Characterisation and antimicrobial activity of active polypropylene films containing oregano essential oil and Allium extract to be used in packaging for meat products.

Cooked ham is more prone to spoilage than other meat products, making preservation a key step in its commercialisation. One of the most promising preservation strategies is the use of active packaging. Oregano essential oil (OEO) and Proallium® (an Allium extract) have previously been shown to be useful in polylactic acid (PLA)-active films for ready-to-eat salads. The present work aims to study the suitability of polypropylene (PP) films containing OEO and Proallium® in the preservation of cooked ham. Concerning the technological features of the studied material, no significant changes in the mechanical or optical properties of PP films containing the active substances were recorded in comparison to the PP film without extracts. However, films containing both active substances were more flexible than the control film and less strong, highlighting the plasticisation effect of the natural extracts. Moreover, physical properties changed when active substances were added to the film. Incorporation of 4% Proallium® affected the transparency of the film to a higher extent compared to 8% OEO, undergoing decreases in transparency of 40% and 45%, respectively. Moreover, only the film containing the highest amount of OEO (8%) significantly decreased the thickness. Both active substances showed antibacterial properties; however, Proallium®-active films seemed to be more effective against Brochothrix thermosphacta than PP films containing OEO, with all percentages of Proallium® killing the bacterial population present in the ham after 60 days. In addition, materials containing the lowest Proallium® content exhibited higher acceptability by consumers in the sensory analyses with 63-100% willing to purchase, better even than the control package (56-89%). In fact, 2% of Proallium® obtained the best results in the odour study performed by the panellists.

Molecular characterisation of a bio-based active packaging containing Origanum vulgare L. essential oil using pyrolysis gas chromatography-mass spectrometry.

BACKGROUND: Environmental, economic and safety challenges motivate shift towards safer materials for food packaging. New bioactive packaging techniques, i.e. addition of essential plant oils (EOs), are gaining attention by creating barriers to protect products from spoilage. Analytical pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) was used to fingerprint a bioactive polylactic acid (PLA) with polybutylene succinate (PBS) (950 g kg(-1) :50 g kg(-1) ) film extruded with variable quantities (0, 20, 50 and 100 g kg(-1) ) of Origanum vulgare EO. RESULTS: Main PLA:PBS pyrolysis products were lactide enantiomers and monomer units from the major PLA fraction and succinic acid anhydride from the PBS fraction. Oregano EO pyrolysis released cymene, terpinene and thymol/carvacrol peaks as diagnostic peaks for EO. In fact, linear correlation coefficients better than 0.950R(2) value (P < 0.001) were found between the chromatographic area of the diagnostic peaks and the amount of oregano EO in the bioplastic. CONCLUSION: The pyrolytic behaviour of a bio-based active package polymer including EO is studied in detail. Identified diagnostic compounds provide a tool to monitor the quantity of EO incorporated into the PLA:PBS polymeric matrix. Analytical pyrolysis is proposed as a rapid technique for the identification and quantification of additives within bio-based plastic matrices. © 2015 Society of Chemical Industry.

Cytotoxicity and mutagenicity assessment of organomodified clays potentially used in food packaging.

Modern food packaging has made great advances as result of global trends and consumer preferences, which are oriented to obtain improved food quality and safety. In this regard, clay minerals, and mainly Montmorillonite (Mt) are attracting considerable interest in food packaging because of the improvements developed in mechanical and barrier properties. Hence, the present work aim to assess the toxicity of four Montmorillonite-based clay minerals, an unmodified clay, Cloisite®Na+ (CNa+), and three modified Mt clays: Cloisite®30B (C30B), a commercial clay, and Clay1 and Clay2, two novel modified organoclays developed by the Packaging, Transport, & Logistics Research Institute (ITENE). First, the cytotoxic effects were studied in the Human Umbilical Vein Endothelial Cells (HUVEC). In addition, the potential mutagenicity of the clays was evaluated by the Ames test. Clay1 did not induce any cytotoxic effects in HUVEC, although it exhibited potential mutagenicity in TA98 Salmonella typhimurium strain. In contrast, Clay2 produced cytotoxicity in endothelial cells but no mutagenicity was recorded. However, CNa+ was not cytotoxic neither mutagenic. And finally, C30B showed positive results in both assays. Therefore, results showed that clay minerals have a different toxicity profile and a case by case toxicity evaluation is required.

