kowal and Krupińska, 1979). Domokos et al. (1995) described a chemotype
from Bulgaria rich in neral (18.7%) and geranial (52.8%). Stahl-Biskup (1991)
defined four chemotypes of T. pulegioides from the Czech Republic according to
the content of thymol, citral, linalyl acetate and fenchone. Two chemotypes,
citral/geraniol and carvacrol of T. pulegioides from Lithuania collected in the Vilnius
region were reported in 1999 by Mockute and Bernotiene. However, the samples
analysed in the reported literature have been characterised only to the species level.
Considering the remarkable variations in the composition of the essential oil of
T. pulegioides from different locations, which was reported in the surveyed literature,
it is of interest to continue research on the chemical polymorphism of this species.
The goal of this study was to examine the chemical composition of the essential oil
 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259 251

of different varieties of T. pulegioides collected from different habitats in Lithuania

and to determine the relationship between the varieties and chemotypes.

2. Materials and methods

Aerial parts of T. pulegioides varieties were collected from 11 different popu-

lations of Lithuania in July of 1996 and 1997. In total, 25 selected plants were
collected at the flowering stage. As the plants are spread all over the country the
habitats were selected randomly to represent as big an area as possible. It was
attempted to collect a higher number of different infra-specific taxa (varieties and
subspecies). Voucher specimens of each taxa have been deposited in the Herbarium
of the Institute of Botany, Vilnius, Lithuania; the numbers of the specimens are from
60,863 to 60,888 and they correspond to sample numbers from 1 to 25 in this study.
The plants were dried at room temperature and stored until analysis inside paper
bags in the dark. The essential oils were isolated by hydrodistillation using European
Pharmacopoeia apparatus for 2 h. The numbers of the samples, their habitat, collec-
tion date and variety are provided in Table 1. Fig. 1 provides the map of Lithuania
with the numbered localities where the samples were collected.
The essential oils were diluted in diethyl ether (20 µl in 1 ml) and analysed with
Fisons 8261 gas chromatograph with flame ionisation detector (FID) on a fused silica
capillary column DB-5, 25 m in length, 0.32 mm i.d., and 0.5 µm film thickness.
Helium was used as the carrier gas with a flow rate of 1.6 ml/min; the detector
temperature was 260 °C, the oven temperature was programmed to increase from
40 to 250 °C at a rate of 4 °C/min. The split injector was heated at 250 °C, the split
ratio was 15:1. Data were processed on a DP 800 integrator.
For identification, essential oils were also analysed on a HP 5890 (II) instrument
equipped with a 5971 series mass selective detector in the electron impact ionisation
mode at 70 eV, and the GC parameters were as follows: split inlet 1:10; helium as
carrier gas at a flow rate of 2 ml/min; fused silica HP5 MS column (Hewlett Packard,
crosslinked 5% phenyl methyl silicone) 30 m length, 0.25 mm id, 0.25 µm film
thickness, temperature program from 40 to 250 °C increasing at 4 °C/min. Identifi-
cation was based mainly on the comparison of retention indices (RIs) (Adams, 1995;
Davies, 1990) and mass spectra (NIST/EPA/NIH Mass Spectral Database NBS75K).
Analytical data for cluster analysis were treated by means of the statistical package
STATISTICA/W release 5.0.

3. Results and discussion

The composition of the 25 selected T. pulegioides plants from 11 growing sites

was determined. In total, 86 compounds were identified by capillary GC and coupled
GC/MS, which are listed in Table 2. Using hierarchical cluster analysis, five main
groups of the samples were observed (Fig. 2), according to the major constituents
in the essential oil. Consequently the following chemotypes of T. pulegioides have
252 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

Table 1
Collection localities and varieties of investigated Thymus pulegioides L. samples

