Postharvest Biology and Technology 57 (2010) 139–148

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Postharvest Biology and Technology

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Review

Recent approaches using chemical treatments to preserve qualityof fresh-cut fruit: A review

Gemma Oms-Oliua, Ma Alejandra Rojas-Graüa, Laura Alandes Gonzálezb, Paula Varelac,Robert Soliva-Fortunya, Ma Isabel Hernando Hernandob, Isabel Pérez Munuerab,Susana Fiszmanc, Olga Martín-Bellosoa,∗

a Department of Food Technology, TPV-Xarta, University of Lleida, Av. Rovira Roure 191, 25198 Lleida, Catalonia, Spainb Department of Food Technology, Universidad Politécnica de Valencia, Camino de Vera 14, 46022 Valencia, Spainc Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Apartado de correos 73, 46100 Burjassot, Spain

a r t i c l e i n f o

Article history:Received 16 July 2009Accepted 3 April 2010

Keywords:Fresh-cut fruitPreservationBrowning

a b s t r a c t

This review covers some recent advances for the maintenance of fresh-cut fruit quality with respectto the use of chemical compounds, including plant natural antimicrobials and antioxidants, as well ascalcium salts for maintaining texture. It focuses especially on the use of natural preservatives, which areof increasing interest because of toxicity and/or allergenicity of some traditional food preservatives. Thedifficulties in the application of these substances on fresh-cut fruit without adversely affecting sensorycharacteristics of the product are reviewed. Edible coatings are presented as an excellent way to carryadditives since they are shown to maintain high concentrations of preservatives on the food surfaces,reducing the impact of such chemicals on overall consumer acceptability of fresh-cut fruit.

Texture

Antibrowning agentsAntimicrobialsCE

© 2010 Elsevier B.V. All rights reserved.

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alcium saltsdible coatings

. Introduction

Lifestyles of modern consumers, along with the desire for nat-ral products that claim health benefits, have been responsible forhe current rise of production and consumption of fresh-cut fruit.owever, mechanical operations during minimal processing dam-ges fruit tissues, which in turn limits the shelf-life of products.uch research is still to be done in order to develop safe fresh-cut

ruit products with high sensory quality and nutritional value. Theevelopment of new processing techniques for preserving fresh-ut fruit needs to overcome some of the hurdles to successfulommercial distribution of such products.

Antimicrobial agents such as plant essential oils have beenntroduced as a novel way to improve microbiological stability ofresh-cut fruit. Interest in the possible use of natural compoundso prevent microbial growth has notably increased in response to

onsumer awareness of the use of chemically synthesized addi-ives in foods. Plants and derived products can represent a sourcef natural alternatives to improve the shelf-life and the safety ofoods. A wide range of volatile compounds such as hexanal, hex-

∗ Corresponding author. Tel.: +34 973702593; fax: +34 973702596.E-mail address: omartin@tecal.udl.cat (O. Martín-Belloso).

925-5214/$ – see front matter © 2010 Elsevier B.V. All rights reserved.oi:10.1016/j.postharvbio.2010.04.001

anol, 2-(E)-hexenal and 3-(Z)-hexenol, important constituents ofthe aroma of tomatoes, tea, strawberry, olive oil, grape, applesand pear, and also plant essential oils, comprised mainly of ter-penoids, have been studied for their antimicrobial activity againstmany microorganisms including several pathogens (Lanciotti et al.,2004). Further investigations are necessary to evaluate the poten-tial of plant essential oils and their active constituents as naturalpreservatives to improve the shelf-life and the safety of minimallyprocessed commodities.

