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Trends in Food Science & Technology 20 (2009) 92e99

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New antimicrobial
active package for 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
bakery products 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,
Laura Gutiérreza, Cristina 1993; Pagani, Lucisano, Mariotti, & Limbo, 2006). Specif-
Sáncheza,b, Ramón Batllea,b and ically, ethanol has been commercially used in the form of
a sachet or incorporated into the packaged material;
Cristina Nerı́na,* however, this option has a negative public perception, raises
the chances to develop a residual flavour and implies regu-
a
Aragon Institute of Engineering Research (I3A), CPS, latory issues (Plastic additives, 2004). Apart from chemical
University of Zaragoza, Marı́a de Luna 3, 50018 preservatives, ultraviolet light and aseptic packaging have
Zaragoza, Spain (Tel.: D34 976761873; fax: D34 been proposed for controlling microbial growth in bread.
976762388; e-mail: cnerin@unizar.es) Nowadays, consumers show preferences for products
b without preservatives but keeping free from microbial
Department of IDDDi, ARTIBAL S.A., Cañada Real
12, 22600 Sabiñánigo, Huesca, Spain growth, toxins and other quality deteriorating factors main-
taining freshness and sensorial qualities. Therefore,
challenge for the food industry is to fulfil these demands
The use of essential oils as food preservatives has gained a great with minimum change in food quality and maximum secu-
deal of attention over the last years. Specifically, the protective rity, without using chemical preservatives (Vermeiren,
potential achieved by their addition to the packaging material Devlieghere, van Beest, de Kruijf, & Debevere, 1999).
has been demonstrated in vitro. Nevertheless, there is very little Active packaging is a very interesting alternative to both
information available concerning their use with real, complex the use of preservatives or modified atmosphere packaging
food and no procedure for quality evaluation is readily available. MAP. It involves the incorporation of agents in the packag-
ing that can either interact directly with the packaged
This manuscript presents a number of alternatives useful for foodstuff or with the atmosphere inside the package. The
active essential oil-based packaging as well as a procedure for development of active materials with properties for enhanc-
sensory evaluation that can be easily implemented in different ing the shelf-life and safety of packaged food is nowadays
food products. The use of a cinnamon-based active package one of the most challenging research activities. Several ap-
has been proven to increase more than three times the product proaches have been proposed but only a few of them have
shelf-life of a complex bakery product with a minimal change become commercially available (Matan et al., 2006; Nerı́n
in the packaging and no additional manipulation steps. The et al., 2006; Suppakul, Miltz, Sonneveld, & Bigger, 2006;
quality and definition characteristics were not altered by the Tovar, Salafranca, Sánchez, & Nerı́n, 2005) and applica-
use of this concept. Product is nowadays in the final steps prior tions to bakery products are very scarce (Nielsen & Rios,
to market introduction. 2005; Suhr & Nielsen, 2005). Some antimicrobial films
have already been evaluated for food-packaging applica-
tions, such as alpha- and beta-cyclodextrin encapsulated
Introduction
allyl isothiocyanate in polylactide-co-polycaprolactone
Fungi are the most common spoilers in bakery products.
films (Plackett & Ghambari-Siahkali, 2007); polymeric
Commonly, a shelf-life around 3e4 days may be expected
films coated that incorporate Enterocin 416K1 (Iseppi
when they are unpreserved. Apart from the repelling sight
et al., 2008); blends of gliadins and chitosan (Fernández-
of visible growth, fungi are responsible for off-flavour for-
Saiz, Lagarón, Hernández-Muñoz, & Ocio-Saiz, 2008) or
mation and the production of mycotoxins and allergenic
hexamethylenetetramine (Devlieghere, Vermeiren, Jacobs,
& Debevere, 2000); polyethylene coated with an antimicro-
* Corresponding author. bial peptide (Miltz, Rydlo, Mor, & Polyakov, 2006) or
0924-2244/$ - see front matter Ó 2008 Published by Elsevier Ltd.
doi:10.1016/j.tifs.2008.11.003
L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99 93

nisin-incorporated cross-linked hydroxypropylmethyl-cel- development, a confidential agreement applies and no


