INNFOO-01394; No of Pages 8

Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

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Innovative Food Science and Emerging Technologies

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

Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging
containing green tea extract
Daniel Carrizo a, Gonzalo Taborda a, Cristina Nern a,, Osvaldo Bosetti b
a
b

Aragon Institute of Engineering Research (I3A), EINA, GUIA Group, Department of Analytical Chemistry, University of Zaragoza, Mara de Luna 3, 50018 Zaragoza, Spain
Goglio S.p.A., R&D Chemical Laboratory, Packaging Division, Via dell'Industria, 7, 21020 Daverio, VA, Italy

a r t i c l e

i n f o

Article history:
Received 29 July 2015
Received in revised form 30 September 2015
Accepted 24 October 2015
Available online xxxx
Keywords:
Active packaging
Green tea extract
Volatile prole
Fatty acid prole
Food
Chocolate derivatives
Antioxidants
Radical scavengers

a b s t r a c t
The performance of an antioxidant multilayer packaging containing green tea extract was studied over a longterm experiment with real commercial food. Two types of food (dark chocolate peanuts and milk chocolate
cereals) were packaged and tested during 16 months. Antioxidant performance of the multilayer packaging
was demonstrated. Volatile prole (hexanal, pyrazines and related compounds) and fatty acids were used as
indicators to monitor the system performance. The volatile compounds wereanalyzed monthly, identied and
quantied by HS-SPME-GC-MS, while the fatty acids were analyzed after derivatization by GC-MS every 3
months. Signicant lower values of hexanal were obtained for the active system during the time experiment
while the values for the fatty acids were higher in the active system compared to the non-active one. Organoleptic tests carried out with a trained panel conrmed the antioxidant effect. The radical scavenger capacity of the
multilayer was also quantitatively evaluated. The new approach of antioxidant behavior as a radical scavenger
of green tea is discussed in detail.
Industrial relevance: This paper describes the performance of a new antioxidant multilayer packaging material,
tested for 16 months with real food and in industrial conditions, as the format, size and material were produced
at an industrial level, as a new prototype, following the indications from previous research. In this material green
tea extract has been grafted in the internal layer of a multilayer material, in which it acts as a radical scavenger,
which means as an antioxidant. The antioxidant behavior of the polymer as well as the performance of the polymer versus real food is described. It is the rst time that such a material is successfully produced at the industrial
level and in fact, the material is nowadays in the market. For all these reasons the paper is of great industrial
relevance. Many industries are and will be interested in it.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
Food oxidation is considered a major cause of quality deterioration,
affecting both the nutritional and sensory quality and safety of foods
and has thus been a challenge for food preservation. Peanuts and cereals
covered by chocolate are dried foods that are susceptible to lipid autoxidation due to a high content of unsaturated fatty acids. Flavor and
quality of peanuts and peanut products are largely functions of lipid
chemistry. Peanuts contain ~50% oil (Cobb & Johnson, 1973). Palmitic
acid (16:0), oleic acid (18:1), and linoleic acid (18:2) are the major
fatty acids in peanuts and may comprise N 90% of the total fatty acids
(Ahmed & Young, 1982). The remaining fatty acids, stearic (18:0),
arachidic (20:0), eicosenoic (20:1), behenic (22:0), and lignoceric
(24:0) acid, normally occur in weight percentages between 0.02 and
4.0%. Free radicals are important chain-carrying intermediates in lipid
autoxidation, which lead to the formation of hydroperoxides, which
Corresponding author. Tel.: +34 976 761873.
E-mail address: cnerin@unizar.es (C. Nern).

