Chemical composition of the volatile oil from different plant parts of Anethum graveolens L. (Umbelliferae) cultivated in Romania

Abstract

The essential oil from dried leaves, flowers and fruits of Anethum graveolens L. (dill) cultivated in Romania was isolated by hydrodistillation and analysed by gaschromatography coupled with mass spectrometry (GC-MS). The main components in leaves were α- phellandrene (62.71%), limonene (13.28%) and anethofuran (16.42%). The main components in flowers were α- phellandrene (30.26%), limonene (33.22%) and anethofuran (22%). Cis-carvone and limonene are the major constituents of seeds volatile oil with 75.2% and, respectively 21.56%.

Full text

FARMACIA, 2010, Vol.58, 5
594
CHEMICAL COMPOSITION OF THE VOLATILE
OIL FROM DIFFERENT PLANT PARTS OF
ANETHUM GRAVEOLENS L. (UMBELLIFERAE)
CULTIVATED IN ROMANIA
VALERIA RĂDULESCU*, MARIA LIDIA POPESCU, DIANA-
CAROLINA ILIEŞ
University of Medicine and Pharmacy “Carol Davila”, Faculty of
Pharmacy, 6 Traian Vuia, 020956, Bucharest, Romania
*corresponding author: valeria_radulescu@yahoo.com
Abstract
The essential oil from dried leaves, flowers and fruits of Anethum graveolens L.
(dill) cultivated in Romania was isolated by hydrodistillation and analysed by gas-
chromatography coupled with mass spectrometry (GC-MS). The main components in
leaves were α- phellandrene (62.71%), limonene (13.28%) and anethofuran (16.42%). The
main components in flowers were α- phellandrene (30.26%), limonene (33.22%) and
anethofuran (22%). Cis-carvone and limonene are the major constituents of seeds volatile
oil with 75.2% and, respectively 21.56%.
Rezumat
Uleiul volatil obţinut din frunze, flori şi fructe uscate de Anethum graveolens L.
(mărar) cultivat în România, a fost izolat prin hidrodistilare şi analizat prin cromatografie
de gaze cuplată cu spectrometrie de masă (GC-MS). Principalii componenţi din frunze au
fost α-felandrenul (62,71%), limonenul (13,28%) şi anetofuranul (16,42%). Principalii
componenţi din flori au fost α-felandrenul (30,26%), limonenul (33,22%) şi anetofuranul
(22%). Cis-carvona şi limonenul sunt principalii compuşi din uleiul volatil obţinut din
seminţe, reprezentând 75,2% şi, respectiv, 21,56%.
Keywords: Anethum graveolens, dill, essential oil, carvone, limonene.
Introduction
Anethum graveolens L. or dill, belonging to Apiaceae
(Umbelliferae) family, is an annual aromatic herb known for culinary and
medicinal use since ancient times. It is cultivated in the most parts of
Europe and the United States of America. A variant called east Indian dill or
sowa (Anethum sowa Roxb.) is cultivated in India, Egipt and Japan. The
chemical composition of the essential oil of the two chimiotypes of dill and
sowa differs mainly by the dillapiole content. The typical flavour of herb
dill oil is due to α-phellandrene, limonene and dill ether (anethofuran). For
flavouring purposes the herb oil with low content of carvone is preferred
[1]. The dill seed oil contains a small quantity of dillapiole up to 3% when
grown in tropical climate [1, 2]. In the east Indian dill (sowa) the content of