lulose (Sebti, Delves-Broughton, & Coma, 2003) . more data about the company will be provided. The product
Although the antimicrobial properties of natural extracts is a complex mixture prone to microbial infestation and
have been known for centuries, only limited publications quality deterioration. Yolk, almonds, nuts, raisins swilled
have reported their use in active packaging (López, in rum as well as other common ingredients are included
Sanchez, Batlle, & Nerı́n, 2007b; Matan et al., 2006; Rodrı́- in the formula. The whole product is covered with a sugar
guez, Nerı́n, & Batlle, 2008). The use of natural extracts, layer.
such as essential oils (EO hereafter) and their constituents, The main desirable characteristics of the product were
categorized as flavourings by the European Union (2002/ defined as tenderness of the inner part while nuts and
113/EC, 2002; 2004/1935/EC, 2004; 89/107/EEC, 1989); almonds kept crunchy and the yolk cannot be visually
and as GRAS (Generally Recognized as Safe) by the US distinguished from the mixture and only the taste can be
Food and Drug Administration, in plastic films to avoid perceived. This product has an additional challenge as the
microbial food spoilage is an attractive option for both requirement of humidity to keep tenderness should not
packaging manufacturers and demanding consumers. influence the crunching and flavouring of the other ingredi-
In these concepts, the active substances are released ents. This product is well balanced just made and the same
from the packaging material to the surface of the product properties are required for its shelf-life. No preservatives
during the whole shelf-life of the packaged foodstuff. Anti- are added during its baking. In fact, this is considered as
microbials incorporated into packaging materials can, a handmade product.
therefore, control microbial contamination by reducing
the growth rate and maximum growth population and/or Headspace-single drop microextraction (HS-SDME)
extending the lag-phase of the target microorganism or by The atmosphere generated inside the active bag was
inactivating microorganism by contact (Quintavalla & sampled using headspace-single drop microextraction (HS-
Vicini, 2002). The usefulness of these packaging alterna- SDME) as described by Romero, López, Rubio, Batlle, and
tives have been demonstrated at a laboratory scale using Nerı́n (2007). Briefly, a 5-mL microsyringe with a 26S type
in vitro conditions (López, Sanchez, Batlle, & Nerı́n, needle (Hamilton 85RN, Bonaduz, Switzerland) containing
2007a; López et al., 2007b) and even some of them have p-xylene as the extractive phase was introduced inside the
been tried with slice bread (Rodrı́guez et al., 2008) but bag. The plunger was then depressed and a 2.5-mL drop
far more information is needed on their performance was exposed to the sample for 5 min. After exposure, the
when working with commercial products. drop was retracted again into the syringe and then, it was
Therefore, this paper has two main aims. First, to test the transferred to the injection port of a GCeMS system.
usefulness of the natural extract-active film developed for
its use with a commercial complex bakery product. Second, Gas chromatographyemass spectrometric
to fully evaluate the sensory characteristics of the proposed (GCeMS) analysis
setup in order to detect potential drawbacks or pitfalls not GCeMS analysis were performed using a Hewlett-Pack-
related with food objective quality but to food subjective ard 6890 chromatograph (Wilmington, DE, USA) equipped
perception. Obviously, these results constitute the final with a 5973 mass selective detector and a HP-5 MS
decision tool for launching into the market the packaged (60 m  0.25 mm, 0.25 mm film thickness) capillary column.
food product. The temperature program for the GC was as follows: initial
temperature 75  C, 10  C/min ramp to 190  C, then
Essential oils and antimicrobial films 20  C/min to 280  C, held for 5 min. The injector tempera-
The essential oil (EO) of Cinnamomum zeylanicum ture was maintained at 300  C and operated in splitless
(cinnamon, Chemical Abstract Service, CAS, number: injection mode with the valve closed for 1 min. Helium
8015-91-6) was supplied by Argolide Quı́mica S.L. (Barce- (99.9999% pure, Carburos Metálicos, Zaragoza, Spain)
lona, Spain). The antimicrobial films were prepared in the was used as the carrier gas at a constant flow of 1 mL per
laboratory by incorporating known concentrations (w/w) min. The interface temperature was 280  C and the ionization
of the EO in films of polypropylene (PP hereafter) (30-mm mode was electron impact (70 eV). The mass selective detec-
thick, supplied by Poligal, S.A., Narón, Spain) suitable for tor was operated in the scan mode between 45 and 400 m/z.
food packaging via an innovative process protected by
a European Patent EP1657181 held by the company Sensory testing
ARTIBAL S.A. (Sabiñánigo, Spain). A trained panel of 12 individuals was used to evaluate
the sensorial properties and possible changes in the bakery
Bakery product product by each different option of packaging under study.
A grand total of 54 independent samples of the bakery This was made in order to verify that the extension in shelf-
product from different batches were used in this study. All life would not affect the quality of the product. The
of them were supplied by a small size company interested technique used in this case was the classification of the
in the study. Because of the commercial interest of this 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 this work is that, in a previous work (López, Huerga,
quality, were qualified. The perception of the tasters Batlle, & Nerı́n, 2006), the release and identification
concerning the magnitude of the differences, the intensity of the active compounds from the active film was
of the descriptors related to taste, flavour, sight, color and determined.
tender were evaluated in a scale ranging from 1 to 8 points, Fig. 1 shows the chromatograms obtained when sam-
being 8 the highest intensity. Three experts (members of the pling the headspace generated inside the bag using the
staff of the manufacturing company) were also included in SDME GCeMS strategy described in the Experimental
the panel as a reference. Section. As can be seen, the very same chemicals were
Testing was conducted over a 28-days period, covering detected with (top graph) (PPM) or without (bottom graph)
