2018

NPC Natural Product Communications Vol. 13

No. 7
Chemical Composition and Antifungal Activity of Lavender 895 - 898
(Lavandula stoechas) Oil
Mehmet Musa Özcana, Mira Starovicb*, Goran Aleksicb, Gilles Figueredoc, Fahad Al Juhaimid and
Jean-Claude Chalchatc
a
Department of Food Engineering, Faculty of Agriculture, University of Selcuk,42031 Konya, Turkey
b
Institute for Plant Protection and Environment, Belgrade, Serbia
c
Laboratoire de Chimie des Huiles Essentielles Universite Blaise Pascal de Clermont,63177 Aubiere Cedex, France
d
Department of Food Science & Nutrition, College of Food and Agricultural Sciences, King Saud University,
Riyadh-Saudi Arabia
miragavranstarovic@gmail.com

Received: November 15th, 2016; Accepted: May 15th, 2018

The essential oil of Lavandula stoechas was examined by GC and GC-MS. Discs (5 mmi.d.) of the tested fungi (Alternaria alternata, Fusarium oxysporum
and Botritys cinerea) were inoculated separately onto each assay plate and incubated at 25oC for 7 days. The oil yield of dried parts (v/dw) obtained by hydro-
distillation was 2.9%. Thirty-two compounds representing 98.3% of the essential oil were determined. Linalool (49.9%), linalyl acetate (14.4%), lavandulyl
acetate (5.7%), α-terpineol (5.6%), terpinene-4-ol (5.1%), lavandulol (3.7%), (E)-β-ocimene (2.6%) and (Z)-β-ocimene (2.4%) were identified as the main
constituents of the oil. In addition, both doses of the lavender oil showed varying levels of inhibitory effects on the mycelial growth of tested fungi used in the
experiment. The results demonstrated the strongest effect on B.cinerea, followed by A.alternata and F.oxysporum. The inhibitory effect is probably dependent
on the concentration of essential oils.

Keywords: Lavender, Essential oil, Chemical composition, Linalool, Antifungal effect.

Lavender, Lavandula stoechas, belong to the family Lamiaceae and
is a perennial herbaceous plant in a number of regions in Turkey. It
is widely distributed in subtropical and moderate regions [1].
Lavender is native plant to the Mediterranean, the Arabian
Peninsula, Russia and Africa. It has been used cosmetically and
medicinally throughout history. In modern times, lavender is
cultivated around the world and the fragrant oils from the flowers
are used in aromatherapy, baking, candles, cosmetics, detergents,
jellies, massage oils, perfumes, powders, shampoo, soap and tea.
Infusion of the aerial parts of Lavandula species are used as an
appetite stimulant as well as for asthma, bronchitis, colic, common
cold and fatigue healing in folk medicine [2]. The chemical
composition of Lavandula oils he been investigated previously [3].
In addition to their antimicrobial properties, a number of herbs and
spices have been used since ancient times for food preservation [4].
Recently, there has been a considerable emphasis on studies
involving derivatives of spices and their constituents’ growth
inhibition of microorganisms [5]. The aim of this study is to
determine the chemical composition and antifungal effects of oil
obtained from Lavandula stoechas flowers.
The flowers of lavender (Lavandula stoechas) were collected in
Konya (Çumra) province in Turkey, and identified by the
Laboratory of Systematic Botany, University of Selçuk. A specimen
(GMB-2007) has been deposited in the Department of Food
Engineering of the Selçuk University.
The fungi used for this assay were obtained from the collections of
Plant Protection Department, Faculty of Agriculture, Selcuk
University. In order to enable microbial growth, potato dextrose
agar (PDA) was used. Czapex-Dox Agar was used for the
determination of fungi toxicity.
The aerial parts of L. stoechas were extracted by using a Clevenger
type apparatus for 3 h. The obtained essential oil (2.9%; v/w) was
dried over anhydrous sodium sulfate, and stored at -18oC in
readiness for GC/MS analysis.