No. of Name of locality Collec- Number Subspecies, varieties

locality tion date

1 Vilnius reg., Bezdonys 1996 1 subsp. pulegioides var. pulegioides

2 Vilnius reg., Žalieji 1996 2 subsp. pulegioides var. pulegioides
ežerai
3 Vilnius reg., Verkiai 1996 7 subsp. silvestris var. conglomeratus
regional park
8 subsp. silvestris var. Conglomeratus
9 subsp. pulegioides var. Ovatus
4 Vilnius reg., Rokantiškės 1996 4 subsp. pulegioides var. pulegioides
5 subsp. pulegioides var. pulegioides
10 subsp. pulegioides var. pulegioides
14 subsp. pulegioides var. glaber
3 subsp. pulegioides var. glaber
12 subsp. pulegioides var. glaber
11 subsp. pulegioides var. adscendens
5 Vilnius reg., Rokantiškės 1996 6 subsp. silvestris var. conglomeratus
13 subsp. silvestris var. conglomeratus
15 subsp. pulegioides var. ovatus
20 subsp. pulegioides var. ovatus
6 Vilnius reg., Balsiai 1997 24 subsp. silvestris var. conglomeratus
25 subsp. silvestris var. conglomeratus
7 Molėtai reg. 1997 16 subsp. silvestris var. silvestris
8 Šiauliai reg., Pakapė for- 1997 17 subsp. pulegioides var. ovatus
estry
18 subsp. pulegioides var. glaber
9 Šiauliai., Salduvė park 1997 19 subsp. silvestris var. conglomeratus
10 Skuodas reg., Šaukliai 1998 21 subsp. pulegioides var. glaber
22 subsp. pulegioides var. pulegioides
11 Utena reg., Panatryčia 1998 23 subsp. pulegioides var. glaber

been defined: (1) linalool (L)—one sample; (2) geranial/geraniol/neral (G/G/N)—11

samples; (3) thymol (T)—two samples; (4) carvacrol/γ-terpinene/p-cymene
(C/γT/pC)—seven samples; (5) thymol/carvacrol/p-cymene/γ-terpinene
(T/C/pC/γT)—four samples.
The range in the content of the major quantitative compounds in the detected
chemotypes is presented in Table 3. The T/C/pC/γT chemotype can be considered
as an intermediate one; there is a biosynthetic correlation between the two major
phenolic compounds, thymol and carvacrol (Russo et al., 1998). The essential oils
from the plants attributed to T, C/γT/pC and T/C/pC/γT chemotypes were rich in p-
cymene and γ-terpinene, which are chemical precursors of thymol and carvacrol.
Variations in the total content of p-cymene+γ-terpinene, and thymol+carvacrol were
not very big in all three chemotypes. However, the content of p-cymene+γ-terpinene
was lower in the C/γT/pC-chemotype than in the T-chemotype. Most likely, the rate
of bioconversion of these compounds in T-chemotype was higher and as a conse-
 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259 253

Fig. 1. Localities of investigated T. pulegioides L. in Lithuania.

quence the mean content of thymol in the T-chemotype in most cases was higher
than the mean content of carvacrol in the C/γT/pC-chemotype plants. In general, T,
C/γT/pC and T/C/pC/γT chemotypes could be gathered together into a larger, phe-
nolic chemotype group.
The T-chemotype can be characterised by a higher content of thymol methyl ether
as compared to C/γT/pC-chemotype. Variations in the content of the main sesquiterp-
enes, β-caryophyllene, germacrene D and β-bisabolene, were similar in all chemo-
types except for L-chemotype, which contained less β-bisabolene. The content of
geranial was higher than the content of neral in all G/G/N-chemotype plant essential
oils. The essential oils of sample nos 9 and 21 contained a rather low percentage of
neral, 1.3 and 0.8%, respectively; that of sample nos 8 and 12 contained a remarkably
lower amount of geraniol (4.9 and 2.5%, respectively) as compared with other
G/G/N-chemotype plants. Therefore, these samples can be attributed to this particular
chemotype with some reservations; more precise distribution of the plants would
indicate two subgroups, namely geranial/geraniol and geranial/neral. Nerol was also
detected in all G-chemotype essential oils, however its content was considerably
lower as compared with its trans isomer, geraniol. It is interesting to note that sample
nos 9 and 21 with lower content of neral contained higher percentage of nerol, 11.7
254 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

