#507_LAVELLI_bozza ! Ital. J. Food Sci., vol 28, 2016 - 542 REVIEW RECOVERY OF WINEMAKING BY-PRODUCTS FOR INNOVATIVE FOOD APPLICATIONS V. LAVELLIa*, L. TORRIb, G. ZEPPAc, L. FIORId and G. SPIGNOe aDeFENS, Department of Food, Environmental and Nutritional Sciences, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy bUniversity of Gastronomic Sciences, Piazza Vittorio Emanuele 9, 12042 Bra, Italy cDISAFA, Department of Agricultural, Forestry and Food Sciences, Università degli Studi di Torino, L. go P. Braccini 2, 10095 Grugliasco, Italy dDepartment of Civil, Environmental and Mechanical Engineering, Università degli Studi di Trento, Via Mesiano 77, 38123 Trento, Italy eInstitute of Oenology and Agro-Food Engineering, Università Cattolica del Sacro Cuore, Via Emilia Parmense 84, 29122 Piacenza, Italy *Corresponding author. Tel.: +39 0250319172; fax: +39 0250316632 E-mail address: vera.lavelli@unimi.it ABSTRACT Winemaking by-products are potential resources for second-generation biorefineries, i.e., biorefineries fed with biowaste to produce added-value products, particularly for the food sector. In fact, winemaking by-products are outstanding sources of oil, phenolic compounds and dietary fibre and possess numerous health benefits and multifunctional characteristics, such as antioxidant, colouring, antimicrobial and texturizing properties. The present review highlights promising developments for the conversion of winemaking by-products into novel food ingredients, as well as their use in innovative foods, focusing on the type of recovered ingredients, dosage, formulation and processing. In addition, the primary benefits of winemaking by-products to new foods are described. Keywords: by-products, dietary fibre, grape phenolics, grape seed oil, winemaking ! Ital. J. Food Sci., vol 28, 2016 - 543 1. INTRODUCTION Food supply chains have significant environmental impacts due to their use of resources and production of emissions, effluents and wastes. According to the European Union (EU) Commission Council Directive 2008/98/EC, “waste” is defined as “any substance or object, which the holder discards or intends or is required to discard”. The importance of food waste stretches from environmental pressures to economic and social impacts, including negative effects on food and nutrition security (OTLES et al., 2015). To meet the overall objective of increasing the sustainability of production chains, waste prevention/minimization is the main priority and best option, followed by reuse, recycling and energy recovery. Alternatively, disposal (the use of landfills or incineration with low energy recovery) must be considered the worst environmental option. Reuse and recycling strategies have drawn attention on the valorisation of by-products. A “by- product” is defined as a product that must originate from a production process without being the main goal of production, be usable in the same production process or in a subsequent production or utilization process and be directly re-usable without further treatment outside normal industrial practices. Moreover, a by-product must have a market value, and the final use should be integral without negatively impacting human health or the environment (GALANAKIS, 2015). Like all agro-food productions, the winemaking process generates a series of by-products that are important from both a quantitative and qualitative point of view and have been considered potential resources for second-generation biorefineries, i.e., biorefineries fed with biowaste to produce added-value products (SCOMA et al., 2016). In fact, grapes are one of the most cultivated fruits worldwide. According to the FAO (www.faostat3.fao.org), 77 Mt of grapes were produced throughout the world in 2013 (most of which were used in winemaking), along with 3.4 Mt of grape pomace (otherwise referred to as grape marc). The latter represents, by weight, the primary winemaking solid by-product (60% on average), followed by lees (approximately 25%) and stalks (approximately 14%). Minor solid by-products mainly include wine filtration residues. On average, depending on the grape variety and winemaking process, 100 kg of processed grapes generates 20-25 kg of pomace, a mixture of skins and seeds, 3-5 kg of stalks and 8- 10 kg of lees (SPIGNO, 2015). All of the aforementioned by-products pose serious environmental concerns because their production is typically concentrated in a limited time frame, and their high organic matter content prevents direct disposal into the soil, except for limited and regulated amounts. Although these materials are re-used for other applications, being correctly considered as by-products, landfill additions and incineration are also conducted, depending on the country. Conventional applications for winemaking by-products include: agronomic use, animal feed production and compost production for all residues; distillation for pomace and lees; tartaric acid manufacturing and the production of colouring additives and nutritional supplements from pomace, lees and filtration residues; oil recovery from seeds. Agronomic use, animal feed production, composting and distillation, a relatively new approach, are not considered remunerative strategies (SPIGNO, 2015). If properly recovered, winemaking by-products show a wide range of potential and remunerative applications in many industrial sectors, including cosmetics, pharmaceuticals, biomaterials and food (BORDIGA, 2015; YU and AHMEDNA, 2013). In fact, grape pomace is a rich source of both dietary fibre and various phenolic compounds (TEXEIRA et al., 2014). The amount of phenolic compounds that remain in the pomace depends on the initial, genetically dependent content of grapes, as well as the processing conditions and skin thickness, which is another genetically dependent parameter that is crucial for the maceration phase (BATTISTA et al., 2015). A study of various cultivars of ! Ital. J. Food Sci., vol 28, 2016 - 544 Vitis vinifera L. has revealed that the content of soluble proanthocyanidins in the skin ranges between 1.16 and 44.6 g/kg d. w., while the content of soluble proanthocyanidins in seeds ranges between 23.1 and 68.5g/kg d. w. (TRAVAGLIA et al., 2011). The total anthocyanin content of red grape skins is in the range of 2.5-132 g/kg d. w. (KAMMERER et al., 2004; SRI HARSHA et al., 2013). The presence of anthocyanins in the red grape seed fraction, due to mash constituents adhering to the seeds, is generally neglected or reported to be low (KAMMERER et al., 2004; LAVELLI et al., 2015a). However, a recent patent was focused on the extraction of anthocyanins from grape seeds, suggesting that their content deserves attention in a full recovery strategy (BI and RUI, 2014). The total flavonol content of grape skins is in the range of 0.3-2.6 g/kg d. w. (SRI HARSHA et al., 2013; SRI HARSHA et al., 2014), whereas these compounds are generally less than 0.1 g/kg d. w. in grape seeds (MAIER et al., 2009b). Compared to the above-mentioned phenolic compounds, phenolic acids and stilbenes are present in considerably lower amounts in winemaking by-products (KAMMERER et al., 2004). Grape seeds contain oil with a high nutritional value. Among various vegetable oils, grape seed oil shows the largest percentage of linoleic acid (C18:2 ≈70%). Other major fatty acids present in grape seed oil include oleic acid (C18:1 ≈15%), palmitic acid (C16:0 ≈7%) and stearic acid (C18:0 ≈3%) (HANGANU et al., 2012; FERNANDES et al., 2013; FIORI et al., 2014). In addition to the interesting fatty acid profile, grape seed oil contains significant amounts of bioactive compounds such as tocopherols and tocotrienols, presenting a total tocol content up to 1208 mg/kg (BEVERIGE et al., 2005; CREWS et al., 2006; FIORI et al., 2014). Winemaking