Browning is also a major concern related to the extension ofshelf-life of fresh-cut fruit, and strongly affects the consumer’s pur-chase decision. Traditionally, sulfites have been used for browningprevention. However, their use on fresh-cut fruit and vegetableswas banned in 1986 by the FDA owing to their potential hazardsto health (Buta et al., 1999). Thus, various alternative approacheshave being studied to minimize visual deterioration in fresh-cutfruit. Reducing agents such as citric acid, ascorbic acid, isoascor-bic acid and sodium erythorbate (Sapers and Miller, 1998; Butaet al., 1999; Dong et al., 2000; Soliva-Fortuny et al., 2002a), thiol-

containing amino acids such as N-acetylcysteine and glutathione(Oms-Oliu et al., 2006; Rojas-Graü et al., 2006), oxalic acid (Son etal., 2001) and 4-hexylresorcinol (Luo and Barbosa-Cánovas, 1997)have been investigated to prevent browning. Calcium treatmentscan maintain or improve tissue firmness and crispness of fresh-

140 G. Oms-Oliu et al. / Postharvest Biology and Technology 57 (2010) 139–148

Table 1Treatments to improve microbial stability of fresh-cut fruit.

Treatment Fruit Antimicrobial substances Reference

Dipping Kiwifruit, melon Carvacrol and cinnamic acid (1 mM) Roller and Seedhar (2002)Mangoes Vanillin (0.12%, w/v) Ngarmsak et al. (2006)Apple Vanillin (0.18%, w/v) Rupasinghe et al. (2006)Fruit mixture (apple, pear, grape,peach, kiwifruit)

Citrus, mandarin, cider, lemon and limeessential oils (0.02%, v/v)

Lanciotti et al. (2004)

Volatile exposition Apple Hexanal (150 �L/L), hexylacetate (150 �L/L),(E)-2-hexenal (20 �L/L)

Lanciotti et al. (2003)

Hexanal (0.225 �L/L)Hexanal (0.030–0.120 mmol/60 g), (E)-hexenal(0.006–0.024 mmol/60 g)

Lanciotti et al. (1999)

Kiwifruit Methyl jasmonate (11.2–22.4 �L/L) Corbo et al. (2000)Pineapple Methyl jasmonate (15 �L/L) Wang and Buta (2003)

Martínez-Ferrer and Harper (2005)

Edible coatings Apple Alginate/apple puree + (0.3–0.6% (w/v) vainillinor 1–1.5% (v/v) lemongrass or 0.1–0.5% (v/v)oregano oil)

Rojas-Graü et al. (2007a)

Alginate + (2.5% MA + 0.7% lemongrass, or 0.3%cinnamon oil)

Raybaudi-Massilia et al. (2008a)

Strawberries Chitosan (1%, w/v) Han et al. (2005)Chitosan (2%, w/v) Park et al. (2005)Chitosan (1%, w/v) Campaniello et al. (2008)

Melon Alginate + (2.5% MA + 0.3% palmarosa oil) Raybaudi-Massilia et al. (2008b)an (0.era (1:an (0.eenan

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Mango ChitosGrapes Aloe vPapaya ChitosBanana Carrag

ut fruit. Calcium chloride has been one of the most frequentlysed salts of calcium although it is reported to impart residualaste to the product. Thus, other calcium salts such as calciumactate, calcium propionate or calcium ascorbate have been inves-igated as alternative sources of calcium (Buta et al., 1999; Dongt al., 2000; Gorny et al., 2002; Alandes et al., 2006; Quiles et al.,007).

Surface treatments involving dipping fruit pieces into aqueousolutions containing antimicrobial agents, antioxidants, calciumalts or functional ingredients such as minerals and vitamins areidely practiced to improve quality of fresh-cut fruit. However, the

ffectiveness of these compounds could be very much improvedith their incorporation into edible coatings. The application of

dible coatings to deliver active substances is one of the recentajor advances made in order to increase the shelf-life of fresh-cut

roduce.

. Microbiological stability of fresh-cut fruit

During the preparatory steps of minimal processing, the naturalrotection of fruit is generally removed and hence, they becomeighly susceptible to microbial spoilage (Martín-Belloso et al.,006). In addition, cross-contamination may occur during cuttingnd shredding operations because sanitation in whole fruit mayot have been carried out properly. Leakage of juices and sug-rs from damaged tissues allows the growth and fermentation ofome species of yeasts such as Saccharomyces cerevisiae and Sac-haromyces exiguous (Heard, 2002). As well, damage to plant tissuencreases susceptibility to attack by pathogenic microorganismsnd contamination with human pathogens.

