Journal of Food Engineering 84 (2008) 313–320

www.elsevier.com/locate/jfoodeng

Behaviour of a new antioxidant active film versus oxidizable

model compounds
C. Nerı́n *, L. Tovar, J. Salafranca
Department of Analytical Chemistry, Aragon Institute of Engineering Research I3A, CPS-University of Zaragoza, Torres Quevedo Building,
Marı́a de Luna St. 3, E-50018 Zaragoza, Spain

Received 16 November 2006; received in revised form 2 May 2007; accepted 11 May 2007
Available online 26 May 2007

Abstract

In this research a new antioxidant active food packaging has been developed and evaluated. It consists of an innovative system in
which several natural antioxidants from rosemary (Rosmarinus officinalis L.) have been immobilized in a polypropylene film. The influ-
ence of concentration of the active compounds in the polymer and the contact system between the active film and simple food simulants
have been studied. L-ascorbic acid, iron (II) and fatty acids from flax seed oil have been used as model compounds, being representative
of the behaviour of different foodstuffs with respect to oxidation. Whereas ascorbic acid has proven to be not adequate for antioxidant
quantitation purposes due to its prooxidant properties, increased protection of up to 30.1% for iron (II) and 62.0% for fatty acids were
found. Among the advantages of this developed system, which extends the shelf life of packaged food, the non contact plastic-food
requirements can be emphasized.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Active packaging; Natural antioxidants; Rosemary; Polypropylene film; L-Ascorbic acid; Iron (II); Fatty acids

1. Introduction resolve food oxidation by adding natural or synthetic anti-

Oxidation is one of the most important degradation
reactions in foodstuffs, which seriously limits their preser-
vation. Fat content of food products, even those with less
than 1% lipids, is affected by degradation reactions, mainly
chemical oxidation (Miller & Krochta, 1997). Oxidation
has negative both nutritional and organoleptic conse-
quences, namely: changes in nutritional value of products
such as the destruction of essential fatty acids and lipid sol-
uble vitamins A, D, E and K; decrease in caloric content;
rancidity which produces off-flavours and odours; colour
changes such as darkening of fats and oils and lightening
of pigments, and flavour loss.
Oxidation actually affects the competitiveness, reducing
the shelf life of food products and consequently decreasing
sales. Both traditional food packaging and producers
*
Corresponding author. Tel.: +34 976 761873; fax: +34 976 762388.
E-mail address: cnerin@unizar.es (C. Nerı́n).
oxidants to the food product, which can be incorporated
during the process or at the end of the production.
Although several synthetic antioxidants are approved
food additives and have the GRAS (generally recognized
as safe) consideration, international regulations tend to
establish more and more restrictions to their use. Nowa-
days, BHA, BHT, TBHQ and propyl gallate are limited
to about 200 mg kg1 food by Codex Alimentarius (FAO/
WHO Food Standards, 2005) as well as European regula-
tions (Directive 2006/52/EC, 2006) and FDA Food Addi-
tive Status List (US Food & Drug Administration, 2006).
Besides, consumers increasingly prefer to avoid synthetic
additives in favour of those perceived as natural due to their
higher safety and lower toxicity (Balaguer, Gallardo,
Infante, Costa-Batllori, & Marzo, 2003; Frost & Sullivan,
2003). Throughout it, the search for antioxidants naturally
occurring in plants as alternative to synthetic antioxidants
is an interest field to researchers (Bera, Lahiri, & Nag,
2006; Cheah & Abu Hasim, 2000; Ladrón de Guevara

