International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 4, Issue 6, Dec 2014, 185-192
TJPRC Pvt. Ltd.

CHEMICAL COMPOSITION AND INSECTICIDAL ACTIVITIES OF ESSENTIAL OILS OF

TWO ALGERIAN ENDEMIC PLANTS: FERULA VESCERITENSIS COSS. ET DUR. AND
THYMUS PALLESCENS DE NOE
OTMANE BENCHABANE
Laboratory of Food Technology and Human Nutrition, Ecole Nationale Suprieure Agronomique (ENSA), Algria

ABSTRACT
The essential oils (E.O.) of two Algerian endemic plants (Ferula vesceritensis Coss.et Dur and Thymus pallescens
de No) were extracted by hydrodistillation and analyzed by GC and GC-MS. The yields were 0.4%, 2.3%, respectively.
Chromatographic analysis lead to the identification of 54 components representing 92.2% of F. vesceritensis oil which was
characterized by shyobunol (18.1%) and -cadinene (14.2%) as the main compounds while 35 components representing
93% of T. pallescens essential oil were identified among which carvacrol (40.2%), thymol (13.5%), -terpineol (7.0%) and
-terpinene (6.5%) were the major constituents. Insecticidal activity against rice weevil (Sitophilus oryzae) by contact and
inhalation methods showed appreciable insecticidal property with the essential oil of F. vesceritensis (LD50= 16.4 l/ml or
0.26 l/cm2; LD90= 53.45 l/ml or 0.84 l/cm2) and potent activity with T. pallescens oil (LD50= 8.76 l/ml or 0.14
l/cm2; LD90= 17.15 l/ml or 0.27 l/cm2). The results suggest that essential oils from the studied plants may be used
against Sitophilus oryzae in grain storage.

KEYWORDS: Essential Oils, Insecticidal Activity, Ferula vesceritensis, Thymus pallescens, Sitophilus oryzae
INTRODUCTION
The genus Ferula, belonging to the family Apiaceae, includes about 170 species occurring from central Asia
westward throughout the Mediterranean region to northern Africa (Pimenov and Leonov, 1993). Out of which only 5
species are identified in Algeria among which 2 are endemic whose F. vesceritensis (Quezel and Santa, 1963; Ozenda,
1958). Ferula species have been known as a rich source of aromatic resins and reputed in the traditional medicine as
natural remedies for a variety of disorders such as anticonvulsant, carminative, antispasmodic, diuretic, aphrodisiac,
antihelmintic etc. At least part of the biological activities of the plants of this genus can be attributed to their essential oils
(Asili et al 2009; Maggi et al. 2009).
Several species of thyme genus (Lamiaceae) grow wild in Algeria, among which Thymus pallescens de No which
is endemic to Algeria. The plants of Thymus genus are among the most popular plants throughout the world, commonly
used as herbal teas, flavoring agents, aromatic, and medicinal plants (Stahl-Biskup and Saez, 2002).
The purpose of the study was the analysis of the essential oils from F. vesceritensis and Thymus pallescens and
the assessment of their insecticidal activities against Sitophilus oryzae.

