Postharvest Biology and Technology 111 (2016) 380389

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Postharvest Biology and Technology

journal homepage: www.elsevier.com/locate/postharvbio

Design of biodegradable bio-based equilibrium modied atmosphere

packaging (EMAP) for fresh fruits and vegetables by using
micro-perforated poly-lactic acid (PLA) lms
Antonis Mistriotisa,* , Demetrios Briassoulisa , Anastasios Giannoulisa ,
Salvatore DAquinob
a
Agricultural University of Athens, Laboratory of Agricultural Structures, 75 Iera Odos, Athens, 11855 Greece
b
National Council of Reserch of ItalyInstitute of Sciences of Food Production (ISPA-CNR), Traversa La Crucca, 3, loc. Baldinca, Li Punti - 07100 Sassari, Italy

A R T I C L E I N F O A B S T R A C T

Article history: Equilibrium modied atmosphere packaging (EMAP) is used for prolonging the shelf life of fresh produce
Received 12 February 2015 such as fruits and vegetables and preserving their quality characteristics such as freshness, colour and
Received in revised form 3 September 2015 aroma. However, the resulting extensive use of plastic packaging lms raises serious environmental
Accepted 14 September 2015
concerns. Replacing conventional plastic lms with novel biobased and biodegradable under composting
conditions materials such as lms made of poly-lactic acid (PLA), offers new sustainable solutions to the
Keywords: problem. In the present work, the gas transport properties of three layer PLA lms have been studied in
EMAP
relation to their potential as EMA packaging materials for fresh produce. A simple one-dimensional
PLA lm
Gas diffusion
model simulating the combined gas diffusion through micro-perforations and the permeable membrane
Micro-perforation was developed as a designing tool for PLA based EMA packaging systems. By using this model, innovative
Horticultural produce packaging biodegradable bio-based EMAP systems were designed specically for selected fruits and vegetables
Bio-based materials using laser microperforated, water vapour (WV) permeable PLA lms. It was shown that the relatively
Bio-based packaging high water vapour permeability of PLA lms compared to conventional OPP lms allows the development
of an optimised biodegradable bio-based EMA packaging for the specic applications.
2015 Elsevier B.V. All rights reserved.

1. Introduction the temperature increases O2 requirements from tissues increase

Fruits and vegetables are two of the main agricultural product
categories in Europe. Packaging is particularly important for
exported fruits as it allows preserving their quality and health
safety and facilitates labelling of the produce. Recently, advanced
techniques such as equilibrium modied atmosphere packaging
(EMAP) for dynamically modifying the in-package atmosphere are
used for extending the shelf life of high value fresh produce
(Sandhya, 2010).
Successful applications of EMAP include a wide variety of
produce, such as broccoli orets, cauliower orets, carrots, baby
carrots, peeled garlic (Lee et al., 1996). However, most of the
published studies for recommended CO2 and O2 concentrations
generally represent the conditions in the temperatures range of 0
5 ! C that maximize the storage life of each commodity (Fernndez-
Trujilio et al., 1988; Malakou and Nanos, 2005; Irtwange, 2006). As
* Corresponding author. Fax: +30 2105294023.
E-mail address: briassou@aua.gr (A. Mistriotis).
too, while tolerance to CO2 decreases (Kader, 1997).
The main disadvantage of EMAP, like any other conventional
food packaging, is the extensive use of plastic lms resulting into
huge quantities of domestic plastic wastes. Global plastic lm and
sheets market is expected to reach 70.9 million tons by 2018
(Markets and Markets, 2014). A large percentage of this quantity is
used for food packaging. Although recycling of these materials
increased during the last decade, only a small part of the generated
amount of plastic waste is nally recycled (Themelis et al., 2011).
Therefore replacing non-degradable conventional plastics based
on fossil oil with sustainable bio-based biodegradable materials is
of great environmental importance. Various bio-based plastics
have been studied already as possible alternatives to conventional
packaging materials (Siracusa, 2008).
Such a promising biodegradable plastic material is poly-lactic
acid (PLA). PLA was introduced as a packaging material and started
being produced at a commercial level about ten years ago. It has
attracted attention mostly because of its sustainable nature: it is
synthesized from processed corn or other naturally produced

http://dx.doi.org/10.1016/j.postharvbio.2015.09.022
0925-5214/ 2015 Elsevier B.V. All rights reserved.
 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389 381

carbohydrates, thus it is a bio-based material, and it biodegrades contribute to a longer shelf-life of the packaged produce (Almenar
after use under industrial composting conditions (Auras et al., et al., 2008; Briassoulis et al., 2013).
2004). The main objective of the present work was the development of
a one-dimensional steady state analytical model of the in-package
PLA can be used for the development of transparent thin lms
atmosphere assuming that WV transport process through the
suitable for food packaging (Auras et al., 2004). PLAs certied
permeable packaging lm obeys Ficks law. Using this analytical
compostability and compliance with the food contact safety
model, packaging bags made of PLA have been optimally designed
regulations (Auras et al., 2004) makes it attractive as packaging
and developed for two selected horticultural produce: cherry
material, since it meets the compostability requirements of
tomatoes and peach, as typical high value vegetables and fruits
EN13432 for packaging (EN 13.432, 2005), thus alleviating the
respectively. These optimized packaging systems were tested
plastic wastes problem. PLA exhibits high water vapour (WV)
through full-scale experiments to conrm the validity of the design
permeability compared to conventional plastic packaging lms,
methodology.
while its permeability to CO2 and O2 is comparable.