0260-8774/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2007.05.027
314 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320
et al., 2002). Among them, phenolic extracts from different
vegetable products and aromatic plants traditionally used
as spices, such as rosemary, are the most used in the food
industry (Ibáñez et al., 2001; Murphy, Kerry, Buckley, &
Gray, 1998).
Traditional packaging systems are reaching their limits
with regard to further extension of shelf life of packaged
food. An innovative approach with enhanced shelf life of
foodstuffs while improving their quality, safety and integ-
rity, consists of active packagings (Dawson, 2002; Nerı́n
et al., 2006; Suppakul, Miltz, Sonneveld, & Bigger,
2003a; Tovar, Salafranca, Sánchez, & Nerı́n, 2005; Vermei-
ren, Devlieghere, van Beest, de Kruijf, & Debevere, 1999),
whose extra functions are obtained by incorporating
actively functional ingredients or materials into packaging
systems (Han, 2002). These components are usually
applied as sachets inserted into the package or as adhe-
sive-bonded label to the inner wall of the package, in the
closure or in the packaging material through dissolution
or dispersion into the plastic material, or by immobiliza-
tion of oxidizing enzymes in the packaging material (De
Kruijf et al., 2002).
Active packagings can be classified into active-releas-
ing systems (emitters) which actively add compounds to
the packaged food such as carbon dioxide, water, antimi-
crobials (López, Huerga, Batlle, & Nerı́n, 2006; López,
Sánchez, Batlle, & Nerı́n, 2005; Suppakul, Miltz, Sonne-
veld, & Bigger, 2003b), aromas or preservatives (Lau &
Wong, 2000), and active-scavenging systems (absorbers),
which remove undesired compounds such as oxygen, rad-
icals, water, ethylene, carbon dioxide, taints and other
specific compounds. Ethylene absorbers and oxygen scav-
engers are the most used and patented of all active pack-
aging technologies (Brody, 2001; Charles, Sánchez, &
Gontard, 2006; Devlieghere, Vermeiren, & Debevere,
2004). Oxygen removers are developed to avoid oxidation
process. However, radicals mainly oxo, hydroxyl and
superoxide are originated from oxygen and they are the
main initiators of oxidation. Thus, a different approach
can be considered: by eliminating radicals as soon as they
are formed, the propagation of the oxidation reaction
cannot take place and consequently, the concentration
of molecular oxygen is not important, but the presence
of radicals.
From this point of view, the antioxidant behaviour
could be achieved by eliminating the radicals instead of
oxygen. Some natural compounds act this way and react
very efficiently with the radicals by trapping them, thus
avoiding further oxidation. In such a case, neither high bar-
rier nor vacuum packaging materials would be required to
avoid oxidation, but only the presence of a radical scaven-
ger to protect the food against the oxidation process.
This paper shows an innovative active food packaging
which greatly inhibits oxidation reactions and behaves as
scavenger of oxygen radicals (Tovar, 2003). According to
literature (Basaga, Tekkaya, & Acikel, 1997; Lee & Shi-
bamoto, 2002) a rosemary extract as antioxidant mixture
has been immobilized in the polymer, consisting of poly-
propylene films. The antioxidant performance of active
films has been measured by using several model com-
pounds – ascorbic acid, iron (II) and flax seed oil with a
high content on linolenic acid – representative of different
foodstuffs, whose oxidation versus time in the presence of
the new packaging material has been measured. The results
obtained are shown and discussed.
2. Materials and methods
2.1. Chemicals and solvents
The following chemicals were used: iron (II) ammonium
sulphate 6-hydrate (for analysis, CAS number 10045-89-3),
L(+) ascorbic acid 99.7% (for analysis ACS-ISO, 50-81-7),
potassium dichromate (for analysis, 7778-50-9), phospho-
ric acid 85% (for analysis 7664-38-2) and sodium chloride
(for analysis ACS ISO, 7647-14-5) were purchased from
Panreac (Barcelona, Spain). Sodium acetate trihydrate
(6131-90-4) and potassium hydroxide 84% (1310-58-3)