For identification purposes, the essential oils were analyzed on an
Agilent gas chromatograph Model 7890, coupled to an Agilent MS
model 5975C, equipped with a DB5 MS column (20 m x 0.18 mm x
0.18 µm). The temperature increased from 50°C (3.2 min) to 280°C
at 8°C/min, with a 5 min hold. Helium was used as carrier gas (1.0
mL/min); injected in split mode (1:120) with the injector and
source temperatures of 280 and 250°C respectively. The MS was
working in electron impact mode at 70eV; mass spectra data were
acquired in the scan mode in m/z range from 33-450. Identification
of the constituents was based on the comparison of their linear
indices relative to the series of n-hydrocarbons, and matching them
against the commercially available spectra (Wiley & NIST).
In order to calculate the correct quantities, the essential oils were
analyzed using an Agilent gas chromatograph Model 6890,
equipped with a DB5 MS column (20m x 0.18mm x 0.18µm),
programmed from 50°C (3.2 min) to 280°C at 10°C/min with a 5
min hold. Hydrogen was used as a carrier gas (1.0 mL/min) injected
in a split mode (1:60); injector and detector temperatures were 280
and 300°C respectively. The essential oil was diluted in hexane in
the ratio 1:30. The relative amounts of individual components were
calculated based on the GC peak areas [6].
The inhibitory effects of essential oils of flowers of lavender were
determined against A. alternata, F. oxysporum and B. cinerea
growth using Czapek-Dox agar medium only. Discs of the tested
fungi (5 mmi.d.), cut from the periphery of a 7 day-old cultures,
were inoculated separately onto each assay plate and incubated at
25°C for 7 days.
896 Natural Product Communications Vol. 13 (7) 2018 Özcan et al.

Table 1: Chemical composition of lavender (Lavandula stoechas) oila. Table 2: The effect of lavender (Lavandula stoechas L) oil concentration on the
RT KI Constituents % mycelial growth (mm) of some fungi per day.
8.39 923 α-thujene 0.1
8.64 930 α-pinene 0.1 Incubation Concentrations F. oxysporum B. cinerea A. alternaria
9.21 945 Camphene 0.3 Days (ppm)
9.35 954 Butyl isobutyrate tr 3 10 16 - 14
9.69 961 Benzaldehyde tr 40 -* - -
10.06 968 Sabinene tr control 24 7 25
10.21 972 β-pinene tr 4 10 26 - 24
10.39 977 Octen-3-ol tr 40 - - -
10.54 981 Octan-3-one 0.7 control 34 12 36
10.72 986 Myrcene 1.4 5 10 32 - 31
10.92 992 Butyl butyrate tr 40 - - -
11.05 995 Octanol tr
control 38 15 42
11.24 1000 α-phellandrene tr
6 10 38 - 38
11.34 1003 α-3-carene tr
11.51 1008 Butyl isobutyrate 0.4 40 11 - -
11.63 1012 α-terpinene 0.3 control 46 16 52
11.73 1015 o-Cymene tr 7 10 45 - 44
11.90 1020 p-Cymene 0.1 40 15 - -
12.06 1025 Limonene 0.6 control 48 18 60
12.15 1027 Eucalyptol tr 8 10 47 - 55
12.38 1034 (Z)-β-ocimene 2.4 40 20 - -
12.73 1045 (E)-β-ocimene 2.6 control 56 19 66
13.07 1055 γ-terpinene 0.2 9 10 54 - 61
13.50 1068 (Z)-Linalool oxide 0.1 40 25 - -
13.73 1075 1-octanol tr control 57 21 68
13.99 1082 Terpinolene 0.3 10 10 60 - 70
14.22 1089 3-methyl-2-(2-methylbutenyl) furan tr
40 30 - 8
14.84 1108 Linalool 49.9
control 65 24 74