by-products are of particular interest for food uses when they are obtained via organic production because consumer preference is positively influenced by information on sustainable production practices (LAUREATI et al., 2013). In particular, a great deal of interest in sustainability issues has been expressed for winemaking (LAUREATI et al., 2014). The purpose of using winemaking by-products in foods may be fortification or enrichment. The distinction between these terms is not always recognized in scientific studies but has been clarified as follows: a fortified product is defined as a food containing additional nutrients, while an enriched product is defined as a food with additional novel nutrients or components not normally found in a particular food (SIRO’ et al., 2008). Tartaric acid, enocyanine (E163) and grape seed oil are classical examples of successful commercial products obtained from winemaking by-products. Additionally, in the last several years, grape seed and grape skin powders have been commercialized by different companies and promoted as highly nutritional ingredients to enrich conventional cereal flours and baked products with fibre, minerals, antioxidants, colour and aroma. The concept of antioxidant dietary fibre was first proposed by SAURA-CALIXTO (1998), who set the criteria that 1 g of antioxidant dietary fibre should possess a free radical scavenging capacity equivalent to at least 50 mg of vitamin E and should contain more than 50% dry matter of dietary fibre from the natural constituents of the material. Whole grape pomace, grape seeds and grape skin generally meet these criteria and are often referred to as antioxidant dietary fibre. Enocyanine and grape antioxidant dietary fibre represent the two basic solutions for the reintroduction of grape pomace into the food chain, including indirect and partial use as concentrated extracts, or direct use as ground, dehydrated and micronized antioxidant dietary fibre. In both of the above-mentioned cases, a new production process must be implemented. Fig. 1 shows a schematic depiction of the basic conventional process for antioxidant dietary fraction production. ! Ital. J. Food Sci., vol 28, 2016 - 545 Figure 1: Scheme of the basic process for the production of grape pomace, skin or seed antioxidant dietary fibre. Pomace collection should follow preliminary care selection to identify the best pomace for the production of food grade ingredients, based on the content of functional constituents (fibre, polyphenols and minerals), as well as possible contaminants (heavy metals, pesticides residuals and mycotoxins) (CORRALES et al., 2010; SOLFRIZZO et al., 2012). Washing and cleaning operations should be required, while the recovery of phenolic compounds would be reduced. If needed, skins and seeds can be separated by sieving, before or after drying. Of course, drying is necessary to obtain a final powder but is also the most common stabilisation treatment. Grape pomace has a high moisture content (greater than 60%) and undergoes rapid fermentation if not properly treated. Low temperature preservation may precede the drying step for logistic and timing reasons. Drying, which is an energy consuming process, should be reduced to a minimum because the thermal degradation of antioxidant compounds is detrimental to the nutritional profile. However, drying allows for the inhibition of enzymatic activity and can be considered a mild sanitization process. The operating temperature should not exceed 60°C to limit the degradation of phenolic compounds (AMENDOLA et al., 2010). In the production of antioxidant dietary fibre from skins and seeds, the seeds can be defatted in a previous step to recover the oil and produce a fibre-rich ingredient with limited rancidity issues. The final milling step must set the ideal particle size, depending on the expected application. If dried skins and seeds are destined for the production of an extract, a particle size range of 0.5 to 2 mm is acceptable. As outlined in the following paragraphs, the particle size should be less than 0.5 mm for use in bakery products or pasta. For applications into fruit-based and dairy products, an even lower particle size is required, which leads to additional energy consumption. Fig. 2 shows the basic process for the production of grape pomace, skin or seed extracts (SPIGNO, 2015). ! Ital. J. Food Sci., vol 28, 2016 - 546 ! Figure 2: Scheme of the basic process for the production of extracts from grape pomace, skins or seeds. As indicated in the production of antioxidant dietary fibre, the operating temperature should be less than 60°C. Different extraction techniques can be applied, such as conventional solvent extraction using food-grade solvents (AMENDOLA et al., 2010), or non-conventional solvents and systems for the development of sustainable and environmentally friendly processes. The ultrasound-assisted extraction has been successfully applied for the extraction of grape pomace phenolics, using water as a solvent and achieving high extraction yield with a short extraction time (MARINELLI et al., 2015). Microwave-assisted solvent extraction using ethanol/water (PEDROZA et al., 2015) and high-pressure extraction using ethanol/water (PAINI et al., 2016), as well as the use of aqueous solutions of organic acids (TZIMA et al., 2015) have also been proposed. In general, all of these systems are characterized by low selectivity, and other compounds (such as sugars, minerals and organic acids) are co-extracted with the phenolic compounds, producing a crude extract. For food applications, the purification of extracts may be omitted without further increasing the production costs. The crude extract can then be simply concentrated to give a liquid extract or dried to give a powder extract. In this case, using a tailored approach (that takes into account the food category of the final target), the addition of suitable carrier materials (e.g., maltodextrins) can be exploited to increase and modify the stability and solubility of phenolic compounds (SPIGNO et al., ! Ital. J. Food Sci., vol 28, 2016 - 547 2013; LAVELLI et al., 2016b). To obtain an extract with a higher purity in total phenolic compounds or within a selected class of phenolic compounds, a purification step is required. Adsorption resins (SOTO et al., 2011) and membranes (ZAGKLIS and PARASKEVA, 2015) are the most commonly investigated purification systems, along with other non-conventional approaches, such as the use of colloidal gas aphrons (SPIGNO et al., 2015). In any case, microfiltration is also suggested as a non-thermal technology to produce crude extracts while promoting microbiological stability. Independent of the production process used to obtain grape pomace antioxidant dietary fibre, phenolic extracts or seed oil, this review describes literature examples of innovative food applications into meat, fish, cereal, fruit-based and dairy products, with a focus on the type of recovered ingredient, dosage level and primary results achieved by the application. 2. PHENOLIC EXTRACTS AND ANTIOXIDANT DIETARY FIBRE FROM GRAPE SKINS AND SEEDS AS INGREDIENTS IN INNOVATIVE FOODS 2.1. Functional effects New ingredients recovered from winemaking by-products have the potential to provide a wide range of food products with numerous health benefits (SAURA-CALIXTO, 1998; TEXEIRA et al., 2014). Moreover, as outlined in the following sections, these by-products possess multifunctional properties and could be used as natural antioxidants, colorants, antimicrobial agents and texturizers. 