Final Report Summary - EASYFRUIT (Active packaging for extended shelf life of peeled and cut fruit) Executive Summary:EASYFRUIT project was focused on extending 5 days the shelf-life of fresh-cut orange and pineapple by means the development of an active packaging solution in combination with a minimal processing treatment.EASYFRUIT project started in January 2013 and ended in December 2014. Different activities dealing with research and technology developments as well as other activities related with dissemination, exploitation of results and management were carried out along these two years project.EASYFRUIT involved the participation of seven partners from different European countries (Spain, Denmark, Serbia, Belgium and Norway). The consortium had the participation of 4 SMEs (2 fresh-cut fruit companies and 2 packaging companies), 2 research centres specialized in food and packaging technologies, and an international retailer representing over 15,000 stores.The combination of these companies was crucial to resolve the current limitations on fresh-cut fruit shelf life. Team with high experience in postharvest/minimal processed treatments and spoilage mechanisms of fruit had the support of experienced group in active and intelligent packaging developments. Furthermore, the commercial perspective from a big retailer was taken into account in the project.Along the project, several technological barriers were encountered in the developing of the new active packaging. In this sense, the high level of expertise provided by the SMEs and Research Centres allowed solving each barrier in a positive way. In conclusion, the cooperation between all the partners enabled to achieve the final EASYFRUIT goal with successful results within the established deadlines.Finally, the project generated 11 technical deliverables at confidential level, 3 dissemination materials developed for the general public and other documents related with the protection and exploitation of the results by the consortium. Project Context and Objectives:Fruits and vegetables are known to be rich in vitamins and minerals, fiber, and other substances such as phenolic acids and flavonols which are important for health. Scientific evidences have proved that eating more fruits and vegetables is important to combat obesity since its consumption reduces the "energy density" in the diet, and in turn, it plays a protective role in heart disease, cancer and diabetes.According to the World Health Organization inadequate intake of fruits and vegetables is one of 10 major risk factors contributing to attributable mortality. Overall, 2.7 million (4.9%) deaths and 26.7 million (1.8%) disability-adjusted life years (DALYs) are attributable to low fruit and vegetable intake. Of the burden attributable to low fruit and vegetable intake, about 85% was from cardiovascular diseases and 15% from cancers.Despite the health benefits of eating fruits and vegetables their intake is decreasing daily in Europe. Statistics indicate that 57% of children do not meet the guidelines of recommended daily fruit consumption. Children prefer other less healthy foods that are easier to eat and are more attracting to them. Additionally, elderly people or people with limited mobility have a limitation when peeling and cutting fruit products. In addition, changes in lifestyle (less time to prepare meals and eating away home) also contributes to the low intake of fruits and vegetables.In order to promote their consumption, fresh-cut fruit products are being placed in the market. However, their short shelf-life is nowadays one of the main problems affecting their commercialization and consumer´s acceptance. Existing technologies such as standard minimal processing, MAP, fruit dipping, edible coating are not able to provide a desired longer shelf-life.In this sense, the rapid expiration of fresh-cut products makes exportation almost impossible and also limits distribution within the same country. Commercialization of fresh peeled and cut fruit is limited to a reduced area surrounding the fruit processing and packaging plant that means an under-exploitation of this business. As an example, a local solution of some supermarkets is to sell peeled and sliced fruit which they prepare daily and throw away the unsold remains. This makes the process highly expensive and waste generating, thus increasing the final price of the product. The EASYFRUIT proposed solution is to combine improved minimal processing treatments with the development of an active package for fresh-cut fruit that extends the shelf life of peeled and cut fruit longer than current technologies. The specific objective of EASYFRUIT is to extend up to 5 days the shelf life of fresh-cut orange and pineapple by means of investigation on deterioration mechanisms and optimization of peeling and cutting methods and also preservation treatments were performed but also by the use of an active packaging able to interact with the peeled and cut fruit, maintaining the quality parameters longer. In this way, food preservatives are included in the packaging materials by different processing techniques that will be released to the fruit due to the mass transfer properties given by the polymeric materials selected. This technology allows making the addition of food preservatives more effective, rather than direct addition to the fruit where higher dose of preservatives are usually need due to their rapid consumption and degradation.Therefore, the specific goals contributing to the main EASYFRUIT project purpose were:- Identification of the main deterioration mechanisms in peeled and cut orange and pineapple and its relation with the minimal processing methods. - Development of a minimal processing treatment to minimize the fruit damage and increase fruit shelf life and quality. - Selection of the most promising agents to be included in the packaging materials - Development of effective active materials with a homogenous distribution of active agents and minimal losses during processing.