are further precursors of secondary oxidation products, such as ketones

and aldehydes. This process may be accelerated by moisture, light,
oxygen or exposure to high temperatures. The presence of secondary
oxidation products (i.e., aldehydes) is associated with changes inodor
and avor of the products, resulting in rancidity. Hexanal is formed
during the oxidation of linoleic acid via the 13-hydroperoxide and it
has an odor described as grassy which contributes to off-avors. It is
easily detected as it has a low odor threshold (in water: 4.5 g/kg)
(Fenaille, Visani, Fumeaux, Milo, & Guy, 2003, Ruiz, Ventanas, & Cava,
2001). Measurements of hexanal and other secondary oxidation products
(e.g., pentanal or octanal) using headspace solid phase microextraction
coupled to gas chromatography mass spectrometry (HS-SPME-GC-MS)
has been widely used to monitor the oxidative deterioration of dry foods
(e.g., peanuts, potato chips) (Sanches-Silva, Rodrguez-Bernaldo de Quirs,
Lpez-Hernandez, & Paseiro-Losada, 2004). Flavor compounds produced
from roasting peanuts are very important for acceptability. The most important compounds produced after roasting the peanuts or cereals are
pyrazines (Ho, Jim, Lee, & Chang, 1961). Loss of peanut avor occurs rapidly
after roasting and is known as avor fade (Reed, Sims, Gorbet, & O'Keefe,

http://dx.doi.org/10.1016/j.ifset.2015.10.018
1466-8564/ 2015 Elsevier Ltd. All rights reserved.

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

2002). Auto-oxidation is a slow, radical process that can take several days
(Ingold, 1961).
Active compounds and ingredients can be incorporated into packaging materials to provide several functions that do not exist in conventional packaging systems. Active packaging may carry antioxidants,
antimicrobial agents and/or nutrients. Due to health concerns of
consumers and environmental problems, current research in this type
of packaging has been focused on the use of natural components and
biodegradable packaging materials (Suppakul, Miltz, Sonneveld, &
Bigger, 2003, Lpez, Snchez, Batlle, & Nern, 2007, Gutirrez, Snchez,
Batlle, Lpez, & Nern, 2009, Rodrguez-Lafuente, Batlle, & Nern, 2010,
Nern et al., 2006). Substances with antioxidant potential are available
from a variety of natural sources or as synthetic chemicals. Among
them green tea extract (GTE) can be mentioned as a rich source of polyphenol antioxidants, particularly catechins (Coln & Nern, 2012) and
has the status of food additive (Roy et al., 2010; Fernandez, Martin,
Gonzalez, & Pablos, 2000). A formula for antioxidant packaging based
on GTE for food packaging applications was developed in a recent
publication by Carrizo, Gullo, Bosetti, and Nern (2014). In this paper a
multilayer antioxidant packaging material has been developed and
evaluated rst in vitro and later in vivo for its antioxidant properties.
The new packaging material was initially produced at laboratory scale
and later at industrial scale.
One of the main difculties when developing an antioxidant material for food protection is the incorporation of the active agent in an
efcient and feasible way, so that the new material can act as an antioxidant without modifying the packaging line or the characteristics of the
packaged product. Several approaches have been proposed but all of
them fail by one or another reason. Extrusion of polymers involves
high temperature and the active agents decompose. Coating systems
affect the sensory properties of the packaged food when essential oils
are used as antioxidants. However, in all these approaches the antioxidant agent is incorporated in the layer in contact with food. In the
present work a very different system has been explored. To understand
the antioxidant performance in this case a clear idea of the oxidation
reaction of lipids is essential. It is known that the oxidation process is
a radical reaction initiated by the presence of OH and O free radicals
(Nern, 2011). It is a chain reaction including initiation, propagation
and termination. When free radicals are removed the reaction does
not take place. If the free radicals can efciently diffuse through the
polymer it is not necessary that the active agent, GTE in this case,
were in the layer in contact with the food. This is the principle applied
to this new active material.
The aim of this work was to evaluate the performance of a new
active multilayer packaging material, based on GTE and produced at industrial scale, in two commercial foods during a long-term experiment
(16 months) as well as to demonstrate the efciency of the new antioxidant material in extending the shelf life of real food.
2. Materials & methods
Dark chocolate roasted peanuts and milk chocolate cereals were
produced by CHOCOLATES LACASA S.A (Zaragoza, Spain). Flexible bags
of a multilayer OPP25 m + OPP25 m packaging material with active
compounds were prepared by GOGLIO, identical in size and materials
used as those used for commercial purposes by LACASA. A special adhesive with the GTE incorporated was used to build the multilayer. The
system is under patent petition. Commercial trays and the same multilayer exible material as that used in the current application without
any active agent were also used as blank for the long-term experiment
(16 months). Two sets of packaging systems were used in the same
format and size as the current packaging (36 plastic bags of 1 kg and
36 trays of 1 kg), half of them (18 bags and 18 trays) with the active
components and the other half without the active components
(Table 1). The plastic bags were made by thermosealing the multilayer
once lled in with the food, while in the trays only the lid was made