2.1.1 Meat products Due to the growing interest in convenience foods, ready-to-eat products such as dehydrated meat, frozen and precooked hamburgers, patties and meatballs have become a major category in the meat industry. The quality and shelf-life of these products is primarily dependent on the inhibition of lipid oxidation, which affects the colour, flavour, odour, texture and nutritional value of foods (FERNANDEZ et al., 1997). Consequently, research efforts have been devoted to the application of winemaking by-products in various meat products to prevent lipid oxidation during precooking and storage under refrigerated or frozen conditions, representing natural alternatives to the use of synthetic antioxidants (Table 1). In addition to auto-oxidation, microbial contamination is another serious factor that affects the quality and shelf-life of ready-to-eat meat products. However, only a few studies have investigated the antimicrobial properties of winemaking by-products in meat products (AHN et al., 2007). Regarding chicken meat, when red grape skin extract powder was added to the dehydrated product at a level of 1 g/kgmeat, the content of hexanal and thiobarbituric acid reactive substances (TBARS) formed during processing and storage at 22°C under air decreased. However, the efficacy was lower than those of rosemary extract and synthetic antioxidants, such as butylated hydroxyanisole and butylated hydroxytoluene (NISSEN et al., 2000). The antioxidant dietary fibre obtained from red grape pomace with particle sizes < 0.5 mm (total dietary fibre: 782 g/kg; soluble phenolics: 49.3 g/kg) has been applied to chicken hamburger, delivering fibre and imparting antioxidant effects during processing and refrigerated storage under air (SAYAGO-AYERDI et al., 2009). Alternatively, the extract obtained from the entire pomace (skins and seeds) has been proven to act as an antioxidant in uncooked and cooked chicken meat at a concentration corresponding to 60 mg phenolics/kgmeat during processing and frozen storage under vacuum (SELANI et al., 2011). ! Ital. J. Food Sci., vol 28, 2016 - 548 Table 1: Applications of winemaking by-products as new food ingredients in meat products. Food product Recovered ingredient Main results and references Chicken meat (dehydrated) Red grape skin extract powder (soluble TP: 1.60 mmol phenol Eq./g) Integration: 1 g/kgmeat Decrease in hexanal and TBARS content during processing and storage at 22°C in aluminized sachets sealed in air. Lower efficacy than rosemary extract. NISSEN et al., 2000 Chicken hamburger (uncooked and pre-cooked) Red grape pomace antioxidant dietary fibre (particle size < 0.5 mm; TDF: 782 g/kg; soluble TP: 49.3 g GAE/kg) Integration: 5-20 g/kgmeat Decrease in TBARS content during processing and storage in polyvinyl chloride bags (OTR: 13.500 cm3/m2d) at 4°C. High antioxidant activity and fibre content. SÁYAGO-AYERDI et al., 2009 Chicken meatballs (uncooked and pre-cooked) Red and white grape pomace extract (soluble TP: 7.8-9.4 g GAE/kg) Integration: 60 mg TP/kgmeat Decrease in TBARS content during processing and storage at -18°C under vacuum. SELANI et al., 2011 Pork patties (pre-cooked) Red grape skin extract powder (soluble TP: 1.60 mmol phenol Eq./g) Integration: 0.2 g/kgmeat Decrease in hexanal and TBARS content during processing and storage at 4°C in polyethylene bags (OTR > 2000 cm3/m2d). Lower efficacy than rosemary extract. NISSEN et al., 2004 Beef and pork patties (pre-cooked) Grape seed extract powder (TP: 980 g/kg) Integration: 0.1-0.2 g/kgmeat Decrease in TBARS content during processing and storage in polyvinyl chloride bags (OTR: 880 cm3/m2d) at 4°C. ROJAS and BREWER, 2007 Pork patties (pre-cooked) Grape seed extract powder (TP: 865 g/kg) Integration: 0.05-1 g/kgmeat Decrease in TBARS content during processing and storage in barrier film packs (OTR: 3 cm3/m2d) under 75% O2 and 25% CO2, at 4°C. CARPENTER et al., 2007 Beef ground meat (pre-cooked) Grape seed extract powder (TP: not specified) Integration: 10 g/kgmeat Reduced numbers of Escherichia coli O157:H7, Salmonella typhimurium, Listeria monocytogenes and Aeromonas hydrophila during storage in bags (OTR: not specified) at 4°C. AHN et al., 2007 Beef sausage (pre-cooked) Grape seed extract powder (TP: 800 - 990 g/kg) Integration: 0.1-0.5 g/kgmeat Decrease in TBARS content during processing and storage in polyvinyl chloride bags (OTR: 880 cm3/m2d) at -20°C. Higher efficacy than ascorbate. KULKARNI et al., 2011 OTR: oxygen transmission rate; TBARS: thiobarbituric acid reactive substances; TDF: total dietary fibre; TP: total phenolics (expressed as GAE: gallic acid equivalents or phenol Eq: phenol equivalents or PAs: proanthocyanidins). Considering pork meat, red grape skin extract powder added at a level of 0.2 g/kgmeat enhanced the oxidative stability of cooked patties during processing and refrigerated storage under air but showed lower efficacy than rosemary extract, as observed for chicken meat (NISSEN et al., 2004). Grape seed extract appears to be more effective than grape skin extract. In fact, grape seed extract powder added at a level of 0.1-0.2 g/kgmeat had better antioxidant effects compared to rosemary oleoresin and oregano water extract during the processing and refrigerated storage under air of cooked beef and pork patties (ROJAS and BREWER, 2007). Grape seed extract powder is also an effective antioxidant in cooked pork patties during processing and frozen storage under a high-oxygen atmosphere, when added at a concentration as low as 0.05 g/kgmeat (CARPENTER et al., 2007). ! Ital. J. Food Sci., vol 28, 2016 - 549 In ground beef, grape seed extract powder has been demonstrated to inhibit the growth of Escherichia coli O157:H7, Salmonella typhimurium, Listeria monocytogenes and Aeromonas hydrophila (AHN et al., 2007) at a level of 10 g/kgmeat during refrigerated storage. However, the required addition of grape seed extract is higher than the effective amount for the inhibition of meat oxidation. In fact, at an extract concentration of 0.5-1 g/kgmeat, grape seed extract powder prevented the oxidation of beef sausage during processing and frozen storage under air, more effectively than ascorbic acid (KULKARNI et al., 2011). 2.1.2 Fish products Fish tissues have a high content of polyunsaturated fatty acids (PUFA), which undergo degradation via auto-oxidation. The use of natural antioxidants has become as an effective strategy for controlling the stability of these products, either during the frozen storage of minced tissue or during the processing and refrigerated storage of pre-cooked fish-based products. For this purpose, winemaking by-products have also been considered (Table 2). Table 2: Applications of winemaking by-products as new food ingredients in fish products. Food product Recovered ingredient Main results and references Atlantic mackerel minced muscle (uncooked) Phenolic fractions of white grape pomace. Integration: 0.1 g monomeric flavonoids or PAs/kgfish Longer induction period for the formation of peroxides and aldehydes during storage in Erlenmeyer flasks under air at -10°C. Maximum protection by PAs with high degree of polymerization and percentage of galloylation. PAZOS et al., 2005 Horse mackerel minced muscle (uncooked) White grape pomace antioxidant dietary fibre (particle size < 0.25 mm; TDF: 760 g/kg; soluble TP: 78 g GAE/kg) Integration: 20 - 40 g/kgfish Inhibition of formation of conjugated dienes and trienes and TBARS during storage in Cryovac BB4L bags (OTR: 30 cm3/m2d) at -20°C. Significant antioxidant activity and high fibre content. SANCHEZ-ALONSO et al., 2008 Chub mackerel minced muscle (uncooked) Red grape seed extract (soluble TP: 66 g GAE/kg) Integration: 20 g/kgfish Inhibition of lipid hydroperoxides and TBARS formation during storage in cartoon trays under air at -20°C. OZEN et al., 2011 Meagre sausage (pre-cooked) White grape skin antioxidant dietary fibre (particle size < 1 mm; TDF: 820 g/kg; soluble TP: 42 g