were from Merck (Darmstadt, Germany). Malondialde-
hyde tetrabutylammonium salt 98% (102-52-3) and hepta-
decanoic acid 99% (506-12-7) were from Fluka (Madrid,
Spain). 2-Thiobarbituric acid (TBA, 98%, 504-17-6) was
purchased from Aldrich (Madrid, Spain). Hexane (for
analysis, 110-54-3) and methanol (for analysis, 67-56-1)
were from Scharlau (Barcelona, Spain). Sodium diphenyl-
amine-4-sulfonate (for analysis, 6152-67-6) was from Dr.
Theodor Schuchardt (Munich, Germany). Sodium phos-
phate (CAS number 68915-31-1) was from Euro-Kurtz
(Barcelona, Spain). Trichloroacetic acid 99.5% (for analy-
sis, 76-03-9) was from Riedel-de-Häen (Madrid, Spain).
Flax seed (Linum usitatissimum) unrefined oil was from
Biosan (Madrid, Spain). Amexol, a commercially available
natural extract of Rosemary (Rosmarinus officinalis L.) reg-
istered as spice, mainly for meat-derived products, was
provided by Laboratorios Amerex S.A. (Madrid, Spain).
Water was obtained from an Elix 5 water purification sys-
tem from Millipore (Madrid, Spain). Helium (C-50 qual-
ity) was supplied by Carburos Metálicos (Zaragoza,
Spain).
2.2. Samples
Base polymer for active films consisted of 20 lm thick-
ness polypropylene film made by Poligal, S.A. (Narón,
Spain). Three different experimental samples designed as
PR1, PR2 and PR3 were laboratory-produced in Artibal
S.A. (Sabiñánigo, Spain), containing the natural rosemary
extract Amexol (1.0, 2.0 and 4.0% w/w, respectively in plas-
tic film). The system used for immobilizing the natural anti-
oxidants in the polymer as well as the specific formulation
are protected by the European Patent ‘‘Antioxidant active
varnish” EP1477519 (A1). The same polypropylene (PP)
film used to prepare the active film was considered as
blank.
 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320
2.3. Apparatus
Equipment used to determine L-ascorbic acid consisted
of a Jasco (Tokyo, Japan) LG-1580 liquid chromatograph
equipped with an UV–VIS detector Jasco 1575 and a Luna
C18 (2) column 250  4.6 mm from Phenomenex (Tor-
rance, CA, USA).
The concentration of thiobarbituric acid-reactive sub-
stances (TBARS) was determined on a Unicam Helios k
(Waltham, MA, USA) UV–VIS spectrophotometer.
Fatty acids after derivatization to methyl esters (FAME)
were identified and quantitated by using a Hewlett-Packard
6890 Series (Palo Alto, CA, USA) gas chromatograph
equipped with a mass selective detector HP 5973.
A refrigerated cabinet J.P. Selecta Medilow S (Barce-
lona, Spain) was used to keep the samples at 40 °C during
test time. When required, samples were concentrated in a
Büchi R-124 (Flawil, Switzerland) rotary evaporator.
2.4. Model compounds
The research carried out involves the use of easily oxi-
dizable model compounds which are used to represent
the behaviour of a wide range of food products with
respect to oxidation processes. These model compounds
were: ascorbic acid, present in almost all foods from plant
origin (Griffiths & Lunec, 2001); iron (II) which represents
the behaviour of inorganic compounds as well as hemo-
derivatives of animal origin, and fatty acids, present in all
kind of foods that contain lipids (Morrissey, Sheehy, Gal-
vin, Kerry, & Buckley, 1998).
2.5. Experimental procedure
Numerous food products are packaged without direct
contact with packaging materials, such as meat fillets and
derivatives, fish or bakery products contained in polysty-
rene trays without physical contact with thermosealed clo-
sure films. In order to be representative of this very
common real packaging, the experimental procedure con-
sisted of hanging a strip of the film under study (PP,
PR1, PR2 or PR3) in the headspace of a glass cell contain-
ing a solution of the model compound at the bottom, as
shows Fig. 1, thus avoiding direct contact polymer-food.
This subject is of main importance, since one of the goals
Screw metal cap
Nylon wire
Active film strip
Glass body
Aqueous analyte solution
Fig. 1. Glass cell used in the study.
315
of the paper is to assess whether the protective effect takes
place by scavenging radicals in gas-phase.
2.6. Ascorbic acid studies
The most important characteristic of L-ascorbic acid is