Table 1
Details for the experiments using active and non-active packaging materials.
Starting date for
the experiment:

24/02/2013

Best before:
Sample size:
Samples n:
Samples types:
Active
Non-active

24/10/2014
1 kg bags
36
18 bags with active material
in the adhesive
18 bags without active
material in the adhesive

1 kg trays
36
18 trays with active material
in the adhesive
18 bags without active
material in the adhesive

with the new antioxidant multilayer. This lid was thermosealed to the
tray after lling. All the products were produced immediately before
packaging. The packaging process was done at pilot plant scale. Room
temperature (23 C) was used for the study, as these chocolate products
are usually kept in the linear of supermarket without refrigeration or
additional care. This temperature was selected as the worst scenario.
The current shelf life of these products is 9 months. The present study
lasted for 16 months.
3. Experimental design
The evaluation of this new active material was based on a long-term
monitoring of key components of the samples involved in the food
quality due to lipid oxidation. The rst step was to evaluate the radical
scavenging properties of the new multilayer produced by GOGLIO.
Once demonstrating the antioxidant performance, this material was
used for packaging real food produced by LACASA. For the dark chocolate peanuts volatile compounds (i.e., hexanal, pyrazines and related
compounds) and the fatty acid prole were used. For milk chocolate cereal samples only the volatile compound prole (i.e., hexanal, pyrazines
and related compounds) was used. These two types of monitored
chemicals are key components in the oxidation process of fatty food.
Volatile analysis prole was performed monthly for all the samples
(e.g., four analysis for four samples, two of each type of product and
each type of material active/non-active), whereas the analysis of
fatty acid prole was performed every 3 months and only for the
dark chocolate peanut samples (two samples, one with the active material and the other with the non-active material and three independent
replicates in each case). The experiment started on 24th February 2013
and lasted for 16 months. The best before date was in all cases 24th
September 2013. The study lasted until the end of May 2014.
3.1. Chemicals and reagents
Green tea extract (GTE) was supplied by Taiyo Green-power Co. Ltd.
(Jiangsu, China). Multilayer active and non- active packaging bags and
the multilayer lm for the lid of the trays were produced by Goglio
S.p.A (Daverio, Italy). Hexanal and the fatty acids methyl ester mixture
(FAME) used as standard for quantication purposes were supplied by
Sigma-Aldrich S.A. (Madrid, Spain). High purity quality solvents such
as dichloromethane, cyclohexane, heptane and boron triuoride were
from Scharlau (Barcelona, Spain). Ultrapure water was obtained from
a Milli-Q system (Millipore, Billerica, MA, USA).
3.2. Antioxidant evaluation
The antioxidant properties of the new polymer were measured by
the free radicals method developed by Pezo, Salafranca, & Nern, 2008.
The method consists of generating an atmosphere enriched in free
radicals from a H2O2 solution nebulized in a quartz tube. The generation
of free radicals is enhanced by a photochemical reaction using UV lamps
on the quartz tube. An inert gas carries the atmosphere through the
plastic bag made from the polymer under evaluation and after this
step the nal gas bubbles into a solution of aqueous salicylic acid (SA).

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

Then, SA is hydroxylated by the free radicals. If the plastic bag acts as a

scavenger the free radicals do not arrive at the SA solution. After 48 h of
continuous testing the SA is analyzed by HPLC with uorescence detector, where the resulting products 2,3 DHB (2,3-dihydroxibenzoic acid)
and 2,5 DHB, formed by hydroxylation of SA, are quantitatively
measured. A detailed description of the method as well as the required
installation can be seen in Pezo et al., 2008. The antioxidant capacity is
then measured as 100% minus the percentage of hydroxylation, where
100% hydroxylation means that the material is not acting as free radicals
scavenger.
3.3. Fatty acid prole by GC-MS
Dark chocolate peanuts samples (50 g. from each bag with and
without active agent) were ground for 2 min with a commercial grinder.
After that, 100 ml of a dichloromethane/cyclohexane mixture (1:1 v/v)
were added while continuously mixing for about 3 min. The mixture
was transferred to a glass beaker and left for 24 h. The supernatant solvent containing the lipids was then transferred to a previous weighted
glass beaker and left to dry for 48 h. The beaker was weighted again to
know the weight of the lipid fraction of the sample. A small part of
this lipid sample (~0.2 g) was further used for fatty acid derivatization
to methyl esters, following the procedure by AOAC Ofcial Method
965.49 (2005).
Fatty acid prole was analyzed after the derivatization reaction
using a 5975B Agilent gas chromatograph connected to a 6890N mass
spectrometer. Chromatographic separation was carried out on a BP20
(30 m 0.25 mm I.D., 0.25 mm lm thickness) from SGE Europe Ltd.

