Gracia López-Carballo

Antifungal properties of gliadin films incorporating cinnamaldehyde and application in active food packaging of bread and cheese spread foodstuffs.

Gliadin films incorporating 1.5, 3 and 5% cinnamaldehyde (g/100g protein) were tested against food-spoilage fungi Penicillium expansum and Aspergillus niger in vitro, and were employed in an active food packaging system for sliced bread and cheese spread. Gliadin films incorporating cinnamaldehyde were highly effective against fungal growth. P. expansum and A. niger were completely inhibited after storage in vitro for 10 days in the presence of films incorporating 3% cinnamaldehyde. Indeed 1.5% cinnamaldehyde was sufficient in the case of P. expansum. The amount of cinnamaldehyde retained in films after storage for 45 days at 20 °C and 0% RH was also sufficient in most cases to prevent fungal growth in vitro. Active food packaging with gliadin films incorporating 5% cinnamaldehyde increased the shelf-life of both sliced bread and cheese spread. Mold growth was observed on sliced bread after 27 days of storage at 23 °C with active packaging, whereas in the control bread packaged without the active film fungal growth appeared around the fourth day. In the cheese spread, no fungi were observed after 26 days of storage at 4 °C when the product was packaged with the active film. However, growth of fungi was observed in control packaged cheese after 16 days of storage. This work demonstrates a noteworthy potential of these novel bioplastics incorporating natural antimicrobial compounds as innovative solutions to be used in active food packaging to extend shelf-life of food products.

Antimicrobial food packaging film based on the release of LAE from EVOH

The aim of this work was to develop antimicrobial films for active packaging applications containing the natural antimicrobial compound LAE (lauramide arginine ethyl ester) in EVOH copolymers with different mol % ethylene contents (i.e. EVOH-29 and EVOH-44). EVOH-29 and EVOH-44 films were made by casting and incorporating 0.25%, 1%, 5%, and 10% LAE in the film forming solution (w/w with respect to polymer weight). Previously, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of LAE against Listeria monocytogenes, Escherichia coli, and Salmonella enterica were determined by a microdilution assay. The antimicrobial activity of the resulting films was tested in vitro against these microorganisms in liquid culture media. The activity of the films was also evaluated over time. The results showed that films containing 5% and 10% LAE produced total growth inhibition and viable counts decreased with 0.25% and 1% LAE. Finally, the effectiveness of the films was tested by applying them to an infant formula milk inoculated with L. monocytogenes and S. enterica and stored for 6 days at 4°C. The application of films with LAE to infant formula milk inoculated with L. monocytogenes reduced at the end of storage period about 4 log in case of 10% LAE and with S. enterica reduced 3.74 log and 3.95 log with EVOH 29 5% and 10%, respectively, and EVOH-44 5% and 10% LAE reduced 1 log and 3.27 log, respectively, at the end of storage. The antimicrobial capacity of EVOH-29 films was greater than that of EVOH-44 films in all the cases tested. In general, the films were more effective in inhibiting the growth of L. monocytogenes than S. enterica, this inhibition being more acute at the end of the storage time.

Photoactivated chlorophyllin-based gelatin films and coatings to prevent microbial contamination of food products

The aim of this work was to develop antimicrobial photosensitizer-containing edible films and coatings based on gelatin as the polymer matrix, incorporating sodium magnesium chlorophyllin (E-140) and sodium copper chlorophyllin (E-141). Chlorophyllins were incorporated into the gelatin film-forming solution and the inhibiting effect of the cast films was tested against Staphylococcus aureus and Listeria monocytogenes. The results demonstrated that water soluble sodium magnesium chlorophyllin and water soluble sodium copper chlorophyllin reduced the growth of S. aureus and L. monocytogenes by 5 log and 4 log respectively. Subsequently, the activity of self-standing films and coatings containing E-140 was assessed on cooked frankfurters inoculated with S. aureus and L. monocytogenes. These tests showed that it was possible to reduce microorganism growth in cooked frankfurters inoculated with S. aureus and L. monocytogenes by covering them with sodium magnesium chlorophyllin-gelatin films and coatings.