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595
dillapiole ranges from 5 to 27% [3].
In recent years the scientific literature reports pharmacological
effects of dill such as antibacterial [4, 5], antimycobacterial [6], antioxidant
[7-10], cancer chemopreventive [11]. The well-known properties of dill
from the traditional medicine, such as carminative, stomachic, diuretic have
been reported [12, 13]. The dill essential oil has hypolipidemic activity and
could be a cardioprotective agent [14]. Many studies showed that dill
essential oil quantity and chemical composition varies depending on the
plant parts and the developing stage of the plant at harvest time [13, 15-17].
The scientific literature data concerning chemical composition of volatile oil
from different plant parts of Anethum graveolens L. are poor and differ from
one author to the other [3, 13-15, 18].
The aim of this paper is to elucidate the chemical composition of the
essential oil from leaves, flowers and fruits of dill cultivated in Romania. In
order to study the complex chemical composition of volatile compound
from plants, advanced analytical GC-MS techniques must be used, these
allowing the identification of compounds even in minute quantities [23, 24].
Materials and methods
Reagents and solvents
All solvents and reagents were purchased from Merck, Darmstadt,
Germany: dichloromethane, supraSolv for gas chromatography, anhydrous
Na2SO4 granulated for organic trace analysis, the C8-C20 and C21-C40 n-
alkanes used for the determination of Kovats retention indices.
Plant material
The raw material consisted of the leaves (harvested during the plant's
flowering), flowers (on the blossom stage), and fruits (at their full maturity)
of Anethum graveolens L., dill (Apiaceae) harvested in 2008 from Arges
county (the southern part of Romania; 500 m altitude). The products were
naturally dried in shadow and stored in controlled laboratory conditions.
Isolation of the essential oil
100 grams of fragmented dried vegetal products were hydrodistilled
with 500 mL water in a Clevenger-type apparatus without organic solvent
for 3h [25]. The essential oil was dried over anhydrous Na2SO4, stored in a
dark glass bottle and kept at 4°C until analysis.

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Gas chromatography-mass spectrometry
GC-MS analyses was performed on a Fisons Instrument GC 8000
equipped with an electron impact quadrupole, MD 800 mass spectrometer
detector. The electron ionisation energy was 70 eV, ion-source temperature
200°C and the interface temperature 280°C.
A fused silica capillary column 5% phenyl-poly-dimethyl-siloxane
(DB-5MS 30 m x 0.32 mm i.d. and 0.25 µm film thickness, J&W Scientific)
was used. The column temperature was programmed as follows: from 40°C
(3 min hold) raised at 4°C/min to 250 °C and finally held at 250 °C for 10
min. A split-splitless injection (split ratio 1:30) at 280 °C was employed.
The carrier gas (helium) flow rate was 2 mL/min. Two µL of sample were
injected. Data acquisition was performed with MassLab software for the
mass range 30 - 600 u with a scan speed of 1 scan/s. The identification of
compounds was performed by comparing their mass spectra with data from
Adams [19], US National Institute of Standards and Technology (NIST,
USA), WILEY 1996 Ed. mass spectra library and a personal library of 600
spectra. The identification of compounds was also based on the Kovats
retention indices.
The Kovats retention indices were calculated using n-alkanes C8-C20
and C21-C40 and the experimental values were compared with those reported
in literature [20, 21].
Results and discussion
The average content in essential oil of Anethum graveolens samples
(3 determinations) was: 12 mL/kg for leaves, 32 mL/kg for flowers and 34
mL/kg for fruits (the results were calculated with reference to the dried
material). These values are comparable with the results mentioned in the
scientific literature about Anethum graveolens [3, 22].
In figure 1 are shown the chromatograms of essential oil from leaves
(a), flowers (b) and fruits (c).

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6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
36
38
0
16.481
9.490
4.034
15.154
9.540
8.772
6.311
4.034
6.178
14.544
16.379
22.752
30.753
c) fruits
8.839
6.328
4.034
7.721
14.536
9.531
31.243
22.743
30.192
38.484
tR
(min)
a) leaves
b) flowers
Figure 1.
Chromatograms of leaves, flowers and fruits essential oil from Anethum
graveolens.
Table I shows the relative content of volatile compounds from
essential oil, expressed as percentage from total area.