The performance of an EMA packaging greatly depends on its
2. Materials and methods
optimised design which has to be tailored to the needs of the
specic packaged produce. Gases are exchanged between the
2.1. Denitions and terminology
package and the environment by a restricted diffusion process
resulting into an EMA in the package. In a typical EMAP the
Before presenting the relevant methodology and the results of
synthesis of the in-package atmosphere is controlled by selecting
the present work, it is important to clarify the denitions related to
the gas transport properties of the packaging lm. The equilibrium
the diffusive gas transport through membranes in order to avoid
(steady state) modied atmosphere (EMA) is obtained when the
confusions, as the relevant terminology is not unique in the
exchange rate of gases through the covering lm is in equilibrium
literature. The gas barrier property of a material is expressed by its
with the production or consumption of gases at the respiring and
gas mass permeability P (kg m m"2 s"1 Pa"1). This is dened as the
transpiring produce. Therefore, the gas transport properties of the
gas mass (kg) which, under steady conditions, crosses unit area
packaging system have to be designed to meet the specic
(m2) and unit thickness (m) of the specimen, in unit time (s) under
requirements of the packaged produce. A technique for adjusting
unit pressure difference (Pa) and at constant temperature. In
the gas transport properties of a packaging lm is perforation. Both
particular, the permeability of a plate or membrane is expressed by
macro-perforation by mechanical means (Van der Steen et al.,
the one-dimensional Ficks law of diffusion as:
2002) and laser made micro-perforation (Ozdemir et al., 2005) are
used for this purpose. In the case of a perforated lm, the diffusion dp
F "PA 1
is not spatially uniform and the combined transmission effect of dx
both perforations and lm has to be investigated. The size and the
where F (kg s"1) is the mass ow rate of the gas, dp=dxis the
number of perforations have to be estimated to meet the specic
gradient of the gas pressure across the specimen (Pa m"1) and A
needs of the packaged fruits or vegetables for a given packaging
(m2) is the area of the membrane. If a gas mixture is considered, gas
conguration. For this reason, the development of a model
pressure, p (Pa) refers to the partial pressure of a gas in the mixture,
predicting the gas transfer through a perforated lm is a necessary
thus it is a measure of the molar fraction, c, of the gas in the
tool for designing the perforation pattern. Such a model has to take
mixture.
into account the combined gas transport through both the
In the case of a gas mixture, the partial pressure gradient can
packaging lm and the perforations (Paul and Clarke, 2002;
also be expressed as the molar fraction gradient of the gas,
Rennie and Tavoularis, 2009; Briassoulis et al., 2013).
dc=dx(m"1). Then the mass permeability is expressed in (kg m
Several attempts have been made to model gas transfer through
m"2 s"1) and the volume permeability in m2 s"1.
a perforated permeable lm for optimizing EMAP (Paul and Clark,
When a thin lm is considered, its gas transfer characteristics
2002; Rennie and Tavoularis, 2009). These models focus on the gas
are described by the permeance, PR. In the Standard ASTM-E96/
diffusion through a perforation (Chung et al., 2003), while the lm
E96-05 (ASTM, 2005) water vapour permeance PR (kg m"2 s"1) is
itself is considered impermeable (barrier lm) (Chung et al., 2003;
dened by the equation:
Gonzalez et al., 2008). Such modelling approaches are based on
Ficks law of diffusion (Chung et al., 2003) or StefanMaxwell law F P
PR 2
(Rennie and Tavoularis, 2009). The simplest model assumes Ficks A Dc L
diffusion in a cylindrical pore lled with stagnant air simulating where L (m) is the thickness of the thin lm.
the perforation. However, an end correction term for the diffusive An additional, practically used measure of the barrier property
path length must be introduced to compensate for end effects at of a lm is its gas transmission rate, TR. In the Standard ASTM-E96/
the mouths of the hole (Chung et al., 2003). Certain models also E96-05 the transmission rate TR (kg m"2 s"1) of a gas through a lm
take into account convection mass transfer in the headspace of thickness L (m), that crosses unit area (m2) of the specimen, in
between the produce and the packaging lm due to temperature
unit time (s) for a specic molar fraction difference Dc, is then
differences (Rennie and Tavoularis, 2009). However, this effect has
dened as:
a limited importance.
F
Relevant to the present work are studies concerning the use of TR PR & Dc 3
A
lms made of other biodegradable materials for food packaging
(Avella et al., 2005; Siracusa et al., 2008). Most of these bio-based
lms are rather permeable in water vapour. The gas transport 2.2. The modied Ficks law for gas diffusion through a hole
properties of these lms are particularly interesting for further
optimizing EMAP systems. The diffusion coefcient D (m2 s"1) of a gas in a gas mixture,
The present work focuses on the use of micro-perforated PLA such as air, is dened as the diffusion volume ow rate FD (m3 s"1)
dc
lms for designing optimized equilibrium modied atmosphere divided by the molar fraction gradient of the gas, dx , per unit area.
packaging (EMAP) for fresh fruits and vegetables. Recent studies The gas diffusion through a single perforation (e.g., an
indicate that PLA is a versatile packaging material, which may impermeable lm with one hole) may be modelled as diffusion
382 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
through a tube of nite length. Diffusion at the mouth of a tube
differs from the diffusion at its interior due to end effects. If the
aspect ratio (thickness/diameter) of the tube (hole) is high then the
diffusion through this hole is described well by Ficks law. On the
other hand, low aspect ratios mean that end effects are important.
An empirical modication of Ficks law (Eq. (4)) incorporating end
effects makes this equation appropriate for describing diffusion
through perforations (Chung et al., 2003):
Dc
FD DA
L kd
In this case the gas volume ow rate FD (m3 s"1) is the diffusion
ow rate through a hole, D (m2 s"1) is the diffusion coefcient of
the considered gas in air, L (m) is the thickness of the lm, Dc is the
molar fraction difference of the gas across the hole, A (m2) is the
area of the perforation, d (m) is the perforation diameter and k is a
phenomenological factor describing the end effects of the
perforation (Chung et al., 2003).
Typical k factor (Eq. (4)) values, representative of the end effects
of the perforation are provided in the literature. They range
between 0.4 and 1.0 (Gonzalez et al., 2008). For the extreme value
of k = 0 this empirical law corresponds to Ficks law.