14.88 1110 1-Octen-3-yl acetate tr
11 10 69 - 78
14.96 1112 (2E)-Hepetenyl acetate tr
15.19 1119 3-Octyl acetate 0.1 40 37 - 13
15.35 1125 (Z)-Menth-2-en-1-ol tr control 74 27 80
15.43 1127 Alloocimene tr 12 10 75 - 83
15.59 1132 Butyl tiglate tr 40 40 - 17
15.87 1141 (E)-menth-2-en-1-ol tr control 76 32 84
16.01 1146 Camphor tr 13 10 76 - 85
16.15 1150 Nerol oxide tr 40 54 - 21
16.62 1165 Lavandulol 3.7 control 77 38 87
16.83 1172 Borneol Tr 14 10 78 - 86
17.13 1181 Terpinene-4-ol 5.1 40 58 - 58
17.26 1185 Cryptone tr control 79 40 88
17.42 1190 Hexyl butanoate tr 15 10 79 - 87
17.59 1196 α-terpineol 5.6 40 61 - 63
17.93 1208 2,6-dimethyl-3,5,7-octatriene-2-ol tr control 80 47 89
18.42 1225 Nerol 0.9
16 10 81 - 88
18.89 1242 Cuminaldehyde tr
19.20 1253 Linalyl acetate 14.4 40 63 - 67
19.25 1255 Geraniol tr control 82 54 90
19.61 1268 Geranial tr 17 10 82 - 89
20.12 1286 Lavandulyle acetate 5.7 40 66 - 74
20.35 1295 p-cymene-7-ol tr control 83 61 90
21.29 1329 Hexyl tiglate 0.1 18 10 84 - 90
21.76 1346 α-Terpinyl acetate tr 40 71 - 77
22.09 1358 Neryl acetate 1.1 control 86 69 90
22.63 1378 Geranyl acetate 0.9 19 10 85 - 90
22.79 1384 Hexyl hexanoate tr 40 73 - 80
23.54 1413 α-(Z)-bergomotene tr control 87 77 90
23.73 1420 β-caryophyllene 0.3 20 10 86 - 90
24.04 1432 (E)-α-bergamotene tr
40 75 - 84
24.32 1443 β-sesquifenchene tr
control 88 83 90
24.53 1452 (E)-β-farnesene 0.6
24.62 1455 α-humulene tr 21 10 89 - 90
25.26 1481 Germacrene D 0.1 40 79 - 87
25.81 1503 Germacrene A tr control 90 86 90
26.04 1512 γ-Cadinene 0.1 22 10 90 - 90
27.71 1581 Caryophyllene oxide tr 40 84 - 90
28.38 1612 Cyclohexylcetone 0.3 control 90 90 90
28.47 1616 1,10-di-epi cubenol tr *100% inhibition
29.08 1643 epi-α-Cadinol 0.2
Total 98.3
a
Compounds listed in the order of elution from a HP-5MS column; b Each compound reported in order of their elution from HP-5MS column. Thirty-two
is a mean of two values. c trace <0.08 components were identified, accounting for 98.3%. The lavender oil
contained mainly monoterpenes, oxygenated monoterpenoids and
Also, both doses (10 and 40 ppm) of the lavender oil exerted sesquiterpenoid hydrocarbons. Linalool (49.9%), linalyl acetate
varying levels of the inhibitory effect on the mycelial growth of the (14.4%), lavandulyle acetate (5.7%), α-terpineol (5.6%), terpinene-
fungi used in this experiment. Each essential oil was added onto the 4-ol (5.1%), lavandulol (3.7%), (E)-β-ocimene (2.6%) and (Z)-β-
sterile disc paper (10 mmi.d.), and put into Petri dishes. The colony ocimene (2.4%) were determined as the major constituents of the
diameter was measured and the percentage mycelial inhibition was oil.
calculated using the formulae [5d]:
Inhibitory effect: The inhibitory effect of the flower oil of lavender
I=C-T/C x 100, where I is inhibition (%), C is the colony diameter (Lavandula stoechas) against tested fungi are presented in Table 2.
of mycelium from a control Petri plate (mm) and T is the colony
diameter of the mycelium from a control Petri plate (mm). I was B. cinerea was the most sensitive fungus at both concentrations.