.1. Microbial growth

Changes in the microbial population in packaged fresh-cut pro-

uce are expected. The high humidity conditions within a packagend the presence of a large area of cut surfaces, which provide a richource of nutrients, create an environment conducive to growth oficroorganisms. The type and growth rate of the microorganisms

resent will be greatly influenced by the product temperature over

5–2.0%, w/v) Chien et al. (2007)3) Valverde et al. (2005)02 g/mL) González-Aguilar et al. (2009)

(0.5%, w/v) Bico et al. (2008)

time, relative humidity, atmosphere and intrinsic factors such aspH, water content and nutrients.

Due to the low pH values of most fruit, the main typicalflora consist of moulds and yeasts. Botrytis cinerea and Aspergillusniger have been found to be important moulds as well as yeastssuch as Candida, Cryptococcus, Fabospora, Kluyveromyces, Pichia,Saccharomyces, and Zygosaccharomyces (Chen, 2002). However,consumption of fresh-cut fruit has been associated with food-borne diseases due to some pathogenic bacteria such as Cyclosporacayetanensis in raspberries, Salmonella spp. in precut watermel-ons, and Shigella spp. in fruit salad, among others (Heard, 2002).In general, pathogens may often be able to grow on some fruitsurfaces such as melon, watermelon, papaya, or avocado becauseof the high pH of the fruit. The number of documented outbreaksof human infections associated with the consumption of raw andminimally processed fruit and vegetables has increased consider-ably over the past decades (Bean et al., 1997; Sivapalasingam etal., 2004). A variety of pathogenic bacteria such as Listeria mono-cytogenes, Salmonella spp. and Shigella spp., Aeromonas hydrophila,Yersinia enterocolitica, and Staphylococcus aureus as well as somepathogenic Escherichia coli strains may be present on fresh fruitand, consequently, in fruit-based salads (Gould, 1992; Breidt andFleming, 1997). Regulatory initiatives specific for fresh-cut prod-ucts are still under development, although the United States andmost European countries already have relevant regulations. Mostof them limit the counts of aerobic microorganisms to 106 cfu/g atthe expiry date of the product. In addition, pathogenic microorgan-isms are not allowed (Salmonella) or are greatly restricted (E. coli, L.monocytogenes) in ready-to-eat meals prepared from raw vegetableproducts (Martín-Belloso et al., 2006).

2.2. Antimicrobials

The search for methods to retard microbial growth is of great

interest to all the sectors involved in production and preservation offresh-cut fruit and many different solutions have been proposed todelay the deterioration. Proliferation of microorganisms on the sur-face of fresh-cut fruit is currently retarded or inhibited by using lowstorage temperature, modified atmosphere packaging, and antimi-

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robial substances (Rojas-Graü and Martín-Belloso, 2008a). Foodntimicrobials are chemical compounds that may delay microbialrowth or cause microbial death when they are incorporated intofood matrix (Davidson and Zivanovic, 2003). The major targets

or antimicrobials are food poisoning and spoilage microorgan-sms, whose metabolic end products or enzymes cause off-odors,ff-flavors, texture changes and discoloration (Davidson, 2001).

Regulations of most countries regard chemical substancesdded as antimicrobials as food additives if the primary purposef the substance is shelf-life extension. However, each country hasts own regulations defining a list of approved additives (ED, 1995;SDA, 2006). For instance, according to US regulations, organiccids including acetic, lactic, citric, malic, propionic, tartaric andheir salts are GRAS for miscellaneous and general purpose usageDoores, 1993). In addition, many plant essential oils are usedidely in the food, health and personal care industries and are

lso classified as generally regarded as safe (GRAS) substances orermitted as food additives (Kabara, 1991).