GAE/kg) Integration: 30 g/kgfish Inhibition of TBARS formation and oxidation during storage in barrier bags (OTR: < 2.1 cm3/m2d) at 2°C. Significant antioxidant activity and high fibre content. Antimicrobial effect on H2S producers and a reduction in total viable counts. RIBEIRO et al., 2013 OTR: oxygen transmission rate; TBARS: thiobarbituric acid reactive substances; TDF: total dietary fibre; TP: total phenolics (expressed as GAE: gallic acid equivalents or sum of monomeric flavonoids or PAs: proanthocyanidins). Fractionated grape pomace phenolic compounds at a concentration of 0.1 g/kgfish have been proposed as inhibitors for fatty fish species, such as the muscle of Atlantic mackerel (Scomber scombrus) during frozen storage under air. The induction period for the formation ! Ital. J. Food Sci., vol 28, 2016 - 550 of peroxides and aldehydes was significantly increased in samples treated with grape phenolic fractions, and the maximum protection was achieved using procyanidins with a high degree of polymerization and percentage of galloylation (PAZOS et al., 2005). Non- fractionated grape seed phenolics also increased the oxidative stability of minced fish during frozen storage under air (OZEN et al., 2011). The antioxidant dietary fibre obtained from white grape pomace with a particle size < 0.25 mm (total dietary fibre: 760 g/kg; soluble phenolics: 78 g/kg) at a concentration of 20-40 g/kgfish can also increase the oxidative stability of the minced muscle of horse mackerel (Trachurus trachurus) during frozen storage under a low oxygen atmosphere (SANCHEZ- ALONSO et al., 2008). Similarly, white grape skin antioxidant dietary fibre with a particle size < 1 mm (total dietary fibre: 820 g/kg; soluble phenolics: 42 g/kg), which was used at a concentration of 30 g/kgfish in precooked meagre (Argyrosomus regius) sausage, showed antioxidant effects as well as antimicrobial effects on H2S producer counts and total viable counts, during refrigerated storage under a low oxygen atmosphere (RIBEIRO et al., 2013). 2.1.3 Bakery products and pasta Bread is a staple food, and fortification with polyphenols and dietary fibre derived from winemaking by-products has been investigated to improve the diet of consumers (Table 3). Grape seed extract powder added at a level of 0.6-2 g/kgbread greatly increased the antioxidant activity of the final product, despite the loss of phenolic compounds during processing due to either thermal treatment or interaction with the food matrix. Interestingly, a decreased amount of N-(carboxymethyl) lysine, an advanced glycation end-product associated with health risks, was observed in bread containing grape seed extract. Moreover, the addition of grape seed extract powder did not significantly affect the hardness of bread but did increase the darkness (PENG et al., 2010). To obtain another vehicle for grape seed phenolics, the entire grape seed can be milled to fine particle sizes (< 0.150 mm). However, upon the addition of this ingredient (total dietary fibre and total phenolic contents not specified) to dough at a level of 25-100 g/kgflour, a decrease in loaf brightness and volume, along with an increase in hardness and porosity, was observed (HOYE and ROSS, 2011). These effects were likely due to the inhibition of yeast activity, which reduced the gassing power. Moreover, phenolics can inhibit the activity of endogenous amylases in dough, leading to inadequate maltose release for yeast activity during proofing (MILDNER-SZKUDLARZ et al., 2011). Because grape phenolics also inhibit mammalian α-glucosidase and α-amylase, white grape skin antioxidant dietary fibre with particle sizes < 0.250 mm (soluble phenolics: 20.0 g/kg) at a level of 100 g/kgflour has been used in functional flat bread for diabetic people (LAVELLI et al., 2016a). Sourdough fermentation improves the textural properties of wheat and rye breads. Hence, the fortification of mixed wheat-rye bread with red grape pomace dietary fibre (total dietary fibre: 593 g/kg; soluble phenolics: 58.9 g/kg) has been investigated (MILDNER- SZKUDLARZ et al., 2011). An increase in hardness, gumminess and springiness was once again observed in wheat-rye mixed sourdough bread, but the cohesiveness and resilience did not change (MILDNER-SZKUDLARZ et al., 2011). To increase the fibre and/or phenolic content, the fortification of brownies and biscuits with winemaking by-products has been achieved. In brownies, upon addition of 150-250 g/kgflour of red grape pomace antioxidant dietary fibre with particle size < 0.589 mm (total dietary fibre and total phenolic contents not specified), hardness and chewiness decreased, while springiness increased. Hence, the fortification of brownies showed an opposite trend compared to bread fortification, likely due to the presence of fat (WALKER et al., 2014). ! Ital. J. Food Sci., vol 28, 2016 - 551 Table 3. Application of winemaking by-products as new food ingredients in bakery products and pasta. Food product Recovered ingredient Main results and references Wheat bread Grape seed extract powder TP: not specified Integration: 0.6-2 g/kgbread Decreased level of N-(carboxymethyl)lysine, an advanced glycated end-product related to health risk. No significant difference in hardness. Increase in darkness. PENG et al., 2010 Wheat bread Grape seed antioxidant dietary fibre (particle size < 0.150 mm; TDF and TP: not specified) Integration: 25-100 g/kgflour Decrease in loaf volume, and increase in hardness and porosity. HOYE and ROSS, 2011 Wheat bread White grape skin antioxidant dietary fibre (particle size < 0.25 mm; soluble TP: 1.5 g monomeric flavonoids/kg and 18.5 g PAs/kg) Integration: 100 g/kgflour Inhibition of mammalian α-glucosidase and α- amylase. LAVELLI et al., 2016a Wheat-rye bread (sourdough) Red grape pomace antioxidant dietary fibre (particle size not specified; TDF: 593 g/kg; soluble TP: 58.9 g GAE/kg). Integration: 40-100 /kgflour Significant increase in TDF and TP. Increase in hardness, gumminess and springiness. No change in cohesiveness and resilience. MILDNER-SZKUDLARZ et al., 2011 Brownies Red grape pomace antioxidant dietary fibre (particle size < 0.589 mm; TDF and TP: not specified). Integration: 150-250 g/kgflour Decrease in firmness and chewiness and increase in springiness. WALKER et al., 2014 Biscuits Grape seed extract encapsulated in mesquite gum, zein and maltodextrin (soluble TP: 21 g GAE/kg) Integration: 0.6 g TP/kgdough Increase in TP content and thermal stability Partial masking of darkness DAVIDOV-PARDO et al., 2012 Biscuits White grape pomace antioxidant dietary fibre (particle size < 0.150 mm; TDF: 509 g/kg; soluble TP: 31 g GAE/kg) Integration: 100 - 300 g/kgflour Significant increase in TDF and TP. Decrease in hardness, brightness and yellowness MILDNER-SZKUDLARZ et al., 2013 Biscuits Red grape marc extract (soluble TP: 2.1 g GAE/L) Integration: 450 mL/kgsemolina Increase in TP and antioxidant activity Increase in compounds derived from the Maillard reaction, except pyrazines, and lipid oxidation PASQUALONE et al., 2014 Pasta Red grape pomace antioxidant dietary fibre (particle size < 0.811 mm; TDF: 689.5 g/kg; TP: not specified). Integration: 25-75 g/kgflour Increase in TP and antioxidant activity Increase in cooking loss SANT’ANNA et al., 2014 Pasta Red grape marc (soluble TP: 4.43 g GAE/kg) Integration: 300 g/kgsemolina Increase in TP and antioxidant activity Decrease in cooking loss MARINELLI et al., 2015 TDF: total dietary fibre content; TP: total phenolic content (expressed as GAE: gallic acid equivalents or sum of monomeric flavonoids or PAs: proanthocyanidins). Regarding biscuits, the addition of white grape pomace antioxidant dietary fibre with particle size < 0.150 mm (total dietary fibre: 509 g/kg; soluble phenolics: 31 g/kg) up