The potential of Carvacrol as

an Active Food Packaging

Agent

Advisor: Shin-Ping, Lin

Student: Anita Chandra Kusuma
ID: MA47110001
“ Introduction
“
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Introduction Unpleasant to eat
and less nutritious

Food spoilage

Serious illnesses

Economic losses

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Garcia and Copetti, 2019
Food Spoilage Prevention
Food processing
enhances the shelf life of food through various ways such
as microorganism control, low-temperature storage,
dehydration, and removal of oxygen

Food additive
Substances that are added to food to maintain or
improve the safety, freshness, taste, texture, or
appearance of food

Food packaging
maintaining the quality of food products
during storage, transportation and
distribution.

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Food Packaging

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Food Packaging
Edible films
produced from agricultural
wastes and/or waste from
industrial food production Intelligent
and thus imparting value
addition
Packaging
utilizes a variety of chemical
sensors or biosensors to
monitor the quality & safety Active packaging
of food products
involves the incorporation
of certain additives inside
the polymer, into packaging
film

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Day, 1989; Kuswandi et al., 2011
Active Packaging

Active Packaging
Incorporation certain components into
packaging system that release or adsorb
substance from or into the packed food or
the surroundings

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Vilela et al., 2018
 Carvacrol
 A monoterpene phenol
 Natural abundant essential oil of oregano, oil
of thyme, oil obtained from pepperwort, and wild
bergamot
 Carvacrol is usually used as a fungicide,
disinfectant, and fragrance in cosmetic
formulations and perfumes
 Has been classified as Generally Recognized As
Safe (GRAS) by the American Food and Drug
Administration

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Ait-Ouazzou et al. 2013; Andersen A., 2006; Ultee et al., 2002
 Carvacrol
 Has antimicrobial activity against fungi
(Fusarium sp., Pseudomonas sp.). Against gram-
positive bacteria (Staphylococcus sp.,
Streptococcus sp. Mycobacterium sp.) And gram-
negative (Salmonella sp., E.coli, Helicobacter sp.)
Due to the presence of the free hydroxyl group,
hydrophobicity, and the phenol moiety
 Has high antioxidant activity by inducing a
significant hepatoprotective and improving the
activity of enzymatic antioxidants

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Ahmad et al., 2011; Kavoosi & Rabiei, 2015; Sharifi‐Rad et al., 2018; Suntres et al., 2015
Aims of seminar
To evaluate the use of carvacrol in
different food packaging applications
1st paper
Introduction
 Packaging acts as a physical barrier, excluding the produce within from
bacterial and/or fungal contaminants in storage environments, hence,
limiting decay development (D’Aquino et al., 2016; Kader et al., 1989).

 Halloysite nanotubes (HNTs) can be used as active carriers for carvacrol,

while exhibiting potent antimicrobial properties and displayed much
higher carvacrol content in the final film (Shemesh et al., 2015).

 PA (Polyamide 6) is characterized by its high melting temperature

(220oC) (Swain and Isayev, 2009), thus PA can withstand in harsh
conditions of melt compounding and film production.

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Aims
To applicate HNTs/carvacrol hybrids
by melt-compounded with PA and
films produced by cast extrusion
Methods
Halloysite HNTs/Carvacrol
Carvacrol
nanotubes hybrids

1:2

Amp 40% at room

temperature

cast extrusion
HNTs/Carvacrol Polyamide 6
hybrids (PA) 16mm using 45 mm
twin-screw screw diameter
25:1 extruder
extruder

150 rpm speed feeding ±50 mm thick films at

rate of 2 kg h-1 at 250oC
250oC
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Film Analysis
 Thermal gravimetric analysis (TGA)
 High-resolution scanning electron microscopy (HRSEM)
 In-vitro antifungal activity
 Fresh product storage
 Carvacrol residue analysis in fresh produce

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Results
 Images of PA/carvacrol(a) and
PA/HNTs/carvacrol(b)

Large holes
and fishnet
structure
Feasible and
High-quality
transparent film

HNTs can protect the carvacrol during high

temperature process
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 HRSEM images of neat HNTs(a),
PA/HNTs/carvacrol(b), higher magnification(c)
PA/HNTs/carvacrol:
HNTs individually dispersed within
the PA matrix

Neat HNTs:
cylindrical Fine adhesion between
shape the individual
nanotubes and the
matrix

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 Carvacrol content in
different films
In vitro antifungal activity
after 21 days incubation

PA/HNTs/Carvacrol retained around 90%

Carvacrol content

PA/HNTs/Carvacrol exhibited high efficiency in

inhibiting fungal growth

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Colony radial growth rates and reduction of
four postharvest molds
exposed to different HNT/carvacrol films