The oven temperature was set at 50 C for 4 min, then increased from
50 to 210 C at 6 C min1 and remained at 210 C for 10 min. Helium
was used as a carrier gas at 1 ml min1. Mass spectra were recorded
in electron impact (EI) mode at 70 eV, scan mode was used for the
acquisition, quadrupole and source temperature were set at 150 and
230 C respectively.
3.4. Volatile prole by HS-SPME-GC-MS
SPME in headspace mode was used for the analysis of the
volatile prole, a fused silica ber coated with a 50/30 m layer of
divinylbenzenecarboxenpolydimethylsiloxane (DVBCARPDMS;
Supelco) was used to extract hexanal and related compounds from
the samples. The ber was exposed to the headspace of the bag or the
tray during 10 min (extraction time) at room temperature (23 C)
(sorption step) and further desorbed in the GC injection port for 2 at
250 C and analyzed by GC-MS. The experimental conditions were
previously optimized (Montero-Prado, Bentayeb, & Nern, 2013; Vera,
Canellas, & Nern, 2014).
A CTC Analytics CombiPal autosampler was coupled to a 5975B
Agilent gas chromatograph (Agilent, Palo Alto, California) and connected to a 6890N mass spectrometer. The selection of the most sensitive
solid phase micro-extraction (SPME) ber and the optimization of the
HS-SPME conditions were carried out in a previous work (Canellas,
Aznar, Mercea, & Nern, 2010; Sanches-Silva et al., 2004). The manual
SPME holder (Supelco, Bellefonte, PA, USA) was used to perform the
experiments. Chromatographic separation was carried out on a BP20
(30 m 0.25 mm I.D., 0.25 mm lm thickness) from SGE Europe Ltd.

Fig. 1. a. Representative chromatogram of HS-SPME-GC-MS analysis of milk chocolate cereals in active packaging b. Representative chromatogram of HS-SPME-GC-MS analysis of milk
chocolate cereals in non-active packaging.

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

The oven temperature was set at 40 C for 5 min; the temperature was
then increased from 40 to 220 C at 10 C min1 and set at the maximum temperature for 5 min. Helium was used as a carrier gas at
1 ml min1. Mass spectra were recorded in electron impact (EI) mode
at 70 eV. Scan mode was used for the acquisition to get all the volatile
compounds sampled. Quadrupole and source temperature were set at
150 and 230 C respectively.
3.5. Specic migration analysis
After 16 months, which was the end of the experiment, chocolate
coated cereals and dark chocolate peanuts were extracted with methanol in an ultrasonic bath. Food samples were ground in an ultraturrax,
the solvent added and continuously mixed for 3 min. After that the
mixture was transferred to a glass beaker, sonicated for 20 min and
left to settle. Then, the supernatant was ltered and transferred to a
new beaker. Both sample extracts (chocolate coated cereals and dark
chocolate peanuts) were concentrated under a gentle stream of nitrogen and transferred to a vial for further UPLC-MS analysis. GTE was
also extracted with methanol and analyzed as control. Possible GTE
components from chocolate coated cereals and dark chocolate peanuts
as well as the GTE extract were analyzed by UPLC-MS-TQ AQUITY
(Waters, Mildford, Massachusetts, USA). UPLC-MS conditions were as
follows: an Acquity UPLC@ BEH C18 1.7 m (2.1 100 mm) was used
(Coln & Nern, 2012). Mobile phases were as follows: eluent A
(methanol with 0.1% formic acid) and eluent B (water with 0.1% formic
acid). Flow rate was 0.3 ml min1 and injection volume of 10 l. The
gradient system was 06 min, 5% A95% B, 68 min, 95% A5% B,

