Trends in Food Science & Technology 20 (2009) 92e99
Viewpoint
New antimicrobial
active package for
bakery products
Laura Gutiérreza, Cristina
Sáncheza,b, Ramón Batllea,b and
Cristina Nerı́na,*
a
Aragon Institute of Engineering Research (I3A), CPS,
University of Zaragoza, Marı́a de Luna 3, 50018
Zaragoza, Spain (Tel.: D34 976761873; fax: D34
976762388; e-mail: cnerin@unizar.es)
b without preservatives but keeping free from microbial
Department of IDDDi, ARTIBAL S.A., Cañada Real
12, 22600 Sabiñánigo, Huesca, Spain
The use of essential oils as food preservatives has gained a great
deal of attention over the last years. Specifically, the protective
potential achieved by their addition to the packaging material
has been demonstrated in vitro. Nevertheless, there is very little
information available concerning their use with real, complex
food and no procedure for quality evaluation is readily available.
This manuscript presents a number of alternatives useful for
active essential oil-based packaging as well as a procedure for
sensory evaluation that can be easily implemented in different
food products. The use of a cinnamon-based active package
has been proven to increase more than three times the product
shelf-life of a complex bakery product with a minimal change
in the packaging and no additional manipulation steps. The
quality and definition characteristics were not altered by the
use of this concept. Product is nowadays in the final steps prior
to market introduction.
Introduction
Fungi are the most common spoilers in bakery products.
Commonly, a shelf-life around 3e4 days may be expected
when they are unpreserved. Apart from the repelling sight
of visible growth, fungi are responsible for off-flavour for-
mation and the production of mycotoxins and allergenic
* Corresponding author.
0924-2244/$ - see front matter Ó 2008 Published by Elsevier Ltd.
doi:10.1016/j.tifs.2008.11.003
compounds. The use of weak organic acids such as pro-
pionic, benzoic, and sorbic, investigation on the packaging
material, or modified atmosphere packaging in the last
years have been the main choice for satisfying the market
demands to extend the shelf-life of bakery products
(Fernández, Vodorotz, Courtney, & Pascall, 2006; Legan,
1993; Pagani, Lucisano, Mariotti, & Limbo, 2006). Specif-
ically, ethanol has been commercially used in the form of
a sachet or incorporated into the packaged material;
however, this option has a negative public perception, raises
the chances to develop a residual flavour and implies regu-
latory issues (Plastic additives, 2004). Apart from chemical
preservatives, ultraviolet light and aseptic packaging have
been proposed for controlling microbial growth in bread.
Nowadays, consumers show preferences for products
growth, toxins and other quality deteriorating factors main-
taining freshness and sensorial qualities. Therefore,
challenge for the food industry is to fulfil these demands
with minimum change in food quality and maximum secu-
rity, without using chemical preservatives (Vermeiren,
Devlieghere, van Beest, de Kruijf, & Debevere, 1999).
Active packaging is a very interesting alternative to both
the use of preservatives or modified atmosphere packaging
MAP. It involves the incorporation of agents in the packag-
ing that can either interact directly with the packaged
foodstuff or with the atmosphere inside the package. The
development of active materials with properties for enhanc-
ing the shelf-life and safety of packaged food is nowadays
one of the most challenging research activities. Several ap-
proaches have been proposed but only a few of them have
become commercially available (Matan et al., 2006; Nerı́n
et al., 2006; Suppakul, Miltz, Sonneveld, & Bigger, 2006;
Tovar, Salafranca, Sánchez, & Nerı́n, 2005) and applica-
tions to bakery products are very scarce (Nielsen & Rios,
2005; Suhr & Nielsen, 2005). Some antimicrobial films
have already been evaluated for food-packaging applica-
tions, such as alpha- and beta-cyclodextrin encapsulated
allyl isothiocyanate in polylactide-co-polycaprolactone
films (Plackett & Ghambari-Siahkali, 2007); polymeric
films coated that incorporate Enterocin 416K1 (Iseppi
et al., 2008); blends of gliadins and chitosan (Fernández-
Saiz, Lagarón, Hernández-Muñoz, & Ocio-Saiz, 2008) or
hexamethylenetetramine (Devlieghere, Vermeiren, Jacobs,
& Debevere, 2000); polyethylene coated with an antimicro-
bial peptide (Miltz, Rydlo, Mor, & Polyakov, 2006) or
 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99
nisin-incorporated cross-linked hydroxypropylmethyl-cel-
lulose (Sebti, Delves-Broughton, & Coma, 2003) .
Although the antimicrobial properties of natural extracts
have been known for centuries, only limited publications
have reported their use in active packaging (López,
Sanchez, Batlle, & Nerı́n, 2007b; Matan et al., 2006; Rodrı́-
guez, Nerı́n, & Batlle, 2008). The use of natural extracts,
such as essential oils (EO hereafter) and their constituents,
categorized as flavourings by the European Union (2002/
113/EC, 2002; 2004/1935/EC, 2004; 89/107/EEC, 1989);
and as GRAS (Generally Recognized as Safe) by the US