to 300 g/kgflour led to a decrease in hardness, brightness and yellowness (MILDNER- ! Ital. J. Food Sci., vol 28, 2016 - 552 SZKUDLARZ et al., 2013). In contrast, the incorporation of grape seed phenolics encapsulated with mesquite gum, zein and maltodextrin (soluble phenolics: 21 g/kg) partially masked the dark colour (DAVIDOV-PARDO et al., 2012). Alternatively, biscuits containing red grape extract (soluble phenolics: 2.1 g/L) at a level of 450 mL/kgsemolina displayed a particular red colour and aromatic profile and possessed greater contents of Maillard-reaction compounds, except pyrazines, as well as higher amounts of compounds derived from lipid peroxidation than the control biscuits (PASQUALONE et al., 2014). The fortification of pasta with winemaking by-products is also promising. Grape pomace antioxidant dietary fibre with particle size < 0.811 mm (total dietary fibre: 689.5 g/kg; total phenolic content not specified) formulated into pasta (25-75 g/kgflour) increased the total phenolic content and antioxidant activity but also caused a slight increase in the cooking loss at high levels of addition. This effect could be attributed to changes in the gluten protein network due to the interference of grape pomace fibre, which reduces the gluten strength and interrupts the overall structure of pasta (SANT’ANNA et al., 2014). However, the incorporation of red grape marc extract (soluble phenolics: 4.43 g/kg) into pasta at a ratio of 1:10, w/v, led to an increase in the phenolic content and antioxidant activity, as well as improved cooking performance. In fact, the cooking loss decreased due to the presence of grape pomace phenolics, which were complexed with proteins around starch granules, encapsulating phenolic compounds during cooking and restricting excessive swelling and amylose diffusion. The fortified sample was also characterized by a low adhesiveness value due to the formation of a stronger gluten network in the presence of phenolics, which entrapped the starch granules, slowing down amylose release during cooking. However, the hardness of fortified pasta was similar to that of the control pasta (MARINELLI et al., 2015). 2.1.4 Fruit-based products The replacement of synthetic additives with natural compounds such as winemaking by- products and the development of functional foods are emerging trends in the fruit processing industry (Table 4). When the phenolic extract of red grape pomace (soluble phenolics: 30 g/kg) was added to a model fruit gel at a concentration of 8.2 g/kgmixture (prior to concentration), a stable red colour and a marked increase in the antioxidant capacity were observed. These effects were maintained, even after storage for 24 weeks at room temperature, likely due to intermolecular associations between pectins and anthocyanins (MAIER et al., 2009a). The formulation of 0.01 g/kgjuice of white grape skin extract (phenolic content not specified) in model fruit juice containing Lactobacillus rhamnosus, Bifidobacterium lactis and Lactobacillus paracasei improved the stability of probiotic bacteria during storage due to the presence of a more stable anaerobic environment (SHAH et al., 2010). Red and white pomace extracts (soluble phenolics: 75-280 g/kg) were added to apple and orange juice to achieve a concentration of 20-100 g/kgjuice of phenolics, and antimicrobial effects toward microbial contaminants were observed, including Zygosaccharomyces rouxii and Z. bailii (SAGDIC et al., 2011). Alternatively, the use of antioxidant dietary fibre with particle size in the range of 0.125-0.5 mm obtained from white grape pomace (total dietary fibre: 505 g/kg; soluble phenolics: 30 g/kg; insoluble phenolics: 139 g/kg) imparted multiple functional effects. When this antioxidant- and fibre-rich ingredient was added to tomato puree at a level of 30 g/kgpuree, the reducing capacity and inhibitory effect of hyperglycaemia-induced damage increased due to the ability of grape phenolics to act as both oxygen-radical and carbonyl-radical scavengers (LAVELLI et al., 2014; TORRI et al., 2015). ! Ital. J. Food Sci., vol 28, 2016 - 553 Table 4: Application of winemaking by-products as new food ingredients in fruit-based products. Food product Recovered ingredient Main results and references Model fruit gel Red grape pomace extract soluble TP: 30 g GAE/kg Integration: 8.2 g /kgmixture (before concentration) Brilliant red colour and strong antioxidant capacity during storage at room temperature. MAIER et al., 2009a Model fruit juice containing probiotic bacteria White grape skin extract TP: not specified Integration: 0.01 g/kgmixture Improved stability of the probiotic bacteria Lactobacillus rhamnosus, Bifidobacterium lactis, and Lactobacillus paracasei during storage. SHAH et al., 2010 Orange and apple juices Red and white pomace extracts (soluble TP: 75-280 g GAE/kg) Integration: 20-100 g/kgjuice Antifungal activity towards Zygosaccharomyces rouxii and Z. bailii with variety-dependent efficacy. SAGDIC et al., 2011 Tomato puree White grape skin antioxidant dietary fibre (particle size in the range of: 0.125-0.5 mm; TDF: 505 g/kg; soluble TP: 30 g flavonoids/kg; insoluble TP: 139 g PAs/kg). Integration: 30 g/kgpuree Increase in reducing capacity and potential ability to inhibit hyperglycaemia-induced damage. LAVELLI et al.,2014; TORRI et al., 2015 Tomato puree White grape skin antioxidant dietary fibre (particle size in the range of 0.125-0.5 mm; TDF: 505 g/kg; soluble TP: 22 g GAE/kg). Integration: 30 g/kgpuree Increase in Bostwick consistency, storage (G) and loss (G) moduli, and complex viscosity (η*). LAVELLI et al., 2015b Apple-based fruit jelly Red grape skin antioxidant dietary fibre (particle size in the range: 0.125 - 0.5 mm; TDF: 600 g/kg; soluble TP: 26 g flavonoids/kg). Integration: 63 g/kgmixture (before concentration) Increase in puncture energy, i.e., stronger texture. Requires less dehydration. High antioxidant and fibre content. CAPPA et al., 2015 TDF: total dietary fibre content; TP: total phenolic content (expressed as GAE: gallic acid equivalents or sum of flavonoids or PAs: proanthocyanidins). In addition, the fibre content increased, which improved the Bostwick consistency, storage and loss moduli and complex viscosity, providing a means to modulate the textural properties of puree (LAVELLI et al., 2015b). A similar antioxidant- and fibre-rich ingredient obtained from red pomace with particle size in the range of 0.125-0.5 mm (total dietary fibre: 600 g/kg; soluble phenolics: 26 g/kg) added at a level of 63 g/kgmixture to apple- based candy modified the textural properties, resulting in a stronger structure that required greater puncture and penetration energy. As a result, during candy processing, the dehydration step could be reduced (CAPPA et al., 2015). 