Surface treatments by spraying with antimicrobial agents or byipping fruit in antimicrobial solutions are widely practiced to pre-ent microbial growth (Table 1). Dipping treatments after peelingnd/or cutting both reduce microbial loads and rinse of tissue flu-ds, and thus reduce the growth of microorganisms (Martín-Bellosot al., 2006). Organic acids are usually applied as a dip. The antimi-robial effect of the addition of organic acids to food is to increasehe proton concentration, thereby lowering the external pH. Therowth of a microorganism is inhibited when the pH falls belowhe range of pH values that allows its development. Citric acid haseen widely used as an effective preservative because it is able toeduce the pH of cut fruit such as oranges (Pao and Petracek, 1997),pples (Rocha et al., 1998) and bananas (Moline et al., 1999).

However, in recent years there has been a considerable pressurerom consumers for a reduction or elimination of chemically syn-hesized additives in foods. The use of antimicrobial agents fromlants and plant products can therefore provide a natural alterna-ive to food additives. These natural products are characterized bywide range of volatile compounds, some of which are importantavor quality factors. They are usually GRAS compounds, and areble to inhibit microbial growth and influence flavor and qualityecause of the presence volatile compounds (Utama et al., 2002).ome natural constituents, such as hexanal, hexanol, 2-(E)-hexenal,nd 3-(Z)-hexenol, responsible for the aroma of some vegetablesnd fruit, provide protective action against microbial proliferationn wounded areas (Gardini et al., 2002).

The precise mechanisms of action of these antimicrobial com-ounds are not yet clear, but passive diffusion across the plasmaembrane is involved (Lanciotti et al., 2004). Lanciotti et al. (2003)

eported significant extensions of the lag phase of E. coli andalmonella enteritidis inoculated at levels of 104–105 CFU/g onresh apple slices treated with hexanal (150 �L/L), hexyl acetate150 �L/L) and (E)-2-hexenal (20 �L/L) and stored at 20 ◦C, whereasor L. monocytogenes a bactericidal effect after 4 d was found underhe same experimental conditions. Hexanal (0.225 �L/L) prolongedhe lag phase of native yeasts for 8 d on sliced apple stored at5 ◦C under a modified atmosphere, whereas mesophilic bacteriarowth was retarded by more than 20 d under the same condi-ions (Lanciotti et al., 1999). Similarly, Corbo et al. (2000) obtainedreduction of spoilage microbial populations on sliced apple storedt 15 ◦C under modified atmosphere using hexanal and (E)-hexenal.anciotti et al. (1999) suggested that the antimicrobial activitiesf hexanal, 2-(E)-hexenal, and hexylacetate are positively influ-

nced by a rise in temperature, since their action is dependent onapor pressure. In addition, other volatile compounds as methylasmonate, a natural derivative of jasmine, have been also reportedo control microbial growth on fresh-cut fruit. Wang and Buta2003) showed that methyl jasmonate at concentrations of 11.2

nd Technology 57 (2010) 139–148 141

and 22.4 �L/L applied as vapor was effective in preventing mouldgrowth on fresh-cut kiwifruit during 3 weeks of storage at 10 ◦C.Likewise, Martínez-Ferrer and Harper (2005) achieved 3 log CFU/greductions of the native microbiota on fresh-cut pineapple after12 d of storage at 7 ◦C with an emulsion of methyl jasmonate ata concentration of 15 �L/L. These authors also reported that thesame concentration of methyl jasmonate applied as vapor was lesseffective in reducing the microbial population.