Food and Drug Administration, in plastic films to avoid
microbial food spoilage is an attractive option for both
packaging manufacturers and demanding consumers.
In these concepts, the active substances are released
from the packaging material to the surface of the product
during the whole shelf-life of the packaged foodstuff. Anti-
microbials incorporated into packaging materials can,
therefore, control microbial contamination by reducing
the growth rate and maximum growth population and/or
extending the lag-phase of the target microorganism or by
inactivating microorganism by contact (Quintavalla &
Vicini, 2002). The usefulness of these packaging alterna-
tives have been demonstrated at a laboratory scale using
in vitro conditions (López, Sanchez, Batlle, & Nerı́n,
2007a; López et al., 2007b) and even some of them have
been tried with slice bread (Rodrı́guez et al., 2008) but
far more information is needed on their performance
when working with commercial products.
Therefore, this paper has two main aims. First, to test the
usefulness of the natural extract-active film developed for
its use with a commercial complex bakery product. Second,
to fully evaluate the sensory characteristics of the proposed
setup in order to detect potential drawbacks or pitfalls not
related with food objective quality but to food subjective
perception. Obviously, these results constitute the final
decision tool for launching into the market the packaged
food product.
Essential oils and antimicrobial films
The essential oil (EO) of Cinnamomum zeylanicum
(cinnamon, Chemical Abstract Service, CAS, number:
8015-91-6) was supplied by Argolide Quı́mica S.L. (Barce-
lona, Spain). The antimicrobial films were prepared in the
laboratory by incorporating known concentrations (w/w)
of the EO in films of polypropylene (PP hereafter) (30-mm
thick, supplied by Poligal, S.A., Narón, Spain) suitable for
food packaging via an innovative process protected by
a European Patent EP1657181 held by the company
ARTIBAL S.A. (Sabiñánigo, Spain).
Bakery product
A grand total of 54 independent samples of the bakery
product from different batches were used in this study. All
of them were supplied by a small size company interested
in the study. Because of the commercial interest of this
93
development, a confidential agreement applies and no
more data about the company will be provided. The product
is a complex mixture prone to microbial infestation and
quality deterioration. Yolk, almonds, nuts, raisins swilled
in rum as well as other common ingredients are included
in the formula. The whole product is covered with a sugar
layer.
The main desirable characteristics of the product were
defined as tenderness of the inner part while nuts and
almonds kept crunchy and the yolk cannot be visually
distinguished from the mixture and only the taste can be
perceived. This product has an additional challenge as the
requirement of humidity to keep tenderness should not
influence the crunching and flavouring of the other ingredi-
ents. This product is well balanced just made and the same
properties are required for its shelf-life. No preservatives
are added during its baking. In fact, this is considered as
a handmade product.
Headspace-single drop microextraction (HS-SDME)
The atmosphere generated inside the active bag was
sampled using headspace-single drop microextraction (HS-
SDME) as described by Romero, López, Rubio, Batlle, and
Nerı́n (2007). Briefly, a 5-mL microsyringe with a 26S type
needle (Hamilton 85RN, Bonaduz, Switzerland) containing
p-xylene as the extractive phase was introduced inside the
bag. The plunger was then depressed and a 2.5-mL drop
was exposed to the sample for 5 min. After exposure, the
drop was retracted again into the syringe and then, it was
transferred to the injection port of a GCeMS system.
Gas chromatographyemass spectrometric
(GCeMS) analysis
GCeMS analysis were performed using a Hewlett-Pack-
ard 6890 chromatograph (Wilmington, DE, USA) equipped
with a 5973 mass selective detector and a HP-5 MS
(60 m  0.25 mm, 0.25 mm film thickness) capillary column.
The temperature program for the GC was as follows: initial
temperature 75  C, 10  C/min ramp to 190  C, then
20  C/min to 280  C, held for 5 min. The injector tempera-
ture was maintained at 300  C and operated in splitless
injection mode with the valve closed for 1 min. Helium
(99.9999% pure, Carburos Metálicos, Zaragoza, Spain)
was used as the carrier gas at a constant flow of 1 mL per
min. The interface temperature was 280  C and the ionization
mode was electron impact (70 eV). The mass selective detec-
tor was operated in the scan mode between 45 and 400 m/z.
Sensory testing
A trained panel of 12 individuals was used to evaluate
the sensorial properties and possible changes in the bakery
product by each different option of packaging under study.
This was made in order to verify that the extension in shelf-
life would not affect the quality of the product. The
technique used in this case was the classification of the
responses given by the tasters in a scale where several
94 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99