- Adjustment of processing parameters of materials and strategy to incorporate active agents into/onto the material to minimize losses of active agent during processing. - Comprehension of the mechanisms of release of the active components from the packaging materials to the fresh cut fruit and adjustment of the adequate control release of active compounds. - Characterization and validation of the active packaging materials demonstrating the performance of the final prototypes in and demonstrating the ability of these materials to improve the shelf life of the fruit- Increase the knowledge regarding the combination of minimal processing and active packaging to evaluate synergies between them. Facilitating the marketing of peel and cut fruit by overcoming the inconvenience generated by their short shelf life; Improving the quality of those already on the market so as to retain their organoleptic and nutritional properties for longer; Facilitating the export and marketing over long distances; Providing a new product to consumers who do not currently consume it; to encourage the consumption of fruit so highly recommended by the WHO (World Health organization) and reducing the waste generated by throwing away expired packaged cut fruits are some of the benefits that are achieved with EASYFRUIT project.Project Results:EASYFRUIT project was divided into three main phases: - Phase 1: Minimally processing of the fruit for shelf life extension- Phase 2: Development of active packaging - Phase 3 Combination of both technologies, minimally processing and active packaging in order to extend the shelf life of peeled and cut orange and pineapple 5 daysMain results obtained in Phase 1Deterioration mechanismsThe first main results obtained in the project were related with deterioration mechanisms in fresh-cut fruit. The main deterioration mechanisms in orange were found to be microbial growth and juice leakage, and in pineapple microbial growth, juice leakage and browning.Minimal processing of fruitFresh-cut fruit were produced by so-called minimal processing. That means to prepare pieces of fresh-like edible fruit flesh, where the cells are alive and in such a way that the product can be stored for some time and still be edible and fresh-like. Peeling and cutting must therefore be done in a smart and precautious way. Diverse ways to peel and cut orange and pineapple were studied. The aim was to minimise undesirable physiological response induced by minimal processing of fruit and to minimise the load of microbes to obtain fresh-like products with a sufficiently long shelf-life. A shelf-life extension from 7 to 12 days was the main goal.Raw fruit qualityThe quality of the raw fruit is essential for the final quality and storability of the cut fruit products. The fruit must be of a suitable cultivar, grown under specified conditions of soil, climate, fertilization, etc. and harvested, postharvest treated, packed and shipped by certified procedures. Harvest time to get optimal ripeness, as well as temperature and time after harvest before minimal processing are the very important factors for final product quality. Other important factors are absence of plant disease, pests and mechanical stress or injury. The physiological state (respiration rate, etc.) is very much dependent on the temperature. Both orange and pineapple are prone to chilling injury when stored at too low temperature. This will result in tissue and cell breakdown when the temperature is increased. At too high temperature metabolic processes will run faster and senescence will therefore go faster and result in a shorter shelf-life of both whole fruit and fresh-cut products. Optimal storage temperatures for whole orange and pineapple have been determined to 2-4 ⁰C and 7 ⁰C, respectively - the exact temperature for orange depending upon cultivar. For cut fruit, both orange and pineapple should be stored around 4 ⁰C. In the distribution of fresh-cut products it can sometimes be higher temperature. Therefore, storage experiments in the project were carried out at 4 ⁰C for 2/3 of the time and 7 or 8 ⁰C for the rest of time, to simulate wholesale including shipping and retail, respectively.Washing and disinfection of whole fruitImmediately before peeling, the fruit should be washed in running cold water to remove possible dust and particles. Washing can also be done by immersing the fruit in cold water with agitating for a certain time, and preferably with rinsing shortly afterwards. Disinfection of fruit can be done similarly by adding a chemical disinfectant to the water or by using hot water. Several disinfections methods and chemical disinfectants have been tested in the project. High temperature water bath and disinfection with hypochloric acid (HClO) and hydrogen peroxide (H2O2) were tested on whole fruit. Irradiation with UV-C has been evaluated. The SMEs participating companies either used disinfection during washing of whole fruit or used just washing without disinfection. Disinfection of whole fruit was investigated in two tests. In the first test the levels of microorganisms were very low in cut fruit after cold storage and the experiment did not show differences between the treatments. This could be explained by either very low microbial load on the fruit skin at start or that the procedure for minimal processing was very hygienic. In the second experiment, only skin samples were analysed before and after disinfection. Orange skin had very low microbial load, below detection limit. Pineapple skin had low loads of psychrotrophic bacteria and moulds, but higher load of yeasts. Disinfection with HClO or H2O2 had no effect on microbial load of pineapple skin. Thus, the low microbial counts of cut fruit in the first disinfection trial were an effect of a high level of hygiene in the minimal processing and not due to effective disinfection of whole fruit before peeling. The results obtained showed that disinfection had little effect.Irradiation with UV-C has a quite well documented effect in killing or inhibiting microorganisms. The main problem appears to be effectiveness of exposure. UV-C has an effect only if the microorganisms are directly exposed on a fairly smooth surface. Then the reduction of organisms is comparable to the effect of HClO or H2O2 treatments (maximum 2-3 log reduction). In practice, the products to be irradiated with UV-C must be exposed from all sides. Peeling of fruitUsing sharp stainless steel knives for manual peeling of oranges and pineapples was investigated. A specific procedure for each fruit was established to achieve a clean and peel-free fruit in a rapid way. There is equipment for machine peeling of oranges and both participating fresh-cut companies use such machines. Peeling machines are rather slow and were not tested in the laboratory. For pineapples, the green top was removed before washing to reduce transfer of wax and other dust. Topless pineapples could be immersed and washed easily. De-coring of pineapples is best done after complete peeling with a cork-bore like tool. Alternatively, de-coring can be done by knife-cutting after splitting the peeled pineapple.Enzymatic peeling of oranges A study was carried out on the efficacy of enzymatic peeling of oranges. Commercial enzyme mixtures containing pectinase were tested. Enzymatic peeling of citrus fruit includes the following steps: 1. Pricking or cutting the skin many places.2. Vacuum infusion of