Plant essential oils (EOs) also have been studied for their antimi-crobial activity against many microorganisms, including severalpathogens (Dorman and Deans, 2000; Delaquis et al., 2002). Inaddition, the action of individual active constituents of these oilshas been exploited to better understand the cell targets of thesemolecules, to identify the most active molecules, and to counter theintrinsic variability of essential oils (Karatzas et al., 2000; Vázquezet al., 2001). The mechanisms of action of EOs are associated withdegradation of the cell wall, damage to cytoplasmic membraneand membrane proteins, leakage of cell contents, coagulation ofcytoplasm and depletion of the proton motive force (Burt, 2004).While in vitro antimicrobial activity is well demonstrated, thereare a limited number of studies reporting the use of EOs to inhibitmicrobial growth on foods. The difficulties in application to foodsis their limited solubility and the impact of these substances onthe organoleptic food properties, variability of their composition,and their variable activity in foods due to interactions with foodcomponents (Gutierrez et al., 2008). Nevertheless, EOs to controlmicrobial growth have been proposed for several products includ-ing fresh-cut fruit. Lanciotti et al. (1999) suggested that the additionof citrus essential oils to a fresh sliced fruit mixture (apple, pear,grape, peach and kiwifruit) inhibited the proliferation of naturallyoccurring microbiota. Lanciotti et al. (2004) also reported that theaddition of 0.02% (v/v) citrus, mandarin, cider, lemon and lime EOsto a minimally processed fruit mix inhibited the proliferation ofthe naturally occurring microbiota and reduced the growth rateof inoculated S. cerevisiae populations, thus extending the shelf-life of the fruit salad without affecting its sensory properties. Inthis way, Roller and Seedhar (2002) demonstrated that carvacroland cinnamic acid (0.015%, v/v) were effective in reducing andinhibiting microbial growth on fresh-cut kiwifruit and honeydewmelon, respectively, without detrimental sensory effects. In fresh-cut kiwifruit, concentrations of carvacrol (0.075–0.225%, v/v) wereeffective in reducing the natural microbiota of the product, butundesirable color and odor changes were observed. Ngarmsak et al.(2006) applied a vanillin dip (0.12%, w/v) to delay the developmentof total aerobic bacteria and yeast and mould populations of fresh-cut mangoes stored at 5 and 10 ◦C for up to 14 and 7 d, respectively.A dip of vanillin (0.18%, w/v) inhibited a 37 and 66% of the microbialgrowth on “Empire” and “Crispin” apple slices, respectively, after19 d of storage (Rupasinghe et al., 2006).

2.3. Edible coatings

As commented previously, dips of antimicrobial solutions arewidely practiced to improve microbial stability of fresh-cut fruit.However, antimicrobial agents rapidly diffuse into the food, thusreducing efficacy due to a decrease in concentration on the fruit sur-face. The incorporation of antimicrobial agents into edible coatingsprovides more inhibitory effects against spoilage and pathogenicbacteria by maintaining effective concentrations of the active com-pounds on the food surfaces (Gennadios and Kurth, 1997). The useof edible coatings with antimicrobial properties or with incorpora-

tion of antimicrobial compounds therefore is a potential alternativeto enhance the safety of fresh-cut produce.

Several types of edible coatings have been used for extendingshelf-life of fresh commodities (Table 1). For instance, chitosan,a film-forming polysaccharide, has been widely used due to its

142 G. Oms-Oliu et al. / Postharvest Biology and Technology 57 (2010) 139–148

Table 2Edible coating formulations to improve quality of fresh-cut fruit.

Fruit EC matrix Stabilizing substances Reference

Apple Apple puree/pectin alginate 0.5% AA + 0.5% CA McHugh and Senesi (2000)2.5% MA + 1% NAC + 1% GSH + 0.7% lemongrass or 0.3%cinnamon oil + 2% CaL

Raybaudi-Massilia et al. (2008a)

WPC 1% AA + 1% CaCl2 Lee et al. (2003)Alginate/gellan alginate/apple puree 1% NAC + 2% CaCl2 Rojas-Graü et al. (2008b)

1% NAC + 2% CaCl2 + 0.3–0.6% vainillin or 1–1.5%lemongrass or 0.1–0.5% oregano oil

Rojas-Graü et al. (2007a)

WPC-BW 1% AA or 0.5% cys Perez-Gago et al. (2006)Alginate/pectin/methylcellulose 1% AA + 0.5% CA + 0.25% CaCl2 Wong et al. (1994)

Melon Alginate 2.5% MA + 0.3% palmarosa oil + 2%CaL Raybaudi-Massilia et al. (2008b)Alginate/pectin 2% CaCl2 Oms-Oliu et al. (2008a)

Papaya Alginate/gellan 1% AA + 2% CaCl2 Tapia et al. (2008)

