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New antimicrobialactive package forbakery products

Laura Gutie´rrez

, CristinaSa´nchez

a,b

, Ramo´n Batlle

a,b

andCristina Nerı´n

Aragon Institute of Engineering Research (I3A), CPS,University of Zaragoza, Marı´a de Luna 3, 50018Zaragoza, Spain (Tel.:

34 976761873; fax:

34976762388; e-mail:cnerin@unizar.es)

Department of I

i, ARTIBAL S.A., Canãda Real12, 22600 Sabinãńigo, Huesca, Spain

The use of essential oils as food preservatives has gained a greatdeal of attention over the last years. Speciﬁcally, the protectivepotential achieved by their addition to the packaging materialhas been demonstrated

in vitro

. Nevertheless, there is very littleinformation available concerning their use with real, complexfoodandnoprocedureforqualityevaluationisreadilyavailable.This manuscript presents a number of alternatives useful foractive essential oil-based packaging as well as a procedure forsensory evaluation that can be easily implemented in differentfood products. The use of a cinnamon-based active packagehas been proven to increase more than three times the productshelf-life of a complex bakery product with a minimal changein the packaging and no additional manipulation steps. Thequality and deﬁnition characteristics were not altered by theuse of this concept. Product is nowadays in the ﬁnal steps priorto market introduction.

Introduction

Fungi are the most common spoilers in bakery products.Commonly, a shelf-life around 3

4 days may be expectedwhen they are unpreserved. Apart from the repelling sightof visible growth, fungi are responsible for off-flavour for-mation and the production of mycotoxins and allergeniccompounds. The use of weak organic acids such as pro-pionic, benzoic, and sorbic, investigation on the packagingmaterial, or modified atmosphere packaging in the lastyears have been the main choice for satisfying the marketdemands to extend the shelf-life of bakery products(Fernańdez, 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, raisesthe chances to develop a residual flavour and implies regu-latory issues (Plastic additives, 2004). Apart from chemicalpreservatives, ultraviolet light and aseptic packaging havebeen proposed for controlling microbial growth in bread.Nowadays, consumers show preferences for productswithout preservatives but keeping free from microbialgrowth, toxins and other quality deteriorating factors main-taining freshness and sensorial qualities. Therefore,challenge for the food industry is to fulfil these demandswith 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 boththe use of preservatives or modified atmosphere packagingMAP. It involves the incorporation of agents in the packag-ing that can either interact directly with the packagedfoodstuff or with the atmosphere inside the package. Thedevelopment of active materials with properties for enhanc-ing the shelf-life and safety of packaged food is nowadaysone of the most challenging research activities. Several ap-proaches have been proposed but only a few of them havebecome commercially available (Matan

et al.

, 2006; Nerı´n

et al.

, 2006; Suppakul, Miltz, Sonneveld, & Bigger, 2006;Tovar, Salafranca, Sańchez, & Nerıń, 2005) and applica-tions to bakery products are very scarce (Nielsen & Rios,2005; Suhr & Nielsen, 2005). Some antimicrobial filmshave already been evaluated for food-packaging applica-tions, such as alpha- and beta-cyclodextrin encapsulatedallyl isothiocyanate in polylactide-co-polycaprolactonefilms (Plackett & Ghambari-Siahkali, 2007); polymericfilms coated that incorporate Enterocin 416K1 (Iseppi

et al.

, 2008); blends of gliadins and chitosan (Fernańdez-Saiz, Lagaroń, Hernańdez-Munõz, & Ocio-Saiz, 2008) orhexamethylenetetramine (Devlieghere, Vermeiren, Jacobs,& Debevere, 2000); polyethylene coated with an antimicro-bial peptide (Miltz, Rydlo, Mor, & Polyakov, 2006) or

* Corresponding author.

0924-2244/$ - see front matter

2008 Published by Elsevier Ltd.doi:10.1016/j.tifs.2008.11.003

Trends in Food Science & Technology 20 (2009) 92

nisin-incorporated cross-linked hydroxypropylmethyl-cel-lulose (Sebti, Delves-Broughton, & Coma, 2003) .Although the antimicrobial properties of natural extractshave been known for centuries, only limited publicationshave reported their use in active packaging (Lo´pez,Sanchez, Batlle, & Nerı´n, 2007b; Matan

et al.