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Table I.
Chemical composition of essential oil from leaves,
flowers and fruits of Anethum graveolens L.
In the essential oil from Anethum graveolens L. leaves 21
compounds were identified, adding up to 97.55% of the total area.
Monoterpenic hydrocarbons were found predominant in the leaves oil
representing 79.14% of the total content, where α-phellandrene (62.71%)
constituted the major compound. This result is in agreement with that
reported by Amin and Sleem [13]. The oxygenated compounds reached up
to 16.56% with 16.42% dill ether (3,9-epoxy-1-p-menthen), which was the
major compound. The n-alkanes from C19-C27were found in small quantities
between 0.05 – 0.19% totalling 1.14%.
Nr
Compounds
tR /min*
Kovats
retention
indices
Area/&%&
Leaves
Flowers
Fruits
1
α
-Tujene
6.186
925
0.14
0.07
-
2
α
-Pinene
6.328
930
1.66
0.90
0.02
3
Sabinene
7.721
974
0.12
0.05
-
4
β
-Myrcene
8.472
994
0.12
0.06
-
5
α
-Phellandrene
8.839
1004
62.71
30.26
0.12
6
p-Cymene
9.406
1024
1.11
0.65
-
7
Limonene
9.531
1028
13.28
33.22
21.56
8
p-Menth-3-en-2-one
14.394
1181
0.07
-
-
9
Dill Ether
14.536
1185
16.42
22.00
-
10
cis-Dihydrocarvone
15.088
1201
-
0.18
0.04
11
trans-Dihydrocarvone
15.154
1204
-
1.88
3.02
12
Carvone
16.481
1249
-
10.29
75.21
13
α
-Copaene
22.752
1378
-
0.18
-
14
Cadinol
27.391
1667
-
0.09
-
15
γ
-Muurolene
22.743
1477
0.29
-
-
16
p-Menth-1,8-dien-6-ol
24.003
1527
0.07
-
-
17
Neophytadiene
31.243
1838
0.42
-
-
18
n-Nonadecane
32.636
1900
0.07
-
-
19
n-Eicosane
34.696
2000
0.05
-
-
20
n-Heneicosane
36.632
2100
0.09
-
-
21
n-Docosane
38.484
2200
0.12
-
-
22
n-Tricosane
40.277
2300
0.19
-
-
23
n-Tetracosane
42.004
2400
0.19
-
-
24
n-Pentacosane
43.664
2500
0.19
-
-
25
n-Hexacosane
45.266
2600
0.16
-
-
26
n-Heptacosane
47.051
2700
0.08
-
-
Identified from total area
97.55
99.83
99.97

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In the flower essential oil 13 compounds were identified adding up
to 99.83% of the total area. The main components of the volatile oil from
flowers are monoterpenic hydrocarbons: α-phellandrene (30.26%) and
limonene (33.22%), total monoterpenic hydrocarbons representing 65.21%.
In flowers essential oil the content of α-phellandrene is only 30.26%
compared with 62.71% in the leaves oil. On the other hand, the content of
limonene and dill ether in flowers oil is higher (33.22% and respectively
22.00%) than the one determined in leaves oil.
In the leaves oil, the ketonic compound carvone was not present but
in the flowers oil its content is 10.29%. In the essential oil of fruits, carvone
is the main compound with 75.21%.
Another ketonic compound absent in the leaves essential oil was
present in the flowers and fruits oils as cis- and trans-dihydrocarvone
isomers. The fruit essential oil was rich in limonene 21.56% and cis- and
trans-dihydrocarvone adding up to 3.06%. The amount of α-phellandrene in
leaves oil and flowers oil was 62.71%, respectively 30.26%, and only 0.12%
in fruits oil.
Conclusions!
The chemical composition of dill volatile oil varies depending on the
plant parts. In the leaves oil monoterpenic hydrocarbons are predominant,
amounting to 79.14% (62.71% α-phellandrene and 13.28% limonene). In
the flowers oil the content of α-phellandrene and limonene is 32.26% and
33.22%, respectively. Anethofuran (dill ether) is present in leaves and
flowers with 16.42% and 22%, respectively, but is missing in the fruit oil.
The main compound in fruits essential oil is carvone (75.21%), while the
content of α-phellandrene is only 0.12% and limonene is 21.56%.
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Manuscript received: December 5th 2009