enzyme solution, normally several pulses.3. Incubation for about 30 min at 30-40 ⁰C.4. Mechanical removal of skin.5. Rinse in citric acid bath (optional)Special equipment for industrial use is needed: peel pricker, vacuum machine, water bath, peeling machine or line for manual removal of peeling. Equipment is not readily available, since very few if any of the fresh-cut industries are using enzymatic peeling presently.A preliminary test was carried out to select optimal vacuum pressure and holding time for fluid uptake by orange. Then, two commercial enzyme preparations were tested in the laboratory. It resulted in nicely peeled oranges largely without albedo, with higher yield than knife-cutting ( ̴70% vs. 50-60%), lower leakage rate, unchanged respiration rate, shelf-life at least similar to knife-cut products, good sensory quality, but a bit less firm than knife-cut orange after storage. Mechanical peeling after enzyme treatment was easy (like ‘ball out of bag’), but threads of albedo had to be removed by gentle rubbing. Cutting equipmentCutting of peeled fruit should be done with very sharp tools to reduce mechanical stress, which can shorten the shelf-life of cut products. Ceramic knives were tested. They are sharp much longer than stainless steel knives, but are fragile and cannot be machine-washed. Both water jet and laser technologies were evaluated from literature. They have large potential for future applications in fully automated minimal processing. The hurdles are initially high costs and lack of commercially suitable equipment. Both technologies might, however, become accurately controlled and, in combination with robotics, replace complex manual operations, and will then probably be able to improve both the quality and shelf-life of cut fruit.Cutting methodCutting was done in various ways. The participating companies cut in different ways according to type of product and requirements of their customers. The effect of fruit chunk size on juice leakage after storage was evaluated in the project. The results showed, as expected, that leakage in percent is larger in small than in large chunks. The sensory quality was, however, not different. Optimization of preservation treatment of cut fruitSeveral treatments can be apply to prolong the shelf-life fresh-cut fruit, including optimization of the storage conditions. The following factors and treatments with a potential for optimization of minimal processing and prolongation of shelf-life were studied or evaluated: temperature level, humidity level, packaging properties, gas composition in modified atmos¬phere packaging (MAP), dipping of cut fruit in some organic acids, coating of cut fruit, and hygiene. Temperature levelCut pineapple are stored at 4ºC although whole pineapple gets chilling injury. Storage tests simulating commercial distribution of products were carried out at 4 ⁰C (2/3 of the time) followed by 7 ⁰C (1/3 of the time) to simulate abuse in the retail market. Humidity level The relative atmosphere humidity at equilibrium becomes 100% in a cut fruit closed packaging.Selection of packaging for storage of cut orange and pineappleThe gas permeability of the fruit packaging should be larger through the top film to avoid anaerobic conditions by normal cold storage or storage at higher (abuse) temperatures during commercial distribution. Therefore, perforation with a small number of holes is necessary, even if pineapple and orange have relatively low respiration rates. Otherwise, anaerobic conditions come into being after about one week. During the project the holes were made with an acupuncture needle. Modified atmosphere compositionDifferent starting gas compositions and presence of small holes in the top film were investigated. Samples were flushed with either of the two gas mixtures: (a) 5% O2 + 5% CO2 + 90% N2 and (b) 5% O2 + 10% CO2 + 85% N2 before the top film was attached. The kinetics of gas composition was then followed and compared with that of air-packed samples. After 1-2 days the gas composition in the packaging approached that of samples packed with air. At equilibrium, the concentration of O2 was in the range of 15-17% and that of CO2 in the range of 5-8%.In this way, psychrotrophic bacteria, yeasts and moulds were very low for all of the treatments. The experiment showed that active modified atmosphere is not an option to prolong the shelf-life of fresh-cut fruit in modified atmosphere packaging with perforation, since the concentrations of O2 and CO2 after a few days of storage approach the concentrations obtained after air packaging, and since these concentrations do not have any effect on neither fruit respiration nor microbial growth nor visual quality.Dipping and coatingPossible chemical food additives that could retard deterioration and increase product quality were considered based on known properties and performance. Only a few were interesting enough to be tested. Calcium chloride (CaCl2) was needed for coating fruit chunks. However, CaCl2 is not approved as a food additive for cut fruit by EFSA.Experiments were done to test effects of organic acids and of one coating formulation. Coating reduced juice leakage by half in cut pineapple, but addition of organic acids had no effect. Surprisingly, the coating increased the leakage in cut orange. Drain of chunks prior to packing reduced transfer of juice to the packed product. The sensory quality after storage for 14 days was good, but coating gave fair, although acceptable odour and flavour. Appearance changed less. Organic acids gave initially acidic flavour, but after storage the acidic flavour was reduced. The respiration rate, acidity and soluble solids (sugar) content were similar or differed little between treatments and after storage. Final counts of bacteria, moulds and yeasts were low and acceptable. Coating had no effect on microbial counts.HygieneBoth oranges and pineapples are usually washed, waxed and treated with fungicides before export. Results from the project showed that the microbial load on whole, raw fruit was low, especially for orange. Disinfection experiments of whole fruit showed no effect on the microorganisms detected (psychrotrophic bacteria, moulds and yeasts). Transfer of microbes from the peel to cut fruit was measurable for pineapple but mostly nil for orange. Separation of peeling and cutting in two operations at different sites - both with allocated, clean equipment - was implemented in the protocol of optimal processing. This was considered the most important improvement to reduce the microbial load on the cut fruit. It is concluded that a high level of hygiene during minimal processing and packing should be prioritised. Disinfection of whole fruit is less important. Disinfection