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Images of films on the development of
molds for 135h incubation

Fungal reduced
development

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 Decay reduction for fruits packed after storage

Higher concentration of carvacrol associate with

phytotoxic effect and tissue sensitivity Carvacrol residue in fruit tissue
following 10 days

Carvacrol residues were

detected in all the studied
fruits 22
Cherry tomatoes images from
fresh produce storage trial
(a) Fresh produce tomatoes packed in film at the beginning of the trial
(b) Tomatoes were packed in neat PA film following 51 days of storage at
10oC/90% RH
(c) Tomatoes were packed in PA/(HNTs/2% carvacrol) film following 51
days of storage at 10oC/90% RH

Higher quality due to

fungal inhibition

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Conclusion

 The carvacrol-containing PA films exhibit in vitro antifungal efficacy

against a wide range of fungal molds

 The carvacrol-containing PA films presented excellent performance of

inhibiting the development of decay and extending shelf life.

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2nd paper
Introduction
 Biodegradable polymers such as starch have been studied as a potential
active film because of their good physical, mechanical, and barrier
properties.

 The popularity of starch relates to its abundance, renewability and relatively

low cost; however, starch alone has limitations related to its hydrophilic
nature and lack of antioxidant and antimicrobial properties (Shi et al., 2007;
Souza, Goto, Mainardi, Coelho, & Tadini, 2013).

 Enriching of starch with natural antioxidant and/or antimicrobial substances

will extend the functional properties of it and provide an bioactive
biomaterial (Perazzo et al., 2014; Souza, Ditchfield, & Tadini, 2010).

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Aims
To investigate the properties of
tapioca and gelatinized tapioca
starch dispersions and film and
evaluate the effects of carvacrol on
their properties
 Methods
Tapioca Carvacrol Citric acid
stored at 4oC
starch for at least 100
days
Water

At 140W for 5 Stirred at 800RPM Vacuum in oven and

Stirred at 70oC until
min at 30oC at 40oC equilibrated in
homogenous
For 12 h desicator

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Starch Dispersion Analysis
 Conductivity and pH of starch dispersion
 Zeta-potential of starch dispersion
 Hydrodynamic particle size of starch dispersion
 Apparent viscosity of starch dispersion
 Antioxidant activity of dispersion
 Antimicrobial activity of dispersion

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Film Analysis
 Mechanical behavior of the film
 Solubility of the film
 Water vapor permeability of the film
 Swelling of the film
 Scanning electron microscopy (SEM) image of the film

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Results
pH, conductivity, Zeta-potential and particle size of tapioca
and gelatinized tapioca starch dispersion incorporated
carvacrol

Carvacrol Lipophilic nature of Carvacrol and starch both Carvacrol has a

phenol ring has carvacrol or change in pH
natural low pH and ionic status of OH
group
have slight negative surfactant-like activity and
charge dispersed well in the
starch solution, decreasing
aggregation of the starch
particles
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Viscosity of tapioca and gelatinized tapioca starch dispersion
incorporated with carvacrol at different shear rate

The increased can be

linked to an increase in
the physical interaction
that prevents the
particles from flowing
freely

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Mechanical behaviour of tapioca and gelatinized tapioca starch
film incorporated with carvacrol

The interactions between starch chains could be replaced with new interactions between starch and
carvacrol. It affected polymer chain organization and modified film structure

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Scanning electron microscope image of tapioca starch films

Due to low solubility and

Tapioca starch Tapioca starch/carvacrol hydrophobic of carvacrol,
it can cause formation of
cravacrol crystal that
increases the coarseness
of film

Gelatinized tapioca Gelatinized tapioca

starch/carvacrol

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Antioxidant activity of tapioca
starch dispersion

MIC and MBC of

tapioca starch dispersion

Carvacrol increase the antioxidant activity due to

electron donating capacity of carvacrol

Carvacrol show potent antimicrobial activity

against both gram positive and gram negative
bacteria
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Conclusion

 Carvacrol incorporation induced noteworthy changes in the conductivity,

pH, zeta-potential, and particle size, viscosity, and antioxidant properties of
the starch dispersions.

 Their improved antioxidant and antimicrobial activity suggests the

potential of the starch dispersions and film for applications such as
bioactive film systems.
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3rd paper
Introduction
 Materials at nanometric scale, such as nanofibers are used as food
preservation technique and intended to extend the shelf life of food
products, while maintaining its nutritional and sensory value.

 nanomaterials from natural polymers have high potential for the

substitution of synthetic polymers, due to their biocompatibility,
biodegradability and low cost (L. Kong et al., 2012)

 Starch is widely applied in the food and food packaging industries

because of its versatility, biodegradability, low cost, and abundance (R.
M. Daudt et al., 2014)

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Aims
To develop a method to encapsulate
carvacrol in electrospun nanofibers
derived from soluble potato starch
 Methods
Formic acid Carvacrol in
75% 20% 30% 40%
Potato
starch 50%
Aged for 24
Electrospinning
h

Stirred for 24 h Stirred for 15 min Electrospun at room temperature

Humidity set at 45±2%
Pumped at a steady flow-rate of 0.60
mL.h−1 and applied
voltage of +25.0 kV and –3.0 kV.