8.1010 min, 5% A95% B. Run time was 10 min; sample temperature

was set at 7 C and column temperature at 35 C. SIM mode was used
for the mass spectra conditions. ESI in negative mode was used for all
the catechins and gallic acid, while for caffeine ESI was used in positive
mode. Selected m/z were 169.02 m/z for gallic acid (GA), 194.08 m/z for
caffeine (CAF), 289.07 m/z for (+)-catechin (C) and ()-epicatechin
(EC), 305.07 m/z for ()- gallocatechin (GC) and ()-epigallocatechin
(EGC), 441.08 m/z for ()-epicatechin gallate (ECG) and ()-catechin
gallate (CG), and nally 457.08 m/z for ()-epigallocatechin gallate
(EGCG) and ()-gallocatechin gallate (GCG). The linear dynamic ranges
obtained were calculated with at least seven calibration points and the
values are expressed as micrograms of standard mixture per gram of
methanolic solution.
3.6. Sensory evaluation
Samples from the two types of foods (peanuts and cereals) with and
without active material were tested every three months (T3, T6, T9 and
T12) applying the optimized procedure above described. A panel constituted by 12 trained panelists evaluated every month the packaged
products. Triangular tests were carried out in which the panelists had
to identify the appearance of rancidity and then the acceptability of
the food product. The panelists tasted blind samples, where the information about the food packaged in active material or in the non-active
one was not known. A scale from 0 to 5 was given for rancidity, the
most important quality of food in this case. Signicance of the evaluation was calculated by applying statistical measurements with all the
values obtained. At the end of the 16 month period the samples were

Fig. 2. a. Montly hexanal prole for active (diamond) and non-active (Square) packaging systems in dark chocolate peanuts 1191 604 (96 96 DPI). b. Montly hexanal prole for active
(diamond) and non-active (Square) packaging systems in milk chocolate cereals 1231 604 (96 96 DPI).

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

also tested by an expert panel from the company producing the chocolate peanuts and cereals.
4. Results and discussion
4.1. Radical scavenger performance
Antioxidant active packaging is usually conceived either as the material supplying antioxidants or acting as oxygen absorber. However,
the concept of free radical scavenger is quite different. Free radicals
have short life, are very reactive and can easily permeate through the
polymers, as has been demonstrated in previous work dealing with
the development of these multilayer antioxidant exible polymers
(Pezo, Salafranca, & Nern, 2007; Pezo et al., 2008; Coln & Nern,
2015; Nern, 2011; Dicastillo et al., 2011). Thus, this antioxidant approach is not a positive migration of antioxidants, but a real scavenger
system. It can be highlighted that the antioxidant performance of catechins is based on their capacity as free radicals scavengers. In these

multilayer lms catechins were not in direct contact with the food,
but in indirect contact, as they were grafted on the internal layer of
the multilayer by the adhesive used to build the laminate. The
quantitative measurement of the antioxidant performance applying
the free radicals method above described provided a 46% with RSD 2%
of antioxidant value while the non active material showed 0% of antioxidant. The results obtained conrmed that free radicals diffuse throughout the polymer and are scavenged by the catechins. Previous studies
carried out with this kind of polymer demonstrated that a grafting phenomenon occurs in the adhesive formulation. Catechins from the GTE
were present in the whole polymer behind the OPP layer that is in contact with the food and they do not migrate, while the behavior as free
radical scavengers is not compromised. These experimental results
open a new concept of antioxidant packaging, in which the positive migration of antioxidants is not required and the oxygen absorption is not
necessary to protect the food versus oxidation processes. As was above
mentioned, all the food samples under test were packed under atmospheric pressure and contained the normal atmosphere, with oxygen

Fig. 3. a. Pyrazine prole for active (blue bars) and non-active (red bars) packaging systems in dark chocolate peanuts. b. Pyrazine prole for active (black bars) and non-active (red bars)
packaging systems in milk chocolate cereals.

