Food Chemistry 330 (2020) 127268

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Review

Essential oils: A promising eco-friendly food preservative T

⁎
Hanen Falleh , Mariem Ben Jemaa, Mariem Saada, Riadh Ksouri
Laboratory of Aromatic and Medicinal Plants, Centre of Biotechnology of Borj Cedria, BP 901 Hammam-lif 2050, Tunisia

A R T I C LE I N FO A B S T R A C T

Keywords: Essential oils (EOs) are natural, volatile and aromatic liquids extracted from special plants. EOs are complex
Antioxidant activity mixture of secondary metabolites (terpenes, phenolic compounds, alcohol). EOs possess a wide range of bio-
Antimicrobial activity logical activities including antioxidant, antimicrobial and anti-inflammatory ones. Particularly, EOs exhibit
Food preservation pronounced antibacterial and food preservative properties that represent a real potential for the food industry.
Safe preservative
Numerous EOs have the potential to be used as a food preservative for meat and meat products, vegetables and
Terpenes
fruits as well as for dairy products. The main obstacles for using EOs as food preservatives are their safety limits,
marked organoleptic effects and possible contamination by chemical products such as pesticides. This review
aims to provide an overview of current knowledge about EOs food preservative properties with special emphasis
on their antibacterial activities and to support their uses as natural, eco-friendly, safe and easily biodegradable
agents for food preservation.

1. Introduction medicinal plants (Falleh et al., 2019). In general, EOs correspond to a

It is undeniable that food is essential to life. Therefore food safety is
a crucial matter for consumers as well as for food industries. Indeed, the
World Health Organization (WHO) estimated that 1 in 10 people fall ill
every year from eating contaminated food and 420 000 die as a result
(World Health Organization, 2015). This important incidence of food-
borne threat associated with the new social and economic implications
leads to an urgent need for safer food through the development of new
and nontoxic preservative agents with important antimicrobial and
antioxidant properties. With this respect, the actual synthetic chemi-
cals, commonly used to control pathogen strain, raise serious pre-
occupations related to human health (Prakash, Kedia, Mishra, & Dubey,
2015; Falleh et al., 2019). Gutiérrez-del-Río and co-workers (2018)
reported that some synthetic antimicrobials, approved by regulatory
agencies and used as food preservatives, were a health menace for the
consumer. For instance, sulfites (a group of sulfur-based compounds
commercially used as a food preservative) have been associated with
some anti-nutritional consequences such as the degradation of thiamine
or vitamin B1 in food (Gutiérrez-del-Río, Fernández, & Lombó, 2018).
As a natural consequence to that, there is an increasing trend of the use
of natural and safer preservatives making food have a natural or
“green/organic” image. Particular attention has focused on the appli-
cations of plant essential oils (EOs) as food preservatives (Burt, 2004;
Dhifi, Bellili, Jazi, Bahloul, & Mnif, 2016).
EOs are secondary metabolites synthesized by aromatic and
very small fraction of plant total composition, approximately less than
5% of the vegetable dry matter (Valderrama & Ruiz, 2018). Essential
oils are volatile, usually liquid and colorless at ambient temperature.
They are poorly soluble in water but highly soluble in alcohol, organic
solvents, and fixed oils. They have a variable refractive index and a very
high optical activity and sometimes a distinctive taste (Adelakun,
Oyelade, & Olanipekun, 2016; Calo, Crandall, O’Bryan, & Ricke, 2014).
Moreover, essential oils exhibit a very characteristic odor and are
therefore responsible for the specific scents that aromatic plants emit
(Dhifi et al., 2016; Valderrama & Ruiz, 2018). Chemically, EOs are a
rich mixture of numerous bioactive chemical components such as ter-
penes, terpenoids, and phenolics (Voon, Bhat, & Rusul, 2012). In
nature, the particular aromatic and chemical characteristics of EOs
achieve many important functions for the plants such as (i) attracting
beneficial insects and pollinators, (ii) shielding the plants from some
environmental stress (heat, cold, etc.) and (iii) protecting the plants
from pests and/or microorganisms (Burt, 2004; Dhifi et al., 2016).
Those bioactive EOs are worldwide known for their proven biological
activities including antimicrobial, antifungal, antioxidant, antiviral,
antimycotic, antiparasitic and insecticidal properties (Burt, 2004; Calo
et al., 2014; Dhifi et al., 2016). Some examples of these activities were
detailed in Table 1. Regarding these activities, hundreds of EOs are used
in cosmetics (perfumes and make-up products), in agriculture (bio-
pesticide and repellent), in sanitary products (fragrances for household
cleaning products) and as natural remedies in aromatherapy (Burt,

⁎
Corresponding author.
E-mail addresses: Hanenfalleh@gmail.com, Hanenfalleh@yahoo.fr (H. Falleh).