calculated as an average of the three repeats of each treatment. The oil of 40 ppm level of lavender oil showed a strong antifungal
activity against mycelial growth of A. alternata compared to the
Chemical composition: The chemical composition of the lavender control until 7 days of incubation. 40 ppm level of oil showed a
(Lavandula stoechas) oil is given in Table 1. Constituents were stronger percentage inhibition than those of 10 ppm level of oil on
Essential oil of Lavandula stoechas
Figure 1: Inhibitory effect of lavender oil on some fungi used in experiment (%).
the growth of F. oxysporium. Each dose of lavender oil exhibited a
different fungistatic activity against F .oxysporum. In addition,
lavander oil showed weak activity after 3 days of incubation on the
same fungi. The most affected fungi from lavender oil was B.
cinerea, followed by A. alternata and F. oxysporum. A moderate
inhibition effect against mycelial growth of F. oxysporum was
provided by 40 ppm dose of oil (Table 2). Consequently, the 40
ppm dose of oil caused a complete inhibition (100%) against
mycelial growth of B. cinerea. The analysis showed that lavender
oil exhibited fungistatic activity at different levels depending on the
doses (Figure 1).
The volatile fraction of the Lavandula stoechas oil differs from
many lavandula species growing in the Mediterranean region. The
oil of L. stoaches contains mainly monoterpenoid alcohols such as
linalool, α-terpineol, terpinene-4-ol as major constituents. The main
constituent of L. stoechas ssp. atlantica oil was camphor (39%)
followed by fenchone (9%), while for L. stoechas ssp. stoechas oil
the main constituents was fenchone (30%) followed by camphor
(18%) [3e].
The analyses of L. stoechas ssp. stoechas oil from Crete, Greece
yielded two chemotypes: a fenchone/camphor-type and a 1,8-
cineole/fenchone-type [3c]. The main components of L. stoechas oil
were linalyl acetate+linalool (50.0-60.0%) [7]. In other studies, the
major components of L. stoechas ssp. Stoechas oil were camphor
(23.0%) and fenchone (11.0%) [7]. The main components of L.
stricta oil was carvone 43.0%, in L. pubescene was carvacrol 69.6%
and in L. dentata were camphor 37.2% and fenchone 21.3% [8].
Lavender was determined as Lavandula latifolia which grows wild
in Croatia but is very rare. Wild growing plants showed the
following oil characteristics: lavender yielded 5.5% essential oil
from dried flowers and the main components were linalool (30.8%),
borneol (18.4%) and 1,8-cineole (17.0%), while the total esters were
only 6.7%, from which 5.6% was linalyl-acetate. Commercial
samples of lavender oil showed the following results: imported
lavender, declared as L. angustifolia, had very low linalyl-acetate
(less than 6.0%) and the major components were 1,8-cineole (16.0
Natural Product Communications Vol. 13 (7) 2018 897
%), linalool (40.0%), borneol (10.0%) and camphor (4.0%), while
the Adriatic lavender (lavandine) had 8.0% linalyl-acetate, with
major components being linalool (47.0%), 1,8-cineole (8.5%),
borneol (8.5%), terpinen-4-ol (7.5%) and only 2.0% camphor.
Earlier results have already indicated that lavender oil inhibits
mycelia growth of the plant fungal pathogens, such as F. oxysporum
and Alternaria alternata, as well as Phomopsis sp. F. solani, F.
sporotrichioides, F. verticilioides [9a], and Verticillium spp. [9].
The results obtained from the cited authors state a minimal
inhibitory concentration (MIC) between 1 to 3 mg/mL. Our results
are consistent with an earlier study [10] where Lavandula stoechas
oil showed high antifungal effect on the Botrytis cinerea (IC50 - 1.64
ppm). Some researchers reported that there is a relationship between
the chemical structure of the most abundant compounds in the tested
essential oils and their antimicrobial activity [3c,e,5d,7]. The
observed differences may be due to a different harvest date and
species factors that can influence the oil composition. The oil of
Lavandula stoechasis is defined by the presence of a relatively high
concentration of linalool. The lavender oil used in this work
exhibited partial inhibitory effect on tested fungi. This could lead to
a potential use of lavender oil as antifungal preservatives in food,
especially as they are largely nontoxic and easily biodegradable.