of cut fruit is difficult and can change the quality, and should therefore not be used.Recommendation on hygiene: Examine carefully the hygienic procedures, especially around the steps of peeling and cutting. Small improvements in the routines here could have a large impact on the quality and disinfection of whole fruit could be omitted. As a basis, a high standard of hygiene for the premises and the personnel must be in place. A sufficient and certified quality management of the supply chains for both raw fruit and fresh-cut products is very important. Protocol for each fruit A protocol for suitable minimal processing of each fruit was prepared for use in further experiments. It is based on the optimal procedures and conditions found, as well as on selected parameters like size of fruit chunks etc. Evaluation of the shelf-life of minimally processed fruit before packaging The optimised peeling, cutting and preservation methods were applied and evaluated in shelf-life experiments. Sensory quality and quality related properties of each fruit were compared before and after storage. A shelf-life increase of the peeled and cut fruit of about five days was defined based on the optimised methods.In the first test, the possible length of shelf-life was investigated by several samplings during 14 days of storage of cut fruit, which was prepared and packed the same day. The experiment showed that microbial counts were below risk limits throughout the storage period. Juice leakage was low for orange, as well as for pineapple treated with edible coating, whereas uncoated pineapple leached more. Sensory scores were high at start, but fair or good after storage for 10-14 days, although still acceptable. Only minor changes in acidity and sugar content were observed. Some translucency was, however, seen in pineapple. It was concluded that a shelf-life of 12-14 days is possible.A second shelf-life test used descriptive sensory analysis by an ISO-certified method with a permanent panel of trained judges to evaluate the samples. The sampling had to be done on the same day for all storage times, since descriptive sensory analysis requires direct comparison of samples with different treatments in a randomised design. Therefore, fruit was cut and packed multiple times before the day of sensory analysis. The storage times for pineapple were 1, 6 and 13 days and for orange 1, 7 and 14 days. Microbial counts were below the risk limits. Juice leakage was low for orange and coated pineapple, and minor or no changes in acidity and sugar content were observed. Sensory analysis was incomplete due to mouth problems of panellists when tasting pineapple. Therefore orange, which was tested after pineapple, was only analysed for appearance and odour attributes. There were significant differences in several attributes for pineapple, but no clear effects of storage time or coating could be seen. Only whiteness showed some differences for orange. For pineapple the observed quality differences could be due to different properties of the raw material, since the country of origin differed between the three cutting days. It was concluded that a shelf-life of 12-14 days is possible. Thus, the experiments showed that a shelf-life of 12-14 days is possible for fresh-cut orange and pineapple produced with optimised procedures. This means that a common shelf-life of one week today can be extended by five days. Main results obtained in Phase 2: Development of active packagingAs a first main result obtained in this phase, a list of potential active agents was drafted in order to test later their antimicrobial effectiveness and their suitability to be incorporated into the final active packaging.In this way, nine antimicrobials were proposed according to the microbial deterioration of the peeled and cut orange and pineapple studied in Phase 1. Effectiveness of the agents in food products (especially in fresh-cut fruits) based on literature reviews, their origin (given preference to natural or those that have been already used in nutrition), their physico-chemical characteristics (thermal stability, state, solubility, etc.) and their compatibility with the food product (trying not to alter the fruit organoleptic properties) were the factors taken into account in this research. In addition, current European legislation on food and food contact materials was also considered for the choice of the active substances.The effectiveness of the selected agents were testes in vitro against the typical microorganisms of the fruit including yeasts such as Saccharomyces cerevisiae and moulds such as Penicillium species and Aspergillus niger. Additionally, other complementary characterisations were performed dealing with the evaluation of the thermal stability as well as their sensory properties on food. As a result, three volatile antimicrobials were selected as the most effective against the tested microorganisms and organoleptically compatible discarding the rest of the compounds. Furthermore, the synergy among these three active agents was studied defining the most effective ratio. This combination was chosen to be incorporated into the polymeric matrix of the lid that will be later validated in fruit.In parallel, three non-volatile antimicrobial compounds to be incorporated in the tray were selected for their antimicrobial properties in fruit based on scientific studies. One of the tested antimicrobials was discarded due to its low solubility in water, the high concentration required to achieve the desired effect, and its high cost. Nevertheless, a combination of the other two compounds was selected to be incorporated in the packaging materials for tray development. For the active materials development, the polymeric matrices were selected based on their compatibility with the active compounds, the respiration rates of the fruit and the suitable packaging processing techniques. In this way, depending on the way the active compounds will be incorporated (coating or extrusion) and the processing technologies required, different polymeric matrices were studied and different active materials were developed at lab and semi-industrial scale.Lab-scale materials• Active lid:Coating technique: Three different polymeric varnishes were formulated with the selected antimicrobial mixture. A polyolefin substrate was coated with the active varnished with an automatic film applicator at lab scale. According to the results obtained, two varnishes were discarded because one of them lost its integrity in contact with the fruit and the other reacted with some of the active compounds selected. Thus, the third varnish was chosen as the most promising one for coating the active lid and scaling the coating application at semi-industrial scale. Extrusion technique: In the case of the extruded lid at lab scale, two polymers were selected to incorporate the active compounds in a monolayer film. These materials were obtained using a lab scale twin extruder coupled with a liquid dosing pump to feed the active compounds. One was discarded due to the difficulty in processing, its low stiffness and the reduced antimicrobial effectiveness against the target microorganisms. The other material was selected to develop the active lid at semi-industrial scale.