https://doi.org/10.1016/j.foodchem.2020.127268
Received 6 January 2020; Received in revised form 2 June 2020; Accepted 5 June 2020
Available online 08 June 2020
0308-8146/ © 2020 Elsevier Ltd. All rights reserved.
H. Falleh, et al.
Table 1
Major biological activities of selected EOs.
Plant family Essential oil
Monimiaceae Peumus boldus
Lamiaceae Mentha piperita
Lamiaceae, Apiaceae, Zingiberaceae Origanum majorana, Coriandrum sativum,
Burseraceae Hedychium spicatum, Commiphora myrrha
Piperaceae Piper nigrum
Asteraceae Artemisia scoparia
Ranunculaceae Nigella sativa
Cyperaceae Cyperus esculentus and Cyperus rotundus
Lamiaceae Peppermint
Rutaceae Lauraceae Citrus Cinnamomum sp.
Apiaceae Ajowan (Trachyspermum ammi)
Rosaceae Cardiopteridaceae Rose citronella
Myrtaceae Clove
Apiaceae Lamiaceae Caraway spearmint
2004).
Essential oils are synthesized by almost all plant organs particularly
the flowers, buds, leaves, seeds, stems, and fruits (Dhifi et al., 2016).
These EOs can be stored in epidermal cells, cavities, secretory cells of
glandular trichomes (Dhifi et al., 2016). Numerous techniques can be
used to extract EOs from these plant parts. Hydrodistillation and steam
distillation are of the oldest, simplest and most commonly used
methods of extracting EOs (Dima & Dima, 2015). Particular methods
can also be applied for specific plants exp. cold pressing (exclusively for
the peel of Citrus fruits) or enfleurage especially applied for rose
(Perricone, Arace, Corbo, Sinigaglia, & Bevilacqua, 2015). In total, over
3000 types of EOs are identified, of which only 300 are of industrial
importance for applications in the food industries often for the flavors
and fragrances market (Bakkali et al., 2008). It should be highlighted
that EOs are generally accepted by consumers thanks to their high
volatility, ephemeral and biodegradable nature.
The purpose of this review is to present an overview of the pub-
lished data on the suitability of EOs for application in foods as eco-
friendly preservatives. A brief description of essential oils chemical
composition is presented followed by the discussion of their anti-
bacterial activity mode of action. Then, this paper focuses on EOs
preservative efficacies in food systems. Finally, EOs safety, as well as
the perspectives and limitations of their uses as food eco-preservatives,
were also presented.
2. Essential oils chemical composition
Essential Oils are a complex mixture of volatile compounds char-
acterized by a strong smell and flavor that are variables depending on
the EO chemical composition (Fig. 1). Generally, EOs are complex
mosaic containing a variable number of polar and non-polar compo-
nents at different concentrations (Raut & Karuppayil, 2014). Yet, only
two or three predominant components are present at fairly concentra-
tions (20–90%), while the other compounds concentrations are gen-
erally significantly lower. Further characterization of the complex mix
of natural compounds within EOs divided it into two fractions resulting
from the different biosynthetic chain: the terpenes group and the aro-
matic and aliphatic one. The terpenes fraction is incontestably the
major group constituting the EOs. Terpenes are characterized by their
basic structural element, (isoprene unit) and are formed by the
Food Chemistry 330 (2020) 127268
Medicinal properties Source
colagogue, hepatoprotective, antioxidant, anti- De Souza et al. (2019)
inflammatory, antifungal, antimicrobial, and
anthelmintic activities
Antimicrobial activities Tassou, Drosinos, and Nychas (1995)
antifungal, antiaflatoxin, antioxidant, and phytotoxic Prakash et al. (2012)
activities
Antibacterial, antifungal, anticancer and Raut and Karuppayil (2014)
antiprotozoal activities
Antioxidant and radical scavenging activity Singh, Mittal, Kaur, Batish, and Kohli
(2009)
Antimicrobial and antioxidant activities Paarakh (2010)
Anti-inflammatory, anti-arthritic, analgesic and Biradar, Kangralkar, Mandavkar,
anticonvulsant activities Thakur, and Chougule (2010)
Antioxidant, antimicrobial, and anti-inflammatory Bhavaniramya, Vishnupriya, Al-
activities Aboody, Vijayakumar, and Baskaran
(2019)
Antimicrobial, antioxidant, and insecticide activities Bhavaniramya et al. (2019)
Antimicrobial, antioxidant, antiaflatoxigenic, and Kedia, Prakash, Mishra, Dwivedy, and
antitermitic activities Dubey (2015)
Antiseptic, antibacterial, and anti-inflammatory Bhavaniramya et al. (2019)
activities
Antiseptic, antibacterial, antifungal, and anaesthetic Bhavaniramya et al. (2019)
activities
Aromatherapy and complementary medicine Bhavaniramya et al. (2019)
condensation of two or more isoprene units (Pavela, 2015). Considering
the number of isoprene subunits they contain, terpenes are classified in
different subgroups, such as hemi- (C5), mono-(C10), sesqui- (C15), di-
(C20), sester- (C25), tri- (C30), tetra- (C40) and polyterpenes (C5)n
(isoprene units > 8) (Pavela, 2015). It should be underlined that the
monoterpenes (C10) are frequently the predominant molecules in EOs,
and they may reach a percentage of up to 90% of the whole EO. Then,
the aromatic and aliphatic group which is the less abundant in EOs, and
its compounds correspond to alcohols, aldehydes, phenols, hetero-
cycles, and methoxy derivatives (Dhifi et al., 2016; Pavela, 2015).
Nevertheless, it should be highlighted that phenolic compounds (exp.
carvacrol, thymol, eugenol) are one of the most important molecules
responsible for the antimicrobial effects of EOs (Burt, 2004). There is
evidence that EOs chemical composition is highly variable, qualita-
tively and quantitatively, depending on countless factors such as har-
vesting seasons/time during the day, geographical sources, plant organ,
plant maturity degree, etc. (Dhifi et al., 2016). For example, coriander
(Coriandrum sativum L.) EOs were chemically different when extracted
from seeds than when extracted from leaves (Delaquis, Stanich, Girard,
& Mazza, 2002). The major compound (linalool) percentage was equal
to 70% in seeds EOs while it was limited to 26% in leaves EOs.
3. Mechanism of action of eos antibacterial activity
EOs are known for harboring a broad spectrum in biological activ-
ities (Burt, 2004; Calo et al., 2014; Dhifi et al., 2016; Valderrama &
Ruiz, 2018). In this paper, special interest will be dedicated to their
antimicrobial potency. The antimicrobial efficacies of EOs and their
components were well established since antiquity. This efficacy of EOs
is either bacteriostatic (EOs inhibit the bacterial growth then the mi-
crobial cells may recuperate their reproductive ability), or bactericide
which means that EOs kill bacterial cells (Voon et al., 2012; Calo et al.,
2014; Prakash, Singh, Kedia, & Dubey, 2012). Indeed, the EOs me-
chanisms of action are not yet formally established partially due to the
important variability of their chemical compounds (Hyldgaard,
Mygind, & Meyer, 2012). Nevertheless, it is most likely that EOs anti-
microbial propriety is not attributable to a single or a specific me-
chanism but it is probably related to many targets in the bacterial cell
(Calo et al., 2014). With this respect, there is a general agreement on
the fact that the EOs hydrophobicity/ lipophilicity allows them to cross
2
H. Falleh, et al.
Fig. 1. Chemical structures of some EO constituents.
the cell cytoplasmic membrane and mitochondria and permeabilizes
their different layers of fatty acids, polysaccharides, and phospholipids
(Burt, 2004). Moreover, Rodriguez-Garcia et al. (2016) explained that
the lipophilic ends of lipoteichoic acids in gram-positive bacteria cell
membrane profits to the penetration of EOs hydrophobic corposants,
whereas the outer membrane surrounding the gram negative-bacteria
cell wall, restricts the flow rate of hydrophobic EOs through the lipo-
polysaccharide layer. That is the main reason explaining that gram
positive-bacteria are to some extent more sensitive to EOs action than