MATERIALS AND METHODS

Plant Material
Thymus pallescens plant was collected in May 2013 from Lakhdaria at 80 Km east of Algiers while Ferula
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vesceritensis was in March 2012 from Dhaya Bendahoua in Ghardaia region (650 Km south of Algiers).
Essential Oil Extraction
The essential oils were isolated from the entire plants dried leaves by hydrodistillation for 3 h using a Clevengertype apparatus according to the European Pharmacopoeia method. The essential oils were stored at - 10 C in the dark until
use.
GC Analysis
The chemical compositions of the essential oils of T. pallescens and F. vesceritensis were examined by GC on a
Hewlett-Packard 6890 GC-FID system using a fused-silica-capillary column with apolar stationary phase HP5MS.
The various parameters fixed for HP5MS column are: 30 m x 0.25 mm x 0.25 m film thickness column; temp. prog, 60C
for 8 min, then 2/min to 280C for 15 min; nitrogen was used as carrier gas at a flow rate 0.5 ml/min in the split mode
1:20, with injection volume 0.2 L. Injection was performed at 250C and flame ionization detection at 320C.
GC-MS Analysis
The essential oils were analyzed by gas chromatography coupled to mass spectrometry (GCMS) (Hewlett-Packard
computerized system comprising a 6890 gas chromatograph coupled to a 5973A mass spectrometer) using the nonpolar
column HP5MS (30 m x 0.25 mm x 0.25 m film thickness). We got hold of GC-MS spectra using the following
conditions: He as carrier gas at flow rate of 0.5 ml /min; split mode (1:20); 0.2 l without dilution as injected volume;
250C as injection temperature. The oven temperature program is described in section Gas chromatography. We used an
ionization mode with electronic impact at 70 eV over a scan range of 30-550 atomic mass units.
Identification of Compounds
The constituents of the oils were identified by comparison of their GC Kovats retention indices (R.I.), determined
with reference to an homologous series of C8-C21 n-alkanes and with those of available authentic standards and with those
of bibliography (Babushok et al, 2011). Then, we tried to confirm such identification by comparing their mass spectral
fragmentation patterns with those stored in the MS database (NIST 2005 and Wiley 7N libraries) and with mass spectra
literature data (Adams, 2007). Components relative concentrations were obtained directly from GC peak areas obtained
with GC-FID.
Insecticidal Activity
Insect Cultures
Colonies of the rice weevil beetle, S. orizae, were maintained in the laboratory without exposure to any
insecticide. They were reared in glass containers (16 cm diameter22 cm height) covered by a fine mesh cloth for
ventilation. Each container contained a mixture of wheat flour and wheat germ (13:6 w/w). The cultures were maintained
in a growth oven at 30 1 C, with a relative humidity (RH) of 705 %. Only adults were used for the test.
Contact Toxicity
Four stocks solutions of each essential oil were prepared with the concentrations of 4, 8, 16 and 32 L/mL in
acetone for F. vesceritensis oil and 6, 12, 24 and 48 L/mL in acetone for T. pallescens oil. 1 mL of each solution or
acetone (control) was uniformly spilt on filter paper (8.0 cm diameter) and placed into a Petri dish (9 cm diameter, 1.5 cm)
which is allowed open to evaporate the solvent at room temperature. After evaporation of solvent, twenty unsexed adults of
Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

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Chemical Composition and Insecticidal Activities of Essential Oils of Two Algerian

Endemic Plants: Ferula vesceritensis Coss. Et Dur. and Thymus pallescens De Noe

S. oryzae were introduced (three replicates for essential oils and control). Mortality was recorded daily during 6 days.
LD50 and LD90 which represent the concentrations causing the death of 50% and 90%, respectively, of individuals of the
same dish after 24 hours of exposure to oil were determined by probit analysis test using the method of Finney (1971).
Fumigant Toxicity
A cotton bud of 2 cm in length was impregnated with 32 L of pure essential oil (corresponding to 0.466 l /cm3)
and glued onto the cap of a glass vial (volume 68.64cm3). The cap was screwed tightly onto the vial containing 20 unsexed
insects. Three replicates were set up for each Thymus oil and control (without oil). The mortality was recorded each 24 h
during 6 days. LT50 which represent the times causing the death of 50% of individuals of the same glass vial were
determined. In both the contact and fumigant toxicity tests, data were transformed using the Abott (1925) formula, given
by:
Mc (%) = 100 (M-Mt)/ (100 - Mt)
Where, Mc is the corrected insect mortality, M the insect mortality in the treated insect population and Mt the
insect mortality in the control.
Data Analysis
Probit analysis (Finney 1971) was conducted to estimate lethal doses (LD50 and LD90) and lethal times (LT50
and LT90) with their 95 % confidence interval by SPSS 12.0 Statistical Software; LD and LT values were considered
significantly different when their respective 95 % confidence interval did not overlap.