, 2006; Rodrı´-guez, Nerıń, & 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 USFood and Drug Administration, in plastic films to avoidmicrobial food spoilage is an attractive option for bothpackaging manufacturers and demanding consumers.In these concepts, the active substances are releasedfrom the packaging material to the surface of the productduring the whole shelf-life of the packaged foodstuff. Anti-microbials incorporated into packaging materials can,therefore, control microbial contamination by reducingthe growth rate and maximum growth population and/orextending the lag-phase of the target microorganism or byinactivating microorganism by contact (Quintavalla &Vicini, 2002). The usefulness of these packaging alterna-tives have been demonstrated at a laboratory scale using

in vitro

conditions (Lo´pez, Sanchez, Batlle, & Nerı´n,2007a; Lo´pez

et al.

, 2007b) and even some of them havebeen tried with slice bread (Rodrı´guez

et al.

, 2008) butfar more information is needed on their performancewhen working with commercial products.Therefore, this paper has two main aims. First, to test theusefulness of the natural extract-active ﬁlm developed forits use with a commercial complex bakery product. Second,to fully evaluate the sensory characteristics of the proposedsetup in order to detect potential drawbacks or pitfalls notrelated with food objective quality but to food subjectiveperception. Obviously, these results constitute the ﬁnaldecision tool for launching into the market the packagedfood 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 Quı´mica S.L. (Barce-lona, Spain). The antimicrobial ﬁlms were prepared in thelaboratory by incorporating known concentrations (w/w)of the EO in ﬁlms of polypropylene (PP hereafter) (30-

mthick, supplied by Poligal, S.A., Naro´n, Spain) suitable forfood packaging

via

an innovative process protected bya European Patent EP1657181 held by the companyARTIBAL S.A. (Sabin˜a´nigo, Spain).

Bakery product

A grand total of 54 independent samples of the bakeryproduct from different batches were used in this study. Allof them were supplied by a small size company interestedin the study. Because of the commercial interest of thisdevelopment, a confidential agreement applies and nomore data about the company will be provided. The productis a complex mixture prone to microbial infestation andquality deterioration. Yolk, almonds, nuts, raisins swilledin rum as well as other common ingredients are includedin the formula. The whole product is covered with a sugarlayer.The main desirable characteristics of the product weredefined as tenderness of the inner part while nuts andalmonds kept crunchy and the yolk cannot be visuallydistinguished from the mixture and only the taste can beperceived. This product has an additional challenge as therequirement of humidity to keep tenderness should notinfluence the crunching and flavouring of the other ingredi-ents. This product is well balanced just made and the sameproperties are required for its shelf-life. No preservativesare added during its baking. In fact, this is considered asa handmade product.

Headspace-single drop microextraction (HS-SDME)

The atmosphere generated inside the active bag wassampled using headspace-single drop microextraction (HS-SDME) as described byRomero, Lo´pez, Rubio, Batlle, andNerı´n(2007). Brieﬂy, a 5-

L microsyringe with a 26S typeneedle (Hamilton 85RN, Bonaduz, Switzerland) containingp-xylene as the extractive phase was introduced inside thebag. The plunger was then depressed and a 2.5-

L dropwas exposed to the sample for 5 min. After exposure, thedrop was retracted again into the syringe and then, it wastransferred to the injection port of a GC

MS system.

Gas chromatography

mass spectrometric(GC

MS) analysis

MS analysis were performed using a Hewlett-Pack-ard 6890 chromatograph (Wilmington, DE, USA) equippedwith a 5973 mass selective detector and a HP-5 MS(60 m

0.25 mm,0.25

mﬁlmthickness)capillarycolumn.The temperature program for the GC was as follows: initialtemperature 75



C, 10



C/min ramp to 190



C, then20



C/min to 280



C, held for 5 min. The injector tempera-ture was maintained at 300



C and operated in splitlessinjection mode with the valve closed for 1 min. Helium(99.9999% pure, Carburos Meta´licos, Zaragoza, Spain)was used as the carrier gas at a constant ﬂow of 1 mL permin.Theinterfacetemperaturewas280



Candtheionizationmodewaselectronimpact(70 eV).Themassselectivedetec-tor was operated in the scan mode between 45 and 400 m/z.