Development of a novel antimicrobial film based on chitosan with LAE (ethyl-Nα-dodecanoyl-l-arginate) and its application to fresh chicken

Chitosan (CS) films incorporating the antimicrobial compound ethyl-N(α)-dodecanoyl-l-arginate (LAE) were developed for food packaging applications. Cast chitosan films were made with 1, 5 or 10% LAE and 20% glycerol in the film forming solution. Optical properties, release of LAE and antimicrobial activity of developed films was determined. The minimum inhibitory concentration (MIC) and the minimum biocide concentration (MBC) of LAE were determined. CS films with LAE were transparent and uniform, without discontinuities or visible particles and no visual differences could be perceived between CS and CS-LAE films. When in contact with an aqueous food simulant, the agent was fully released following a Fickian behavior in a few hours at 4 and 28°C. Antimicrobial activity of films against mesophiles, psychrophiles, Pseudomonas spp., colifoms, lactic acid bacteria, hydrogen sulfide-producing bacteria, yeast and fungi, was evaluated at two, six and eight days for its application on chicken breast fillets. Films were active against bacteria, yeasts and fungi in liquid and solid media. CS films evidenced antimicrobial activity in the range 0.47-2.96 log reductions, while CS-5%LAE film produced 1.78-5.81 log reduction. Results highlighted that LAE incorporation in a chitosan-based packaging structure may provide a relevant antimicrobial activity that could improve the stability of fresh poultry products.

Silver Ions Release from Antibacterial Chitosan Films Containing in Situ Generated Silver Nanoparticles

This study aims to develop antimicrobial films consisting of chitosan and silver nanoparticles that are homogeneously distributed throughout the polymer matrix. Nanoparticles were generated in situ during the neutralization of the chitosan acetate film with sodium hydroxide. The temperature of neutralization and the concentration of silver in the film were crucial determinants of the shape and size of the nanoparticles. Neutralized films exhibited antimicrobial activity against Escherichia coli and Staphylococcus aureus in liquid growth media. However, the effectiveness of the films was considerably greater in diluted growth media. Furthermore, no significant differences were found either in the antimicrobial capacities of films incorporating different amounts of silver or in the amount of silver that migrated into the liquid media after 18 h of immersion of the film. Neutralized films maintained their activity after 1 month of immersion in deionized water, which can be attributed to the slow sustained release of silver ions and thus efficacy over time.

Active films based on cocoa extract with antioxidant, antimicrobial and biological applications.

Novel films of ethylene-vinyl alcohol copolymer (EVOH) containing flavonoid-rich cocoa were developed. To understand their potential application as active packaging material, antioxidant and antimicrobial properties of the films were determined as well as the antioxidant activity of the release compounds in Caco-2 human epithelial colorectal adenocarcinoma cells. Exposure of the films to aqueous food simulant showed antioxidant capacity. The release of cocoa extract components was dependent on the antioxidant concentration incorporated in the film and on temperature. Cocoa extract and the fraction obtained after in vitro gastrointestinal digestion presented antioxidant activity against oxidative stress induced by hydrogen peroxide in Caco-2 cells. Films with 10%, 15%, and 20% cocoa extract produced bactericidal effect against Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Salmonella enterica. The application of films to an infant milk formula, previously inoculated with L. monocytogenes, inhibited the growth of bacteria 1.5 log units the first day and showed sustained release, inhibiting 0.52 and 0.76 log units, respectively, by the sixth day, while cocoa powder added directly did not produce any effect.