its capacity of oxidation to L-dehydroascorbic acid
(e0dehydroascorbic acid=ascorbic acid ¼ 0:390 V). Two different cell
sizes were used for the studies. One of them with about
134 mL of oxygen (contained in 640 mL of air) and the
other with 30 mL of oxygen (143 mL of air). Three different
concentrations of ascorbic acid (100, 500 and 1200 lg g1)
in a sodium phosphate buffer (5 mM, pH 3.5) solution to
avoid interferences due to the possible ascorbic acid disso-
ciation were analyzed. Samples of 20 mL were introduced
at the bottom of the glass cells and one strip of 40 cm2 of
the active film was hung at the top. The cell was hermeti-
cally closed and exposed to cool white-type fluorescent
light and temperature (40 °C) for a period of time. Mea-
surement of the content of the remaining ascorbic acid
was carried out by HPLC with an UV–VIS detector by fol-
lowing a well established procedure already described
(Romero, Vázquez, López, & Simal, 1992; Vázquez, Váz-
quez, López, Simal, & Romero, 1994). The isocratic mobile
phase consisted of aqueous sodium phosphate buffer
(5 mM, pH 3.5) and methanol (80:20 v/v). The flow rate
was 1 mL/min and detection wavelength was set at
263 nm. Quantification was done by interpolation in a 5-
level calibration graph. All the experiments were performed
in triplicate.
2.7. Iron (II) studies
The value of the standard reduction potential of the sys-
tem ðe0FeðIIIÞ=FeðIIÞ ¼ 0:77 VÞ means that iron (II) is an impor-
tant reductor and therefore easily oxidizable. Two different
cell sizes were used for the studies. One of them contained
116 mL of oxygen (550 mL of air) and the other 69 mL of
oxygen (325 mL of air). Two different concentrations of
iron (II) in aqueous solution were also analyzed: 160 and
670 lg g1. Iron (II) ammonium sulphate 6-hydrate was
used as source of iron (II). In all cases, 20 mL of aqueous
solution of iron (II) were introduced at the bottom of the
glass cell and a strip of 40 cm2 of the active film was placed
at the top avoiding direct contact with the aqueous solu-
tion. The cell was hermetically closed and exposed to fluo-
rescent light for a period of time, after which the samples
were analyzed. A redox titrimetry (potassium dichromate
as titrant agent in acid medium and diphenylaminesulpho-
nate as indicator) was used to measure the concentration of
iron (II) before and after the oxidation tests. Aliquots
of 10 mL were taken, solution was acidified with 0.20 mL
of sulphuric acid (98%) and 0.15 mL of phosphoric
acid to improve colour development. Finally diph-
enylaminesulphonate was added to detect the colour change
of the solution from colourless to violet when reached the
equivalence point. Each test was replicated four times.
316 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320
Two different sets of experiments were carried out. In
the first one, samples of 670 lg g1 of iron (II) were intro-
duced in the big cells (116 mL of oxygen) and exposed to
light for 8, 12, 19, 22, 26, 29 and 32 days. In the second
set, samples of 160 lg g1 of iron (II) were introduced in
both size cells, in order to analyze the possible differences
between the two amounts of oxygen in contact with the
iron solution. Exposure time ranged from 5 to 12 days.
2.8. Fatty acids studies
As commented in the introduction, the oxidative deteri-
oration of lipids is a great concern in the shelf life of foods,
leading to development of undesirable off-flavours and
decrease of the acceptability of foods (Rossell, 1989). To
prevent and retard lipid oxidation, antioxidants are
required and this is the case of the new antioxidant film.
Only one cell size was used for these studies that con-
tained 325 mL of air, which means 69 mL of oxygen. To
study the lipid oxidation, flax seed oil was selected due to
its high content of both linoleic and linolenic polyunsatu-
rated fatty acids which are highly susceptible to oxidation
(Del Nobile, Bove, La Notte, & Sacchi, 2003; Lukaszewicz,
Szopa, & Krasowska, 2004). They were analyzed by two
different tests: 2-thiobarbituric acid test (TBA) and gas
chromatographic determination of fatty acids.
The assessment of lipid oxidation in flax seed oil samples