2.3. Experimental methods and materials used for investigating the
gas permeability of PLA lms
Water vapour (WV), O2, and CO2 are the most relevant gases in
EMAP for horticultural produce. For this reason, the transport
properties of PLA lms with respect to these gases have been
investigated. In the experimental investigation three-layer PLA
lms of 30 mm thickness produced by Taghleef Industries
(NATIVIA) were used. In these lms, the core PLA is coated on
both sides by a thin layer of sealable PLA.
The WVTR was gravimetrically measured using the water
method described in the ASTM-E96/E96-05 Standard (ASTM,
2005). Specially designed dishes of 80 mm diameter able to tightly
hold a sample lm at their open top, were developed. In each test
dish, 20 ml of distilled water were added leaving a distance of
approximately 8 mm between the water surface and the lm. The
dishes were placed in a controlled environment room with
temperature 23 ( 0.5 ! C and RH equal to 50 ( 5% and their weight
was measured once per day by a balance (model OHAUS Ts4kd)
with accuracy of 0.01 g. The decrease of the weight of each dish was
found linear with time, and the regression slope was used for
estimating the WV mass ow rate F (kg s"1) through the lm. The
WVTR (kg m"2 s"1) was calculated from Eq. (3).
Fig. 1. The dish model: (a) Geometry of a dish model covered by a perforated membrane, (b) dish model mesh.
A ask of volume equal to 2 L was sealed by the studied PLA lm.
An amount of CO2 was blown in the ask resulting to an initial CO2
molar fraction with respect to air in the ask equal to co. The molar
fraction, c(t), of the CO2 was measured using an IR CO2 sensor
(QUBIT S147, QUBIT Systems Inc.) as a function of time. Due to the
transmission of CO2 through the PLA lm, its molar fraction
decreased exponentially with time. Then the CO2 permeability P
(kg m m"2 s"1) was calculated from Eq. (5):
ct co e"
PA
rVLt 5
4
2 3
where: A (m ) is the surface of the lm, V (m ) is the volume of the
ask, r (kg m"3) is the density of CO2, and L (m) is the thickness of
the lm.
Oxygen permeability was measured by a similar method. An
amount of nitrogen was blown into a ask to reduce the molar
fraction of oxygen with respect to air. The volume of the ask was
equal to 2 L. Then the oxygen molar fraction was measured using a
paramagnetic O2 sensor (QUBIT S147, QUBIT Systems Inc.). The
oxygen permeability, P (kg m m"2 s"1), was calculated from Eq. (6):
PA
ct catm " co e" rVLt 6
where: r (kg m"3) is the density of O2,co is the initial molar fraction
of O2 in the ask, catm (%) is the oxygen atmospheric molar fraction,
and catm-co (%) is the oxygen molar fraction in the ask at time 0.
Both measurements of CO2 and O2 permeability were
performed in a controlled chamber environment where the
temperature was 23 ( 0.5 ! C and the RH was 50 ( 5%.
2.4. Experimental methods and materials used for investigating the
gas permeability through perforations
The value of k of Eq. (4) was experimentally determined for the
specic material studied in this work. Since the factor k is
independent of the gas properties, its value can be estimated by
studying the diffusion of any typical gas through a perforation in a
barrier lm. In the present work, water vapour was used as the
tracer gas for validating Eq. (4). The WVTR of holes with diameter
in the range between 0.16 mm perforated in a typical 30 mm thick
PLA lm (Taghleef IndustriesNATIVIA) was measured.
The gas transport through a lm with multiple perforations was
also studied. Water vapour was used as the tracer gas. The WVTR
was measured by using the Standard testing method ASTM-E96/
E96-05. In this case an OPP lm was used instead of PLA for
covering the test dishes in order to investigate the gas transport
through multiple perforations in an impermeable lm. The
analysed cases had 1, 2, 3 and 4 perforations of equal diameter
 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
of 100 mm. The holes were perforated along a line at 1 cm distance
from each other.
2.5. Numerical modelling of gas transport through perforations on
lms
Gas diffusion through perforations was studied by 3D numeri-
cal simulations using the commercial software ANSYS CFX 13.0. A
rigid dish simulating the equipment used in the ASTM-E96/E96-
05 Standard was numerically modelled. The dish had 40 mm radius
and 8 mm height, and it was covered at the top by an impermeable
membrane perforated at its centre. The elements along the
circumference of the perforation had edges of a minimum length
equal to 0.2 mm (Fig. 1). As the grid expanded to the membrane
surface away from the perforation the elements became larger
(maximum edge length 2 mm). In this way the accuracy of the
solution is higher close to the perforation where the gas exchange
occurs. The entire test dish model was discretized using
220.000 elements and 41.000 nodes. The model was also
discretized through ner grids but the results showed no
sensitivity.
All surfaces of the test dish model were considered imperme-
able to gas exchange except for the bottom surface and the
perforation at the upper surface of the model. The diffusion
coefcient, D, of the WV in air was set equal to 2.5 & 10"5 m2 s"1 as
given in literature (Massman, 1998). The lower surface was
modelled as an inlet (simulating the water surface) with constant
RH value of 100% while the perforation was treated as an opening
(exchanging WV with the environment). The pressure of the
external environment of the dish (and the hole) was set equal to
zero (WV expands to a signicantly larger area) and the RH was
assumed to be equal to 50% (ambient conditions). Convergence
was reached when the WV mass fraction residual was smaller than
10"7.
The WVTR as a function of the perforation diameter was
analysed by numerical simulations. Gases were transported
through a single perforation located at the centre of an
impermeable membrane covering the top of a test dish model
(Fig. 1). The simulation was performed for different perforation
diameters and the diffusion gas volume ow rate through the
single hole was measured. The smaller perforation diameter in the
numerical model was set equal to 2.5 mm. Other perforations
tested had size equal to 2, 4, 6, 8 and 20 times the initial diameter.