gram-negative ones (Dhifi et al., 2016). Furthermore, EOs are also
capable to disintegrate bacterial cell wall as well as their cytoplasmic
membrane structures through disturbing the conformation of their
different fatty acids, polysaccharides, and phospholipid layers by
raising their permeability (Fig. 2).
Disturbances to these two structures imply the reduction of the
membrane potential, important ion and other cell contents leakage, a
reduction of the ATP pool, a collapse of the proton pump, a fragilization
of the cellular membrane and finally a loss of macromolecules
(Gutiérrez-del-Río et al., 2018). Every one of these negatives effects is
the main cause of cell essential processes damage and inevitably to cell
lysis. EOs are also able to coagulate the cytoplasm and to inhibit several
enzyme systems such as the ones responsible for energy regulation and
the synthesis of structural components (Burt, 2004).
3
Food Chemistry 330 (2020) 127268
According to several studies, EOs exhibiting the highest anti-
bacterial activities against foodborne pathogens are generally rich in
phenolic compounds like thymol, carvacrol or eugenol (Dhifi et al.,
2016, Ben Jemaa et al., 2017; Ben Jemaa et al., 2018a; Gutiérrez-del-
Río et al., 2018). These terpene phenols targeted the proteins amine and
hydroxylamine groups in the bacterial membrane to alter their per-
meability leading to the bacteria death (Adelakun et al., 2016;
Hyldgaard et al., 2012). Moreover, evidence demonstrates that EOs
minor components possess an outstanding role in EOs antimicrobial
efficiencies, probably by exhibiting a synergistic effect with the other
EOs components. One known example is the synergistic interaction
between p-cymene and carvacrol. ρ-cymene barely inhibits microbial
growth and carvacrol had a proven antimicrobial efficiency against a
wide bacteria range. According to Rattanachaikunsopon and
Phumkhachorn (2010), microbial growth was significantly inhibited
when p-cymene and carvacrol were added at the same time to the
sample medium. Interestingly, this bacterial inhibition was significantly
weaker when each terpene acted separately in the same medium. It has
been detailed that p-cymene play the role of a substitutional impurity in
the bacterial membrane, moderately disturbing the membrane potential
of intact cells. This condition eased the carvacrol activity and conse-
quently lowers the effective concentrations of each component
(Rattanachaikunsopon & Phumkhachorn, 2010). Infrequently,
H. Falleh, et al.
Fig. 2. Principal targets of EOs in bacterial cells and some mechanisms of antimicrobial activity: damage to the cell wall; degradation to the cell membrane;
coagulation of the cytoplasmic protein; leakage of the cell contents and reduction of the proton-motive force (Prakash et al., 2015).
antagonistic effects may arise when the antibacterial activity of two (or
more) compounds is less important when they are combined than when
assessed individually (Dhifi et al., 2016). Indeed, Galluci and coworkers
(2009) stipulated that combining carvacrol and thymol lowered their
Fractionary Inhibitory Concentrations (FIC) against S. aureus, B. cereus,
and E. coli while a synergetic interaction appears against the same pa-
thogens when geraniol and menthol were mixed.
It is well known that bacterial strains, including foodborne ones,
develop many mechanisms of resistance to synthetic antimicrobial
drugs (intrinsic, acquired, or transfer of resistance genes) given rise to
multi-drugs resistance bacteria called superbugs. In this context, it
seems very important to highlight that as far as our literature can as-
certain, there are no specific resistances or bacterial adaptations against
EOs or their components reported. This fact is of crucial importance in
favor of the utilization of EOs as a natural preservative, especially in the
agro-food industry.
4. Mechanism of action of EOs antioxidant activity
Oxidation is one of the most important causes of food degradation.
It is a multilateral reaction that causes undesirable changes in food
value, organoleptic criteria, nutritional quality and the creation of po-
tentially toxic molecules. Food undergone extensive oxidation has
major defects and no consumer acceptability (Prakash et al., 2015).
Oxidation during food processing and/or storage may be revealed
through color changes and the appearance of off-flavors, whereas the
variation of its principal components, such as lipids is not always
marked (Prakash et al., 2015). Antioxidant compounds can prevent,
alter and even end oxidative reactions at relatively low concentrations.
With this respect, EOs and their constituents play a key role in exerting
antioxidant activity. Rodriguez-Garcia et al. (2016) reported that the
EOs have several modes of direct or indirect actions including pre-
vention of chain initiation and free-radical scavenging activity. In
previous work, O. majorana, C. sativum, H. spicatum, C. myrrha, and C.
odorata EOs were tested for their antioxidant activities through DPPH
free radical scavenging and the β-Carotene/linoleic acid bleaching
assay (Prakash et al., 2012). Four of the tested EOs exhibited important
antioxidant activity (expressed by low IC50 values) while the inferior
activity of H. spicatum was attributed to its low phenolic content in
4
Food Chemistry 330 (2020) 127268
comparison to the other EOs supporting the earlier view that phenolic
compounds may play crucial roles in EOs antioxidant activity (Prakash
et al., 2012). Indeed, and according to De Souza et al. (2019), phenolic
compounds, like thymol, eugenol, and carvacrol, are the main potent
antioxidant agents in EOs as they can donate hydrogen atoms to free
radicals and transform them to more stable products. The other com-
ponents such as certain alcohols, ethers, ketones, aldehydes, and
monoterpenes: linalool, 1,8-Cineole, geranial/neral, citronellal, iso-
menthone, menthone, and some monoterpenes also play a key role in
the antioxidant properties of EOs (Rodriguez-Garcia et al., 2016).
5. Preservative efficacy of EOs in food systems
Increased health consciousness by consumers has fueled negative
perceptions about synthetic food additives. Therefore a considerable
number of natural and safe EOs started to be used in the food industry
under the perception of ‘organic or green strategy’ representing a new
barrier to guarantee the elimination of pathogens from a specific food
matrix (Table 2). The target is that these safe and efficient EOs will
replace the actual conventional additives (Ben Jemaa et al., 2018a;
Falleh et al., 2019). Accordingly, many EOs and EO components were
approved by the European Commission (EC) as well as by the United
States Food and Drug Administration, known as FDA, that classified
these naturals components and mixtures as Generally Recognized As
Safe (GRAS) to be used as flavorings and/or preservatives in food
products (Hyldgaard et al., 2012; Ben Jemaa et al., 2018b). According
to the FDA website, the crude essential oils labeled GRAS comprise
lavandin, menthol, rose, sage, oregano, cinnamon, basil, clove, cor-
iander, nutmeg, ginger, and thyme EOs. Similarly, the registered EOs
component included thymol, carvacrol, eugenol, linalool, carvone, va-
nillin, cinnamaldehyde, citral, and limonene, all of which are con-
sidered safe under the limitations on the accepted daily intake (Code of
Federal Regulations, title 21, part 182). At first, it must be underlined
that EOs effects on food preservation depend on their source and
composition. For instance, Ocimum basilicum and Rosmarinus officinalis
EOs acted differently as a food preservative. According to Karoui and
Hassoun (2017), when tested at the same concentration (1%), basil EO
was more efficient at delaying food spoilage (strong antimicrobial ac-
tivity) while rosemary EO was more potent in preventing lipid
H. Falleh, et al. Food Chemistry 330 (2020) 127268