In vivo evaluation of activities and expression of antioxidant enzymes in Wistar rats exposed for 90 days to a modified clay.

Although clays are wildly used in a range of applications, the toxicity assessment of these new materials is still scarce. In the present study, oxidative stress induced by Clay 1, a novel clay, was determined in rats after 90 d of oral exposure. The activities of antioxidant enzymes, namely, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST), were examined. In addition, genetic expressions of SOD and CAT and relative protein abundance of CAT were also determined. Data showed that most of the biomarkers assayed remained unaltered. Only CAT activity, as well as its genetic and protein expressions, appeared enhanced in the kidney. Therefore, further studies are needed to clarify the relevance and consequences of these findings to ensure the safety of this clay.

Cytotoxicity and mutagenicity studies on migration extracts from nanocomposites with potential use in food packaging.

Clays are used in the food packaging industry to obtain nanocomposites. The use of these new materials is a concern, because they could reach consumers by oral exposure through possible migration, and potential toxic effects could be derived. In the present study, several in vitro basal cytotoxicity and mutagenicity tests on migration extracts obtained from a nanocomposite material with poly (lactic) acid (PLA) and two modified clays, Clay1 and Clay2, are shown. Migration extracts in distilled water showed values of 0.1 ± 0.2mg/dm(2) in all samples. Also, the content of characteristic metals of the clays structure (Al, Ca, Mg, Fe, Si) was studied and no statistical differences were observed. For the cytotoxicity assays, the human intestinal Caco-2 and human liver HepG2 cells were selected. Cells were exposed to concentrations between 2.5% and 100% extracts determining three different biomarkers of cellular viability. No significant differences were observed in the cytotoxicity assays. Finally, mutagenicity was evaluated by the Ames test and resulted in the absence of mutagenic response at all the concentrations assayed. Taking in account all above mentioned, these new materials show a good profile for their use in food packaging although further research is still needed.

Cytotoxicity and morphological effects induced by carvacrol and thymol on the human cell line Caco-2.

Essential oils used as additives in the food industry due to its flavour, antimicrobial and antioxidant properties. Therefore, human can be exposed orally to these compounds through the ingestion of foods. In this sense, the present work aims to assess toxicological effects of oregano essential oil on the digestive tract. In concrete, the cytotoxic effects of two components of the oregano essential oils, carvacrol and thymol, and their mixture, on the intestinal cells line Caco-2 after 24 and 48 h of exposure are studied. The basal cytotoxicity endpoints assayed (total protein content, neutral red uptake and the tetrazolium salt reduction) and the annexin/propidium iodide staining indicated that carvacrol and the mixture carvacrol/thymol induced toxic effects. Moreover, a morphological study was performed in order to determine the ultrastructural cellular damages caused by these substances. The main morphological alterations were vacuolated cytoplasm, altered organelles and finally cell death. In addition, although no cytotoxic effects were recorded for thymol at any concentration and time of exposure, ultrastructural changes evidenced cellular damage such as lipid degeneration, mitochondrial damage, nucleolar segregation and apoptosis.

Toxic effects of a modified montmorillonite clay on the human intestinal cell line Caco-2.

The incorporation of the natural mineral clay montmorillonite into polymeric systems enhances their barrier properties as well as their thermal and mechanical resistance, making them suitable for a wide range of industrial applications, e.g., in the food industry. Considering humans could easily be exposed to these clays due to migration into food, toxicological and health effects of clay exposure should be studied. In the present work, the cytotoxic effects induced by two different clays (the unmodified clay Cloisite(®) Na(+) , and the organically modified Cloisite(®) 30B) on Caco-2 cells were studied after 24 and 48 h of exposure. The basal cytotoxicity endpoints assessed were total protein content, neutral red uptake and a tetrazolium salt reduction. Our results showed that only Cloisite(®) 30B induced toxic effects. Therefore, the effects of subcytotoxic concentrations of this clay on the generation of intracellular reactive oxygen species, glutathione content and DNA damage (comet assay) were investigated. Results indicate that oxidative stress may be implicated in the toxicity induced by Closite(®) 30B, in regards of the increases in intracellular reactive oxygen species production and glutathione content at the highest concentration assayed, while no damage was observed in DNA. The most remarkable morphological alterations observed were dilated cisternae edge in the Golgi apparatus and nucleolar segregation, suggesting impairment in the secretory functions, which could be related to inhibition in the synthesis of proteins.