Pear Methylcellulose 1% AA + 0.1% PS + 0.25% CaCl2 Olivas et al. (2003)Alginate/pectin 0.75% NAC + 0.75 GSH + 2% CaCl2 Oms-Oliu et al. (2008b)

Pineapple Alginate 1% CA + 1% AA + 2% CaCl2 Montero-Calderón et al. (2008)

B cium lp

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W: beeswax, AA: ascorbic acid, MA: malic acid, CaCl2: calcium chloride, CaL: calotassium sorbate, WPC: whey protein concentrates.

bility to inhibit growth of many pathogenic bacteria and fungiRomanazzi et al., 2002). Chien et al. (2007) reported the effective-ess of chitosan in maintaining quality and extending shelf-life ofliced mango. Assis and Pessoa (2004) and Han et al. (2005) alsoroposed chitosan for extending the shelf-life of sliced apples andresh strawberries, respectively. Park et al. (2005) reported a reduc-ion of 2.5 and 2 log CFU/g in the initial counts of Cladosporiumpp. and Rhizopus spp., respectively, on strawberries coated withchitosan-based edible film. A reduction in the counts of aerobic

nd coliforms microorganisms was also observed during storage.ther edible coatings with antimicrobial properties have been usedn fresh-cut produce. Recently, some authors have proposed these of Aloe vera gels as antimicrobial coatings for fruit and vegeta-les because of their proved antifungal activity (Martínez-Romerot al., 2003). Valverde et al. (2005) and Martínez-Romero et al.2006) proposed A. vera gel-based edible coatings for preventing

oisture loss, reducing texture decay, and controlling respiratoryate of table grapes and sweet cherries, while reducing microbialroliferation.

Several compounds have been investigated for incorporationnto edible coatings, including organic acids (acetic, benzoic, lac-ic, propionic, sorbic), fatty acid esters (glyceryl monolaurate),olypeptides (lysozyme, peroxidase, lactoferrin, nisin) and plantOs (cinnamon, oregano, lemongrass) (Franssen and Krochta,003). Nevertheless, there is not much literature on the use ofdible coatings as carriers of antimicrobials to prevent microbialrowth on fresh-cut fruit (Table 2). Lee et al. (2003) extended thehelf-life of refrigerated apple slices by more than 2 weeks, whensing a carrageenan coating containing ascorbic, citric and oxaliccids. Rojas-Graü et al. (2007a) reported the effectiveness of algi-ate and gellan edible coatings to incorporate EOs (lemongrass,regano oil and vanillin) with antimicrobial properties to prolonghelf-life of fresh-cut apples. Similarly, Raybaudi-Massilia et al.2008a) observed that the addition of cinnamon, clove or lemon-rass oils or their active compounds into an alginate-based coatingeduced E. coli O157:H7 populations by more than 4 log CFU/g andxtended the microbiological shelf-life of fresh-cut ‘Fuji’ apples fort least 30 d. Raybaudi-Massilia et al. (2008b) also evaluated thefficacy of malic acid and EOs (cinnamon, palmarosa and lemon-

rass) into an alginate-based coating to improve the shelf-life andafety of fresh-cut melon. According to their results, incorpora-ion of 0.3% palmarosa oil into an alginate coating was promising,ince it inhibited the growth of the native microbiota and reducedhe population of inoculated S. enteritidis, while maintaining fruit

actate, CA: citric acid, GSH: glutathione, NAC: N-acetylcysteine, cys: cysteine, PS:

quality properties and the overall acceptance of the product. Chenet al. (1999) developed a methylcellulose edible coating as a car-rier of benzoic acid in order to inhibit the growth of osmophilicyeasts on a Taiwanese-style fruit preserve. Likewise, García et al.(1998) observed a reduction in the microbial growth and extensionof the shelf-life of fresh strawberries using a starch-based coatingcontaining potassium sorbate.