2.1.5 Dairy products The addition of fruit phenolics into food products has increased due to the emerging popularity of functional foods, including phenolic addition in the dairy sector (O’CONNELL and FOX, 2001). The fortification of phenolics into dairy products may increase the heat and foam stability of milk and enhance nutritional benefits (O’CONNELL and FOX, 2001). However, many phenolics are bitter and astringent (VIDAL et al., 2004), and humans instinctively reject bitter substances (DREWNOWSKI ! Ital. J. Food Sci., vol 28, 2016 - 554 and GOMEZ-CARNEROS, 2000). Although phenolic compounds interact with proteins during the cheesemaking process, these interactions are dependent on the pH and molar ratio and molecular properties of the polyphenols and involve both hydrophobic and hydrophilic bonds (FELIX DA SILVA et al., 2015). The application of winemaking by- products to the dairy sector is shown in Table 5. Table 5: Application of winemaking by-products as new food ingredients in dairy products. Food product Recovered ingredient Main results and references Cheese Single phenolic compounds and whole grape extract Integration: 0.5 mg single phenolic or GAE/mLmilk Slightly reduced hydration capacity of cheese curd network. No variation in gel strength. Less smooth and less dense internal structure of cheese curds. More granular outer surfaces than control cheese by SEM microstructure analysis. HAN et al., 2011a Cheese Single phenolic compounds and whole grape extract Integration: 0.5 mg single phenolic or GAE/mLmilk High retention of grape phenolics in the curd. High radical scavenging activity. Increase in gel-formation rate due to a slight decrease in pH. HAN et al., 2011b Cheese Red and white grape pomace antioxidant dietary fibre before and after distillation (particle size < 0.25 mm; TP: 3.64-16.0 g GAE/kg). Integration: 8 and 16 g/kgcurd Highest radical scavenging activity and TP content, yielding 16 g/kg of grape powder after distillation. No effect on lactic bacteria or proteolysis. MARCHIANI et al., 2015 Yogurt and salad dressing Red grape pomace antioxidant dietary fibre (particle size < 0.18 mm; TDF: 613.2 g/kg, TP: 67 g GAE/kg). Integration: 10-30 g/kgyogurt; 5-10 g/kgsalad dressing Increase in TDF, TP and radical scavenging activity (but slight decrease in TP during storage at 4 °C). Decrease in peroxide values for both yogurt and salad dressing. Stable value for lactic acid percentage and syneresis during 3 weeks of storage at 4 °C for yogurt. TSENG and ZHAO, 2013 Yogurt Grape seed extract TP: 76 - 150 g GAE/kg Integration: 50-100 mg TP/kgyogurt Increase in TP (slight decrease in TP during storage at 4 °C). No change in pH or Lactobacilli counts. CHOUCHOULI et al., 2013 Yogurt Red and white grape skin antioxidant dietary fibre (particle size < 0.25 mm; TDF: 345-481 g/kg). Integration: 60 g/kgyogurt Increase in acidity, total phenolic content and antioxidant activity with respect to control, but lower pH, syneresis and fat. Lactic acid bacteria, phenolic content and antioxidant activity were stable during 3 weeks of storage. MARCHIANI et al., 2016 Milk Grape seed extract powder TP: 842-927 g/kg Integration: 2 g/Lmilk Phenolic content equal to that of one serving of fresh apple. Increase in potential health benefit. AXTEN et al., 2008 TDF: total dietary fibre content; TP: total phenolic content (expressed as GAE: gallic acid equivalents). ! Ital. J. Food Sci., vol 28, 2016 - 555 HAN et al. (2011a,b) studied the effect of single phenolic compounds (catechin, epigallocatechingallate, tannic acid, homovanillic acid, hesperetine and flavone) and natural extracts, such as whole grape extract, green tea extract and dehydrated cranberry powder as functional ingredients in cheese at a level of 0.5 mg of total phenolics/mLmilk. Three gel-forming parameters were evaluated, including Tlag (the lag-time before the beginning of milk coagulation), Vmax (the maximum rate of gel formation) and TVmax (the time required to reach the maximum rate of gel formation). The results demonstrated that the addition of phenolic compounds to milk affected all of the gel-forming parameters. In fact, the change in pH due to the addition of phenolics resulted in faster gel formation in cheese samples fortified with single phenolic compounds than those fortified with natural extracts. In addition, several phenolic sources improved the antioxidant properties of cheese products. Among the tested single phenolic compounds and natural extracts, the greatest radical scavenging activity was achieved using whole grape extracts, likely due to the improved retention of grape phenolics in the curd, representing a promising utilization of winemaking by-products. MARCHIANI et al. (2015) used antioxidant dietary fibre obtained from three grape pomaces (Barbera, Chardonnay prior to distillation and Chardonnay after distillation) with particle size < 0.25 mm (soluble phenolics: 3.64-16.0 g/kg) in semi-hard and hard cheeses (Italian Toma-like and Cheddar) to increase the content of phenolic compounds. Powders were added at two different concentrations (8 and 16 g/kgcheese), and the results showed that the amount and type of powder did not significantly affect the physicochemical parameters of cheese, except the pH. Cheeses containing Chardonnay powder after distillation showed the highest phenolic content and radical scavenging activity at the end of ripening. Proteolysis and microbial counts did not show significant differences between fortified and control cheeses. TSENG and ZHAO (2013) applied grape pomace antioxidant dietary fibre with particle size < 0.18 mm (total dietary fibre: 613.2 g/kg; soluble phenolics: 67 g/kg) at a level of 10- 30 g/kg to yogurt and 5-10 g/kg to salad dressing to enhance the nutritional value and improve the storability. The authors demonstrated that this ingredient can be used as an alternative source of antioxidants and dietary fibre to delay the oxidation of lipids during refrigerated storage. However, the total phenolic content and radical scavenging activity of fortified yogurts decreased slightly during storage, likely due to interactions between proteins and phenolic compounds. Consequently, further research is necessary to determine the mechanism of the long-term retention of grape phenolics and radical scavenging activity of the aforementioned products. CHOUCHOULI et al. (2013) used grape seed extracts from two grape varieties (50-100 mg of phenolics/kgyogurt) to fortify full-fat and non-fat yogurt. The addition of the extract did not affect the pH or Lactobacilli count, while fortified yogurts showed higher antioxidant and antiradical activity than control samples, even after 3-4 weeks of cold storage. However, the phenolic content and radical scavenging activity decreased during storage. The fortification of yogurt with up to 60 g/kgyogurt of red and white grape skin antioxidant dietary fibre with particle size < 0.25 mm (total dietary fibre: 345-481 g/kg) was performed by MARCHIANI et al. (2016). In this case, a reduction in the total phenolic content did not occur during refrigerated storage, but the radical scavenging activity decreased. Grape addition did not affect the growth of lactic acid bacteria. Moreover, UHT low fat milk was fortified with different grape seed extract powders to obtain a concentration of 2 g/Lmilk of phenolics in the final product, representing the amount of phenolics ingested when consuming one serving of fresh apple (AXTEN et al., 2008). ! Ital. J. Food Sci., vol 28, 2016 - 556 2.2. Sensory effects Understanding the impact of new ingredients on consumers’ perception is considered a key step in new product development (VERBEKE, 2006; TUORILA, 2007). Therefore, including a sensory- and consumer-based approach in product innovation is strategic for determining the properties that have the greatest effect on consumer preference, as well as relevant attributes that drive product optimization (TORRI et al., 2016). 