Botryris cinerea is a very dangerous pathogen known to damage
over 230 plant species worldwide [11]. The ability of B. cinerea to
quickly develop tolerance and resistance to new fungicides can
compromise the long term fungicide use. In this context, a lavender
oil’s fungistatic effect on the most serious pathogens, as
demonstrated in this research, shows a promising future
development.
Acknowledgments - The authors extend their appreciation to the
International Scientific Partnership Program ISPP at King Saud
University for funding this research work through ISPP# 0015.
Author would like to thank Mrs Svetlana Bajic-Raymond from
Cotham School, England, for valuable comments and suggesting the
text corrections.
898 Natural Product Communications Vol. 13 (7) 2018 Özcan et al.

References
[1] (a) Koca F, Alan S. (2002) Morphologic and anatomic properties of Lavandula stoechas L. ssp. stoechas cultivated in Turkey.(Türkiye’deyetişen
Lavandula stoechas L. ssp. stoechas’ın morfolojik ve anatomic özellikleri). 14.Bitkiselİlaç Hammaddeleri Toplantısı,19-31,Mayıs,
Eskişehir,Türkiye; (b) Akhondzadeh S, Kashani L, Fatouhi A. (2003) Comparison of Lavandula angustifolia Mill. Tincture and imipramine in the
treatment of mild to moderate depression: a double-blind, randomized trial. Progress in Neuro-psychopharmacology & Biological Phychiatry, 27,
123-127.
[2] (a) Baytop T. (1984) Treatment with plants in Turkey. İstanbul University, 3255, İstanbul, Turkey; (b) Dunn C, Sleep J, Collett D. (1995) Sensing
an improvement: an experimental study to evaluate the use of aromatherapy, massage and periods of rest in an intensive care unit. Journal of
Advanced Nursing, 21, 34-40; (c) Diego MA, Jones NA, Field T, Hernandez-reif M, Schanberg S, Kuhn C, Galamaga M, McAdam V, Galamaga
R.. (1998) Aromatherapy positively affects mood, EEG patterns of alertness and math computations. International Journal of Neuroscience, 96,
217-224; (d) Kokkalou E. (1988) The constituents of the essential oil from Lavandula stoechas growing wild in Greece. Planta Medica, 47, 58-59;
(e) Gray SG, Clair AA. (2002) Influence of aromatherapy on medication administration to residential care residents with dementia and behavioral
challenges. American Journal of Alzheimer's Disease and Other Dementias, 17, 169-174; (f) Büyükokuroğlu ME, Gepdiremen A, Hacımüftüoğlu
A, Oktay M. (2003) The effects of aqueous extract of Lavandula angustifolia flowers in glutamate-induced neurotoxicity of cerebellar cell culture
of rat pups. Journal of Ethnopharmacology, 84, 91-94; (g) Graham PH, Browne L,Cox H, Graham J. (2003) Inhalation aromatherapy during
radiotherapy: results of a placebo-controlled double-blind randomized trial. Journal of Clinical Oncology, 21, 2372-2376.