descriptors, defined according their relevance in the final
quality, were qualified. The perception of the tasters
concerning the magnitude of the differences, the intensity
of the descriptors related to taste, flavour, sight, color and
tender were evaluated in a scale ranging from 1 to 8 points,
being 8 the highest intensity. Three experts (members of the
staff of the manufacturing company) were also included in
the panel as a reference.
Testing was conducted over a 28-days period, covering
a time-span longer than the required shelf-life, established
by the manufacturing company in 10 days. Currently the
shelf-life is no longer than three days. The following
check-points were scheduled (reported as days from
manufacturing): 0, 3, 6, 10, 14, 21, and 28. As a reference,
recently just made products were introduced in the trials.
Packaging options
The bakery samples were introduced in a bag made
using all the PP packaging materials and different options
evaluated. Then, the plastic bags were thermosealed in all
cases. The following options were evaluated in the tests.
1 Paraffin paper. This is the traditional packaging
material used to cover this specific product by the
manufacturing company (Blank).
2 PP non-active nor micro perforated (PPno).
3 PP non-active micro perforated with a density of 16
holes per dm2 (PPnoM)
4 PP active (4% w/w) micro perforated with a density of
16 holes per dm2 (PPM).
5 PP active (2% w/w) partially micro perforated (3 rows
of 5 cm each) (3 row).
6 PP active (2% w/w) partially micro perforated (2 rows
of 5 cm each) (2 row).
7 Same as 5 including a polyethylene (PE)-coated board
piece to separate the bottom of the product from the
board secondary package (coated board).
8 Same as 6 where the PE-coated board piece has been
metalized and introduced into the active PP bag (alumi-
num tray).
In every test, fresh, just made products in the traditional
packaging material were used as blank samples. All options
were kept inside the cardboard box used for the company as
secondary packaging for commercial and marketing
purposes.
Development
In a preliminary step, the suitability of the packaging
material intended to come into contact with food was
evaluated. As has been described in the introductory
section, no regulatory issues arise from the use of essen-
tial oils as preservative. Nevertheless, the first stage
consisted of measuring the volatile compounds released
by the active material to get an idea about the potential
impact characteristics of the product. The rationale for
this work is that, in a previous work (López, Huerga,
Batlle, & Nerı́n, 2006), the release and identification
of the active compounds from the active film was
determined.
Fig. 1 shows the chromatograms obtained when sam-
pling the headspace generated inside the bag using the
SDME GCeMS strategy described in the Experimental
Section. As can be seen, the very same chemicals were
detected with (top graph) (PPM) or without (bottom graph)
(PPnoM) active film and they are included in the character-
istic flavour of the product. Nevertheless, when the active
packaging (PPM) was analysed in the absence of the bakery
product, a clear signal corresponding to cinnamaldehyde
(chemical responsible of the cinnamon characteristic
flavour) was obtained. Therefore, it is hypothesized that
this chemical is readily absorbed by the product and as
a consequence, it was included in the flavour descriptors
as an undesirable characteristic.
Different groups of descriptors were identified and
used in the sensory evaluation. They can be roughly
divided into five groups: taste, odour (flavour), texture,
general appearance and presence of moulds. Every group
was fully divided into more specific indicators, which
were included into the evaluation sheet. These indicators
were as follows.
1 Taste: the following individual markers were defined
and included.
 Characteristic taste, as a positive factor. It repre-
sents the similarity obtained by the individual
between the test and the blank items.
 Cinnamon taste. This factor can be considered
either positive or negative, since acceptance of
this specific taste is very subjective. Nevertheless,
since it represents an alteration of the expected
taste, it was included as negative.
 Board taste. This is a negative factor and repre-
sents the alteration due to the board material
used as secondary packaging.
2 Flavour.
 Board flavour. This is also a negative factor and
represents alteration due to the board material
used as secondary packaging.
 Cinnamon flavour. As in the previous paragraph, it
was considered as negative factor.
3 Texture.
 Tenderness. Positive factor, representing the appro-
priate resistance of the product when chewed,
relative to the blank item.
 Crunchiness and crispiness. Positive factor, rela-
tive to the consumer’s perception of the nuts
when compared to the blank item.
 Elasticity rubber-like. Negative factor, referring to
the change in the product overall texture. It was
defined by the expert members of the sensory eval-
uation board as a clear indicative of aging.
 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99 95