2.2.1 Meat products The sensory effects of grape by-products on meat products have been thoroughly analysed in the literature. For chicken meat, grape skin extract added to dehydrated products (1 g/kgmeat) prevented changes in sensory attributes due to oxidation (hot wash, boiled chicken, subcutaneous fat and rancidity), with comparable efficacy to synthetic antioxidants and other natural antioxidants, such as rosemary, coffee and tea (NISSEN et al., 2000). In chicken hamburger, the use of grape antioxidant fibre (5-20 g/kgmeat) increased the lipid stability without affecting the desirability of the odour, flavour, and tenderness during 5 days of storage. Only the colour was modified by the addition of grape skin extract (15-20 g/kgmeat), which did not affect the acceptability of samples. The sensory test comparing the effect of four different addition levels revealed that hamburgers preferred by consumers had the highest extract content (15-20 g/kgmeat) (SÀYAGO-AYERDI et al., 2009). The incorporation of grape pomace extracts in raw and cooked chicken meatballs (60 mg of phenolics/kgmeat) also provided satisfactory results in terms of odour and flavour properties during frozen storage, which were not different from those observed using synthetic antioxidants (SELANI et al., 2011). Regarding pork meat, the addition of grape skin extract (0.2 g/kgmeat) to cooked patties was not sufficient to reduce the intensity of rancid and linseed odours and flavours correlated to lipid oxidation indexes (TBARS and hexanal content); thus, acceptable sensory properties for consumption could not be guaranteed (NISSEN et al., 2004). Nevertheless, grape seed extract (0.1-0.2 g/kgmeat) has been proven to have a positive effect on both cooked pork and beef patties and was even more effective than rosemary oleoresin and oregano water-extracts. In fact, sensory evaluation performed over eight days of storage at 4°C demonstrated the efficiency of grape seed extract at controlling several negative sensory characteristics associated with a warmed-over flavour, such as rancidity, wet cardboard (for beef patties) and grassy (for beef and pork patties) odour descriptors (ROJAS and BREWER, 2007). Accordingly, the addition of grape seed extract (0.05-1 g/kgmeat) did not significantly affect the average scores obtained for cooked pork patties for any of the quality parameters evaluated (colour, flavour, texture, juiciness and off-flavour) over four days of storage at 4°C under a modified atmosphere (CARPENTER et al., 2007). In beef patties, grape seed extract (0.1-0.2 g/kgmeat) led to visual green discoloration (ROJAS and BREWER, 2007). This effect was also observed in precooked beef sausage containing grape seed extract (0.3-0.5 g/kgmeat). Moreover, grape seed extract improved the persistence of fresh cooked beef odour and flavour during the storage of fortified products with respect to the control group and prevented the formation of a rancid odour (KULKARNI et al., 2011). 2.2.2 Fish products Very little information is available on the sensory effects of grape by-products in fortified fish products. In fact, only one paper out of those cited in the present review included a sensory evaluation of fortified fish sausage. The sensory description of this product ! Ital. J. Food Sci., vol 28, 2016 - 557 showed that samples containing grape skin antioxidant dietary fibre (30 g/kgfish) were significantly darker, less elastic, cohesive, succulent and oily and possessed a more unpleasant texture and flavour than the control product. The unpleasant odour and flavour has been described as a sour note, but a rancid aroma or flavour did not develop after 98 days of storage at refrigerated temperatures (RIBEIRO et al., 2013). 2.2.3 Bakery products and pasta Among the food categories considered in this review, cereal products have been the most commonly investigated from a sensory perspective. By adding different amounts of grape seed extract to white bread (0.6-2 g/kgbread), significant alterations in quality attributes (sweetness, porosity, astringency and stickiness) were not detected (PENG et al., 2010). The recommended replacement of hard red spring flour with grape seed antioxidant dietary fibre in bread was 50 g/kgflour. Above this threshold, low sensory acceptance of astringency, sweetness, bitterness and overall liking was observed (HOYE and ROSS, 2011). Similarly, the overall acceptance for sourdough mixed rye bread decreased as the content of red grape pomace antioxidant dietary fibre increased from 40 to 100 g/kgflour, indicating that a maximum of 60 g/kgflour could be used to prepare acceptable products. Higher levels were associated with a decrease in the volume, porosity and typical aroma of freshly baked breads and an increase in hardness, acidity and alcoholic, sharp and fruity notes (MILDNER-SZKUDLARZ et al., 2011). Regarding brownies, fortification with red and white grape pomace antioxidant dietary fibre at levels higher than those acceptable for bread, up to 150 g/kgflour, did not impact the sensory properties and acceptability of the products (WALKER et al., 2014). The acceptance of biscuits enriched with white grape pomace antioxidant dietary fibre was also dependent on the level of addition: 100 g/kgflour incorporation in wheat flour was adequate, while concentrations of 200 or 300 g/kgflour induced a fruity-acidic note and an intense brown colour, which was undesirable to consumers (MILDNER-SZKUDLARZ et al., 2013). However, microencapsulation of grape seed extracts prevented a decrease in the likeability ratings by consumers (DAVIDOV-PARDO et al., 2012). Biscuits enriched with a red grape pomace extract (450 mL/kgsemolina) have been described by a trained panel of assessors as having a more intense colour, fruity odour and sour taste and lower friability than control samples. Moreover, consumers were able to discriminate among biscuits samples based on both colour and taste. However, neither the modifications in sensory profiles nor the differences perceived during the affective test influenced the acceptability or willingness of the subjects to buy anthocyanin-enriched biscuits. Regarding both the colour and taste, in a global evaluation, the number of consumers who preferred enriched biscuits was not significantly different than those who preferred control biscuits (PASQUALONE et al., 2013). In fettuccini pasta formulated with red grape pomace antioxidant dietary fibre (25-75 g/kgflour), the overall liking and acceptance of aroma, aftertaste, flavour and appearance decreased, regardless of the concentration of the ingredient (SANT’ANNA et al., 2014). However, upon addition of red grape pomace extract (300 g/kgsemolina), the sensory properties of fortified, fresh-extruded, fresh-pasteurized and dry spaghetti pasta were as acceptable as control products (MARINELLI et al., 2015). 2.2.4 Fruit-based products Information regarding the effect of grape by-products on the sensory properties of fruit- based products is scarce. The incorporation of different granulometric fractions of white grape skins into either smooth or rough tomato puree (30 g/kgpuree) induced a clear increase ! Ital. J. Food Sci., vol 28, 2016 - 558 in the textural attributes (crispiness and granularity), a decrease in the perceived homogeneity and a change in the vegetable odour notes (spicy hay). The intensity of these effects depended on the fraction particle size, which also influenced consumers’ preferences. A cluster of subjects was found to significantly prefer the smallest particle size fraction (< 0.125 mm), especially when combined with smooth tomato puree, while another group of consumers showed opposite preferences, preferring the largest particle size (0.250-0.500 mm) and rough tomato puree cells (LAVELLI et al., 2014; TORRI et al., 2015). 