oxidation due to its important antioxidant potential. On a practical

Dwivedy, Prakash, ChanotiyC, Bisht,

level, the application of EOs as food preservatives requests precise data

Raybaudi-Massilia et al. (2006)

Skandamis and Nychas (2000)
Smith-Palmer et al. (2001) about the effect of food matrix components on EOs antimicrobial
properties. Several studies reported that EOs efficacy can be modified

Ben Jemaa et al. (2017)

while interacting with certain food components (intrinsic factors) such

Tassou et al. (1995)

Falleh et al. (2019)

Kedia et al. (2015)

as proteins, fats, water, carbohydrates, antioxidants, and salt as well as

and Dubey (2017)

the food pH which is also an important parameter to be considered
(Smith-Palmer, Stewart, & Fyfe, 2001; Calo et al., 2014; Adelakun et al.,
2016). Besides, extrinsic factors are also able to affect (one way or the
Source

other) EOs antimicrobial activity in a specific food matrix. The main

examples of extrinsic factors are the packaging process, gaseous com-
position, temperature, and microorganisms’ nature and initial con-
centrations (Rattanachaikunsopon & Phumkhachorn, 2010; Hyldgaard
et al., 2012; Rodriguez-Adelakun et al., 2016; Rodriguez-Garcia et al.,
Tzatziki (cucumber and yogurt salad),
taramosalata (fish roe salad) and pâté

2016). Previous works stipulated that bacteria susceptibility to EOs

increases at acidic food pH, as at this condition (low pH) EO behaves in
full-fat and low-fat soft cheeses

a more hydrophobic way enhancing its ability to dissolve in the bacteria

membrane (Burt, 2004; Hyldgaard et al., 2012; Perricone et al., 2015).
semi-skimmed UHT milk

As well, it was observed that the presence of important content of fat

wheat and chickpea

and/or protein in foods matrix hamper the EO action against bacteria

eggplant salad

(Smith-Palmer et al., 2001; Calo et al., 2014; Perricone et al., 2015;

Food matrix

Fruit juices

Adelakun et al., 2016). In fact, in high-fat food, EOs will dissolve in the
UHT Milk

dry fruits

lipid fraction which facilitates its antioxidant activity and help to avoid
lipid oxidation or rancidification. Nevertheless, this hydrophobicity will
make EO relatively unavailable to react with the bacterial strains in the
aqueous fraction. With this respect, Tassou and coworkers published in
Staphylococcus aureus, Bacillus licheniformis,

1995 an interesting study describing how Mentha piperita EO displayed

Enteritidis, Escherichia coli, and Listeria

different antimicrobial efficacy depending on the fat amount in food.

According to these authors, M. Piperita demonstrated moderate anti-
Salmonella enteritidis and Listeria

Salmonella enteritidis and Listeria

bacterial activity against Listeria monocytogenes and Salmonella en-

teritidis in high-fat foods (pâté and fish roe salad) albeit the same EO
Escherichia coli O157:H7
andEnterococcus hirae,

was remarkably more efficient against the same bacteria in yogurt salad
and cucumber which are low-fat food (Skandamis & Nychas, 2000).
microorganisms

Aspergilus flavus
Aspergilus flavus
Escherichia coli

Additionally, the interaction between carvacrol (EO component) and

monocytogenes

monocytogenes

food proteins negatively influenced its antibacterial propriety against

Salmonella
Overview of previous studies dealing with the antimicrobial activity of EOs in different food matrix.

innocua

Bacillus cereus in milk as well as against Salmonella enteritidis in cheese

(Smith-Palmer et al., 2001). Also, food physical structure can interfere
with the EOs antibacterial potencies. It seems that EO minimum in-
Main constituent

Cinnamaldehyde

hibition concentration (MIC) may be quite lower in broth medium than

Cinnamldehyde

Cinnamldehyde

in agar one. As demonstrated by Skandamis and Nychas (2000), the

Carvacrol
carvacrol
Camphor
α-pinene

oregano EO activity against Salmonella typhimurium was considerably

menthol

Carvone
Eugenol

Eugenol

Eugenol
Cymene
thymol

limited in the gelatine gel matrix than in a broth medium, probably

–

because of the restraint diffusion in the gel structure.