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

in contact with the food inside the packaging. As has been demonstrated,
in these conditions the antioxidant performance was efcient to protect
the food and extend the shelf life of the packaged products.
4.2. Volatile prole
The dark chocolate peanuts and the milk chocolate cereals analysis
showed initially few differences in the major volatile compounds, either
in the active or in the non-active packaging systems. An increasing trend
in the hexanal concentration and also over some aldehydes over time
was observed for the two sets of samples, but with a higher rate in the
non-active material. A chromatogram of the volatile prole of milk
chocolate cereals packaged in active and non-active material at T16 obtained using SPME is shown in Fig. 1a and b respectively. Fig. 2a and b
shows the hexanal prole for the dark chocolate peanuts experiments
(active and non-active) and for the milk chocolate cereals, respectively.

As can be seen, higher values of hexanal were obtained for the nonactive samples, which means that the oxidation process took place in
a larger extension than in the active samples. It can be pointed out
that the increase in hexanal is much higher from month 11 on in the
non- active samples. This period coincides with the expected shelf life
of about 910 months for the non-active samples. Thus, no signicant
differences were expected in this period. From month 10th to 16th signicant differences were found, demonstrating the antioxidant effect of
the active packaging material. The increase is also more important for
cereals, as expected, as they are even more prone to oxidation than
peanuts. Cereals were covered by milk chocolate, which supplies less
antioxidants than the dark chocolate present in the peanuts samples.
Other related aldehydes (e.g., pentanal, heptanal, octanal and decanal)
showed an unclear trend over time, probably because of the lower
concentration values, which made more difcult to monitor the
changes.

Fig. 4. Fatty acid prole for the dark chocolate peanuts over the time experiment (16 months) using the active packaging material (A) and the non-active material (B).

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

Fig. 5. UPLC-MS chromatograms of a mix standard (10 mg kg1, (ppm)) and the dark chocolate peanut active packaging sample for the migration test at 16 months of the beginning of the
experiment.

the compounds in the active packaging system samples compared to

the non-active one, but no high differences were found between
them. These results were in agreement with the volatile prole, where
hexanal has an inverse relationship and the pyrazines have a direct relationship with the lipid prole (i.e., linoleic acid). It is worth mentioning that the usual shelf life of these packaged products is 9 months, and
the desired shelf life using the active packaging is about 18 months. The
results obtained in this work encouraged the company to use this active
packaging for extending the shelf life.

Pyrazines are typical volatile compounds in roasted cereals and

peanuts and the odor can be easily perceived as characteristic of
the roasting process. However, pyrazines are degraded with time
and the odor changes signicantly. For this reason, one of the critical
indicators for quality control of roasted peanuts and cereals is the
concentration of pyrazines. Several pyrazines such as 1methylpyrazine, 3-methylpyrazine, 2,5-dimethylpyrazine, 2,3,5trimethylpyrazine, ethylpyrazine and tetramethylpyrazine were identied among the volatile compounds found in the headspace of the
packaging, in equilibrium with the packaged product. Higher values
were found in the active packaging material compared to the nonactive material, what conrms the antioxidant effect of the active packaging material. Fig. 3a and b shows the pyrazine prole obtained for the
active and non-active packaging material for the dark chocolate peanuts
and for the milk chocolate cereals, respectively. As can be seen, clear differences were found between the active and non-active samples, as expected, which agrees with the previous data found of hexanal
production.

4.4. Migration of catechins

Determination of active compounds (gallic acid, caffeine and
catechines) released from the active lm was carried out by determining the specic migration into the food (i.e., dark chocolate peanuts)
samples in the 16th month of the experiment. Specic migration is
the amount of a specic component that migrates from the food contact
material to the food in contact with it. Food contact materials must not
transfer their components into the foods in unacceptable quantities
(migration), according to the frame Regulation 1935/2004/CE. Food
samples were extracted following the procedure described in the experimental section. Three replicates of samples were spiked with pure
catechins and the analytical procedure was applied to these samples
to calculate the recoveries and to establish the limits of detection
(LOD) and limits of quantication (LOQ) in the real samples. Fig. 5
shows the chromatogram obtained for this experiment. No migration
of any component (gallic acid (GA), catechins (GC, EGC, C, EGCG, GCG,
EC, ECG and CG) and caffeine (CAF)) was found in the migration experiment after 16 months of storage. This behavior was also expected, as
the migration from these active materials using simulants gave also