3. Recent approaches to control enzymatic browning

Color is a critical quality property of fresh-cut fruit suchas pear, apple and banana, since cutting operations may oftenlead to enzymatic browning. The key enzyme in enzymaticbrowning is polyphenol oxidase (EC 1.14.18.1; PPO). This copper-containing enzyme catalyzes the hydroxylation of monophenolsto o-diphenols (monophenolase or cresolase activity) and the oxi-dation of o-diphenols to o-quinones (diphenolase or catecholaseactivity). These o-quinones condense and react nonenzymaticallywith amino acids and proteins, leading to brown melanin pigments(Zawistowski et al., 1991). Another important family of oxidativeenzymes is peroxidases (EC 1.11.1.7; POD). Peroxidase may alsocontribute to enzymatic browning, oxidizing hydrogen donors inthe presence of hydrogen peroxide (H2O2) (Richard-Forget andGauillard, 1997). It accepts a wide range of hydrogen donors,including polyphenols such as hydroxycinnamic derivatives andflavans, the main phenolic structures implicated in enzymaticbrowning of fruit (Robinson, 1991; Nicolas et al., 1994). The possiblerole of POD in enzymatic browning has remained questionable fortwo main reasons: the high affinity of PPO for its natural substrateand the very low H2O2 levels in vegetable tissues (Richard-Forgetand Gauillard, 1997; Cantos et al., 2002; Degl’Innocenti et al., 2005).However, generation of H2O2 during oxidation of some phenoliccompounds catalyzed by PPO might suggest a possible synergisticaction between PPO and POD, which implies involvement of PODin browning processes (Subramanian et al., 1999).

During minimal processing of fruit and vegetables, tissueintegrity is damaged and the compartmentalization of thecell begins to fail (Soliva-Fortuny et al., 2001). As a con-

sequence, phenolic compounds come into contact with PPOand/or POD (Degl’Innocenti et al., 2005). Some of the commonsubstrates of these enzymes are catechins, chlorogenic acid, 3,4-dihydroxyphenylalanine and tyrosine (Sapers, 1993). The browningrate does not seem to be limited by enzymes associated with

G. Oms-Oliu et al. / Postharvest Biology and Technology 57 (2010) 139–148 143

Table 3Dipping treatments to maintain color or/and firmness of fresh-cut fruit.

Fruit Stabilizing treatment Reference

Apple 0.5% CaL Alandes et al. (2006)0.001 M HR + 0.5 M IAA + 0.05 M CaP + 0.025 M cys Buta et al. (1999)7% CaA Fan et al. (2005)0.01% HR + 0.5% AA Luo and Barbosa-Cánovas (1997)1% AA + 0.2% CA or 0.5% NaCl Pizzocaro et al. (1993)4% CaP Quiles et al. (2007)1% NAC+ 1% GSH + 1% LCa Raybaudi-Massilia et al. (2007)0.75% AA + 0.75% CaCl2 Rocha et al. (1998)1% AA + 0.5% CaCl2 Soliva-Fortuny et al. (2001)0.05% kojic acid Son et al. (2001)0.5% AA + 1% CaCl2 + 0.1% PA Varela et al. (2007)

Banana 0.5 M CA + 0.05 M NAC Moline et al. (1999)

Kiwifruit 1% CaCl2 or 2% CaL Agar et al. (1999)

Peach 2% AA + 1% CaL Gorny et al. (1999)

Pear 2% AA +0.01% HR +1% CaCl2 Arias et al. (2008)0.01% HR + 0.5% AA + 1% CaL Dong et al. (2000)2% AA + 1% CaL + 0.5% cys Gorny et al. (2002)0.75% NAC + 0.75% GSH Oms-Oliu et al. (2006)4% NaE + 0.2% CaCl2 + 100 ppm HR Sapers and Miller (1998)1% AA + 0.5% CaCl2 Soliva-Fortuny et al. (2002c)

Mango 0.001 M HR + 0.5 M IAA González-Aguilar et al. (2000)3% CaCl2 Souza de et al. (2006)

Melon 2.5% CaL Luna-Guzmán and Barrett (2000)1% AA + 0.5% CaCl2 Oms-Oliu et al. (2007)

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Watermelon 2% CaCl2

A: ascorbic acid, CaA: calcium ascorbate, CaCl2: calcium chloride, CaL: calcium lasoascorbic acid, NAC: N-acetylcysteine, PA: propionic acid, NaCl: sodium chloride,

rowning or phenolic substrate concentration (Cantos et al., 2002).n the other hand, these authors have suggested that membrane

tability is potentially a major factor controlling the rate of brown-ng.