Pimenta racemosa, Clove (Syzygium aromaticum), cinnamon (Cinnamomum verum,

5.1. Meat and meat products

palmarosa (Cymbopogon martini), clove (Eugenia caryophyllata), cinnamon
Clove (Syzygium aromaticum), cinnamon (Cinnamomum verum), Lavender

According to Burt (2004), incorporation of specific EOs (exp. or-

egano, coriander, thyme, and clove) as preservative agents in meat
(Cinnamomum zeylanicum), lemongrass (Cymbopogon citrates)

eliminate many pathogens and autochthonous spoilage flora. Once in-

gested, EO compounds, particularly thymol and carnosic acid, may be
accumulated in the animal muscle at enough concentrations to exhibit
(Lavandula stoechas), myrtle (Myrtus communis)

effective antimicrobial activity. For instance, the incorporation of Sa-

tureja horvatii EO (major compounds are p-cymene and thymol) to pork
meat significantly limited Listeria monocytogenes growth in comparison
with controls following 96 h of storage (Kuorwel, Cran, Sonneveld,
Miltz, & Bigger, 2011). Moreover, only 5 to 20 µl.g−1 of clove, cor-
iander, thyme or oregano EOs were sufficient to inhibit Listeria mono-
Ajowan (Trachyspermum ammi)

cytogenes, Aeromonas hydrophila and autochthonous spoilage flora in

thyme (Thymus vulgaris)

Oregano (Origanum vulgare)

Thyme (Thymus capitatus)

some meat products thanks to a noticeable early reduction in the re-

coverable cells number (Burt, 2004).
Mentha cardiaca
Mentha piperita

5.2. Vegetables and fruits

Essential oil

Alfalfa seeds were protected from Salmonella contamination (6 dif-

Table 2

ferent serotypes) by cinnamaldehyde and thymol (Burt, 2004). Eggplant

salad resisted to E. coli O157:H7 contamination when treated with 7 to

5
H. Falleh, et al.
21 µl.g−1 of oregano EO; Rice seeds were sheltered by less than
0.75 µl.g−1 of carvacrol against B. cereus and Kiwi natural bacterial
flora was considerably reduced under the effect of 0.15 µl.g−1 carvacrol
and cinnamaldehyde (Burt, 2004). Considering fruit juice, Raybaudi-
Massilia, Mosqueda-Melgar, and Martin-Belloso (2006) reported that
Lemongrass EO and geraniol were able to control Escherichia coli, Lis-
teria sp., and Salmonella sp. in melon, pear and apple juices.
5.3. Dairy products
Ben Jemaa et al. (2017) reported that thyme EO (Thymus capitatus)
and its nanoemulsion positively fought gram+ bacterial growth in
contaminated milk (Staphylococcus aureus, Bacillus licheniformis, and
Enterococcus hirae) and ameliorated its physicochemical quality too.
Similarly, Smith-Palmer et al. (2001) founded that the addition of 0.5%
of cinnamon, bay, clove or thyme EOs to low-fat cheese resulted in a
statistical reduction of Salmonella enteritidis although all these addition
altered cheese organoleptic properties.
Regarding the published data concerning EOs use as a food pre-
servative, Burt (2004) proposed the following EOs ranking (organized
from the most antibacterial activity to the less active EO): clove/or-
egano/ cinnamon /coriander > thyme > mint > rosemary >
mustard > sage/cilantro. Similarly, an estimated large ranking of EO
components may be as follow (same order than EOs): eugenol >
cinnamic acid/carvacrol > basil methyl chavicol > microbial contamination generally occurs, providing efficient protec-
cinnamaldehyde > geraniol/citral.
6. Safety assessment of plant EOs
Confirmation of EOs safety taking into account their tremendous
chemical variability has become more important for the consumer
(Raut & Karuppayil, 2014). As a result, the characterization of EOs
chemical and toxicological properties when used in the food industry
has been exponentially assessed (Dima & Dima, 2015). According to
Dima and Dima (2015), notions like a ‘long history of safe use’ and the
‘principle of self-limitation’ are adopted to judge EOs as ‘safe under
intended condition of use.’ Taking into consideration these two con-
cepts, health organizations, including FDA, define regulatory restric-
tions on the accepted daily intake of EOs or their components. Im-
portant international organisms like FDA, the International
Organization of Flavor Industries (IOFI), Food Chemical Codex (FCC),
Manufacturers Association (FEMA), Codex Alimentarium and the
Council of Europe (CoE) have developed specific protocols for EOs
chemical and toxicological analyses, approved guides for processing
EOs, and the safety restrictions for the volatile component's content in
EOs as the synergistic and antagonistic effect may arise for specific
essential oils (Dima & Dima, 2015). Consequently, each EO proposed as
a food preservative should be validated for its safety limit on the
mammalian system (Ben Jemaa et al., 2018b). Toxicity/safety tests
were variable and different. One of the most used methods for EOs
safety assessment is the acute oral test which allows the determination
of LD50 or Median Lethal Dose value (Nakavuma, Matasyoh, Wagara,
Kalema, & Alinaitwe, 2016). The higher is the EOs LD50 value the
stronger is their suitability as food preservatives. For instance, table 3
displays the safety limit profile (expressed as LD50) of numerous EOs
(experiences conducted on mice). Practically, since the 1st April 2018,
EOs of Cinnamomum verum (cinnamon), Syzygium aromaticum (clove),
Thymus capitatus (thyme), Rosmarinus officinalis (rosemary) and 156
other aromatic plants are recognized by the Code of Federal Regula-
tions Title 21 as safe under the regulatory limitations. A rising aspect of
EOs safety is their possible contamination by synthetic chemicals such
as pesticides. According to Di Bella and co-workers (2006), consider-
able concentrations of organophosphorus (exp. Ethyl chlorpyriphos and
Methidathion) and organochlorine (exp. Tetradifon and Chloropar-
affins) pesticides residues were found in mandarin and lemon EOs
grown with biological methods.
6
Food Chemistry 330 (2020) 127268
Concerning the allergic risk of consuming EO in food, it is important
to highlight that this risk is very limited. According to a study con-
ducted by Fischman et al. (2004) to assess the irritation potential of an
EO-containing mouth rinse in objectively documented xerostomia
(hyposalivation) population, the oral irritation potential of the EO
mouth rinse was minimal and very limited.
7. Perspectives and limitations in application of EOs in food
systems
As explained in this paper, it is advisable to the extent of food shelf-
life using EOs as an eco-friendly preservative that can also be a valuable
flavoring and aromatizing agent, exp. Citrus EO is used as aromatizers
for tea, liqueurs, sweets and candied fruits (Di Bella et al., 2006). Al-
though the selection of potent EOs that does not alters food taste, aroma
and/or flavor is one of the main limitations of their use as food pre-
servatives. In fact, toward consumers and the food industry, food safety
is as essential as its sensory quality, and EOs exhibit intense aroma that
negatively affects concerned food organoleptic characteristics (Karoui &
Hassoun, 2017; Ben Jemaa et al., 2018b). This hurdle may be overcome
using several strategies. Hyldgaard et al. (2012) suggested that EOs may
be used in active packaging when low EOs concentrations may be in-
corporated in food packaging films. These active packaging slowly re-
leases the volatile antimicrobial into the product surface, where the
tion without affecting food acceptability (Hassoun & Çoban, 2017;
Hyldgaard et al., 2012). One other confirmed strategy to minimize the
EOs organoleptic effects is their encapsulation into micro- or nanoe-
mulsions. Indeed, Ben Jemaa et al. (2017) displayed that the en-
capsulation of EOs, such Thymus capitatus one; in a nanoemulsion de-
livery system may reduce significantly their organoleptic impact on the
food matrix. This method raises EO stability, lower its interaction with
the food matrix (exp. Fat and proteins) and higher its antimicrobial
potency through the increase of the passive cellular uptake (Ben Jemaa
et al., 2017; Hassoun & Çoban, 2017). Another technique to reduce EO
doses in food without compromising their activity is the use of their
synergistic interaction with other antimicrobial agents. With this re-
spect, Dhifi et al. (2016) reported that the combinations of 1,8-cineole
with aromadendrene exhibited an important protective effect against
the methicillin-resistant Staphylococcus aureus (MRSA), the vanco-
mycin-resistant enterococci (VRE) and the pathogen Enterococcus fae-
calis. Also, Burt (2004) reported confirmed synergism between oregano
EO with sodium nitrite (against Clostridium botulinum) as well as be-
tween nisin with either thymol or carvacrol (against Bacillus cereus).
Another valid approach is also actually discussed is the combined
treatment process. For example, the efficacy of the combined action of
carvone and mild heat treatment against Listeria monocytogenes; clove,
oregano and coriander EOs associated with vacuum-packed meat
against spoilage microorganisms were reported (Burt, 2004). Finally, in
several cases, the substitution of EO with its major chemical component
may preserve the same activity without the organoleptic disadvantages,
as this change eschews some compounds that modify food sensorial
properties (Smith-Palmer et al., 2001). As a final point, it should be
underlined that the significant potential of EOs as food eco-preservative
may be a serious menace of the ecological biodiversity causing losses of
specific aromatic plants. This challenge may be avoided by adopting a
tissue culture technique to obtain industrial biomass of the desired
plant.
8. Conclusion(s)
Essential oils exhibit high potential to be used as a natural anti-
microbial and antioxidant agents. Exploring these bioactive mixtures by
providing adequate scientific evidence, contribute to increasing the
chances of developing potent, ecological, and safer food preservative
agents that can be good alternatives to synthetic chemicals. Being
H. Falleh, et al. Food Chemistry 330 (2020) 127268

