Industrial Crops and Products 19 (2004) 1317

Essential oil composition of Helianthus annuus L. leaves and

heads of two cultivated hybrids Carlos and Florom 350
L. Ceccarini
a,
, M. Macchia
a
, G. Flamini
b
, P.L. Cioni
b
, C. Caponi
b
, I. Morelli
b
a
Dipartimento di Agronomia e Gestione dellAgroecosistema, via S. Michele Degli Scalzi, 2-56124 Pisa University, Italy
b
Dipartimento di Chimica Bioorganica e Biofarmacia, Facolt di Farmacia, via Bonanno, 33 56124 Pisa University, Italy
Received 23 October 2002; accepted 6 June 2003
Abstract
The composition of essential oils from leaves and owers of two hybrids (Carlos and Florom 350) of Helianthus annuus
cultivated in Tuscany (Italy) was investigated. The compounds were identied using gas chromatography (GC)/mass spectrometry
(MS) analyses. Sixty-nine compounds were identied in the essential oils of leaves and owers of sunower plants harvested
in July. Signicant percentage variations were recorded between the leaves and owers oil content. The monoterpenes were
the major compounds present in both essential oils examined. -pinene content was higher in owers (72.6%) than in leaves
(28.6%). The content of sabinene was 2 times higher in leaves than in owers. There were no signicant differences between
the essential oil composition of the oils obtained from the same organs of the two hybrids.
2003 Elsevier B.V. All rights reserved.
Keywords: Helianthus annuus (L.); Leaves; Flowers; Chemical composition; Essential oil
1. Introduction
The cultivated sunower (Helianthus annuus L.) is
counted among most important oil crops on the world
(Putt, 1978; Jonic et al., 2000). Sunower oil has ex-
cellent nutritional properties. It is practically free of
signicant toxic compounds and has a high concentra-
tion of linoleic acid. This polyunsaturated fatty acid
is an essential fatty acid not synthesised by humans,
and is a precursor of gamma linolenic and arachidonic
acids (Dorell, 1978).
The sunower is valuable from an economic, as
well as from an ornamental point of view. Every

Corresponding author. Fax: +39-050-540633.

E-mail address: lceccari@agr.unipi.it (L. Ceccarini).
part of the plant may be utilised for some economic
purpose.
Sunower oil is used for cooking, margarine, salad
dressings, lubrication, soaps, and illumination. A
semi-drying oil is used with linseed and other drying
oils in paints and varnishes. Decorticated press-cake
is used as a high protein food for livestock. Kernels
are eaten by humans raw, roasted and salted, or made
into our. Poultry and cage birds are fond of raw ker-
nels. Flowers yield a yellow dye. Plants are used for
fodder, silage and green-manure crop. Hulls provide
ller in livestock feeds and bedding.
The Indians of North America also had several
non-food uses for the sunower. The plant was cred-
ited with medicinal value (Putt, 1978; Duke and Wain,
1981). Medicinally, seeds are diuretic, expectorant,
0926-6690/$ see front matter 2003 Elsevier B.V. All rights reserved.
doi:10.1016/S0926-6690(03)00076-1
14 L. Ceccarini et al. / Industrial Crops and Products 19 (2004) 1317
and used for colds, coughs, throat, and lung ailments.
The owers and seeds are used in folk remedies for
cancer in Venezuela, often incorporated in white wine
(Hartwell, 1982).
In recent years, increasing interest has been de-
veloping on the non-food applications of renewable
resources. Sunower oil constitute a source of fatty
molecules that can be used as valuable reagents
for chemical modications (Rouilly et al., 2000;
Bourdillon et al., 2000; Girardeau et al., 2000; Leyris
et al., 2000).
The sunower heads are known to contain a strong
smelling essential oil (0.2%) (Marechal and Rigal,
1999), but its composition has never been previously
studied.
The aim of this study was to evaluate the quantity
and the composition of the essential oil obtained from
different organs of the plants of H. annuus L. grown
in the coastal region of Tuscany (Italy).
2. Materials and methods
2.1. Plant material
H. annuus L. was cultivated in a eld plot
within the Centro Interdipartimentale di Ricerche
Agro-Ambientali Enrico Avanzi of Pisa Univer-
sity. Soil chemicalphysical properties were as fol-
lows: sand 29.3%; silt 37.6%; clay 33.1%; pH 8.5;
organic matter (Lotti method) 1.66%; total N (Kjel-
dahl method) 1.23; assimilable P (Olsen method)
4.75 ppm; exchangeable K (Internal Method (Barium
Chloride and Tea)) 175.3 mg/kg.
On eld plots, deep ploughing was performed in
January 2001. Soil fertilisation was carried out be-
fore sowing by 124, 96 and 96 kg/ha of N P
2
O
5
and
K
2
O, respectively. Sunower hybrids Florom 350 and
Carlos were sown in May 2001 by a precision drill
to obtain an 8 plants/m
2
crop density. Pre-emergence
herbicide Duasol (Metolaclor + Metobromuron) was
sprayed at the rate of 4 l/ha.
In summer (27/7/2001), during reproductive stages
at the beginning of anthesis (stage R5) (Schneiter
and Miller, 1981), a representative sample of hy-
brid Carlos and Florom 350 were sacriced. Each
sample plant was separated into two parts: inores-
cences and leaves. In order to allow us to individually
characterise the oil composition of each sample the
different organs were taken from different plant posi-
tions, dried in the shade below 40