Fig. 1. Typical gas-chromatograms obtained inside the PP bag in the final commercial disposition with packaged product. Top panel, active film used
(PPM, option 4); bottom panel, raw PP used (PPnoM, option 3).
96 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99

4 General appearance. ONeOFF test, concerning the final acceptance of the

 Freshness. Positive, description of the overall
perception of the product when compared to the
blank item.
 Fat stains. Negative, with the original packaging, it
was observed that stains appeared with time on the
bottom of the board secondary package. They were
attributed to the fat migration from the packaged
product, and severely detract from marketability.
5 Presence of moulds. Zero-tolerance was defined for this
descriptor, meaning that it was considered as an
different packaging options. Nevertheless, for clarity
purposes, the relative abundance of the moulds onto
the packaged foodstuffs was represented by a numerical
value: 1 represents total absence of visible growth,
whereas 7 indicates complete contamination by moulds.
Fig. 2AeC illustrates the evaluation results of the sen-
sory attributes obtained when using the active package
approaches (three independent replicates were analyzed
for each control point). As can be seen, a clear

3 Days
A 9
8
7
6
Fresh Appearance
Score

5
4 Crunchy and Crispy

3
Characteristic Taste
2
1
0
Blank PPNo PP NoM PP M 2 Row 3 Row C. Board A. Tray
Sample

7 Days
B 8

5
Score

Fresh Appearance
4
Crunchy and Crispy
3

2 Characteristic Taste

0
Blank PPNo PP NoM PP M 2 Row 3 Row C. Board A. Tray
Sample

10 Days
C 8

5
Score

Fresh Appearance
4
Crunchy and Crispy
3
Characteristic Taste
2

0
Blank PPNo PP NoM PP MA 2 Row 3 Row C. Board A. Tray
Sample

Fig. 2. Results obtained in the evaluation of the diverse active packages. For package identification, please proceed to the experimental section. All
the results represent the average of three replicates.
 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99 97