Table 3
The safety limit of some EOs expressed as the median lethal dose (LD50).
Essential oil origin Plant part Safety assessment method LD50 value (mice/rat) References

Aegle marmelos L. Correa Leaves acute oral toxicity 23.66 g/kg Singh et al., 2009
Peumus boldus Leaves acute toxicological evaluation 42.18 mg/kg De Souza et al., 2019
Mentha cardiaca Shoots acute oral toxicity 7.13 g/kg Dwivedy et al. (2017)
Nepeta cataria L. Shoots acute oral toxicity 3.16 g/kg Zhu et al., 2009
Cymbopogon citratus Leaves acute oral toxicity 8.1 g/kg Nakavuma et al., 2016
Nigella sativa L. Seeds Intraperitonial injection 0.1 g/kg Paarakh, 2010
Cyperus esculentus Leaves acute oral toxicity 5 g/kg Biradar et al., 2010
Ocimum gratissimum Leaves acute oral toxicity 11.62 g/kg Prakash et al., 2012
Monanthataxis littoralis Leaves acute oral toxicity 13.08 g/kg Nakavuma et al., 2016
Zanthoxylum alatum Seeds acute oral toxicity 6.12 g/kg Prakash et al., 2012
Thymus capitatus Shoots acute oral toxicity > 2 g/kg Ben Jemaa et al., 2018b
Rosmarinus officinalis Leaves acute oral toxicity 5.94 g/kg Nakavuma et al., 2016
Callistemon lanceolatus Aerial part acute oral toxicity 14.63 g/kg Prakash et al., 2012
Angustifolium saligna Leaves acute oral toxicity 23 g/kg, Nakavuma et al., 2016

naturally used in food with minimal known “side-effects” on human 1016/j.ifset.2017.08.017.