a time-span longer than the required shelf-life, established (PPnoM) active film and they are included in the character-
by the manufacturing company in 10 days. Currently the istic flavour of the product. Nevertheless, when the active
shelf-life is no longer than three days. The following packaging (PPM) was analysed in the absence of the bakery
check-points were scheduled (reported as days from product, a clear signal corresponding to cinnamaldehyde
manufacturing): 0, 3, 6, 10, 14, 21, and 28. As a reference, (chemical responsible of the cinnamon characteristic
recently just made products were introduced in the trials. flavour) was obtained. Therefore, it is hypothesized that
this chemical is readily absorbed by the product and as
Packaging options a consequence, it was included in the flavour descriptors
The bakery samples were introduced in a bag made as an undesirable characteristic.
using all the PP packaging materials and different options Different groups of descriptors were identified and
evaluated. Then, the plastic bags were thermosealed in all used in the sensory evaluation. They can be roughly
cases. The following options were evaluated in the tests. divided into five groups: taste, odour (flavour), texture,
general appearance and presence of moulds. Every group
1 Paraffin paper. This is the traditional packaging was fully divided into more specific indicators, which
material used to cover this specific product by the were included into the evaluation sheet. These indicators
manufacturing company (Blank). were as follows.
2 PP non-active nor micro perforated (PPno).
3 PP non-active micro perforated with a density of 16 1 Taste: the following individual markers were defined
holes per dm2 (PPnoM) and included.
4 PP active (4% w/w) micro perforated with a density of  Characteristic taste, as a positive factor. It repre-
16 holes per dm2 (PPM). sents the similarity obtained by the individual
5 PP active (2% w/w) partially micro perforated (3 rows between the test and the blank items.
of 5 cm each) (3 row).  Cinnamon taste. This factor can be considered
6 PP active (2% w/w) partially micro perforated (2 rows either positive or negative, since acceptance of
of 5 cm each) (2 row). this specific taste is very subjective. Nevertheless,
7 Same as 5 including a polyethylene (PE)-coated board since it represents an alteration of the expected
piece to separate the bottom of the product from the taste, it was included as negative.
board secondary package (coated board).  Board taste. This is a negative factor and repre-
8 Same as 6 where the PE-coated board piece has been sents the alteration due to the board material
metalized and introduced into the active PP bag (alumi- used as secondary packaging.
num tray). 2 Flavour.
 Board flavour. This is also a negative factor and
In every test, fresh, just made products in the traditional represents alteration due to the board material
packaging material were used as blank samples. All options used as secondary packaging.
were kept inside the cardboard box used for the company as  Cinnamon flavour. As in the previous paragraph, it
secondary packaging for commercial and marketing was considered as negative factor.
purposes. 3 Texture.
 Tenderness. Positive factor, representing the appro-
Development priate resistance of the product when chewed,
In a preliminary step, the suitability of the packaging relative to the blank item.
material intended to come into contact with food was  Crunchiness and crispiness. Positive factor, rela-
evaluated. As has been described in the introductory tive to the consumer’s perception of the nuts
section, no regulatory issues arise from the use of essen- when compared to the blank item.
tial oils as preservative. Nevertheless, the first stage  Elasticity rubber-like. Negative factor, referring to
consisted of measuring the volatile compounds released the change in the product overall texture. It was
by the active material to get an idea about the potential defined by the expert members of the sensory eval-
impact characteristics of the product. The rationale for 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 different packaging options. Nevertheless, for clarity
perception of the product when compared to the purposes, the relative abundance of the moulds onto
blank item. the packaged foodstuffs was represented by a numerical
 Fat stains. Negative, with the original packaging, it value: 1 represents total absence of visible growth,
was observed that stains appeared with time on the whereas 7 indicates complete contamination by moulds.
bottom of the board secondary package. They were
attributed to the fat migration from the packaged Fig. 2AeC illustrates the evaluation results of the sen-
product, and severely detract from marketability. sory attributes obtained when using the active package
5 Presence of moulds. Zero-tolerance was defined for this approaches (three independent replicates were analyzed
descriptor, meaning that it was considered as an 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


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
Fig. 4. Combination of micro perforation and active concentration: market in the near future.
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
Acknowledgments
partially micro perforated (2 Row, option 6).
This work has been financed by the Research Projects
Cal03-080 from INIA and FEDER; AGL2004-07545 from
Conclusions the Spanish Ministry of Education and University and
This work has illustrated the successful design, develop- FEDER and INTERREG IIIA-5-326 C. L.G. acknowl-
ment and testing of an active package based on the use of edges the Spanish Ministry of Education and University
cinnamon essential oil in combination with micro perfo- for a grant (BES-2005-10186). R.B. expresses his grati-
rated polypropylene. The final design provided an increase tude to the former Spanish Ministry of Science and
L. Gutiérrez et al. / Trends in Food Science & Technology 20 (2009) 92e99 99

Technology for personal funding through the Ramón y Matan, N., Rimkeeree, H., Mawson, A. J., Chompreeda, P.,
Cajal program. Haruthaithanasan, V., & Parker, M. (2006). Antimicrobial activity of
cinnamon and clove oils under modified atmosphere conditions.
International Journal of Food Microbiology, 107, 180e185.
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