FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fragr. J. 2001; 16: 353–355

DOI: 10.1002/ffj.1010

Composition of the essential oil of Erechtites hieracifolia

from Bolivia
Daniel Lorenzo,1 Gloria Saavedra,2 Ingrid Loayza2 and Eduardo Dellacassa1Ł
1 Catedra de Farmacognosia y Productos Naturales, Facultad de Quimica, Universidad de la República, Avda. General Flores 2124,
CP-11800 Montevideo, Uruguay
2 Centro de Tecnologı́a Agroindustrial — Programa Agroquı́mico, Facultad de Ciencias y Tecnologia, Universidad Mayor de San

Simon, PO Box 992, Cochabamba, Bolivia

Received 21 December 2000

Revised 20 March 2001
Accepted 24 March 2001

ABSTRACT: Analysis by GC and GC–MS of the essential oil obtained from aerial parts of Erechtites
hieracifolia revealed 22 components (93% of the total composition). The major constituents were found to
be ˛-pinene, myrcene, (E)-ˇ-ocimene, (E)-caryophyllene and germacrene D; the enantiomeric distribution of ˛-
pinene, sabinene, ˇ-pinene, limonene and linalool was studied by multidimensional HRGC–HRGC. The major
component was ˛-pinene (48%) with an enantiomeric purity of 100% for (1R)-(C)-˛-pinene. Copyright  2001
John Wiley & Sons, Ltd.

KEY WORDS: Erechtites hieracifolia (L.) Raf. ex DC; Clavelı́n; Asteraceae; enantioselective gas chromatogra-
phy; ˛-pinene

Introduction in folk medicine as a blood depurative.7 The roots also

The phytochemistry of some species of the genus Ere-
chtites, E. valerianaefolia (Link ex Spreng) DC,1,2 E.
quadridentata DC3 and E. hieracifolia,4,5 has been stud-
ied previously. E. hieracifolia has been analysed for
volatile compounds by different research groups. Bohl-
mann4 found a syringyl alcohol derivative and sesquiter-
penes from Et2 O–petroleum extracts from roots. Later,
Lemos6 studied the essential oil composition obtained
from fresh leaves of E. hieracifolia collected in Ceara,
Brazil. Analysis of the volatile fraction showed that ˛-
phellandrene (41.3%), p-cymene (22.2%), cis-ascaridol
(10.2%) and (E)-caryophyllene (7.4%) composed 71.1%
of the essential oil.
Some Erechtites species grow wild in Bolivia, in parti-
cular Erechtites hieracifolia (L.) Raf. ex DC (‘Clavelin’)
is a native plant in the Chapare region, Carrasco Pro-
vince, at an altitude of 300 m. This species is an annual
herb 1–3 m in height. The leaves and flowers are used
*Correspondence to: E. Dellacassa, Catedra de Farmacognosia y Pro-
ductos Naturales, Facultad de Quı́mica, Universidad de la República,
Avda. General Flores 2124, UR-11800 Montevideo, Uruguay.
Contract/grant sponsor: Universidad de la República de Uruguay, Pro-
grama Contrapartida de convenios, Uruguay; Contract/grant number:
205 de la Ley 16462.
Contract/grant sponsor: Programa de Desarrollo de las Ciencias Basi-
cas (PEDECIBA), Uruguay; Contract/grant number: PNUD Project
URU/97/016.
are reported to be effective in cardiac disease.7
The purpose of this study was to characterize the
essential oil isolated from the leaves and stems of
E. hieracifolia and provide information about the exis-
tence of a chemotype within the species.
Experimental
Plant Material and Isolation of Volatile
Constituents
Fresh E. hieracifolia plants were collected in the Chi-
moré area, Senda D, located in the Chapare Province,
situated at 16° 370 S latitude, 65° 150 WO longitude and
300 m altitude, during the full flowering period at the
spring season of 1999. The samples were representa-
tive of the species and its geographic area of distri-
bution. Voucher specimens of the plant were identified
and deposited at the Herbarium of the National Flora
Reserve ‘Martı́n Cárdenas’ in Cochabamba (M. Zárate,
BOLV 340).
The oil was obtained by hydrodistillation with a
Clevenger-type apparatus immediately after the collec-
tion of the plant material. The isolation procedure was
continued until the material was exhausted (6 h) and the
yield obtained was 0.39 ml/100 g of fresh plant. The oil

Copyright  2001 John Wiley & Sons, Ltd.