Table 2
Composition of essential oil of different Thymus pulegioides L. chemotypes, GC area % (one selected
sample from each chemotype)

Componentsa T (17)b T/C/pC/γT (1)b G/G/N (2)b C/γT/pC L (19)b

(11)b

α-Thujene 1.6 1.0 – 1.0 –

α-Pinene 0.6 0.4 – 0.5 –
Camphene – 0.1 – 0.4 –
Sabinene – tr – 0.1 –
β-Pinene – 0.1 – 0.2 –
1-Octen-3-ol 5.3 0.5 – 0.5 0.8
3-Octanone 0.5 0.3 0.7 0.2 –
Myrcene 1.5 1.3 - 1.4 0.2
3-Octanol 0.8 1.0 1.9 tr 0.2
α-Phelandrene 0.3 0.4 – 0.2 –
δ-3-Carene – 0.1 – 0.1 –
α-Terpinene 2.0 2.3 – 2.3 –
p-Cymene 10.2 14.3 – 16.7 0.1
1,8-Cineole – – 0.1 – –
Limonene 0.3 1.1 - 1.2 0.1
(Z)-β-Ocimene 0.2 0.4 0.2 0.2 tr
(E)-β-Ocimene 1.0 tr 1.5 tr 0.1
γ-Terpinene 9.9 16.3 – 21.4 tr
cis-Sabinene hydrate 0.4 0.6 – 0.3 –
p-Mentha-3,8-diene – – – – 0.1
Terpinolene – 0.1 – tr 0.1
trans-Sabinene hydrate – – – tr –
Linalool – 0.4 0.4 0.2 80.3
allo-Ocimene – tr – tr –
Camphor – tr 0.1 – –
iso-Borneol – – 0.3 – –
Borneol – 0.2 0.3 0.7 –
Terpinen-4-ol 0.4 0.1 0.4 0.1 –
α-Terpineol – tr 0.4 0.1 0.4
Nerol – – 5.3 – –
Thymol methyl ether 11.8 2.5 – 0.4 –
Neral – – 7.8 – –
Carvacrol methyl ether 4.3 4.3 – 7.1 0.1
trans-Sabinene hydrate acetate – – – – 4.7
Geraniol 0.3 – 31.2 – –
Geranial – – 15.1 – –
Bornyl acetate – – – 0.1 –
Thymol 26.1 11.9 – 0.2 –
Carvacrol ethyl ether – – – 0.6 –
Geranyl formate – – 0.3 – –
Carvacrol 1.5 22.6 – 24.9 –
Methyl geranate – – 0.1 – –
α-Elemene – 0.3 – 0.6 –
4-Terpineol acetate – – – 0.4 –
α-Terpinyl acetate – tr – 0.2 –
α-Cubebene – 0.1 – 0.5 –
Carvacrol acetate – tr – 0.1 –
(continued on next page)
 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259 255

Table 2 (continued)