A similar test dish with impermeable surfaces, of the
dimensions shown in Fig. 1 was simulated numerically to
investigate the effect of multiple perforations. The grid of the
numerical model consisted of tetrahedral elements with similar
dimensions to the previous test. Elements and corresponding
nodes were slightly increased around each additional perforation
to improve mesh quality. Boundary conditions were the same as in
the numerical simulation of a lm with a single perforation.
Convergence was set again to 10"7 of the WV mass fraction
residual. All perforations had the same diameter, equal to 5 mm
and the lm thickness was 1 mm. Four perforations were
Table 1
Physiological properties and targeted gas molar fractions for the EMAP applications.
Produce Respiration activitya
(mg kg"1 s"1)
Cherry tomatoes 58
Peaches 1121
a
Measured at the laboratory of CNR-ISPA.
383
positioned at the vertices of a square of a 5 cm edge. Simulations
were performed when 1, 2, 3 and 4 perforations were open.
2.6. Experimental set-up for testing PLA-based EMAP design for
horticultural produce
Two full-scale experiments were performed on cherry toma-
toes and peaches respectively to demonstrate the validity and the
performance of the proposed model for designing PLA based EMAP
systems. The targeted in-package CO2 and relative humidity for
cherry tomatoes and peaches, reported in Table 1, were determined
from experiments conducted in ISPA-CNR laboratory (HORTIBIO-
PACK, 2011) and from results reported in the literature (Crisosto
and Kader, 2004; Crisosto and Valero, 2008; Ramina et al., 2008;
Saltveit, 2005; Sargent and Moretti, 2004)
The equipment used in these experiments included:
1 A data logger measuring humidity and temperature inside the
bags (LogTag Humidity & Temperature Recorder, Auckland, New
Zealand).
2 A 3300 10 Varian gas-chromatograph equipped with a ame
ionization detector (FID) and a packed column 11 (Unibeds1,
Alltech, Milan) to determine ethylene. Helium was used as
carrier gas at a ow rate of 7 mL s"1. The temperatures of
injector, oven and detector were 150 ! C, 60 ! C, and 200 ! C,
respectively.
3 A combined CO2/O2 analyzer for CO2 determination, a (Combi
Check 9800-1, PBI-Dansensor 14 A/S, Denmarck).
4 A hand-held analyzer (Check Point, PBI-Dansensor Italia Srl,MI,
Italy) for determining in-package CO2 and O2 concentrations.
5 All data were statistically analysed using Stagraphics Centurion
XV (Statpoint, Herndon, VA).
2.6.1. Cherry tomatoes
Two packaging congurations were designed for the cherry
tomatoes named PLA80 and PLA90. The targeted in-package CO2
was 34 kPa, while the targeted in-package RH was 80% (PLA80)
and 90% (PLA90) respectively (Table 1). These experimental
packaging bags made of micro-perforated PLA lms were designed
following the methodology presented in Sections 3.2 and 3.3.1.
Fifteen bags were used for each EMAP design and each bag
contained 20 cherry tomatoes (cv Creative), whose total weight
was around 250 g. OPP (30 mm thick) bags with a total area of
650 cm2 and four 6-mm macro-perforations were used as a
conventional packaging system. Additionally, 200 tomatoes divid-
ed into 10 unwrapped trays (20 fruits per tray) served as control.
Since pathogens growth affects fruit respiration and ethylene
production rates, in-package gas composition was monitored in
bags containing tomatoes treated with a fungicide. For this
purpose, the fruits of ve bags per packaging conguration
(PLA, OPP) were dipped before packaging for 60 s in a fungicidal
solution at 20 ! C containing 600 mg/L udioxonil (Scholar,
Syngenta Crop Protection, Milan, Italy). In all cases, tomatoes
were laid directly into the bags without any tray (Fig. 2a). Inside
Transpirationa rate Targeted CO2 molar fraction (%) Targeted RH
(mg kg"1 s"1) (%)
3570 34 8090
3546 13 8590
384 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
Fig. 2. (a) A typical PLA80 bag used in the validation experiments containing 250 g of cherry tomatoes. (b) Typical PLA bags used in the validation experiments containing
700 g of peaches; indicative measurement of CO2 and O2 in one of the bags is shown.
three of the bags containing fruits treated with the fungicide, a data
logger was placed recording temperature and relative humidity
every four hours.
The fruits of the remaining ten bags per packaging congura-
tion (PLA, OPP), which were not treated with udioxonil, were
stored in a storage room where temperature and RH were set at
20 ! C and 6065% RH, respectively. Three data loggers were also
placed in different points of the storage room to verify if the xed
conditions would be respected. The fruits were stored for 11 days
and inspected every 23 days for the presence of decay, expressed
as the percentage of rotten fruit. Weight loss was also determined
as the percentage reduction of the initial weight. Fruit appearance
was evaluated on a hedonic scale ranging from 1 to 9, where
9 corresponds to fruits as fresh as at harvest; 5 corresponds to
marketability limit and 14 indicate unmarketable fruits. In-
package CO2 and O2 concentrations were determined by the hand-
held analyzer whose needle was inserted through the same hole
done for ethylene sampling.
2.6.2. Peaches
The experiment was conducted on Dixired peaches harvested
from a local commercial orchard in Sardinia, Italy at commercial
maturity. The fruits were sealed in PLA bags, whose in-package
targeted conditions were 13% CO2 and 85% RH (Table 1). Each bag
was containing 4 peaches with a total weight around 700 g and it
was designed following the methodology presented in Sections 3.2
and 3.3.2. The experiment included the following congurations:
(1) unwrapped fruits (CNT), which served as control; (2) fruits
sealed in PLA bags, and (3) fruits sealed in OPP bags (OPPmacro) of
the same size as PLA bags but with four macro-holes of 6 mm
diameter as a conventional packaging system. Twenty bags were
prepared for each of the considered packaging conguration.