354 D. LORENZO ET AL.

was dried with anhydrous sodium sulphate and kept in columns, determined in relation to a homologous series
a sealed flask at 12 ° C until the moment of analysis. of n-alkanes, with those from pure standards or reported
in literature. Comparison of fragmentation patterns in
the mass spectra with those stored on databases8,9 and
GC Analysis
MS data of our collection was also performed. The
Capillary gas chromatography was carried out using a quantification of the components was performed on the
Shimadzu 14 B gas chromatograph equipped with a FID basis of their GC peak areas on SE-52, except those
and a Shimadzu data processor software EZ-Chrom, using indicated in Table 1 which were obtained on Carbowax
two capillary columns. The first was a SE-52 (Mega, 20M, without corrections for response factors.
Legnano, Italy) cross-linked fused-silica capillary col-
umn (25 m ð 0.32 mm i.d.), coated with 5% phenyl-
Chiral Analysis
polymethylsiloxane (0.40–0.45 µm phase thickness); col-
umn temperature, 60 ° C (8 min) to 180 ° C at 3 ° C/min,
Enantiomeric ratios of ˛-pinene, sabinene, ˇ-pinene,
180–250 ° C at 20 ° C/min, 250 ° C (10 min); injector tem-
limonene and linalool were obtained by multidimen-
perature, 250 ° C; detector temperature, 280 ° C; injection
sional gas chromatography, using a developmental mo-
mode, split; split ratio, 1 : 30; volume injected, 0.2 µl of
del10 set up with two GC ovens. The first was equipped
the oil. The carrier gas was hydrogen, 55 kPa.
with a column coated with SE-52 and the second with a
The second column was a Carbowax 20M (Ohio Val-
chiral column coated with a derivatized ˇ-cyclodextrin,
ley, USA) bonded fused-silica capillary column (25 m ð
a hot interface, a rotary switching valve and a system
0.32 mm i.d.), coated with polyethylene glycol (0.25 µm
to maintain a constant flow during the transfer. With
phase thickness); column temperature, 40 ° C (8 min) to
this system a heart-cut of the relevant fractions can be
180 ° C at 3 ° C/min, to 230 ° C at 20 ° C/min; injector tem-
perature, 250 ° C; detector temperature, 250 ° C; injection
mode, split; split ratio, 1 : 30; volume injected, 0.2 µl of Table 1. Percentage composition of the essential oil
of Erechtites hieracifolia (L.) Raf. and linear retention
the oil. The carrier gas was hydrogen, 30 kPa. indices (LRI) of its components
LRIŁŁŁ
GC–MS Analysis CompoundŁ (%)ŁŁ SE-52 CW 20M
˛-Pinene 48.0 937 1013
GC–MS analysis were conducted using a Shimadzu Camphene 0.2 951 1051
QP 5050 equipped with reference libraries,8,9 using Sabinene 1.8 973 1109
two capillary columns. The first was a SE-52 (Mega, ˇ-Pinene 0.9 979 1092
Legnano, Italy) cross-linked fused-silica capillary col- Myrcene 13.7 992 1164
˛-Phellandrene 0.7 1000
umn (25 m ð 0.25 mm i.d.), coated with 5% phenyl p-Cymene 0.5 1025 1257
polymethylsiloxane (0.25 µm phase thickness); column Limonene 1.0 1029 1184
temperature, 60 ° C (8 min) to 180 ° C at 3 ° C/min, to (Z)-ˇ-Ocimene 0.4 1034 1228
(E)-ˇ-Ocimene 13.9 1251
230 ° C at 20 ° C/min; injector temperature, 250 ° C; injec- -Terpinene 0.1 1051
tion mode, split; split ratio, 1 : 40; volume injected, 0.2 µl Linalool 0.3 1100 1548
of the oil. Helium was used as the carrier gas, using 4-Terpineol 0.2 1182 1592
˛-Terpineol 0.1 1191 1662
122.2 kPa (51.6 cm/s); interface temperature, 250 ° C; ˛-Copaene 1.6 1376 1469
acquisition mass range 40–400; solvent cut, 2 min. ˇ-Elemene 0.3 1388
The second column was a BP 20 (SGE, Australia) (E)-Caryophyllene 4.1 1420 1572
˛-Humulene 1.6 1456 1642
bonded fused-silica capillary column (25 m ð 0.25 mm Germacrene D 2.0 1481 1683
i.d.), coated with polyethylene glycol (0.25 µm phase υ-Cadinene 0.6 1517 1732
thickness); column temperature, 40 ° C (8 min) to 180 ° C (E)-nerolidol 0.8 1561 2037
˛-Cadinol 0.5 1655 2219
at 3 ° C/min, to 230 ° C at 20 ° C/min; injector temper-
Total 93.3
ature, 250 ° C; injection mode, split; split ratio, 1 : 40;
Identified components
volume injected, 0.2 µl of the oil. The carrier gas Grouped components (%)
was helium 92.6 kPa (55.9 cm/s); interface temperature, Monoterpene hydrocarbons 81.2
250 ° C; acquisition mass range, 40–400; solvent cut, Oxygen-containing monoterpenes 0.7
Sesquiterpene hydrocarbons 10.2
2 min. Oxygen-containing sesquiterpenes 1.3
Ł The components are reported according their elution order on SE-52.
ŁŁ
Identification and Quantification These percentages were obtained on SE-52 except for those of sabinene and
ˇ-pinene which were obtained on Carbowax 20M.
ŁŁŁ
Peak identifications are based on comparison of their linear retention indices
The components of the essential oil were identified by (LRIs) on the two columns with those from pure standards or reported in
comparison of their linear retention indices on the two literature and on MS comparison with file spectra.