RESULTS AND DISCUSSIONS

Chemical Composition
The chromatographic analysis lead to the identification of 54 and 35 components in the essential oils of F.
vesceritensis and T. pallescens, respectively (Table 1). Ferula oil is characterized by shyobunol (18.1%), -cadinene
(14.2%), aristolene (6.2%), t-cadinol (8.1%) and -cadinol (9.3%) as main constituents. This result is different with that
previously reported for another sample from the same region which showed viridiflorol (13.4 %), -cadinene (10.1 %) and
farnesol (8.1 %) as major constituents (Benchabane et al, 2012). Dehak-Oughlissi et al, (2014) found for a sample collected
from Sebseb that aristolone (14.1%) and aristolne (9.6 %) were the main compounds. These results suggest that this
Ferula species is characterized by a great variability in its chemical composition. T. pallescens essential oil showed that
-terpinene (%), -terpineol (%), thymol (%) and carvacrol (%) are the main prominent constituents. This result is in
accordance whith previous data on the essential composition of this Thymus species (Hazzit et al, 2009).
Insecticidal Activity
Mortality of S. oryzae by the oils of F. vesceritensis and T. pallescens in contact test was dose dependent after 6
days (144 h) of topic application at 0.754 l/cm2 for T. pallescens (Figure 1) and 0.503 l/cm2 for F. vesceritensis
(figure 2). These results demonstrate that the efficiency of the oils was directly related to concentration. Oil from T.
pallescens was the most efficient, causing 90% mortality at the lower lethal concentration (LD 90=17.15 l/ml ; 0.27
l/cm2) (Table 2).

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Table 1: Percentage Composition of Ferula vesceritensis and Thymus pallescens Essential Oils
N
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53

Compound

-Thujene
-Pinene
Camphene
Sabinene
-Pinene
1-Octen-3-ol
-Myrcene
-Phellandrene
-Terpinene
p-Cymene
Limonene
- Phellandrene
1,8-Cineole
(E)--Ocimene
-Terpinene
cis-Sabinene hydrate
Terpinolene
Linalool
Isoamyl isovalerate
n-Amylisovalerate
Borneol
4-Terpineol
-Terpineol
Fenchyl acetate
Thymol methyl ether
Bornyl acetate
Thymol
Carvacrol
-Cubebene
-Copaene
-Cubebene
-Elemene
-Gurjunene
Aristolene
-Gurjunene
-Caryphyllene
Aromadendrene
-Bergamotene
-Humulene
trans--Farnesene
-Muurolene
Germacrene D
-Curcumene
epi-Bicyclosesquphellandrene
-Selinene
Valencene
Bicyclogermacrene
-Muurolene
-Bisabolene
-Cadinene
-Cadinene
-Sesquiphellandrene
Cadina-1,4-diene

Impact Factor (JCC): 4.3594

KI

925
935
948
972
976
982
992
1005
1017
1026
1029
1030
1033
1048
1061
1069
1087
1104
1106
1108
1167
1177
1194
1227
1235
1285
1302
1318
1346
1372
1386
1391
1403
1413
1426
1415
1437
1435
1450
1453
1472
1477
1479
1485
1487
1491
1493
1495
1505
1511
1518
1525
1526

Essential Oils
F. vesceritensis T. pallescens
0.6
0.6
5.6
1.6
0.2
0.1
0.2
0.5
1.8
0.4
0.1
0.4
0.4
2.0
0.5
1.4
0.1
5.1
2.2
0.3
0.1
0.3
0.2
0.2
6.5
0.2
0.1
0.2
2.0
0.6
0.2
0.1
0.1
7.0
5.4
3.4
0.3
13.5
40.2
0.1
0.2
0.1
0.2
0.1
6.2
0.9
0.1
1.6
0.1
0.1
0.2
0.2
0.2
0.1
0.4
0.6
0.4
0.1
0.2
0.3
0.7
0.1
1.2
0.1
3.4
1.2
14.2
0.3
0.1
0.3
-

Identification
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS
RI, MS, E
RI, MS
RI, MS, E
RI, MS, E
RI, MS
RI, MS
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS, E
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS, E
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS