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 Nanofibers Analysis
 Rheological measurement of the fiber-forming solutions
 Thermal stability
 Fourier-transform infrared (FT-IR)
 X-ray diffraction (XRD) and differential scanning calorimeter
analysis (DSC)
 Thickness and mechanical properties of nonwovens
 Antioxidant activity
 Antimicrobial analysis of the electrospun nonwovens

02/17/2023 PRESENTATION TITLE 42

Results
 Morphology and size distribution of soluble
potato starch, carvacrol, and starch nanofibers
No carvacrol: ribbon
shaped and a fused
morphology due to low
stretching

Homogeneous, cylindrical, and randomly oriented, suggesting that the

addition of carvacrol increased the volatilization of the solvent

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Rheological parameters of soluble potato starch fiber-forming
solutions loaded with carvacrol

Viscosity
Viscosity

Carvacrol increase the Not significant The elastic modulus

cohesion force between represents the material
molecule stiffness

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 TGA(a) and DTG(b) soluble
potato starch, carvacrol and
starch nanofibers
Weight loss

Carvacrol has been successfully

encapsulated in the starch fibers
and influences the nanofibers’
First derivatives decomposition temperatures as
well as their weight loss

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 FTIR of soluble potato starch,
carvacrol, and starch nanofibers

No new bands were found in carvacrol incorporation. The

material were mixed together physically without any
chemical reaction 47
XRD of soluble potato starch and
starch nanofibers with carvacrol
Antioxidant activity of
carvacrol and soluble
potato starch nanofibers

Carvacrol addition increase

the antioxidant activity which
is suitable for active packages
Carvacrol addition increase
the crystallinity

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 Antimicrobial activity against the foodborne pathogen of
carvacrol in different concentrations

Can promote
gradual release

The electrospun nonwovens presented the same antimicrobial activity of the 1 st - 7th - 15th day with
clear zones of inhibition with 25mm diameter reducing to 19mm diameter in the 30th day

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Conclusion

 The results showed that the electrospun nonwovens are still releasing the
encapsulated carvacrol up to 30 days of storage

 The starch nanofibers loaded with carvacrol presented in this study

exhibited great antioxidant and antimicrobial activities, which may be
useful in applications such as active food packaging systems to extend the
shelf life and ensure the safety of food products.
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Evaluation
Paper 1 Paper 2 Paper 3
Carvacrol
In HNTs - In starch solution
encapsulation

methods Melt-compounded PA Starch dispersion electrospun nanofibers

Anti-bacteria with MIC

Anti-fungal up to 89% Anti-bacteria up to 89%
Anti-microbial and MBC at least 125
growth reduction growth reduction
µg/mL
ABTS up to 167 µg/mL
Anti-oxidant - Up to 83.1%
and DPPH 152 µg/mL
Food aplications Fruit packaging - -

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Summary
Carvacrol can be applied in different food
packaging methods and effectively used
as antimicrobial and antioxidant
scavenger active agent.
Thank you
For your attention!
MIC and MBC
Minimum Bactericidal Concentration (MBC)
Minimum bactericidal concentration (MBC) is the lowest concentration of antibiotics that
kills 99.9% of the inoculum

Minimum Inhibitory Concentration (MIC)

Minimum Inhibitory Concentration (MIC) is the lowest concentration (expressed as mg/L or
μg/μL) of an antimicrobial agent that inhibits the visible in-vitro growth of microorganisms.

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Carvacrol max. dose per day
NHPD (Natural Health Products Ingredients Database)
Acceptable safe limit of carvacrol as a medicinal ingredient to 189 mg (2.7
mg / kg body weight / day) in an average 70 kg healthy adult.

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100 days of stabilization
Dispersions are unstable from the thermodynamic point of view
However, they can be kinetically stable over a large period of time, which
determines their shelf life. The kinetic process of destabilisation can be rather
long (up to several months or even years for some products) 

the dispersed phase maintains a consistent particle size over time and remains
in suspension

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TGA and DTG
TGA (thermogravimetric analysis): the amount of materials removed in
% against temperature

DTG (first derivatives): tells about the rate at which these materials are
removed in %/min

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