C, until constant
weight.
Percentage data were evaluated as arcsin(

% trans-
formed) using the ANOVA statistical package. In both
cases, means were separated on the basis of the LSD
test only when the F-test of the ANOVA per treatment
was signicant at the 0.05 or 0.01 probability level
(Gomez and Gomez, 1984).
2.2. Essential oils analyses
The plant material was water distilled the next day
in a Clevenger-type apparatus for 2 h.
The gas chromatography (GC) analyses were ac-
complished with a HP-5890 Series II instrument
equipped with HP-WAX and HP-5 capillary columns
(30 m 0.25 mm, 0.25 m lm thickness), work-
ing with the following temperature program: 60

C
for 10 min, ramp of 5

C/min up to 220

C; injector
and detector temperatures 250

C; carrier gas nitro-

gen (2 ml/min); detector dual FID; split ratio 1:30;
injection of 0.5 l). The identication of the com-
ponents was performed, for both the columns, by
comparison of their retention times with those of
pure authentic samples and by mean of their linear
retention indices (LRI) relative to the series of n-
hydrocarbons.
The relative proportions of the essential oil con-
stituents were percentages obtained by FID peak-area
normalisation, all relative response factors being taken
as one.
Gas chromatography/electron impact mass spec-
trometry (EIMS) analyses were performed with a
Varian CP-3800 gas chromatograph equipped with
a DB-5 capillary column (30 m 0.25 mm; coating
thickness 0.25 m) and a Varian Saturn 2000 ion
trap mass detector. Analytical conditions: injector
and transfer line temperatures 220 and 240

C, re-
spectively; oven temperature programmed from 60 to
240

C at 3

C/min; carrier gas helium at 1 ml/min;