by the TBA method (Djenane, Sánchez-Escalante, Beltrán,
& Roncalés, 2003) is based on malondialdehyde generation
during the oxidative degradation of lipids. This compound
reacts with 2-thiobarbituric acid giving a red compound
which is spectrophotometrically determined by measuring
absorbance at 531 nm (Rossell, 1989). The TBARS values
were calculated from a standard curve of malondialdehyde.
In this set of experiments, 20 lL of flax seed oil were intro-
duced at the bottom of the glass cell and a strip of 40 cm2
of the active film was placed at the top. The cell was her-
metically closed and exposed to fluorescent light at 40 °C
for 11, 15, 20, 25 and 29 days. After each period of time
the samples were opened and analyzed. After the addition
of 10 mL of 10% (w/w) aqueous trichloroacetic acid and
7 mL of 20 mM thiobarbituric acid, samples were incu-
bated at 97 °C for 20 min, cooled to room temperature
and the absorbance was measured at 531 nm against a ref-
erence blank containing the TBA reagent. All the tests were
performed in triplicate.
With respect to chromatographic determination of fatty
acids, they were extracted from 20 lL flax seed oil samples
using 20 mL hexane by sonication for 15 min. Then, sam-
ples were filtered through 0.22 lm PTFE membrane filters
and the solvent was completely evaporated using a rotary
evaporator. Lipid fraction was then derivatized to methyl
esters (FAME) according to the standard procedure
described in ISO 5509 (2000). The extracted fatty acids
were saponified at room temperature with 1 mL of KOH
(1 M in methanol). After the addition of 50 mg of hepta-
decanoic acid as internal standard, samples were incubated
in a water bath at 100 °C for 10 min and later cooled under
running tap water. FAME were then extracted using 4 mL
hexane solution and 8 mL of distilled water. After centrifu-
gation, the organic phase was poured into a vial and a sec-
ond extraction was performed on the remaining material
with a new 4 mL hexane fraction. The obtained organic
phases were combined and filtered through a Whatman
1-PS filter. The extract was then concentrated under nitro-
gen stream until 1.00 g under gravimetric control and fil-
tered through a 0.22 lm PTFE membrane filter and
encapsulated. Then, FAME were separated on a Sugelabor
(Madrid, Spain) SGL-5,5% phenylmethylsiloxane capillary
column (60 m  0.25 mm, 0.25 lm film thickness). In the
GC-MS system, oven temperature was programmed to be
increased from 180 °C (held for 3 min) to 270 °C at
10 °C min1 held for 2 min, up to 290 °C at 10 °C min1
held for 15 min. The injector temperature was 290 °C and
the carrier gas was helium at a flow rate of 1 mL min1,
being the split ratio 1:40.
3. Results and discussion
3.1. Ascorbic acid
Atmospheric oxygen inside the cell under the light can
generate radicals which are the responsible for the oxida-
tion processes; in this particular case, ascorbic acid can
be oxidized. Since in a previous study (Tovar et al., 2005)
based on migration measurements, the active packaging
under study demonstrated negligible behaviour as active-
releasing system, inhibition of oxidation process to enhance
shelf life for foodstuffs should take place through a radical
scavenging mechanism.
Table 1 shows the results obtained in the presence of a
solution containing 1200, 500 and 100 lg g1 of ascorbic
acid, respectively. As can be seen, in the case of 1200 lg g1
significant differences were obtained in the case of PR1 and
PR2 when compared to the blank. However, none protec-
tion was observed with PR3, which theoretically contained
the highest concentration of antioxidants. When solutions
of 500 lg g1 of ascorbic acid were studied no protection
at all was obtained, since all the active samples were more
oxidized than PP used as reference. Finally, when studying
100 lg g1 solutions total oxidation was found in PP and
PR1 after 90 h and only the most concentrated samples
offered a slight protection.
Table 1
Measured concentrations (lg g1) of ascorbic acid of the different samples
in contact with the active film after 90 h test
Sample Ascorbic acid concentration (lg g1)