Componentsa T (17)b T/C/pC/γT (1)b G/G/N (2)b C/γT/pC L (19)b

(11)b

α-Copaene – 0.1 tr 0.1 -

Geranyl acetate – tr 1.1 – –
β-Bourbonene 0.2 0.1 0.9 0.1 0.4
β-Elemene – – 0.1 tr 0.1
iso-Caryophyllene – – – tr –
β-Caryophyllene 10.1 5.1 14.0 8.3 5.7
β-Gurjunene – 0.1 0.2 0.1 0.1
γ-Elemene – tr 0.1 tr –
(Z)-β-Farnesene – 0.1 tr tr –
E-iso-Eugenol – tr 0.1 tr tr
α-Humulene 0.5 0.2 0.7 0.3 0.2
allo-Aromadendrene – tr – – –
γ-Muurolene 0.6 0.4 tr 0.2 –
Germacrene D 2.2 1.2 tr 1.2 2.7
cis-β-Guaiene – – – tr –
Bicyclogermacrene – 0.6 – – 0.6
α-Muurolene – 0.6 – 0.1 –
trans-β-Guaiene 0.3 – 0.2 0.1 –
(Z)-α-Bisabolene 0.6 – – – –
Germacrene A – – – 0.1 –
β-Bisabolene 3.3 4.7 8.3 5.1 1.4
α-Cadinene 0.5 0.5 – 0.1 –
δ-Cadinene – 0.9 0.2 0.4 tr
Cadina-1,4-diene 0.9 0.1 – tr –
α-Cadinene – 0.1 – 0.1 –
cis-Muurol-5-en-4-β-ol – – tr – –
Germacrene B – – tr – –
Geranyl butyrate – - 0.4 – –
Spathulenol – – 0.1 – –
Caryophyllene oxide 1.2 0.3 3.3 0.6 0.5
epi-α-Cadinol – 0.1 0.1 0.1 –
Total 99.4 97.9 99.1 99.9 98.9
Monoterpene hydrocarbons 27.6 37.9 1.7 45.7 0.7
Oxygenated monoterpene 28.7 35.8 63.3 27.2 85.4
Sesquiterpene hydrocarbons 19.2 15.2 27.9 17.4 11.2
Oxygenated sesquiterpenes 1.2 0.4 3.5 0.7 0.5
Other 22.7 8.6 2.7 8.9 1.1

a
The following compounds were additionally identified in different T. pulegioides samples: perilene,
camphor, trans-verbenol, p-mentha-1,5-dien-8-ol, neryl acetate and α-cadinol (G chemotype); cis-p-ment-
2-en-1-ol (C, T/C, G and T chemotypes), italicene (C chemotype), viridiflorol (G, C and T chemotypes).
tr: tracesⱕ0.05%.
b
Sample number as in Table 1.
256 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

Fig. 2. Two-dimensional dendrogram obtained in the cluster analysis of the essential oils of individual
plants of T. pulegioides L.: 1, linalool chemotype; 2, geranial/geraniol/neeral chemotype; 3, thymol chemo-
type; 4, carvacrol/γ-terpinene/p-cymene chemotype; 5, thymol/carvacrol/p-cymene/γ-terpinene chemotype.

and 18.9%, respectively. One G/G/N-chemotype plant sample, no. 12, was exception-
ally rich in cis-p-menth-2-en-1-ol.
Essential oil composition of L-chemotype of T. pulegioides was the simplest one
in that it contained an extremely high content of linalool (⬎80%), while the percent-
age of this compound in the other samples did not reach 1%. It is worth mentioning
that a very high content of linalool (91.4%) was determined in the L-chemotype of
T. zygis samples by Sáez (1995).
The results show that in terms of a chemical composition the same varieties of T.
pulegioides collected from the same growing sites are rather heterogeneous. Earlier,
Salgueiro et al. (1997) found similar results within the populations of T. camphoratus
collected in Portugal.
Table 4 shows the distribution of different chemotypes between investigated sub-
species and varieties. It can be observed that there is no relationship between varieties
and chemotypes. For instance, four different chemotypes were selected in the subsp.
silvestris Schreb. var. conglomeratus Lyka, three chemotypes in the subsp. pule-
gioides var. pulegioides and var. ovatus (Mill.) Lyka. The G/G/N chemotype was
found in almost all varieties except for subsp. pulegioides var. adscendens Wimm.
et Grab. The C/γT/p -chemotype was found in subsp. silvestris Schreb. var. conglo-
meratus Lyka, subsp. pulegioides var. pulegioides, var. glaber (Mill.) Lyka and var.
adscendens Wimm. et Grab.
Table 3
Variation intervals in the content of the main constituents in various Thymus pulegioides L. chemotypes