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 353–355

Table 2. Enantiomeric ratios for ˛-pinene, sabinene, ˇ-pinene, limonene and linalool in
Erechtites hieracifolia (L.) Raf. ex DC essential oil
Compound ˛-Pinene Sabinene
Enantiomer 1R-(C) 1S -() 1R, 5R-(C) 1S , 5S -() 1R-(C) 1S -() 4R-(C) 4S -() 3S -(C) 3R-()
100.0 tr 28.2 71.8
Cut-times (min) 8.52–8.77 10.65–10.90
made and these fractions transferred from the non-chiral
column to the chiral column according to the experimen-
tal conditions previously described.11
Results and Discussion
The components of the oil examined are reported in
Table 1. Twenty-two components, which represent 93%
of the total composition, were identified. The oil was
characterized by a high content of ˛-pinene (48%),
myrcene (14%), (E)-ˇ-ocimene (14%) and (E)-caryophy-
llene (4%). The oxygenated fraction is almost absent
when compared with the hydrocarbon fraction.
The obtained composition showed a different nature
than those reported in the literature.6 The HRGC–MS
analysis revealed only traces of ˛-phellandrene and
p-cymene, while the ˛-phellandrene epoxide, the ascari-
dol isomers and the ketones were absent.
In order to better characterize this oil, the enantiomeric
ratio of five components was determined by subsequent
transfers during the same analysis using authentic sam-
ples of each enantiomer to determine their order of
elution. Table 2 reports the enantiomeric ratios of the
components analysed. The most noticeable value was
observed for (1R)-(C)-˛-pinene, the main component,
which showed an enantiomeric excess of 100%.
It can therefore be concluded that the examina-
tion carried out on the essential oil of E. hieracifolia
allowed us to identify compounds that characterize
Copyright  2001 John Wiley & Sons, Ltd.
ESSENTIAL OIL OF ERECHTITES HIERACIFOLIA 355
ˇ-Pinene Limonene Linalool
10.3 89.7 tr 100.0 75.9 24.1
10.95–11.20 13.72–13.97 18.45–18.70
this oil with respect to those reported from the same
species. Furthermore, the enantiomeric distribution of
the selected compounds allowed us to characterize
chemotaxonomically the E. hieracifolia oil from Bolivia.
A more detailed investigation studying the variability
between the populations of Erechtites hieracifolia must
be performed to define taxonomic implications, both
botanical and chemical.
Acknowledgements—This research was supported by the Universidad
de la República del Uruguay, Programa Contrapartida de Convenios
(Art. 205 de la Ley 16462). D. Lorenzo would like to thank the
Programa de Desarrollo de las Ciencias Basicas (PEDECIBA, PNUD
Project URU/97/016) for a Master’s grant.
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