Initial (t = 0) 1187.8 ± 25.9 497.6 ± 8.3 102.3 ± 3.1
PP 265.9 ± 35.2 66.1 ± 12.2 Not detectable
PR1 475.3 ± 45.7 44.1 ± 9.5 Not detectable
PR2 401.4 ± 32.8 54.2 ± 14.6 7.1 ± 1.8
PR3 239.3 ± 31.0 18.3 ± 5.9 6.6 ± 2.5
Confidence interval: 95%, n = 3.
 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320
The most likely explanation to this behaviour can be that
although ascorbic acid is very popular due to its antioxidant
properties (Arrigoni & de Tullio, 2002; Elliot, 1999), it can
act as antioxidant or as pro-oxidant depending on the con-
centration in the solution (Yen, Duh, & Tsai, 2002), being
this fact observed in the present study. On the other hand,
rosemary extracts exhibit a powerful antioxidant activity
and they are widely used in the food industry, as mentioned
by Djenane, Sánchez-Escalante, Beltrán, and Roncalés
(2002) when considering the effectiveness of rosemary for
lowering oxidation processes in food. In fact, carnosic acid
has been identified as the compound with the highest anti-
oxidant activity followed by carnosol, rosmarinic acid,
rosmanol and rosmadial, all of them present in rosemary
extracts (Damechki, Sotiropoulou, & Tsimidou, 2001).
Ascorbic acid alone has been also considered for extend-
ing the shelf-life of different foods (Majchrzak, Mitter, &
Elmadfa, 2004), but it can work as antagonist when used
in combination with other antioxidants, by eliminating
their antioxidant effects or as synergist improving their
antioxidant properties (Ahn & Nam, 2004). In fact, both
ascorbic acid and rosemary extracts have been used
together for preventing food oxidation (Djenane et al.,
2002, 2003). In this work, no significant correlation
between protection and oxidation has been observed with
ascorbic acid, concluding that this compound is not appro-
priate to show the antioxidant properties of the active film,
since both antioxidants are in competence each other and a
clear behaviour cannot be attributed to the presence of the
active packaging.
Anyway, other oxidation mechanisms or oxygen radi-
cals can be involved in the whole process that, at the
moment, cannot be directly measured. That is why another
750
700
650
Fe(II), µg·g-1
600
550
500
450
0.1 8 12
Fig. 2. Effect of different concentrations of natural antioxidants in an active film on the oxidation processes using a 670 lg g1 iron (II) solution and cells
of 325 mL. Error bars corresponding to n = 4, 95% confidence level.
317
technical evaluation of the antioxidant behaviour of this
new active packaging is required.
3.2. Iron (II)
In both set of experiments the presence of the active film
in the cell showed an efficient protection against oxidation
and the concentration of iron (II) remained constant for
longer time than in absence of the active film.
Fig. 2 shows an example of this behaviour when a solu-
tion of 670 lg g1 of iron (II) was used as model com-
pound. As expected, higher protection when increasing
natural antioxidant content of the active film is observed,
being PR3 the sample which showed a higher protection
rate of about 13.4% between 19 and 22 days. A significant
decrease of the iron (II) concentration was observed during
the first 12 days, after which the concentration remained
almost constant depending on the samples, as a demonstra-
tion of the non linear trend of the oxidation process.
Besides, it was noticed that protection effect expired in all
cases after 26 days, being this performance similar in most
of the tests carried out.
No significant differences in statistical terms (n = 4,
95%) within the active film samples were found, concluding
that the amount of antioxidants immobilized in the film
was probably enough in PR1 and an increase of their con-
centration had no critical influence on the antioxidant
behaviour.
Fig. 3 depicts the results obtained when big cells were
used, where the quantity of atmospheric oxygen inside
was considerably higher than in the small ones. A very evi-
dent protection effect against oxidation produced by the
active film can be here observed. Although no significant
PP
PR1
PR2
PR3
19 22 26 29 32
Time (days)
318 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320