Index Copernicus Value (ICV): 3.0

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Chemical Composition and Insecticidal Activities of Essential Oils of Two Algerian

Endemic Plants: Ferula vesceritensis Coss. Et Dur. and Thymus pallescens De Noe

54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71

-Cadinene
-Copaen-11-ol
-Bisabolene
Elemol
Elemicin
Nerolidol
Germacrene D- 4-ol
Spathulenol
Caryophyllene oxide
Viridiflorol
-Eudesmol
t. Cadinol
t.Muurolol
-Eudesmol
-Cadinol
Cadalene
Shyobunol
Phytol
Total (%)
Monoterpene hydrocarbons
Oxygenated monoterpenes
Sesquiterpene hydrocarbons
Oxygenated Sesquiterpenes
Others

Table 1: Contd.,
1534
0.3
1537
0.4
1540
1549
2.3
1554
0.3
1563
0.2
1568
4.8
1570
0.5
1573
0.4
1594
0.1
1618
0.3
1635
8.1
1636
1644
0.5
1650
9.3
1672
0.4
1695
18.1
2121
0.1
92.2
11.7
6.0
28.8
45.2
0.5

0.3
0.2
0.1
93
19.2
66.4
6.7
0.3
0.4

Components listed in order of their elution on the non polar column HP5MS ;

C8-C21 n-alkanes;

RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS, E
RI, MS
RI, MS
RI, MS, E
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS
RI, MS, E

retention indices relative to

Identification: E, comparison with authentic compounds (etalons); MS, comparison of mass spectra

with MS libraries; RI, comparison of retention index with bibliography

T. pallescens oil was also the most toxic in fumigant test (Figure 3) with a lethal time lower than that of Ferula oil
(LT=100.15 h vs LT=198.85 h) (Table 3). The potent insecticidal activity of T. pallescens may be related to its richness in
carvacrol which may take the most important part. Indeed, carvacrol has broad insecticidal activity and acts as a fumigant
against agricultural and stored-product insects (Ahn et al.1998 ; Isman, 2000).
In general, the complexity of the chemical composition of most of the volatile oils gives them low specificity
(Bakkali et al, 2008), because biological activity is not assigned to a single mechanism of action, since the wide variety of
chemical groups allows for multiple targets in the cell (Burt, 2004).

Figure 1: Mortality (%) of S. oryzae by Thymus pallescens with

Various Doses and Various Times of Exposure in Contact Test
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Table 2: Lethal Doses of Essential Oils of T. pallescens and F. vesceritensis in Contact Test
Lethal Doses
LD50
LD90

Ferula vesceritensis
16.4 l/ml
0.26 l/cm2
53.45 l/ml
0.84 l/cm2

Thymus pallescens
8.76
0.14 l/cm2
17.15 l/ml
0.27 l/cm2

Figure 2: Mortality (%) of S. oryzae by Ferula vesceritensis Essential Ol

in Contact Test for Various Concentrations and Various Times
Table 3: Lethal times of T. pallescens and F. vesceritensis Essential Oils
Lethal Time (Hour)

Ferula vesceritensis

Thymus pallescens

LT50

107.84 1.3

73.23 0.30

LT90

198.85 0.9

100.15 0.21

Figure 3: Mortality (%) of S. oryzae By Essential Oils of T. pallescens and F. vesceritensis in Fumigant Test

CONCLUSIONS
The potential use of essential oils as fumigants to control stored grain pests has been the subject of several studies.
The results obtained in this work demonstrate that the essential oils tested can be used to control stored grain pests and to
support further studies. We also found variation in toxicity depending upon the application method of the essential oils.
The essential oil of T. pallescens can be used as effective control for stored grain pests, and they may be applied either by
contact or by fumigation. The essential oil of Ferula vesceritensis here tested for the first time for its insecticidal activity
showed appreciable efficiency.

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

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Chemical Composition and Insecticidal Activities of Essential Oils of Two Algerian

Endemic Plants: Ferula vesceritensis Coss. Et Dur. and Thymus pallescens De Noe

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