Additionally, 80 peaches divided into 20 unwrapped trays (4 fruits
per tray) served as control. Both the sealed and the unwrapped
fruits were stored at 20 ! C and 6065% RH for 3, 6 or 9 days. At each
Table 2
Gas transport properties of PLA lms.
Permeability (kg m m"2 s"1)a Flexible lm
Permeance (kg m"2 s"1) 30 mm (current experiments)
WV Volume permeability 1.4 ( 0.3 & 10"9
CO2 Volume permeability 4.4 ( 0)1 &10"12
O2 Volume permeability 5.3 ( 0)4 & 10"12
WV Volume permeance 6.4 ( 1 &10"5
CO2 Volume permeance 2.0 ( 0)1 &10"7
O2 Volume permeance 1.7 ( 0)1 &10"7
a
Variations are due to the variability of the properties (thickness, coatings) of the lms used in these measurements.
inspection time weight loss, appearance and decay incidence were
determined.
In-package CO2, O2 and C2H4 were determined on additional
ve PLA and ve OPP bags (Fig. 2b) containing fruit treated with
udioxonil to prevent pathogens growth. The same bags were used
to monitor temperature and relative humidity in four-hour
intervals (three data loggers per packaging conguration). All
measurements and analyses were carried out following the same
methods used for cherry tomatoes.
3. Results and discussion
3.1. Analysis of the gas transport properties of perforated PLA lms
As a necessary rst step for developing an analytical model for
designing an optimal bio-based EMA packaging system, the gas
transport properties of perforated PLA lms were investigated by
experimental and numerical methods. In particular the overall
transmission rate due to gas diffusion through a permeable lm
with multiple perforations was investigated:
1 The gas transport properties of a typical commercial PLA
packaging lm used in the current experiments were measured.
2 The gas diffusion through a single perforation was studied. The
gas transmission rate, TR, dependence on the perforation
diameter was experimentally and numerically determined. In
this way, the modied Ficks law (Eq. (4)) was conrmed and the
value of the empirical parameter k was determined for the
packaging material used in the present work.
3 The effect of multiple perforations on the overall gas transmis-
sion rate of the lm was examined by both experimental and
numerical techniques.
4 The interaction of neighbouring perforations and the resulting
combined diffusion ow rate were numerically investigated as a
function of the distance between the holes.
Bulk material (Auras et al., 2004) Flexible lm
30 mmtypical values (Taghleef Industries, 2011)
1.8"2.0 & 10"9
1.8"2.8 & 10"12
3.1 &10"12
4.0 & 10"5
2.7 & 10"7
 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
3.1.1. Gas transport properties of PLA lm
PLA lms offer selective permeability with respect to the three
gases which are important in fresh produce packaging, namely O2,
CO2 and H2O (WV).
The measured gas transport properties of the three-layer PLA
lms used in these experiments (Section 2.3) are shown in Table 2
and compared to the corresponding values found in the literature.
The high WV permeability of PLA lms compared to CO2 and O2
(1000 times larger), allows for regulating in-package RH by
selecting a suitable surface area of the permeable to WV packaging
lm without interfering with the control of in-package CO2 molar
fraction. The CO2 and O2 molar fractions are controlled indepen-
dently through the micro-perforation design.
3.1.1. Gas transport through a single perforation of various diameters
As Eq. (4) indicates, the diffusion ow rate of a gas volume
through a hole is not proportional to its area. Actually, the
corresponding transmission rate (TR) decreases as the hole
diameter increases due to end effects.
The WVTR through a single perforation, measured with the
experimental set-up of Section 2.4, with the hole diameter in the
range between 0.1 and 6 mm perforated in a typical 30 mm thick
PLA lm, is shown in Fig. 3. Using the following Eq. (7), which is
derived from Eq. (4), k was estimated equal to 0.43 by a linear
interpolation of the experimental data of Fig. 3.
DA Dc
" L kd
FD
The numerically determined value of k was found equal to 0.30,
which is smaller than the experimentally measured value, 0.43
(Fig. 3). The experimental result agrees better with previous
experimental research works (Gonzalez et al., 2008) that deter-
mined parameter k in the range between 0.4 and 1.0. The difference
between the experimentally determined value of k and the one
obtained by numerical simulations is a result of the geometric
details of the perforation edge. Manually perforated or laser made
perforations have rougher edges compared to an ideal numerically
modelled hole. Therefore, the edge effects are weaker than reality
in the case of the numerical simulation.
3.1.2. Gas transport through multiple micro-perforations
The experimental and numerical results on the effect of
multiple perforations on a packaging lm are presented in
Fig. 4a. The results are presented in terms of the overall TR
through a perforated lm normalized by the TR of a single
perforation. Both experimental and numerical data show that the
7.0E-03
6.0E-03
5.0E-03
D.A.c/FD -L=kd (m)
4.0E-03
y = 0.30x
R = 0.999
3.0E-03
y = 0.43x
R = 0.98
2.0E-03
1.0E-03
0.0E+00
0 0.005 0.01 0.015 0.02
Perfora"on Diameter d (m)
Fig. 3. Estimation of the factor k based on laboratory experiments and numerical
simulation for PLA lm.
385
TR increases proportionally to the number of the perforations. The
interaction between neighbouring perforations was also studied
numerically (Fig. 4b). It was shown that the total transmission rate
through a pair of perforations decreases with the distance between
the two holes due to their interaction. However, this effect
becomes negligible when the ratio L/d is smaller than 5, as it is
shown in Fig. 4b. This result explains the small deviation of the
numerical results from the exact linear relationship observed in
Fig. 4a.
The combined outcome of the gas transport experiments and
analyses through a single and multiple perforations indicates that
multiple perforations result into higher TR than a single hole of
equal total open area. Moreover, the results presented in Fig. 4
show that there is a linear relationship between the TR of a
perforated lm and the number of the perforations provided that
their distance is not smaller than 5 times their diameter. This fact
simplies the EMA package modelling and design.