Sensory testing

A trained panel of 12 individuals was used to evaluatethe sensorial properties and possible changes in the bakeryproduct 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. Thetechnique used in this case was the classiﬁcation of theresponses given by the tasters in a scale where several

L. Gutie ´ rrez et al. / Trends in Food Science & Technology 20 (2009) 92

descriptors, defined according their relevance in the finalquality, were qualified. The perception of the tastersconcerning the magnitude of the differences, the intensityof the descriptors related to taste, flavour, sight, color andtender were evaluated in a scale ranging from 1 to 8 points,being 8 the highest intensity. Three experts (members of thestaff of the manufacturing company) were also included inthe panel as a reference.Testing was conducted over a 28-days period, coveringa time-span longer than the required shelf-life, establishedby the manufacturing company in 10 days. Currently theshelf-life is no longer than three days. The followingcheck-points were scheduled (reported as days frommanufacturing): 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 madeusing all the PP packaging materials and different optionsevaluated. Then, the plastic bags were thermosealed in allcases. The following options were evaluated in the tests.1 Parafﬁn paper. This is the traditional packagingmaterial used to cover this speciﬁc product by themanufacturing company (Blank).2 PP non-active nor micro perforated (PPno).3 PP non-active micro perforated with a density of 16holes per dm

(PPnoM)4 PP active (4% w/w) micro perforated with a density of 16 holes per dm

(PPM).5 PP active (2% w/w) partially micro perforated (3 rowsof 5 cm each) (3 row).6 PP active (2% w/w) partially micro perforated (2 rowsof 5 cm each) (2 row).7 Same as5including a polyethylene (PE)-coated boardpiece to separate the bottom of the product from theboard secondary package (coated board).8 Same as6where the PE-coated board piece has beenmetalized and introduced into the active PP bag (alumi-num tray).In every test, fresh, just made products in the traditionalpackaging material were used as blank samples. All optionswere kept inside the cardboard box used for the company assecondary packaging for commercial and marketingpurposes.

Development

In a preliminary step, the suitability of the packagingmaterial intended to come into contact with food wasevaluated. As has been described in the introductorysection, no regulatory issues arise from the use of essen-tial oils as preservative. Nevertheless, the first stageconsisted of measuring the volatile compounds releasedby the active material to get an idea about the potentialimpact characteristics of the product. The rationale forthis work is that, in a previous work (Lo´pez, Huerga,Batlle, & Nerıń, 2006), the release and identificationof the active compounds from the active film wasdetermined.Fig. 1shows the chromatograms obtained when sam-pling the headspace generated inside the bag using theSDME GC

MS strategy described in the ExperimentalSection. As can be seen, the very same chemicals weredetected 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 activepackaging (PPM) was analysed in the absence of the bakeryproduct, a clear signal corresponding to cinnamaldehyde(chemical responsible of the cinnamon characteristicflavour) was obtained. Therefore, it is hypothesized thatthis chemical is readily absorbed by the product and asa consequence, it was included in the flavour descriptorsas an undesirable characteristic.Different groups of descriptors were identified andused in the sensory evaluation. They can be roughlydivided into five groups: taste, odour (flavour), texture,general appearance and presence of moulds. Every groupwas fully divided into more specific indicators, whichwere included into the evaluation sheet. These indicatorswere as follows.1 Taste: the following individual markers were definedand included.



Characteristic taste, as a positive factor

. It repre-sents the similarity obtained by the individualbetween the test and the blank items.



Cinnamon taste

. This factor can be consideredeither positive or negative, since acceptance of this speciﬁc taste is very subjective. Nevertheless,since it represents an alteration of the expectedtaste, it was included as negative.



Board taste

. This is a negative factor and repre-sents the alteration due to the board materialused as secondary packaging.2 Flavour.



Board ﬂavour

. This is also a negative factor andrepresents alteration due to the board materialused as secondary packaging.



Cinnamon ﬂavour

. As in the previous paragraph, itwas 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 nutswhen compared to the blank item.



Elasticity rubber-like

. Negative factor, referring tothe change in the product overall texture. It wasdeﬁned by the expert members of the sensory eval-uation board as a clear indicative of aging.

L. Gutie ´ rrez et al. / Trends in Food Science & Technology 20 (2009) 92