Preparation and characterization of chitosan/HP-β-cyclodextrins composites with high sorption capacity for carvacrol

The aim of this work was to design new polymer-based systems exhibiting an adjustable loading capacity of carvacrol depending on the film formulation. For this purpose, biocomposite films were developed employing chitosan (CS) as the polymer matrix and hydroxypropyl-β-cyclodextrins (HP-βCDs) as an adjuvant to improve the sorption of carvacrol in the polymer matrix. The morphology, optical, mechanical and barrier properties of the resulting films were investigated, and the sorption capacity of carvacrol evaluated. Biocomposites resulted highly transparent with higher mechanical resistance and moisture barrier properties. Sorption of carvacrol was greatly affected by the humidity (RH) and glycerol (G) content of the biocomposites. The highest sorption values were achieved for composites incorporating 35% glycerol and conditioned at 75% these composites retained 216% carvacrol (g/100 g dry matter). These results indicate that inclusion of carvacrol in the films could be occurring by mechanisms other than formation of inclusion complexes.

Antimicrobial packaging of chicken fillets based on the release of carvacrol from chitosan/cyclodextrin films

Chitosan/cyclodextrin films (CS:CD) incorporating carvacrol were obtained by casting, and conditioned at 23°C and 75% relative humidity prior to being immersed in liquid carvacrol until they reached sorption equilibrium. In a previous work, the in vitro antimicrobial activity of these films was studied. In this work, active films were used to inhibit microbial growth in packaged chicken breast fillets. Samples of CS:CD films loaded with carvacrol, of different sizes and thus with different quantities of antimicrobial agent, were stuck to the aluminium lid used to seal PP/EVOH/PP cups containing 25g of chicken fillets. These samples were stored for 9days at 4°C. The packages were hermetically sealed and it was confirmed that they provided an infinite barrier to carvacrol. The partition of the antimicrobial agent within the food/packaging system was analysed. The antimicrobial devices rapidly released a large percentage of the agent load, amounts that were gained by the adhesive coating of the lid and especially by the chicken fillets. The latter were the main sorbent phase, with average concentrations ranging between 200 and 5000mg/Kg during the period of storage. The microbiota of the packaged fresh chicken fillets - mesophiles, psychrophiles, Pseudomonas spp., enterobacteria, lactic acid bacteria and yeasts and fungi - were analysed and monitored during storage. A general microbial inhibition was observed, increasing with the size of the active device. Inhibition with a 24cm(2) device ranged from 0.3 log reductions against lactic acid bacteria to 1.8logs against yeasts and fungi. However, the large amount of antimicrobial that was sorbed or that reacted with the fillet caused an unacceptable sensory deterioration. These high sorption values are probably due to a great chemical compatibility between chicken proteins and carvacrol.

Active antimicrobial food and beverage packaging

Abstract: Active antimicrobial packaging can be defined as a packaging system that interacts with the food product or the surrounding headspace either to kill the microorganisms that might be present in the food product or food package, or to reduce, inhibit or retard their growth. This chapter reviews: (a) the diverse families of antimicrobial compounds, their mode of action and their aptitude to be processed and be part of a packaging system; (b) the diverse techniques used to include the agents into the packaging materials, reduce their degradation and facilitate their action and the issues to be considered in packaging design; and (c) the diverse methods which can be applied to determine the activity of the wide range of antimicrobial packaging items.

Contact probe electrochemical characterization and metal speciation of silver LLDPE nanocomposite films

A contact probe methodology, based on the voltammetry of immobilized microparticle approach, is used for characterizing silver species present in linear low density polyethylene (LLDPE) films with different Ag(I)/Ag(0) ratios and silver nanoparticle features usable as food contact polymers. The films displayed characteristic voltammetric features in contact with aqueous acetate buffer, in particular signals for the stripping oxidation of nanoparticulate Ag systems. Significant differences between the studied films were also observed by means of electrochemical impedance spectroscopy and detected at the nanoscopic scale using electrochemical scanning microscopy. Differences in optical and thermal properties of the studied films are associated with the presence of silver nanoparticles. The silver oxidation state as well as nanoparticle size also had influence on the oxidative resistance of the LLDPE films; indeed, films containing cationic silver showed the lowest oxidation induction time value.