injection of 0.2 l (10% hexane solution); split ra-
tio 1:30. Identication of the constituents was based
on comparison of the retention times with those of
authentic samples, comparing their linear retention
indices relative to the series of n-hydrocarbons, and
on computer matching against commercial (NIST 98
L. Ceccarini et al. / Industrial Crops and Products 19 (2004) 1317 15
and ADAMS) and home-made library mass spectra
built up from pure substances and components of
known oils and MS literature data (Stenhagen et al.,
1974; Massada, 1976; Jennings and Shibamoto, 1980;
Swigar and Silverstein, 1981; Davies, 1990; Adams,
1995). Moreover, the molecular weights of all the
identied substances were conrmed by gas chro-
matography/chemical ionisation mass spectrometry
(CIMS), using MeOH as CI ionising gas.
3. Results and discussion
Sixty-nine compounds accounting for 96.398.4%
of the whole essential oil were identied in the four
samples. The essential oil yields varied between 0.12
and 0.4%. The highest yields were obtained in the
leaves samples (Table 1).
In both the essential oils obtained from the leaves
of the two cultivars Florom 350 and Carlos, monoter-
penes were the major compounds, accounting for
about 80% of the whole oil; also sesquiterpenes were
Table 1
Essential oil composition
a
of leaves and capitula of the two cultivars of Helianthus annuus
LRI
b
Leaves Capitula
Florom 350 Carlos Florom 350 Carlos
(E)-3-Hexen-1-ol 852 tr
c
tr 0.1 0.1
(E)-2-Hexen-1-ol 861 0.1
Tricyclene 928 0.2 0.5
-Thujene 933 0.3 tr 0.4 0.4
-Pinene 940 28.2 28.9 74.5 70.7
Camphene 955 4.7 5.4 0.9 0.7
Thuja-2,4(10)-diene 959 0.1 tr
Sabinene 977 23.5 23.2 11.2 12.1
-Pinene 981 4.0 4.4 1.7 1.3
Myrcene 992 0.4 0.5 tr tr
2,3-Dehydro-1,8-cineole 993 0.1 tr
Pseudolimonene 1007 tr
-Terpinene 1020 0.1 0.1 0.4 0.5
p-Cymene 1028 0.1 tr tr tr
Limonene 1032 11.1 12.3 1.4 0.8
-Phellandrene 1033 0.7 0.3 0.2 0.1
1,8-Cineole 1035 tr
Phenylacetaldehyde 1044 tr 0.1
(E)-Ocimene 1051 tr 0.1
-Terpinene 1063 0.3 0.3 0.6 0.8
cis-Sabinene hydrate 1070 0.1 0.1 0.1
Terpinolene 1089 0.1 0.1 0.2 0.3
numerically well represented, even if their percentage
was small. These essential oils, from a qualitative and
quantitative point of view, resulted very similar: the
main compounds identied were -pinene, sabinene,
limonene, germacrene D, isobornyl acetate, camphor
and -pinene (Table 1).
Also, the essential oils obtained from capitula of
both cultivars showed similar compositions; with re-
spect to the leaves, the principal differences were due
to -pinene, still the main compound, but with percent-
ages that rose to 74.5% in Florom 350 and to 70.7%
in Carlos, while the amounts of sabinene was about
halved and those of limonene and germacrene D were
about 10 and 8 times less, respectively (Table 1).
The data about some of the oil constituents, sub-
mitted to ANOVA test, showed signicant statistical
differences relatively to the interaction between treat-
ments examined (Table 2).
There were signicant differences in the oil content
of different plant organs examined: the leaf oil con-
tent (20%) was signicantly higher than that extracted
by the ower (15%). On the contrary, no statisti-
16 L. Ceccarini et al. / Industrial Crops and Products 19 (2004) 1317
Table 1 (Continued )
LRI
b
Leaves Capitula
Florom 350 Carlos Florom 350 Carlos
trans-Sabinene hydrate 1099 tr 0.1 0.1
cis-p-Mentha-2-en-1-ol 1125 tr 0.1
trans-p-Mentha-2,8-dien-1-ol 1126 tr
-Campholenal 1127 tr tr 0.1
trans-p-Mentha-2-en-1-ol 1142 0.3 0.3