In vitro toxicological assessment of clays for their use in food packaging applications.

Montmorillonite based clays have a wide range of applications that are going to contribute to increase human exposure to these materials. One of the most promising uses of clays is the development of reinforced food contact materials that results in nanocomposites with improved barrier properties. Different organoclays have been developed introducing modifiers in the natural clay which is commercially available. However, the toxicological aspects of these materials have been scarcely studied so far. In the present study, the cytotoxic effects of a non-modified clay (Cloisite Na+) and an organoclay (Cloisite 30B) have been investigated in the hepatic cell line HepG2. Only Cloisite 30B showed cytotoxicity. In order to elucidate the toxic mechanisms underlying these effects, apoptosis, inflammation, oxidative stress and genotoxicity biomarkers were assayed. Moreover, a morphology study with light and electron microscopy was performed. Results showed genotoxic effects and glutathione decrease. The most relevant ultraestructural alterations observed were mitochondrial degeneration, dilated endomembrane systems, heterophagosomes formation, fat droplets appearance and presence of nuclear lipid inclusions. Cloisite 30B, therefore, induces toxic effects in HepG2 cells. Further research is needed to assess the risk of this clay on the human health.

Monitoring of chicken meat freshness by means of a colorimetric sensor array.

A new optoelectronic nose to monitor chicken meat ageing has been developed. It is based on 16 pigments prepared by the incorporation of different dyes (pH indicators, Lewis acids, hydrogen-bonding derivatives, selective probes and natural dyes) into inorganic materials (UVM-7, silica and alumina). The colour changes of the sensor array were characteristic of chicken ageing in a modified packaging atmosphere (30% CO(2)-70% N(2)). The chromogenic array data were processed with qualitative (PCA) and quantitative (PLS) tools. The PCA statistical analysis showed a high degree of dispersion, with nine dimensions required to explain 95% of variance. Despite this high dimensionality, a tridimensional representation of the three principal components was able to differentiate ageing with 2-day intervals. Moreover, the PLS statistical analysis allows the creation of a model to correlate the chromogenic data with chicken meat ageing. The model offers a PLS prediction model for ageing with values of 0.9937, 0.0389 and 0.994 for the slope, the intercept and the regression coefficient, respectively, and is in agreement with the perfect fit between the predicted and measured values observed. The results suggest the feasibility of this system to help develop optoelectronic noses that monitor food freshness.

Typical diffusion behaviour in packaging polymers - application to functional barriers.

When plastics are collected for recycling, possibly contaminated articles might be recycled into food packaging, and thus the contaminants might subsequently migrate into the food. Multilayer functional barriers may be used to delay and to reduce such migration. The contribution of the work reported here is to establish reference values (at 40 degrees C) of diffusion coefficients and of activation energies to predict the functional barrier efficiency of a broad range of polymers (polyolefins, polystyrene, polyamide, PVC, PET, PVDC, [ethylene vinyl alcohol copolymer], polyacrylonitrile and [ethylene vinyl acetate copolymer]). Diffusion coefficients (D) and activation energies (Ea) were measured and were compiled together with literature data. This allowed identification of new trends for the log D=f(molecular weight) relationships. The slopes were a function of the barrier efficiency of the polymer and temperature. The apparent activation energy of diffusion displayed two domains of variation with molecular weight (M). For low M (gases), there was little variation of Ea. Focusing on larger molecules, high barrier polymers displayed a larger dependence of Ea with M. The apparent activation energy decreased with T. These results suggest a discontinuity between rubbery and glassy polymers.