4.3. Fatty acid prole

In order to compare the fatty acid prole for the active and nonactive packaging systems, an initial sample of fatty acid prole was
analyzed before the experiment started (T0). The major fatty acids
identied were: palmitic acid, stearic acid, oleic acid, linoleic acid,
arachidic acid, eicosenoic acid and behenic acid. A decreasing concentration prole of all the components was found for both packaging,
but with a lower trend in the active packaging system after 16 months
(Fig. 4). The two sets of dark chocolate peanut samples have a similar
fatty acid prole over time, with a trend of higher concentration for all

Table 2
Rancidity tests of chocolate peanuts and chocolate cereals in the packaging materials by a trained (experts) panel.
Food

12th month

16th month

Chocolate peanuts
Active chocolate peanuts
Chocolate cereals
Active chocolate cereals

1
0
2
1

2
0
4
2

Rancidity scale

Commercialization

0
no
Optimum

1
starting
Possible

2
+
Acceptable

3
++
Less recommendable

4
+++
Not recommendable

Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018

D. Carrizo et al. / Innovative Food Science and Emerging Technologies xxx (2015) xxxxxx

negative results (Coln & Nern, 2015). Linear range varied from 0.001
to 75.1 g g1 depending on the compound, so a LOD of 0.001 g g1
(=1 ppb) and LOQ of 0.003 g g1 (=3 ppb) was found. The analytical
method performance was validated through the participation in an
international inter-calibration exercise (Exercise I of the Dietary
Supplements Laboratory Quality Assurance Program, NIST Interagency
Report (IR7955) (see http://nvlpubs.nist.gov/nistpubs/ir/2013/NIST.IR
.7955.pdf). A blank sample (non-active dark chocolate peanuts) was
spiked with the GTE standard mixture to check for method performance
and a recovery of 85+/2% was found.
4.5. Sensory evaluation
All the evaluations carried out by the panelists conrmed what the
chemical analysis showed. This means, that the active material inhibits
or delayed the oxidation process. At the end of the 16 month period,
the panel from the company conrmed again the results obtained by
both the chemical analysis and the sensory analysis carried out by
the other panelists. Table 2 shows the results obtained by the trained
panelists in the food company together with the scale of rancidity
used. The differences found were statistically signicant (p b 0.05).
5. Conclusions
The study carried out demonstrates that the concept of free radical
scavenger to protect the food versus oxidation processes is a real
issue. Catechins, known as efcient antioxidants because of their capacity as free radical scavengers, were incorporated in the adhesive used for
building the laminates, in indirect contact with the food in a multilayer
structure. The new multilayer material demonstrated its good antioxidant performance in the objective measurements of radical scavenging
method. Food (peanuts and cereals covered by chocolate) was
protected for a long term period (16 months of testing) under atmospheric pressure, in the presence of oxygen. This means that free radicals cross the polymeric layer, diffuse through the polymer and arrive
at the catechins position, where they are trapped and consumed. In contrast, catechins did not migrate to the food, what means that they were
grafted into the adhesive formulation and act from there as efcient antioxidants/free radical scavengers. The complete absence of migration
indicates that catechins do not diffuse through the polymer. As result,
sensory analysis demonstrated that the packaged food was not affected
by GTE, but it was well protected against oxidation, signicantly reducing the rancidity and this way extending the shelf life of packaged food.
These facts open the window for a different concept of food protection
under the frame of active packaging. As a whole, the results obtained
conrm the good performance of the multilayer active component
application with real food during a long-term experiment.
Acknowledgements
This research has been nanced by the EU Project SAFEMTECH
(IAPP-Marie Curie Actions, 20010-2015, GA 251382) and by the project
OTRI-2013-005. Thanks are given to LACASA for supplying the food
samples. Mauro Fedeli is gratefully acknowledged for preparing the
sample bags and laminates at Goglio S.p.A.
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Please cite this article as: Carrizo, D., et al., Extension of shelf life of two fatty foods using a new antioxidant multilayer packaging containing green
tea extract, Innovative Food Science and Emerging Technologies (2015), http://dx.doi.org/10.1016/j.ifset.2015.10.018