Compounds Chemotypes

Thymol Carvacrol/γ- Thymol/carvacrol/p- Geranial/geraniol/ Lina-lool

terpinene/p-cymene cymene/γ-terpinene neral

Myrcene 1.1–1.5 0.1–1.6 1.1–1.3 0.0–1.1 0.2

p-Cymene 10.2–11.4 7.5–27.4 13.4–16.0 0.0–0.1 0.1
γ-terpinene 8.7–9.9 11.1–30.6 8.0–20.7 0.0–1.4 0.0
Linalool 0.0–0.1 0.0–0.7 0.0–0.6 0.1–0.7 80.3
cis-p-Menth-2-en 1-ol 0.0–2.8 0.0–3.9 0.0–5.1 0.0–10.8 –
Thymol methyl ether 4.9–11.8 0.0–1.5 2.5–3.0 – –
Carvacrol methyl ether 2.7–4.3 3.7–8.6 3.0–4.3 – 0.1
Nerol – – – 0.8–18.9 –
Neral – 0.0–2.0 – 0.8–17.4 –
Geraniol 0.3–1.4 0.0–1.2 0.0–2.8 2.5–43.8 –
Geranial – – – 11.4–29.4 –
Thymol 26.1–30.9 0.0–9.8 11.8–14.2 0.0–0.2 –
Carvacrol 0.7–1.5 5.9–32.8 12.4–22.6 0.0–1.9 –
β-Caryophyllene 10.1–10.3 5.2–12.0 5.1–8.0 2.2–15.8 5.7
Germacrene D 0.8–2.2 0.3–3.1 0.6–1.2 0.4–5.6 2.7
K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

β-Bisabolene 3.3–4.7 2.6–6.4 4.7–9.0 3.5–12.8 1.4

Caryophyllene oxide 1.1–1.2 0.6–1.6 0.3–1.2 1.0– 4.9 0.5
257
258 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259

Table 4
The number of varieties and growing localities of T. pulegioides L. and their chemotypes

Subspecies Varieties Number of Number of Chemotypes and their number (in the
samples localities brackets)

silvestris Conglomeratus 7 4 G/G/N (4), T (1), C/γT/pC (1), L (1)

Silvestris 1 1 G/G/N (1)
pulegioides Pulegioides 6 4 G/G/N (3), T/C/pC/γT (2), C/γT/pC (1)
Ovatus 4 3 G/G/N (1), T (1), T/C/pC/γT (2)
Glaber 6 4 G/G/N (2), C/γT/pC (4)
Adscendens 1 1 C/γT/pC (1)

G/G/N, geranial/geraniol/neral; T/C/pC/γT, thymol/carvacrol/p-cymene/γ-terpinene; C/γT/pC, carvacrol/γ-

terpinene/p-cymene; T, thymol; L, linalool.

Acknowledgements

The authors wish to thank Dr R. Plaušinaitis from the National Nutritional Center,
Vilnius, Lithuania, for his kind assistance with GC/MS analysis of the samples.