2.2.5 Dairy products A limited number of applications of grape pomace have been investigated in dairy products, especially in terms of sensory effects; however, yogurt fortified with winemaking by-products has been investigated. Fortification with grape seed extract amounts corresponding to 50-100 mg of total phenolics/kgyogurt did not result in any major defects in sensory properties (consistency, colour and flavour) compared to control samples (CHOUCHOULI et al., 2013). On the contrary, higher levels of addition strongly modified the sensory properties of yogurt, which was perceived as too sour by consumers, possessing an unpleasant flavour and grainy/sandy texture. However, significantly different hedonic scores were observed using by-products from different grape varieties, suggesting that Chardonnay was more suitable than Pinot noir and Muscat (MARCHIANI et al., 2016). The fortification of yogurt with red grape pomace in the range of 10 to 20 g/kgyogurt provided satisfying overall likeability values. Nevertheless, lower likeability scores for flavour and texture were observed for the sample with the highest concentration of grape pomace (TSENG and ZHAO, 2013). In salad dressing, the addition of red grape pomace antioxidant dietary fibre at a level of 10 g/kgmixture was best received by consumers (TSENG and ZHAO, 2013). Fortification with grape skin antioxidant dietary fibre clearly influenced the sensory properties of soft cow milk cheeses, especially the appearance and texture. In particular, the marbling aspect, granularity, sandiness, sourness and astringency (due to the presence of fibre and polyphenols from grape skin) negatively impacted the overall likeability of the cheese when the amount of Barbera and Chardonnay grape was greater than 8 and 16 g/kgcurd, respectively (TORRI et al., 2016). The addition of grape seed extracts to low fat UHT milk had a significant effect on the product sensory attributes, tending to suppress sweetness and UHT odour and flavour and to increase the perception of bitterness, sourness, astringency, odours and flavours (fresh raisin, honey, inka, ashy and tobacco), as well as chalkiness (AXTEN et al., 2008). 3. GRAPE SEED OIL EXTRACTION BY GREEN TECNOLOGIES Grape seed oil extraction can be applied in parallel with the recovery of both antioxidant dietary fibre and phenolic extracts from grape skins and defatted grape seeds, thus making the overall recovery strategy more sustainable. With the recent technological advancements, “green technologies” have been proposed to replace the traditional oil recovery process by solvents. The recovery of grape seed oil requires the preliminary separation of seeds from other grape pomace constituents, including skins and stalks. Separation occurs by mechanical devices, usually after grape pomace drying. The conventional “non-green” solvent extraction allows for nearly complete oil recovery. Non-polar solvents are used for oil extraction, including n-hexane and petroleum ether, which are the most common extracting solvents and have the lowest cost. After extraction, ! Ital. J. Food Sci., vol 28, 2016 - 559 the oil must be separated from the solvent in which it is dissolved, and the solvent may be recycled. Separation is achieved by evaporating off the solvent. For n-hexane, evaporation occurs at 69 °C (the boiling point of n-hexane at ambient pressure). High quality oil is obtained by mechanical extraction performed at ambient temperature. Unfortunately, mechanical extraction does not afford a high yield of oil, particularly for grape seeds, whose woody texture makes them mechanically resistant and, more importantly, whose oil content is reduced to the range of 4 to 17% (FERNANDES et al., 2013; FIORI et al., 2014). To increase the oil recovery, mechanical extraction can be performed at higher temperatures: however, at increased extraction temperatures, some of the noble oil constituents, which are thermally unstable, tend to degrade. An emerging solvent for use in the food industry is high pressure CO2, more precisely termed supercritical CO2 (SC-CO2) (DUBA and FIORI, 2015a). A fluid is in the supercritical state when the actual temperature and pressure are higher, respectively, than the critical temperature (Tc) and pressure (Pc) of the fluid. For CO2, Tc and Pc are 31°C and 73 bar, respectively. Therefore, SC-CO2 can be used while operating at temperatures only slightly higher than ambient temperature, making SC-CO2 particularly interesting for thermally unstable compounds, which is often the case in the food sector. An important advantage in the use of supercritical fluids as solvents is the extreme ease in separating the solvent and solute after extraction: separation occurs by simple depressurization. When using SC-CO2 for extracting grape seed oil, after flowing SC-CO2 is contacted with a static bed of milled grape seeds, the mono-phase stream consisting of SC- CO2 and extracted and dissolved grape seed oil is expanded through a back-pressure valve. After expansion, low pressure gaseous CO2 separates from grape seed oil, which is recovered solvent-free. Solvent-free defatted grape seeds can also be recovered and used for further food applications (LAVELLI et al., 2015a). The process and corresponding equipment for an industrial-scale plant for SC-CO2 extraction of grape seed oil have been fully explained in the literature (FIORI, 2010; FREITAS et al., 2013). The effect of process parameters on the extraction kinetics and yield of the SC-CO2 extraction of grape seed oil has been recently outlined (DUBA and FIORI, 2015b). Other obvious advantages of using SC-CO2 in the food sector are represented by the peculiarities of CO2, which is non-toxic, non-flammable and inexpensive. Unfortunately, the proposed process includes drawbacks of an economic nature. The solubility of grape seed oil in SC-CO2 is lower than 10 goil/kgCO2 for pressures lower than 350 bar (DUBA and FIORI, 2016). To achieve relatively high solubility values for oil in SC-CO2, the equipment must be operated at not less than 400 bar, preferably at 500-600 bar, which translates into high investment costs for the facility, where the extractors must be employed at high pressure (FIORI, 2010). 4. CONCLUSIONS The studies summarized in the present review demonstrate an increase in interest in potential food applications of winemaking by-products and provide a new production scenario for winemakers. Winemaking by-products can be processed into various food ingredients, including antioxidant dietary fibres, crude phenolic extracts or encapsulated extracts, and applied to produce new foods. Grape seed oil can be recovered using SC-CO2 extraction. However, economic and regulatory factors prevent these applications from achieving large-scale application. First, the proposed applications imply new production cycles across winemaking and other food sectors. The establishment of these connections demand improved logistic organization, including appropriate technologies for the collection, storage, transportation and processing of grape pomace. Investment costs for ! Ital. J. Food Sci., vol 28, 2016 - 560 new processes are sometimes high. Second, although food legislation strongly promotes the recycling of by-products, a recovery strategy based on value addition and by-product use in functional food production results in additional regulatory issues. Actually, the application of by-products in foods generally leads to novel foods, which opens the debate regarding safety. Fortification/enrichment with the appropriate amount of grape antioxidant dietary fibre only allows for the labelling of foods as fibre-rich, while other possible health claims must be substantiated by specific studies. Thus, further scientific research is necessary to surpass economic and regulatory barriers and achieve significant advance towards the establishment of a biorefinery fed with winemaking by-products. ACKNOWLEDGEMENTS This research was supported by AGER (project number 2010-2222). REFERENCES Ahn J., Grun I.U. and Mustapha A. 2007. Effects of plant extracts on microbial growth, color change, and lipid oxidation in cooked beef. Food Microbiol. 24: 7. Amendola D., De Faveri D.M. and Spigno G. 2010. Grape marc phenolics: extraction kinetics, quality and stability of extracts. J. Food Eng. 97:384. Axten L.G., Wohlers M.W. and Wegrzyn T. 2008. Using phytochemicals to enhance health benefits of milk: Impact of polyphenols on flavor profile. J. Food Sci. 73: H122. 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