3.2. Modelling of PLA-based EMAP systems for horticultural produce
Based on the above experimental and numerical results an easy
to use 1D model was developed for designing the PLA based EMAP
system. The basic assumptions of this model are the following:
1. Gas transport through a perforated membrane takes place
through both the lm and the perforations.
2. Gas transport through a non-perforated lm is governed by
7 Eq. (2). The WV and CO2 permeance PR (kg m"2 s"1) of a PLA lm
of 30 mm thickness is taken equal to 6.4 & 10"5 kg m"2 s"1 and
2.0 & 10"7 kg m"2 s"1 respectively based on the data presented
in Table 2.
3. Gas transport through a hole of the lm (perforation) is
governed by the modied Ficks law (Eq. (4)), where k is taken
equal to 0.5 as approximately measured (Fig. 3).
4. Gas diffusion volume ow rate through multiple perforations is
the superposition of ow rates of separate holes assuming that
the distance between holes is at least larger than 5 times their
diameter.
The design of the EMAP system has to take into account that
two independent physiological processes of the fruits need to be
regulated, namely the respiration and the transpiration. Transpi-
ration is a more intense process producing 20 times larger volume
of WV compared to the CO2 volume produced through respiration.
Considering that diffusion coefcient, D, of WV in air is 25 & 10"6
m2 s"1 compared to 14 & 10"6 m2 s"1 of CO2 (Massman, 1998), the
perforation pattern cannot be designed for regulating simulta-
neously both respiration and transpiration. Therefore, one of these
two gases has to be exchanged through a selectively permeable
covering material such as PLA lm.
The basic design concept is the following: If perforation is
designed to regulate in-package CO2 molar fraction at the desired
level, its effect on the in-package RH is negligible due to the much
higher transpiration rate compared to respiration. However, PLA is
a hydrophilic material with WV permeability much higher than its
Experiment CO2 permeability. Therefore, when PLA lm is used as a packaging
Numerical material in EMAP systems, the perforation pattern can be designed
for controlling only the CO2 molar fraction, while the area of the
packaging lm surface can be designed for controlling the in-
package RH. In this way, the in-package atmosphere can be
adjusted to the required composition with respect to all relevant
0.025
gases that are important for a specic horticultural produce.
Since the O2 permeability of PLA is similar to CO2, oxygen molar
fraction can also be efciently regulated through the perforation
together with CO2 molar fraction.
386 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389

Fig. 4. (a) The overall WVTR through multiple perforations as a function of the number of perforations; (b) Transmission rate through a pair of perforations normalized by
twice the transmission rate of a single perforation as a function of their relative distance L normalized by their diameter d.

3.3. Optimised design of PLA-based EMAP for specic horticultural
produce
The 1D analytical model described above was applied to two
case studies, namely cherry tomatoes and peaches. Optimal EMAP
bags made of micro-perforated PLA lm were designed for these
two high value horticultural produce in order to validate the
model. The design of the optimised EMAP systems was based on
the respiration and transpiration ow rates and the targeted in-
package RH and CO2 molar fraction which are presented in Table 1.
3.3.1. Cherry tomatoes
The transpiration rate of cherry tomatoes was measured in the
laboratory to be in the range 3570 mg of WV kg"1 s"1, under
storage conditions 20 ! C temperature and 85% RH. Accordingly, a
typical transpiration rate was considered approximately equal to
58 mg of WV kg"1 s"1 at the selected conditions. Similarly,
respiration rate of cherry tomatoes was measured to be between
5.2 and 7.9 mg CO2 kg"1 s"1. Based on the targeted headspace
values shown in Table 1, the surface of the packaging material and
the number of perforations were determined to t the targeted RH
(8090 %) and CO2 molar fraction (34%) respectively.
PLA lms of 30 mm thickness were used with laser perforations
of 200 mm diameter. The CO2 transmission rate of the micro-
perforations of 200 mm diameter were calculated by using the
modied Ficks law (Eq. (4)). By applying this equation for such a
hole on a lm of 30 mm thickness, the CO2 transmission rate of a
single hole was calculated equal to 0.94 mg/h if the targeted CO2
molar fraction is 4%. Therefore, 2021 holes per kilogram of cherry
tomatoes will generate a steady state EMAP with CO2 molar
fraction equal to 4%. Hence, 5 holes of 200 mm are necessary for
obtaining the optimal EMA for a package of 250 g cherry tomatoes
in terms of CO2 and O2.
A package of 250 g of cherry tomatoes at 20 ! C and RH 85%
which is within the targeted EMAP range approximately produces
14 mg s"1 of WV due to transpiration (Table 1). The RH in the
storage room was set equal to 65%. Two different values of the in-
package RH were targeted, 80% (PLA80) and 90% (PLA90). These RH
values at 20 ! C correspond to molar fraction differences equal to
0.0036 and 0.0060 respectively, calculated from the psychrometric
diagram. Using Eq. (3), the WVTR of a PLA lm of 30 mm thickness
was calculated from the permeance shown in Table 2, equal to
0.22 mg m"2 s"1 for the case (PLA80), and 0.35 mg m"2 s"1 for the
case (PLA90). Assuming that in equilibrium the WV ow rate
through the total area of the packaging lm has to be equal to the
transpiration rate of the total packaged produce, the area of the PLA
lm required for obtaining the necessary WV removal rate for a
250 g package of cherry tomatoes was calculated equal to 650 cm2
and 400 cm2, respectively for 80% (PLA80) and 90% (PLA90) in-
package RH at 20 ! C.
3.3.2. Peaches
The respiration and the transpiration rates of peaches are
shown in Table 1. The respiration rate of peaches was measured to
vary between 11.4 and 20.8 mg CO2 kg"1 s"1. For designing the
perforation pattern, the respiration rate of peaches was selected
equal to 18 mg CO2 kg"1 s"1. The transpiration rate was measured in
the range of 3546 mg of WV kg"1 s"1 in an environment of 20 ! C
temperature and RH = 85% (Table 1).