health, they are accepted to be safe for consumption. Nowadays, most
EOs are classified as “generally recognized as safe or GRAS” by the
United States Food and Drug Administration (FDA) and are therefore
permitted as food additives/preservatives. Based on the available lit-
erature, EO organoleptic impact in food products limits seriously their
use mainly to spicy foods habitually associated with spices or herbs.
Active packaging, modern encapsulation technologies, and combined
strategies may, therefore, be assessed to minimize the organoleptic ef-
fects and facilitate the use of EOs in a larger range of food products
without compromising their properties. To date, the clinical effects of
EOs in “in vivo” conditions as well as interactions in food systems are
scare. These aspects need to be further studied to estimate correctly the
potential efficiencies of EOs as antimicrobial and antioxidant agents
and to verify the possibility of acute or chronic effects. Deeper in-
vestigations should thus be realized before a regulatory approval for
their use as natural preservatives for food industry applications.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
This study was supported by the Tunisian Ministry of Higher
Education, Scientific Research, Information and Communication
Technologies (Tunisia) within the project of “Encouragement des
Jeunes Chercheurs” coded as 18JEC 08-03.
References
Adelakun, O.E., Oyelade, O.J., & Olanipekun, B.F. (2016). Use of Essential Oils in Food
Preservation. In: Preedy VR. Essential Oils in Food Preservation, Flavor, and Safety.
Academic Press is an imprint of Elsevier. Chapter 7: 71-84. ISBN: 978-0-12-416641-7.
Bakkali, F., Averbeck, S., Averbeck, D., & Idaomar, M. (2008). Biological effects of es-
sential oils - a review. Food and Chemical Toxicology, 46, 446–475. https://doi.org/10.
1016/j.fct.2007.09.106.
Bhavaniramya, S., Vishnupriya, S., Al-Aboody, M. S., Vijayakumar, R., & Baskaran, D.
(2019). Role of essential oils in food safety: Antimicrobial and antioxidant applica-
tions. Grain & Oil Science and Technology. 2, 49–55. https://doi.org/10.1016/j.gaost.
2019.03.001.
Jemaa, M., Falleh, H., Neves, M. A., Isoda, H., Nakajima, M., & Ksouri, R. (2017). Quality
preservation of deliberately contaminated milk using thyme free and nanoemulsified
essential oils. Food Chemistry, 217, 726–734. https://doi.org/10.1016/j.foodchem.
2016.09.030.
Ben Jemaa, M., Falleh, H., Saada, M., Oueslati, M., Snoussi, M., & Ksouri, R. (2018 a).
Thymus capitatus essential oil ameliorates pasteurized milk quality. Journal of Food
Science and Technology, 55(9), 3446–3452. DOI: 10.1007/s13197-018-3261-4.
Ben Jemaa, M., Falleh, H., Serairi, R., Neves, M. A., Snoussi, M., Isoda, H., ... Ksouri, R.
(2018b). b). Nanoencapsulated Thymus capitatus essential oil as natural preservative.
Innovative Food Science and Emerging Technologies, 45, 92–97. https://doi.org/10.
Biradar, S., Kangralkar, V. A., Mandavkar, Y., Thakur, M., & Chougule, N. (2010). Anti-
inflammatory, anti-arthritic, analgesic and anticonvulsant activity of Cyperus es-
sential oils. International Journal of Pharmacy and Pharmaceutical Sciences,
2(975–1491), 54296812.
Burt, S. (2004). Essential oils: Their antibacterial properties and potential applications in
foods—a review. International Journal of Food Microbiology, 94, 223–253. https://doi.
org/10.1016/j.ijfoodmicro.2004.03.022.
Calo, J. R., Crandall, P. G., O’Bryan, C. A., & Ricke, S. C. (2014). Essential Oils as
Antimicrobials in Food Systems–A Review. Food Control, 54, 111–119. https://doi.
org/10.1016/j.foodcont.2014.12.040.
Code Of Federal Regulations. title 21-food and drugs chapter 1-food and drug adminis-
tration department of health and human services subchapter b-food for human
consumption part 182-substances generally recognized as safe.
De Souza, W. F. M., Mariano, X. M., Isnard, J. L., De Souza, G. S., De Souza Gomes, A. L.,
De Carvalho, R. J. T., ... Moreira, R. F. A. (2019). Evaluation of the volatile com-
position, toxicological and antioxidant potentials of the essential oils and teas of
commercial Chilean boldo samples. Food Research International. https://doi.org/10.
1016/j.foodres.2018.12.059.
Delaquis, P. J., Stanich, K., Girard, B., & Mazza, G. (2002). Antimicrobial activity of in-
dividual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils.
International Journal of Food Microbiology, 74, 101–109. https://doi.org/10.1016/
S0168-1605(01)00734-6.
Dhifi, W., Bellili, S., Jazi, S., Bahloul, N., & Mnif, W. (2016). Essential Oils’ Chemical
Characterization and Investigation of Some Biological Activities: A Critical Review.
Medecines, 3, 1–16. https://doi.org/10.3390/medicines3040025.
Di Bella, G., Serrao, L., Salvo, F., Lo Turco, V., Croce, M., & Dugo, G. (2006). Pesticide and
plasticizer residues in biological citrus essential oils from 2003–2004. Flavour and
Fragrance Journal, 21, 497–501. https://doi.org/10.1002/ffj.1663.
Dima, C., & Dima, S. (2015). Essential oils in foods: Extraction, stabilization, and toxicity.
Current Opinion in Food Science, 5, 29–35. https://doi.org/10.1016/j.cofs.2015.07.
003.
Dwivedy, A. K., Prakash, B., ChanotiyC, S., Bisht, D., & Dubey, N. K. (2017). Chemically
characterized Mentha cardiaca L. essential oil as plant based preservative in view of
efficacy against biodeteriorating fungi of dry fruits, aflatoxin secretion, lipid perox-
idation and safety profile assessment. Food and Chemical Toxicology, 106, 175–184.
https://doi.org/10.1016/j.fct.2017.05.043.
Falleh, H., Ben Jemaa, M., Djeblai, K., Abid, S., Saada, M., & Ksouri, R. (2019).
Application of the mixture design for optimum antimicrobial activity: Combined
treatment of Syzygium aromaticum, Cinnamomum zeylanicum, Myrtus communis, and
Lavandula stoechas essential oils against Escherichia coli. Journal of Food Processing and
Preservation. https://doi.org/10.1111/jfpp.14257.
Fischman, S. L., Aguirre, A., & Charles, C. H. (2004). Use of essential oil-containing
mouthrinses by xerostomic individuals: Determination of potential for oral mucosal
irritation. American Journal of Dentistry, 17(1), 23–26.
Gallucci, M. N., Oliva, M., Casero, C., Dambolena, J., Luna, A., Zygadlo, J., & Demo, M.
(2009). Antimicrobial combined action of terpenes against the food-borne micro-
organisms Escherichia coli, Staphylococcus aureus and Bacillus cereus. Flavour Fragrance
Journal, 24, 348–354. https://doi.org/10.1002/ffj.1948.
Gutiérrez-del-Río, I., Fernández, J., & Lombó, F. (2018). Plant nutraceuticals as anti-
microbial agents in food preservation: Terpenoids, polyphenols and thiols.
International Journal of Antimicrobial Agents, 52, 309–315. https://doi.org/10.1016/j.
ijantimicag.2018.04.024.
Hassoun, A., & Çoban, O. E. (2017). Essential oils for antimicrobial and antioxidant ap-
plications in fish and 1 other seafood products. Trends in Food Science & Technology,
68, 26–36. https://doi.org/10.1016/j.tifs.2017.07.016.
Hyldgaard, M., Mygind, T., & Meyer, R. L. (2012). Essential oils in food preservation:
Mode of action, synergies, and interactions with food matrix components. Frontiers in
microbiology, 3, 1–24. https://doi.org/10.3389/fmicb.2012. 00012.
Karoui, R., & Hassoun, A. (2017). Efficiency of rosemary and basil essential oils on the
shelf-life extension of atlantic mackerel (Scomber Scombrus) fillets stored at 2 °C.
Journal of AOAC International, 100, 335–344. https://doi.org/10.5740/jaoacint.16-
0410.