cis-Verbenol 1143 0.3 0.3 0.7 0.6
trans-Verbenol 1145 0.1
Pinocarvone 1164 tr tr tr 0.1
p-Mentha-1,5-dien-8-ol 1168 0.2 0.3
Borneol 1169 0.4 0.3
4-Terpineol 1179 0.6 0.3 1.3 1.8
p-Cymen-8-ol 1185 tr 0.1
-Terpineol 1190 0.2 0.3
Myrtenol 1195 tr 0.1
Safranal 1201 tr tr
Decanal 1205 tr
Verbenone 1206 0.2 0.2
2-Methyl-2-nonen-4-one 1215 0.2 0.4
trans-Carveol 1219 tr 0.1
-Cyclocitral 1223 tr tr
Isobornyl acetate 1286 8.0 7.8 0.9 1.1
2-Undecanone 1293 tr
trans-Pinocarvyl acetate 1298 0.2 0.1 tr tr
methyl geranate 1324 tr
-Copaene 1377 tr tr
-Bourbonene 1385 tr 0.1
-Cubebene 1391 tr 0.1
-Elemene 1392 tr tr 0.1 0.1
-Caryophyllene 1419 0.7 0.7 0.1 0.1
-Gurjunene 1433 0.9 1.2 0.6 1.2
trans--Bergamotene 1441 0.2 0.1 tr
(E)-Geranyl acetone 1453 tr tr
-Humulene 1456 0.4 0.4 tr tr
Germacrene D 1481 8.2 8.8 1.1 1.2
Bicyclogermacrene 1496 0.4 0.3 tr tr
-Muurolene 1502 tr
trans--Cadinene 1514 0.4 0.2
-Cadinene 1523 0.1 tr tr tr
trans-Nerolidol 1565 0.4 0.3 tr tr
Dendrolasin 1574 tr 0.1
Spathulenol 1575 tr 0.1
Germacrene D-4-ol 1576 0.9 0.6
Caryophyllene oxide 1582 0.2 0.2
T-cadinol 1641 0.1 tr tr tr
Desmethoxy encecalin 1647 0.6 tr
epi-13-Manoyl oxide 2011 0.1
Essential oil yields (% w/w) 0.37 0.40 0.12 0.13
Total identied 97.0 98.4 97.7 96.3
a
Percentages obtained by FID peak-area normalisation, all relative response factors being taken as one (HP-5 column).
b
Linear retention indices (HP-5 column).
c
tr < 0.01%.
L. Ceccarini et al. / Industrial Crops and Products 19 (2004) 1317 17
Table 2
Effects of the organ of the plant on some components of the
essential oil in Helianthus annuus
Compound Leaves Flowers
Florom
350
Carlos Florom
350
Carlos
-Pinene 32.4 C 32.8 C 60.0 A 57.1 B
Camphene 12.8 JK 13.2 J 5.2 OP 4.9 P
Sabinene 29.0 D 28.7 D 19.8 FG 20.2 EF
-Pinene 11.8 L 12.0 KL 7.2 M 6.8 MN
Limonene 19.2 G 20.8 E 6.9 MN 5.1 P
Isobornyl acetate 16.2 HI 16.1 I 5.2 OP 6.0 NO
Germacrene D 16.8 HI 17.1 H 6.0 NO 6.3 N
Each value is the mean of three replicates. Means followed by
the same letters are not signicantly different at the 0.01 (capital
letters) probability level according to LSD test.
cally signicant differences were observed between
the essential oil composition of the oils obtained
from the two hybrids analysed (Florom 350 and
Carlos).
The hybrid Florom 350 had the highest -pinene
content in owers (60%) with respect to hybrid Carlos.
In the leaves, the amount of -pinene decreased to
32.6% and both the hybrid showed the same amount
of -pinene.
Among the examined organs, there were signicant
differences between the quantity of sabinene: leaves
contained 28.9% while the owers showed the lowest
amount (20%).
The limonene content showed signicant differ-
ences relative to all the excited treatments. The high-
est value was found in the leaves of the hybrid Car-
los (20.8%), while the owers contained the lowest
amounts (6.9 and 5.1% in the hybrid Florom 350 and
Carlos, respectively).
Isobornyl acetate and germacrene D were present
principally in the leaves (16.6%) and the owers con-
tained signicantly less amount of both compounds
(5.9%).
There were no signicant statistical differences
between the mean -pinene content of Carlos and
Florom 350 hybrid; noteworthy differences were
found between the leaves (11.9%) and owers, where
-pinene was in lower amounts (7%).
The observed differences in the -pinene content of
the two oils could be due to a different physiological
or ecological role in the two plant organs.
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