differences were detected between PP and PR1, the higher
antioxidant content of PR2 and PR3 was critical in this
case. Specifically, PR3 inhibited the oxidation extent up
to 30.1% when compared to PP at 15 days.
When comparing both series of experiments, it is clear
that the presence of the active packaging is essential for
preventing the oxidation. It must be emphasized that the
amount of oxygen inside the cells is much higher than the
usual one in real packages. As this protection is not depen-
dent on physical contact between the active film and the
oxidizable compound, the tests carried out demonstrate
that the protection mechanism takes place in the vapour
phase, which is one of the most important achievements
of this study.

180
PP
170 PR1
PR2
160 PR3

150
Fe(II), µg·g-1

140

130

120

110

100
0.1 5 12 15 18
Time (days)

Fig. 3. Effect of different concentrations of natural antioxidants in an active film on the oxidation processes using a 160 lg g1 iron (II) solution and cells
of 550 mL. Error bars corresponding to n = 4, 95% confidence level.

450

400

350 PP

PR 1
300
PR 3
Malondialdehyde (µmol·g-1)

250

200

150

100

50

0
0 11 15 20 25 29

Time (days)

Fig. 4. TBARS (lmolmalondialdehyde g1 solution) versus time in different solutions in contact with the new active packaging (PR1, PR3) and PP used as
control film. Error bars corresponding to n = 3, 95% confidence level.
 C. Nerı́n et al. / Journal of Food Engineering 84 (2008) 313–320
12000000
11000000
10000000
9000000
Abundance
8000000
7000000
6000000
Methyl
5000000
palmitate
4000000
3000000
2000000
1000000
0
8.00 10.00 12.00
Fig. 5. GC-MS chromatogram of FAME in flax seed oil after 19 days exposed to active packaging sample PR1 (A) and PP used as control (B).
3.3. Fatty acids
Values of TBARS (expressed as lmol of malondialde-
hyde per gram of solution) are shown in Fig. 4. As can
be seen, malondialdehyde formation was strongly inhibited
in the presence of the new antioxidant active packaging,
showing significant differences with respect to control sam-
ples. However, this effect was only evident after 15 days of
testing. Maximum protection was reached at 20 days,
where 38.0% and 42.2% of the PP value were achieved by
PR1 and PR3, respectively. In any case, no significant dif-
ferences in statistical terms (95%, n = 3) were found
between both active samples, thus concluding that the 1%
of antioxidant is quantity enough to get notorious protec-
tion. Future work will be made with lower concentrations
of Amexol in the film.
A semiquantitative peak area-based determination by
GC-MS was carried out as screening procedure to study
the behaviour of the antioxidant active packaging with
respect to fatty acids, by selecting 19 days as exposure time.
As can been in Fig. 5A corresponding to PR1, only two
compounds can be observed after this period: methyl palm-
itate (tR: 9.5 min) and methyl linolenate (tR: 11.1 min), rec-
ognized to be two of the major compounds of flax seed oil.
Nevertheless, all of them were notoriously oxidized in
absence of the active film, as shows Fig. 5B, which shows
the absence of peaks in the control package (PP)
chromatogram.
In conclusion, the active film containing natural antiox-
idants turned out to be effective also with these oxidizable
compounds. Additional studies have been performed with
real food to demonstrate the efficiency of this innovative
system in different situations (Nerı́n et al., 2006).
From all the studies carried out here, there is a clear evi-
dence of the antioxidant properties of the new active film.
319
Methyl linolenate
(A)
(B)
14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00
Time (min)
It must be emphasized that these properties occur in
vapour phase, as no direct contact between the active film
and the model compounds is allowed. This performance
suggests that the film acts in fact as a radical scavenger,
then preventing the oxidation process by trapping the rad-
icals responsible for this. The hypothesis has been con-
firmed and quantitative evidence of the antioxidant
capacity of the new active film can now be measured (Pezo,
Salafranca, & Nerı́n, 2006).
Acknowledgements
This research has been financed by the Project COOP-
ERA – Innovaragón 2003–2004 from Gobierno de Aragón
(Spain) and INIA-CAL03-080 C4 from the Ministry of Sci-
ence and Technology, Spain. Artibal S.A. is acknowledged
for providing the active films samples.
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