7
H. Falleh, et al.
Kedia, A., Prakash, B., Mishra, K. P., Dwivedy, A. K., & Dubey, N. K. (2015).
Trachyspermum ammi L. essential oil as plant based preservative in food system.
Industrial Crops and Products, 69, 104–109. https://doi.org/10.1016/j.indcrop.2015.
02.013.
Kuorwel, K. K., Cran, M. J., Sonneveld, K., Miltz, J., & Bigger, S. W. (2011). Essential oils
and their principal constituents as antimicrobial agents for synthetic packaging films.
Journal of Food Science, 76, R164–77. https://doi.org/10.1111/j.1750-3841.2011.
02384.x.
Nakavuma, J. L., Matasyoh, J. C., Wagara, I. N., Kalema, J., & Alinaitwe, L. (2016).
Toxicity Studies on Anti-fungal Essential Oils Extracted from Selected Aromatic
Plants from Mabira and Kakamega Forests, East Africa. European Journal of Medicinal
Plants, 14(2), 1–14. https://doi.org/10.9734/EJMP/2016/25185.
Paarakh, P. M. (2010). Nigella sativa L.A comprehensive review. Indian Journal of Natural
Products and Resources, 1, 409–429.
Pavela, R. (2015). Essential oils for the development of eco-friendly mosquito larvicides:
A review. Industrial Crops and Products, 76, 174–187. https://doi.org/10.1016/j.
indcrop.2015.06.050.
Perricone, M., Arace, A., Corbo, M. R., Sinigaglia, A., & Bevilacqua, A. (2015). Bioactivity
of essential oils: A review on their interaction with food components. Frontiers in
microbiology, 6, 1–7. https://doi.org/10.3389/fmicb.2015. 00076.
Prakash, B., Kedia, A., Mishra, P. K., & Dubey, N. K. (2015). Plant essential oils as food
preservatives to control moulds, mycotoxin contamination and oxidative deteriora-
tion of agri-food commodities e Potentials and challenges. Food Control, 47, 381–391.
https://doi.org/10.1016/j.foodcont.2018. 01.018.
Prakash, B., Singh, P., Kedia, A., & Dubey, N. K. (2012). Assessment of some essential oils
as food preservatives based on antifungal, antiaflatoxin, antioxidant activities and in
vivo efficacy in food system. Food Research International, 49, 201–208. https://doi.
org/10.1016/j.foodres.2012.08.020.
Rattanachaikunsopon, P., & Phumkhachorn, P. (2010). Assessment of factors influencing
antimicrobial activity of carvacrol and cymene against Vibrio cholerae in food. Journal
of Bioscience and Bioengineering, 110, 614–619. https://doi.org/10.1016/j.jbiosc.
2010.06.010.
Raut, J. S., & Karuppayil, S. M. (2014). A status review on the medicinal properties of
essential oils. Industrial Crops and Products, 62, 250–264. https://doi.org/10.1016/j.
indcrop.2014.05.055.
Raybaudi-Massilia, R. M., Mosqueda-Melgar, J., & Martin-Belloso, O. (2006).
Food Chemistry 330 (2020) 127268
Antimicrobial activity of essential oils on Salmonella enteritidis, Escherichia coli, and
Listeria innocua in fruit juices. Journal of food protection, 69, 1579–1586. https://doi.
org/10.4315/0362-028X-69.7.1579.
Rodriguez-Garcia, I., Silva-Espinoza, B. A., Ortega-Ramirez, L. A., Leyva, J. M., Siddiqui,
M. W., Cruz-Valenzuela, M. R., ... Ayala-Zavala, J. F. (2016). Oregano essential oil as
an antimicrobial and antioxidant additive in food products. Critical Reviews in Food
Science and Nutrition, 56, 1717–1727. https://doi.org/10.1080/10408398.2013.
800832.
Singh, H. P., Mittal, S., Kaur, S., Batish, D. R., & Kohli, R. K. (2009). Chemical composition
and antioxidant activity of essential oil from residues of Artemisia scoparia. Food
Chemistry, 114, 642–645. https://doi.org/10.1016/j. foodchem.2008.09.101.
Skandamis, P. N., & Nychas, G. J. E. (2000). Development and evaluation of a model
predicting the survival of Escherichia coli O157:H7 NCTC 12900 in homemade egg-
plant salad at various temperatures, pHs and oregano essential oil concentrations.
Applied and Environmental Microbiology, 66(4), 1646–1653. https://doi.org/10.1128/
AEM.66.4.1646-1653.2000.
Smith-Palmer, A., Stewart, J., & Fyfe, L. (2001). The potential application of plant es-
sential oils as natural food preservatives in soft cheese. Food Microbiology, 18,
463–470. https://doi.org/10.1006/fmic.2001.0415.
Tassou, C. C., Drosinos, E. H., & Nychas, G. J. E. (1995). Effects of essential oil from mint
(Mentha piperita) on Salmonella enteritidis and Listeria monocytogenes in model food
systems at 4 °C and 10 °C. Journal of Applied Bacteriology, 78(6), 593–600. https://doi.
org/10.1111/j.13652672.1995. tb03104.x.
Valderrama, F., & Ruiz, F. (2018). An optimal control approach to steam distillation of
essential oils from aromatic plants. Computers and Chemical Engineering, 117, 27–31.
https://doi.org/10.1016/j.compchemeng.2018.05.009.
Voon, C. H., Bhat, R., & Rusul, G. (2012). Flower extracts and their essential oils as
potential antimicrobial agents for food uses and pharmaceutical applications.
Comprehensive Reviews in Food Science and Food Safety, 11, 34–55. https://doi.org/10.
1111/j.1541-4337.2011.00169.x.
WHO, 2015. World Health Organization, foodborne disease burden epidemiology re-
ference group 2007-2015. December 2015. Geneva.
Zhu, J. J., Zeng, X. P., Berkebile, D., Du, H. J., Tong, Y., & Qian, K. (2009). Efficacy and
safety of catnip (Nepeta cataria) as a novel filth fly repellent. Medical and Veterinary
Entomology, 23, 209–216. https://doi.org/10.1111/j.1365-2915.2009.00809.x.