• Active tray:Coating technique: Preliminary assays were carried out at laboratory scale with an automatic film applicator in order to set the optimal variables for coating application at lab scale. For this purpose, solubility of the active agents was studied at several concentrations in three different polymeric varnishes. Those varnishes in which the active agents were not soluble were discarded. In addition, water soluble varnished were also discarded. Therefore, only one polymer was chosen for further semi-industrial scale trials.Extrusion technique: With the purpose to evaluate the dispersion of the active agents in the polymeric matrices selected for the development of an extruded tray and evaluate initial tests were performed at lab scale. In this case, the dispersion of the selected active agents in two polymers with different polarities and their mixture in different proportions were studied. In this way, pellets were obtained from a microcompounder and moulded by using a hot plate press obtaining small round specimens. The processing conditions were selected for each polymer based on the technical datasheets supplied by polymer producers. After verifying the feasibility of these materials, the development of an active tray at semi-industrial level was designed and characterized in a further step.Semi-industrial materials• Active lid:Coating technique: A pilot scale coating machine was used to apply the polymeric active varnish selected by gravure technique. The optimal parameters to obtain a stable and homogeneous material with the desired amount of active compounds were adjusted. However, the incompatible sealing between the material selected for the tray and the material selected for the coating and the need of a pattern design for sealing was the reason coating was not selected as a suitable technique to develop the active lid.Extrusion technique: Initially, two different films with same thickness and different concentration of active agents were developed by compounding followed by cast-coextrusion where the inner layer (in contact with) contained a mixture of volatile agents to be released to the head space of the packaging. Nevertheless, a final thinner lid was later produced and evaluated in order to have a more realistic packaging material used in the industry. For all the materials, processing parameters such as temperature, pressure, screw speed, residence time, etc., were optimized to ensure a low thermal damage to the active compounds. • Active trayCoating technique: In order to develop the most appropriate antimicrobial varnish, different blends of the selected polymeric varnish were scaled-up were scaled-up in an Atex pilot plant with a 20 L steel reactor vessel. After that, the varnish solutions were transferred to a pilot scale spraying machine, which uses an air-assisted system, to spray the injected-moulded trays. Extrusion-thermoforming technique: With the aim to improve the compatibility and release of the active compounds from the material, a mixture of the polymers tests at lab scale were produced obtaining a two-polymer blend active compound. The composite processing was performed in a twin screw extruder, with gravimetric main feeder, lateral feeder (for solid feeding), and liquid dosing port. Then, to obtain a bilayer sheet containing an external layer and two-polymer blend internal layer with active compounds, a co-extrusion line was used. Lastly, once the sheets were obtained, a converting step was carried out to obtain the final trays by thermoforming using a table top vacuum thermoforming machine.It should be mentioned that the semi-industrial extruded tray contained higher concentration of active agents than the coated tray due to the limitations of the spraying technique. However, this tray was initially not discarded because could show a better release of the active agents than the extruded tray.Characterisation of materials at lab and semi-industrial scale was performed in terms of physical, chemical and mechanical properties and also the kinetics release of the active compounds was evaluated. The results showed that no relevant differences which could affect the performance of the packaging material were found. Relating to migration kinetics of the active compounds, the volatile agents incorporated into the lid were slightly released in the studied conditions of high relative humidity and low temperature. However, higher temperatures are usually reached during the transport of products between supermarket and home and during their storage by consumer. This fact would prompt the release and therefore, the amount of active in the head-space of the packaging would increase. On the other hand, the release kinetics of the active compounds from sprayed trays was very fast, the equilibrium was reached after 8 h of contact with exposed medium in which they were tested, whereas, the co-extruded and thermoformed trays were able to control the release of both active agents.In addition the antimicrobial properties of these materials were also evaluated in vitro showing significant microbial reductions against the tested yeast and moulds. From the characterisation results and the antimicrobial properties observed, the following processed materials were selected as the most promising and were tested to validate their capability to prolong the shelf-life of fresh-cut fruits:o Semi-industrial extruded lid containing a mixture of volatile active compounds.o Semi-industrial coated active tray containing a mixture of solid antimicrobial compounds. o Semi-industrial extruded and thermoformed tray containing a mixture of solid antimicrobial compounds. In relation to the integrity of the active packaging during the sealing and storage of the fruits, it was observed that the active lid needed higher temperature of sealing in comparison with the lid control but it still allowed the right sealing of the materials. In addition, no presence of leaks, fractures or other signs of deterioration were observed either in the active or the control packaging during the storage of the fruit products. Microbial counts in fruits packed with the active lid and the active tray separately were evaluated. Eventually, the