improvement with respect to the traditional package was

detectable even at the initial stages (day 3) of the evaluation
was demonstrated. It is worth to highlight that the punctu-
ations obtained were consistently high for all the active so-
lutions at day 7 and that the decrease was only detected at
day 10, varying among the various active solutions. Taking
into account the scale used for the different descriptors and
the expectancies, an average value of 5 was defined as an
acceptable product. Only the aluminum tray packaging
(A. Tray, option 8) gave these results at day 10. To explain
in detail these achievements, it is interesting to go deeper
into the results. The next set of spider plots illustrate in
0_Parafina
detail the results obtained for each packaging combination
at day 10. Spider plot was selected since it provides a very
intuitive and direct view of the results; factors were
arranged so that parameters considered as negative are at
the top and the positive attributes can be seen at the bottom
of the figures. Every spider plot represents the average of
three independent replicates.
As can be seen in Fig. 3, when the traditional package
(top) was changed to raw PP (middle graph) significant wors-
ening was obtained for the elasticity descriptor, whereas the
other factors remained somehow unchanged with only slights
improvement for characteristic taste and fat stains. As a first
modification, a physical change in the packaging material
(micro perforation, as described in the materials and methods
section) was then evaluated (bottom graph) and proven only 1_PP non active no microperforated
successful in reduction of the rubber-like perception. As can 2_PP non active microperforated
be seen, very low numbers (2 or 3) were obtained for the
positive factors, whereas very high values were obtained
for the negative ones, reflecting the clear fact that the product
does not fit the required quality standards.
No significant improvement in the positive factors other
than the tenderness (2 to 3) was found. Considering these
results together with the ones described in the previous
paragraph it seems clear that the combination of different mi-
cro perforation densities to reduce elasticity rubber-like and
active packaging to reduce presence of moulds could consti-
tute a better approach. Three different densities were tested
and the results are displayed in Fig. 4. In the top graph,
a reduction of presence of moulds from 7 without active
packaging to 1 for the active option was observed. Middle Fig. 3. Non-active packaging options: spider plots obtained after sen-
sory evaluation at day 10. Top, traditional (blank, option 1) package;
and bottom graphs gave best results for cinnamon perception middle, PPno (option 2); bottom, micro perforated PPnoM (option 3).
while maintaining the antifungal capabilities required.
Moreover, these combinations increased the scores in the
positive factors, especially in characteristic taste, as could success since a reduction in scores for these factors was
be expected. The last challenge was to reduce the high scores obtained as well as a significant increase in positive factors
obtained by the factors that can be related with the secondary such as tenderness and freshness with no worsening of any
package-board interaction such as board taste and flavour, as desirable performance. In order to achieve a final improve-
well and fat stains. To do so, the best active plastic package ment of the package, the PE-coated piece was metalized
results were combined with different secondary package (package option 8) to further minimize the interaction
combinations and are illustrated in Fig. 5. First, a board between the secondary package and the product. As depicted
tray coated with PE layer (package option 7) was introduced in Fig. 5 (bottom), the results obtained were very good,
between the secondary package and the product in order to providing the lowest scores for the negative factors and the
minimize the transference of board characteristics to the maximum scores for the positive ones (with the only excep-
product. As shown in Fig. 5 (top), this option was a clear tion of the freshness which provides a score of 7 instead of 8).
98 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99

Elasticity
Rubber
Board 10.00 Like
Taste 9.00
8.00 Presence
Board Flavour 7.00 of
6.00 Molds
5.00
4.00
3.00
2.00
Fat Stains 1.00 Cinnamon
Taste

Tenderness Characteristic
Taste

Fresh
Appearence Crunch and Crispy

Elasticity
Rubber
Board 10.00 Like
Taste 9.00
8.00 Presence
Board Flavour 7.00 of
6.00 Molds
5.00
4.00
3.00
2.00
Fat Stains 1.00 Cinnamon
Taste

Tenderness Characteristic
Taste

Fresh
Appearence Crunch and Crispy

Fig. 5. Different alternatives tested to minimise secondary packag-

ing-food alterations: spider plots obtained after sensory evaluation
at day 10. Top, use of PE-coated board (option 7); bottom, metalized
PE-coated board (option 8).

in shelf-life from 3 to 10 days with maximum quality and

Fig. 4. Combination of micro perforation and active concentration:
spider plots obtained after sensory evaluation at day 10. Top, PP
(4%) active micro perforated (PPM, option 4); middle, PP (2%) active
partially micro perforated (3 Row, option 5); bottom, PP (2%) active
partially micro perforated (2 Row, option 6).
Conclusions
This work has illustrated the successful design, develop-
ment and testing of an active package based on the use of
cinnamon essential oil in combination with micro perfo-
rated polypropylene. The final design provided an increase
safety, as demonstrated by the sensory evaluation results.
The evaluation procedure described (including factor defi-
nition and evaluation) can be easily adapted to any bakery
product and could be established as a standard for this
development. Further work is currently under way for
industrial scale-up. Hopefully, the product will be in the
market in the near future.
Acknowledgments
This work has been financed by the Research Projects
Cal03-080 from INIA and FEDER; AGL2004-07545 from
the Spanish Ministry of Education and University and
FEDER and INTERREG IIIA-5-326 C. L.G. acknowl-
edges the Spanish Ministry of Education and University
for a grant (BES-2005-10186). R.B. expresses his grati-
tude to the former Spanish Ministry of Science and
 L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99
Technology for personal funding through the Ramón y
Cajal program.
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