The same PLA lms of 30 mm thickness were used as the
packaging material with laser perforations of diameter equal to
200 mm. A lower CO2 molar fraction compared to cherry tomatoes
(Table 1), specically 13%, was targeted for peaches. In the
case of peaches 100 holes of 200 mm are required for obtaining
the targeted value of 2% CO2 for a typical package of 700 g of
peaches.

Fig. 5. Changes in weight loss (a) and decay incidence (b) of cherry tomatoes cv Creative during storage. Vertical bars represent the standard deviation (n = 10 for weight loss
and 200 for rotten fruit).
 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
The area of the packaging lm had to be adjusted accordingly.
The transpiration rate of peaches at 20 ! C and RH in the range of
8590%, which is the targeted EMAP value, was measured equal to
3546 mg of WV kg"1 s"1 (Table 1). Assuming as a typical case a
transpiration rate equal to 35 mg of WV kg"1 s"1, a typical package
of 700 g of peaches produces 24 mg s"1 WV at the targeted in-
package RH equal to 85%. The RH in the storage room was set equal
to 65%. The WVTR of the packaging lm for this RH difference was
calculated equal to 0.27 mg m"2 s"1 from the data of Table 2 using
Eq. (3) with the help of the psychrometric diagram. Considering
that in equilibrium the WV ow rate through the total area of the
packaging lm has to be equal to the transpiration rate of the total
packaged produce, the area of the PLA lm required for obtaining
the necessary WV removal rate for a 700 g package of peaches was
estimated equal to 900 cm2.
3.4. Experimental resultsperformance of PLA-based EMAP for
horticultural produce
3.4.1. Cherry tomatoes
During the full scale experiment, the average ambient
temperature in the storage room was 19.5 ( 1.5 ! C. The RH in the
storage room was 65 ( 4%, showing marked uctuations. As a
result, RH also uctuated inside the packages, although at a lesser
extent. The average package RH was 79.8 ( 2% and 85 ( 2% in the
PLA80 and PLA90 bags, respectively, values which are very close to
the expected ones, especially in PLA80. In contrast, in OPP bags RH
was always higher than 90%, with peaks of 9598%, despite the four
6-mm holes per bag, while the presence of condensation inside the
packages was frequently detected.
Weight losses of unwrapped fruit after the eleven days of
storage were 7.7% (Fig. 5a), with an average weight loss per day of
6.9 g kg"1. Fruit sealed in PLA80 and PLA90 bags at the end of
storage lost 5.09% and 4.66% of their initial weight respectively,
which is equivalent to 4.6 and 4.2 g kg"1 per day. This measured
transpiration rate is within the range of 3570 mg kg"1 s"1
reported in Table 1. The weight loss in OPP bags were only 3.3%,
equivalent to 3 g kg"1 per day (Fig. 5a).
The results presented in Fig. 5a also show that the standard
deviation of weight losses is much smaller in both PLA and OPP
packages compared to unwrapped fruits. This indicates that the
PLA packaging generated a stable in-package atmosphere offering
a predictable control.
As shown in Fig. 5b, no rotten fruits were detected over the
11 days of storage when they were kept unpackaged. In contrast in
packaged fruit decay developed from day 8 and further increased
after 11 days of storage (Fig. 5b). Decay incidence was affected by
Fig. 6. Fungi infection of cherry tomatoes at high relative humidity.
387
the type of the package: for fruits sealed in PLA80 packages there
was no decay after 8 days and only 4% of rotten fruit were detected
after 11 days. Slightly higher levels of decay developed in PLA
90 packages, while considerably higher losses occurred in OPP
packages.
The stem end scar is generally the starting site of decay in
cherry tomatoes (see Fig. 6), however if the scar is dry it is difcult
for pathogens to start the infection. As a result, in unpackaged fruit
no decay occurred over the storage time. In contrast, the higher
humidity inside the packages enhanced pathogen growth, and the
effect was more pronounced in OPP packages due to the higher
humidity and the presence of condensation. The presence of
condensed water prevented the drying process of the stem end
scar, but also caused cracks of the peel starting from the end scar,
thus increasing fruit susceptibility to decay.
Therefore, the high in-package humidity in OPP packages which
reduced weight loss also resulted into a higher susceptibility of the
fruits to decay. On the other hand, the PLA packaging offered a
better optimized, balanced EMA which combined a considerable
reduction in weight loss with a slower decay due to pathogens.
Thus, if in one hand high levels of humidity strongly suppress
transpiration and weight loss, on the other hand may favour
growth of spoilage microorganisms (Kader et al., 1989). As shown
by Boylan-Pett (1986), the appearance of moulds, which were
associated with high levels of in-package humidity, resulted in a
shorter shelf-life of red-ripe tomatoes packaged in an EMAP of low
density polyethylene (LDPE) designed for optimal levels of O2 and
CO2 compared to tomatoes stored in a ow-throw system with the
same gas molar fractions.
The PLA bags offer an optimal packaging solution by reducing
the decay incidences compared to the OPP bags while keeping
weight loss limited. In fact, the appearance of fruit sealed in PLA
bags did not show differences with respect to those sealed in OPP
macro bags, despite the higher reduction in weight (the score was
in all cases higher than 8); while a signicant deterioration of the
appearance (average score 6.2) was detected only in unwrapped
fruits.
After 4 days of storage in-package CO2 in the PLA bags was
3.5 kPa, but declined to 2.51 kPa after 11 days, while in-package O2
increased from 17.6% to 18.7%, with no signicant difference
between PLA80 and PLA90. In contrast, air composition in OPP
bags, as expected, was slightly different than air and did not change
over storage (CO2 * 0.02 kPa; O2 * 20.5 kPa).