References

Adams, R.P., 1995. Identification of Essential Oil Components by GC/MS. Carol Stream Publishing Corp.,
Carol Stream, IL.
Davies, N.W., 1990. Gas chromatographic retention indices of monoterpenes and sesquiterpenes on methyl
silicone and Carbowax 20M phases. J. Chromatogr. 503, 1–24.
Domokos, J., Héthelyi, É., Dános, B., Pálinkás, J., Szirmai, S., 1995. The essential oil of a Hungarian
taxon of Thymus pulegioides. In: Başer, K.H.C. (Ed.), Proceedings of 13th International Congress of
Flavour, Fragrances and Essential Oils, Vol. 1. AREP Publ, Istanbul, 75.
Gudžinskas, Z., 1999. Vascular Plants of Lithuania (Lithuanian). Institute of Botany, Vilnius.
Jalas, J., 1947. Zur Systematik und Verbreitung der Fennoskandischen Formen der Kollektivar Thymus
serpyllum L. em. Fr. Acta Bot. Fenn. 39, 1–92.
Kowal, T., Krupińska, A., 1979. Antibacterial activity of essential oil from Thymus pulegioides L. (in
Polish) Herba Polonica 25, 303–310.
Kustrak, D., Martinis, Z., Kuftinec, J., Blazevic, N., 1990. Composition of the essential oils of some
Thymus and Thymbra species. Flav. Fragr. J. 5, 227–231.
Lekavičius, A.A., Jaskonis, J.A., 1968. The genus of thyme (Thymus L.) in Lithuania (2. Infraspecific
forms and some biochemical properties of Thymus pulegioides L.). (in Russian) Proceedings of the
Academy of Sciences of the Lithuanian SSR 46, 19–24.
Lekavičius, A.A., Jaskonis, J.A., 1969. The genus of thyme (Thymus L.) in Lithuania (3. Key for the
thyme taxon determination and some biometrical data concerning the representatives of the genus).
(in Russian) Proceedings of the Academy of Sciences of the Lithuanian SSR 48, 31–37.
Ložienė, K., 1997. Morphological characteristics and essential oil analysis of Thymus pulegioides L. in
the habitat of hilly landscape of Vilnius vicinities. (in Lithuanian) In: Proceedings of the Lithuanian
Young Botanists. Institute of Botany, Vilnius, pp. 145–151.
Ložienė, K., Vaičiūnienė, J., Venskutonis, P.R., 1998. Chemical composition of the essential oil of creep-
ing thyme (Thymus serpyllum s. l.) growing wild in Lithuania. Planta Medica 64, 772–773.
Ložienė, K., Vaičiūnienė, J., 1999. Intraspecific diversity of Thymus pulegioides L. and characteristics of
habitats. Botanica Lithuanica 5, 27–40.
 K. Ložienė et al. / Biochemical Systematics and Ecology 31 (2003) 249–259 259

Lundgren, L., Stenhagen, G., 1982. Leaf volatiles from Thymus vulgaris , T. serpyllum, T. praecox, T.
pulegioides and T. x citriodorus (Labiatae). Nord. J. Bot. 2, 445–452.
Lyka, K., 1927. Thymian Quendel. In: Hegi, G. (Ed.), Illustrierte Flora von Mitteleuropa, Vol. 5 (4),
München., pp. 2306–2327.
Mártonfi, P., 1992. Polymorphism of essential oil in Thymus pulegioides subsp. chamaedrys in Slovakia.
J. Essent. Oil Res. 4, 173–179.
Mockutė, D., Bernotienė, G., 1999. The main citral-geraniol and carvacrol chemotypes of the essential
oil of Thymus pulegioides L. growing wild in Vilnius district (Lithuania). J. Agric. Food Chem. 47,
3787–3790.
Russo, M., Galletti, G.C., Bocchini, P., Carnacini, A., 1998. Essential oil chemical composition of wild
populations of Italian oregano spice (Origanum vulgare subsp. hirtum (Link) Ietswaart). A preliminary
evaluation of their use in chemotaxonomy by cluster analysis. 1. Inflorescences. J. Agric. Food Chem.
46, 3741–3746.
Sáez, F., 1995. Essential oil variability of Thymus zygis growing wild in southeastern Spain. Phytochem.
40, 819–825.
Salgueiro, L.R., Vila, R., Tomi, F., Tomas, X., Cañigueral, S., Casanova, J., Proença da Cunha, A.,
Adzet, T., 1997. Composition and infraspecific variability of essential oil from Thymus camphoratus.
Phytochem. 45, 1177–1183.
Senatore, F., 1996. Influence of harvesting time on yield and composition of the essential oil of a thyme
(Thymus pulegioides L.) growing wild in Campania (Southern Italy). J. Agric. Food Chem. 44,
1327–1332.
Stahl-Biskup, E., 1986. The essential oil from Norwegian Thymus species. II. Thymus pulegioides. Planta
Medica 3, 233–235.
Stahl-Biskup, E., 1991. The chemical composition of Thymus oils: a review of the literature 1960–1989.
J. Essent. Oil Res. 3, 61–82.