[3] (a) Tanker T, Şarer E, Başan V. (1977) Pharmacognozic researches of Lavandula stoechas L essential oil Lavandula stoechas L bitkisini
nuçucuyağıüzerin de farmakognozi karaştırmalar. Journal of Faculty of Pharmacy of Ankara University, 7, 61-66; (b) Valentin IG, Arnold N,
Bellomaria B. (1993) Etude chimique comparative des huil esessentielles de quatre populations de Lavandula stoechas L. subsp. Stoechas
spontanees de Chypre. Plantes medicinales et phytotherapie, 224, 289-299; (c) Skoula M, Abidi C, Kokkalou E. (1996) Essential oil variation of
Lavandula stoechas L.ssp. stoechas growing wild in Crete (Greece). Biochemical Systematics and Ecology, 24, 255-260; (d) Partl A, Brekalo M,
Blajevic N, Stilinavic B. (2002) Comparisan of essential oil composition and antibacterial activity between some Crauti an wild growing plants and
commereially available oils. 33rd International Symposium on Essential Oils, 4-7 September, Lisboa-Portugal, 152; (e) Zrira S, Benjilali B. (2003)
The constituents of the oils of Lavandula stoechas L. ssp. atlantica Br.-Bl. And L. stoechas ssp. stoechas from Morocco. Journal of Essential Oil
Research, 15, 68-69; (f) Kırmızıbekmez H, Demirci B, Yeşilada E, Başerb HC, and Demirci F. (2009) Chemical Composition and Antimicrobial
Activity of the Essential Oils of Lavandula stoechas L. ssp. stoechas Growing Wild in Turkey, Natural Product Communications, 4, 1001-1006; (g)
Tschiggerl C, Bucar F. (2010) Volatile fraction of lavender and bitter fennel infusion extracts. Natural Product Communications, 5, 1431-1436.
[4] (a) Zaika L. (1988) Spices and herbs: their antimicrobial activity and its determination, Journal of Food Safety,9, 97-118; (b) Özcan M, Erkmen O.
(2001) Antimicrobial activity of the essential oils of Turkish plant spices. European Food Research and Technology, 212, 658-660.
[5] (a) Baratta TM, Damien Dorman HH, Deans SG, Figueiredo CA, Barroso GJ, Ruberto G. (1998) Antimicrobial and antioxidant properties of some
commercial oils. Flavor and Fragrance Journal, 13, 235-244; (b) Bullerman B, Lieu FY, Seier SA. (1977) Inhibiton of growth and aflatoxin
production by cinnamon, and clove oils, cinnamic aldehyde and eugenol. Journal of Food Science, 42, 1107-1109; (c) Özcan M. (1998) Inhibitory
effects of spice extracts on the growth of Aspergillus parasiticus NRRL 2999 strain. Zeitschrift fuer Lebensmittel-Untersuchung und-Forschung A –
Food Research and Technology, 207: 253-255; (d) Deans SG, Svoboda KP. (1990) The antimicrobial properties of marjoram (Origanum majorana
L.) volatile oil. Flavor and Fragrance Journal, 5, 187-190.
[6] Adams R. (2001) Essential oil components by Quadrupole GC/MS. Allured Publishing Corp., Carol Stream, IL, USA.
[7] TankerM, Tanker N, Şarer E, Atasü E, Şener B,Kurucu S, Meriçli F. (1993) Results of certain investigations on the volatile oil containing plants of
Turkey. Essental oils Perfumery and Flavours, Başer KHC, Güler N.(eds), 16-32,İstanbul .
[8] Albar AH, Youssef MH. (2002) Chemical constituents on essential oil in three species of Lavandula. 2nd Conference on Medicinal & Aromatic
plants of Southeast European Countries, 29 September-30 October, Chalkidiki-Greece: 94.
[9] (a) Stević T, Berić T, Šavikin K, Soković M, Gođevac D, Dimkić I, Stanković S. (2014) Antifungal activity of selected essential oils against fungi
isolated from medicinal plant. Industrial Crops and Products, 55, 116-122; (b) Soković MD, Van Griensven LJLD. (2006) Antimicrobial activity of
essential oils and their components against the three major pathogens of the cultivated button mushroom, Agaricus bisporus. The European Journal
of Plant Pathology, 116, 211-224.
[10] Behnam S, Farzaneh M, Ahmadzadeh M, Tehrani AS. (2006) Composition and antifungal activity of essential oils of Mentha piperita and
Lavendula angustifolia on post-harvest phytopathogens. Communications in Agricultural and Applied Biological Sciences, 71, 1321-1326.
[11] Elad Y. (2007) Responses of plants to infection by Botrytis cinerea and novel means involved in reducing their susceptibility to infection.
Biological Reviews, 72, 381-422.