final active packaging system (lid + tray) was also tested up to 12 days at 4/8 °C (representing abuse conditions). Other quality parameters were measured in both fruits such as head-space gas composition, leakage of juice, pH and total soluble solids were evaluated along the storage period. The active packaging system consisting of the extruded active lid and the extruded active tray demonstrated to have higher effect in maintaining the freshness of fresh-cut pineapple and orange in terms of microbiological and physicochemical stability in comparison with the active lid and trays tested individually. In this sense, microbial growth was significantly inhibited by the active system in both fruits observing a pineapple reduction of 1-3 logs CFU whereas a slighter reduction was found for orange (1-2 logs CFU1).Concerning physico-chemical parameters of fruits packed in active packaging (pH and total soluble solids) and head-space composition no relevant changes were observed throughout the storage period. On the contrary, soluble solids of fruit packed with control materials underwent a decrease throughout the time probably associated with deterioration processes. In addition, fruit packed with active system showed higher leakage of juice, more pronounced in pineapple than in orange. Moreover, respiration rate and deterioration processes were slowed down when active packaging was employed, showing a high potential for preserving fresh-cut pineapple and orange.Sensory analysis was also performed in order to assess the acceptability of the product by consumers. In general terms, the overall acceptance of the peeled and cut fruit packed with active agents presented a decreasing tendency compared to the control, which was more significant for fresh-cut pineapple than for orange. Usually the liking scores were higher for the fruits packed in the control packaging than in active packaging irrespective of storage time. Attributes such as smell and taste of fruits presented the greatest significant differences between the control and the active packaging, while attributes such as texture and appearance were not highly affected by the type of packaging. The compliance under the Framework Regulation (EC) Nº 1935/2004 on materials and articles intended to be in contact with food and the Regulation (EC) Nº 450/2009 on active and intelligent materials to be in contact with food of the new active materials developed was evaluated. The evaluation of the global migration of the lid and tray was performed in order to evaluate the inertness of the material. In this sense, plastic materials and articles shall not transfer their constituents to food simulants in quantities exceeding 10 mg of total constituents released per dm2 of food contact surface (mg/dm2). According to Regulation (EC) No 450/2009, the overall migration from active releasing materials can exceed the overall migration limits (10 mg/dm2) as long as the levels transferred to the food comply with restrictions in the existing food law. The results of the Overall Migration analysis shown that, for the simulants testes, the overall migration values of the lid were far away of the limits for OM stated in the Regulation (EU) No 10/2011 (10 mg/dm2). These results point that the materials tested have a great inertness in the simulants tested. In contrast, the OM for the active tray widely exceed the OML (10 mg/dm2). Nevertheless, Regulation (EC) No 450/2009 clarifies that the transfer of the active substances should not be included in the calculation of the overall migration limit because they are incorporated in the packaging material to be liberated to the food. Because of that, the quantity corresponding to the active compounds was subtracted from the overall migration results, complying therefore with the OML established in the plastic Regulation (10 mg/dm2)Specific migration tests of the active substances incorporated in lid and the tray were evaluated. The results for the Specific Migration analysis was far below the limit established.Non-intentionally added substances analysis (NIAS) was also carried out in order to detect any intended or potential unintended or breakdown products originated during the manufacturing process, or the storage of the active samples and that could be present in the materials. A total of 16 chromatographic peaks were detected and tentatively identified in the active film and up to seven chromatographic peaks were detected in the tray. Main results obtained in Phase 3 The final phase of this project consisted in evaluating the effect of the combination of the most suitable protocol of the fruit processing developed in Phase 1 and the most promising active packaging system selected in Phase 2, and validation of the developed active packaging both in terms of design and industrial up-scaling of the packaging and in success commercially. With this purpose, the shelf life of the fruit processed according to the established protocol and packed with the active packaging system was studied. In addition the packaging performance throughout the shelf life of the fruit was evaluated. Furthermore, in order to validate the industrial active packaging, two fundamental criteria were taken into account, consumers´ acceptance by means of a consumer test and validation at retail stage taking into account the distribution cycle. Shelf-life evaluation of fruitIn order to assess the shelf-life of cut fruits and thus determine their final commercial life, a 14 days storage test was carried out with both passive and active packaging. Sensory descriptive analysis of fruit, microbial analysis (psychrophilic bacteria, moulds, yeasts) of fruit, concentrations of O2 and CO2 in modified atmosphere, leakage of juice from fruit, titratable acidity, pH and soluble solids (sugar) contents in fruit, concentrations of volatiles (active compounds) in modified atmosphere and appearance of fruit by calibrated photography were evaluated.The main conclusion arising from the shelf-life test was that optimised minimal processing and passive packaging was enough for storing the fruit up to 14 days with a sensory quality similar to freshly cut fruit, and with acceptable microbial load and juice leakage. Nevertheless, by using a combination of active packaging and optimised minimal processing the appearance of the fruit was significantly improved and its microbial growth reduced compared with the use of passive packaging and optimised minimal processing, keeping the quality of the fruit much better.Packaging evaluationIn order to ensure that the packaging system kept all the requirements required by the fruit throughout their shelf life, the Integrity and sealing, visual aspect and active compounds evolution (both in the tray and in the lid) were evaluated using different analytical techniques. The integrity and sealing of the active packaging system (lid+tray) was evaluated by means of a peelability assay. This test represents the conventional way that consumers use to open the food packages. Sealed trays were kept in horizontal position and the film of the lid was pulled under controlled speed.In relation to the aspect of the packaging materials, conventional and active packaging systems were monitored throughout the shelf life of both fruits with a high-resolution digital camera.At the same time, the residual content of the compounds in the active lid during the storage period was determined by thermogravimetric analysis (TGA) and by chemical solvent extraction and subsequent chromatographic analysis. While, the active compounds remaining in the tray were evaluated using a muffle furnace and chromatographic analysis were carried out to determine their content in the fruit.From the evaluation of the active packaging it was concluded that either the active or the conventional packaging could not be considered as a peelable seal since the lid was broken before peeling. Therefore, there was a need to improve the peelability of the system based on the use of incompatible resins. In this sense, the combination of a non-polar polymer with different concentration of a polar polymer should be evaluated to provide improved sealing and peelability behaviour. No visual changes were observed in the active films that were in contact with orange and pineapple during the shelf-life of the fruit. However, shadows appeared in the active tray due to the loss of the active compounds released from the tray into the fruit. To improve the tray appearance, a white food contact colorant was added to the active trays at the following industrial scaled. Volatiles concentration from the lid decreased faster in pineapple (up to 58%) than in orange (up to 66%) as a consequence of the higher antimicrobial activity. On the contrary, concentration of the active compounds from the tray decreased over time for both orange and pineapple being the content remaining in the tray around 80% of the initial concentration.The remaining concentration of the active compounds both in the tray and the lid at the end of the shelf-life of the fruits pointed out that there is a possibility to reduce the initial active compounds concentration but taking into account the diffusion coefficient and the release kinetics of these compounds which will be decisive for the effectiveness of the active materials on the fruit.Up-scalingBlend processing was considered in a streamlined approach. This was based on the consideration of different parameters and equipment for the processing. Processing steps was divided in development of compound/masterbatches, film and sheet processing, thermoforming and sealing. To carry out the composite processing, a twin screw extruder, with gravimetric main feeder, lateral feeder, and liquid dosing port was used and different masterbatches and compounds were processed. Different masterbatches were processed, based on solid active compounds and talc mixed with a polymeric blend. In addition volatile compounds were also incorporated in a masterbatch using gravimetric dosing system.During sheet processing, molten polymer was extruded through a slit die onto a polished metal roll (chill roll), which serves to quench the hot melt. From the quench roll, the film passes around a series of other rolls designed to guide it and keep it wrinkle-free at wind-up. Film and sheet processing was based on cast film co-extrusion. Cast film extrusion process is widely used for making thermoplastic films for subsequent processing such as calendaring, coating, or lamination. During trials, temperature profile on the two extruders, transfer lines, feed-block and die was established and was kept, as processing conditions were stable and reproducible, being below degradation temperature of materials.Materials were run without major problems, providing a bilayer structure by closing one of the manifolds of the feedblock. This processing step is readily applicable at industrial sites as is based on compounding steps (with up scalable equipment), and coextrusion lines follow the same layout and properties of applied equipment.Sheets were thermoformed to trays using moulds developed for the project. These trays have been used for characterization and packaging trials. During the project, it has been evaluated the samples from two approaches, a pilot thermoforming unit and application of developed materials in an industrial line at industry facilities. Different batches of these materials were used for shelf life evaluation and consumer panels, as well as for validation at retail stage.Commercial validationThe last stage of the project dealt with the commercial validation of the EASYFRUIT developments. In this sense, semi-industrial prototypes were used for acceptability of the product by consumer and validation at retail stage. A test to better understand consumers’ acceptance of oranges and pineapples stored in active packs and to explore consumer’s perception of the concept of “active packaging” was carried out with Norwegian consumers at NOFIMA Institute. Parameters evaluated on the samples were: overall liking, appearance liking, odour liking in 9-point hedonic scales, and willingness to taste in a 5-point categorical scale. In addition, a concept test was also performed, through a word association exercise, where the stimuli were the concept of “packed cut fruit” and the concept of “cut fruit in active pack”. The results obtained showed that consumers generally showed lower acceptability ratings than passive packed fruits; this was mainly because of the artificial odour generated by the active compounds. The effect was more evident in pineapples than in oranges. Regarding the concept of «active pack with natural substances» consumers perceived it as healthy but they were somehow sceptical and saw it as «complicated» and «marketing» related. More research would be needed if this information would be used in communication to consumers.A validation from a commercial point of view of the fresh-cut orange and pineapple packed in two types of packaging (passive and active) representing the storage conditions that take place at the selling point was conducted by Carrefour supermarket. The test was performed with the participation of experts from different areas (Quality Platforms, Sales point, Traditional Fresh Product, Sustainable Development, etc.). The tests were conducted under the conditions specified in the sensory analysis standards UNE 87-005-92 and ISO 5495:1983 “Sensory analysis - Methodology - Paired comparison test” in order to determine whether there exists a perceptible sensory difference or a similarity between samples of two products.Packages and fruits were evaluated at day 2 and 9 in terms of packaging and product appearance (1 to 5 scale, in growing satisfaction order), overall rating (1 to 9 scale, in growing satisfaction order), preference between active or passive packaging, shelves section in which this type of product will be placed (Fruits, ready-to-eat, free service product (FSL), and other), and if Carrefour as a retailer will included this product in their supermarkets. Moreover, the willingness to buy the active and the passive packaging was also checked. Visual inspection was also conducted after 12 and 16 days of storage. The results pointed out that only at the beginning of the storage period (day 2) passive packaging was preferred with respect to active packaging. Orange and pineapple in active packaging maintained optimal visual appearance after 16 days of storage in selling point conditions (presenting pineapple in active packaging a lower browning). However, pineapple and orange in active packaging presented a higher amount of exudate. On the other hand, a significant citrus smell was detected in all samples, being more noticeable in those ones with active packaging which could affect its acceptability. Willingness to buy was significant and positive concerning active packaging, and Carrefour will accept this type of product which will be included to be sold in the fruits section.Potential Impact:The main objective of the EASYFRUIT project was to develop a minimal processing protocol in combination with the development of an active package for the extension of shelf-life of peeled and cut orange and pineapple for 5 days. In particular, the final results obtained were:- Minimal processing treatment for peeled and fresh-cut orange and pineapple- Active film- Active tray - Active packaging for pineapple - Active packaging for orangeTaking into account that those results will extend the shelf-life of the product increasing its availability for the consumer, the following impacts would be obtained at European level: Social: Increasing the consumption of fresh fruits among the populationStatistics indicate that 57% of children do not meet the guidelines of recommended daily fruit consumption. Also, elderly people or people with limited mobility have a limitation when peeling and cutting fruit products. In addition, the changes in lifestyle include modified eating habits and more time away from home. Therefore, convenience – or lack thereof – is a major factor in consumers' fruit purchase decisions. The new fruit packaging in a peeled and sliced format with longer shelf life will increase the consumption of fruit, which are really difficult to peel (especially pineapples).Health and political: Decreasing cardiovascular diseases and obesity, and reducing the cost associated with the health treatmentsAccording to the World Health Organization inadequate intake of fruits and vegetables is one of 10 major risk factors contributing to attributable mortality. Up to 1.7 million lives could be saved, each year, if global consumption of fruits and vegetables was enough. In this sense, obesity is one of the greatest public health challenges and its prevalence has tripled in many countries of the European Region since the 1980s particularly among children. Fruits and vegetables are known to be rich in vitamins and minerals, fibre, and other substances that are important for good health; therefore, fruit and vegetable intake is associated with improved health and reduced risks for chronic diseases and to combat obesity. On top of this, the cost associated to obesity is growing each day. For example, the cost of obesity treatment in Spain and its impact on health promotion accounts for 7% of total health cost. This sums up to 7% of 64,000 million euros i.e. nearly 5,000 million euros. With these developments, the consumption of healthy peeled and cut fruits will be reinforced, diminishing the diseases associated with the lack of fruit consumption and the cost associated to the related diseases.Commercial: fresh fruit lost reduction and capitalization of new export marketsThe implementation of a new packaging solution able to extend the shelf-life of a peeled and cut product will allow the commercialization from one European country to another and still have a shelf-life remaining. Therefore, by extending the shelf-life of the commercialised fruit the competitiveness will grow, increasing the sales and the turnovers for this specific sector.If the extension of the shelf-life achieved in the project is good enough, even commercialization out of Europe would be possible, contributing to the economic growth and market expansion of European fruits. Environmental: waste reductionBy improving the minimally processing techniques of fruit and combining them with an active packaging, the shelf-life of the fruit will be increased, therefore reducing waste. Also, if the product is peeled and cut, treatment that generates great amounts of waste, this benefit is even greater. Moreover, the waste generated from peeling and cutting fruit could be reused for other purpose using it as a by-product. As a conclusion, the commercialization of an easy to eat healthy fruit product with prolonged shelf-life presented in a convenient format could help to combat health issues, waste generation, and increase competitiveness. List of Websites:http://www.easyfruit.eu/Cooperativa BenaguasilMr. Miguel Ángel MartíManager Cooperativa, SN46180, BENAGUASIL VALENCIAESPAÑA Tel: +34 962730286Email:mamarti@ruralfruit.comSlice Fruit A/S Henriette Holm KristiansenQuality Department /Product DevelopmentGejlhavegaard,15 6000 Kolding Denmark Tel: +45 75569399 Email: Henriette.Holm.Kristiansen@flensted.dkPreduzece za proizvodnju, konsalting i promet Spektar doo Mr. Petar Djordjevic Managing Director Ul Rudnicka 32300 Gornji Milanovac Serbia Tel: +38163601993 Email: petar.djordjevic@spektar.com Omniform S.A Mr Alain Wambeke General Director Avenue Franklin Zoning Industriel 1301Wavre Belgium Tel: +32 10 226955 Email: alain.wambeke@omniform.be Instituto Tecnológico del Embalaje, Transporte y Logística Ms. Carmen Sánchez Reig Deputy Director Parque Tecnológico C/ Albert Einstein, 146980 Paterna Valencia Spain Tel: +34 961820081 Email: itneur@itene.com Nofima AS Mrs. Anne Risbrathe Project Controller EU Muninbakken, 9-13 9291 Tromso Norway Tel: +4764970326 Email: anne.risbraathe@nofima.no Centros Comerciales Carrefour S.A. Mr. Rafael Herrero Responsible for sustianable developmentC/Campezo, 16 28022 Madrid SPAIN Tel: +34 913331308Email: Rafael_Herrero_Gonzalez@carrefour.com