The initial CO2 (after 4 days of packaging) molar fraction
measured in PLA bags compared well with the value, 4%, predicted
by the analytical model. The reduction of CO2 with storage time in
the case of PLA packaging is not a weakness of the model, but a
consequence of the decline in respiratory activity during storage. In
fact, respiration rate decreased from the initial value of 5.4 mg CO2
kg"1 s"1 to the nal value of 2.6 mg CO2 kg"1 s"1.
Cherry tomatoes are harvested when full ripe, after the
climacteric phase, thus, unless fruit are subjected to physical
stress (mechanical impacts, wounds, chilling temperatures) or
invaded by pathogens, low rates of ethylene production can be
expected. Accordingly, ethylene rates of unwrapped samples never
exceeded 2.6 & 10"4 mg kg"1 s"1, while in-package ethylene con-
centration in PLA bags ranged between 0.5 and 0.8 mL L"1.
The result is an environmental friendly packaging offering a
longer shelf time with better quality and safer cherry tomatoes.
3.4.2. Peaches
The temperature in the storage room varied between 18.9 and
21.3 ! C. In-package temperature was always between the varia-
tions range of the storage room. Average room RH was 64.7%,
ranging between 60.47 and 67.66%. Average RH in PLA85 bags was
87%, a value in good agreement with the target RH of the designed
388 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
Fig. 7. Changes in weight loss (a) and decay incidence (b) of peaches cv Dixired during storage. Vertical bars represent the standard deviation (n = 20 for weight loss and
80 for rotten fruit).
PLA package, while in OPP macro-perforated (OPPmacro) bags
average RH was 98%. In both types of packages RH showed slight
changes over time, but in OPPmacro bags condensation was
constantly present.
Weight losses of unwrapped fruits after three days of storage
were 3.4% and increased to values higher than 14% at the end of
storage, with an average weight loss per day of 11 g kg"1 (Fig. 7a).
This high value of transpiration rate might be due to the large
variability in daily weight loss shown by the individual fruits,
which had a standard deviation of 1.7 g kg"1. Packaging not only
reduced weight loss, but also eliminated the variability of
transpiration among fruits. In fact, fruits sealed into PLA lm
experienced only 1.04% weight loss after 3 days and was less than
4% after 9 days, which is equivalent to 3.46 g kg"1 per day, while in
OPPmacro bags the nal weight loss were less than 1%. The high
transpiration of unwrapped fruit signicantly affected fruit
appearance, which had an average score of 5.2 since day 6 of
storage, but with a high percentage of fruit below the limit of
marketability. In contrast the score of wrapped fruit was always
higher than 8 and differences between PLA85 and OPPmacro were
not signicant.
After 3 days of storage no rotten fruits were observed neither
among unwrapped nor among packaged ones of both types of
lms. Six days later, decay developed in all packaging cong-
urations with about 8, 12 and 18% of rotten fruits in unwrapped,
PLA85 and OPPmacro treatments, respectively. After 9 days the
percentage of rotten fruit did not change signicantly in
unwrapped and PLA85 bags, while increased to 57% in OPP bags
(Fig. 7b).
As observed in the case of cherry tomatoes, humidity levels
lower than 90% is a good compromise to reduced transpiration
rate, preserve overall appearance and delay pathogens growth. In
contrast higher levels of humidity, associated with temperature
uctuations during storage may cause the formation of condensa-
tion inside the packages and lead to a rapid decay.
The CO2 molar fraction in the PLA bags was between 2.0 and
3.0 kPa in good agreement with the targeted value as presented in
Table 1 despite the high variability of respiration activity detected
over storage among the ten unwrapped fruits. In-package average
ethylene concentration in PLA bags was slightly below 2 mL
C2H4 L"1 during the rst three days, then it increased progressively,
with nal values of about 6 mL C2H4 L"1, showing that the fruit
were in their ripening stage during storage, as conrmed by
increasing rates measured in unwrapped fruit. In contrast, in
OPPmacro bags both CO2 and O2 were slightly different than air,
and ethylene ranged between 0.3 and 0.8 mL C2H4 L"1.
Despite the high level of ethylene present inside the PLA bags,
the fruits did not show detectable differences in appearance and in
their ripening stage with respect to those sealed in OPP lms,
denoting no evident effect of the high level of ethylene inside PLA
bags, whose potential physiological impact on fruit tissue might be
counteracted by the higher levels of CO2 in PLA85 bags (Burg,
2004).
4. Conclusions
An analytical, user friendly, one-dimensional model was
developed for designing a biodegradable bio-based EMAP system
for selected fruits and vegetables using PLA lms. The model was
developed simulating the combined gas diffusion through micro-
perforations and the permeable PLA membrane of the EMA
packaging system. It was shown that the relatively high water
vapour (WV) permeability of PLA lms compared to conventional
OPP lms allows the development of an optimised biodegradable
bio-based EMA packaging system for the specic applications. The
parameters of this model were determined through a series of
laboratory and numerical experiments.
Two lab experiments were performed under real conditions for
validating the model. Cherry tomatoes and peaches were used as
typical representatives of vegetables and fruits for testing the
newly designed EMA packaging systems. The model predictions
compared well against the corresponding measurements con-
rming its validity.
PLA was shown to be a versatile material which allows through
a proper design, for a better control of the in-package atmosphere
than conventional materials due to its higher permeability to water
vapour combined with its barrier properties with respect to CO2
and O2. This PLA property in combination with the laser micro-
perforation technology makes possible the development of
optimal EMAP systems tting the exact needs of specic high
value horticultural products. In this way, their quality and safety is
preserved for a longer shelf life period by using a bio-based and
compostable material.
Acknowledgement
This work was supported by the European Research for SMEs
project HORTIBIOPACK (FP7-SME-2008-1-232551) Development
of innovative biodegradable packaging system to improve shelf
life, quality and safety of high-value sensitive horticultural fresh
produce.
 A. Mistriotis et al. / Postharvest Biology and Technology 111 (2016) 380389
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