.1. Antibrowning treatments

A dip treatment after peeling and/or cutting is the most commonay to control browning phenomena in fresh-cut fruit (Table 3).

his can either affect the enzyme or their substrates. Most strate-ies to control browning have focused on theoretical approaches toodulate PPO enzyme activities (Martínez and Whitaker, 1995).

.1.1. Ascorbate and calciumThe most frequent alternative to sulfites is ascorbic acid (AA),

hich is recognized as a GRAS substance by the U.S. Food and Drugdministration (FDA) for its use to prevent browning of fruit andegetables. AA is used to control PPO enzyme activity through itsbility to reduce the o-quinones back to their phenolic substratesHsu et al., 1988; McEvily et al., 1992). Dips of AA have long beenpplied in combination with organic acids and calcium salts to pre-ent enzymatic browning of fruit (Pizzocaro et al., 1993; Gornyt al., 1998a; Soliva-Fortuny et al., 2001, 2002a,b). A formulationontaining calcium and AA acts partly to prevent cell and mem-rane breakdown and also modulates PPO activity in damagedells, where loss of compartmentalization has occurred alreadyToivonen and Brummell, 2008). However, this treatment is notompletely effective to control enzymatic browning of fresh-cut

ruit, since once the AA is completely oxidized to dehydroascor-ic acid, o-quinones are no longer reduced and darkening mayccur (Nicolas et al., 1994). In addition, AA may cause importantxidative damage in fresh-cut ‘Fuji’ apples (Larrigaudière et al.,008).

Mao et al. (2005)

CaP: calcium propionate, cys: cysteine, CA: citric acid, HR: 4-hexylresorcinol, IAA:odium eritorbate.

3.1.2. Thiol-containing compoundsSome thiol-containing substances such as N-acetylcysteine and

reduced glutathione are natural compounds with antioxidant prop-erties, and have been proposed as browning inhibitors to preventdarkening on apple, potato and fresh fruit juices (Molnar-Perl andFriedman, 1990a,b; Friedman et al., 1992; Rojas-Graü et al., 2006).Thiol-containing antibrowning additives react with o-quinonesformed during the initial phase of enzymatic browning reactionsto yield colorless addition products or to reduce o-quinones to o-diphenols (Richard et al., 1991). This suggests that thiol-containingantibrowning additives do not inhibit PPO enzymes per se althoughthey have some inhibitory activity due to their ability to conjugatewith primary oxidation products formed in the reaction (Richard-Forget et al., 1992; Billaud et al., 2004). In addition, PPO enzymeshave differential sensitivities to thiol treatments, according to theplant source of the enzyme (Sapers and Miller, 1998; Billaud etal., 2004). Dips in aqueous solutions containing sulfur-containingamino acids such as N-acetylcysteine and/or glutathione at con-centrations around 0.75% have been shown to inhibit browningof fresh-cut pears and apples (Rojas-Graü et al., 2006; Oms-Oliuet al., 2006). Gorny et al. (2002) also reported minor changes inthe surface color of ‘Barlett’ pear slices treated with 2% AA + 1%calcium lactate + 0.5% cysteine provided that this latter chemicalwas added at pH 7, otherwise the appearance of pinkish-red col-ored compounds occurred (Richard-Forget et al., 1992). Among theantioxidants tested by Moline et al. (1999) in banana slices, it wasconcluded that N-acetylcysteine (0.05 M) and citric acid (0.5 M)provided the best results in inhibiting browning during 1 week ofstorage.

3.1.3. Carboxylic acidsCarboxylic acids have been widely used commercially due to

their antibrowning activity. Citric acid exerts a double inhibitoryeffect by reducing pH and chelating copper in the active site ofPPO and, therefore, inactivating the enzyme (Son et al., 2001). Opti-

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um PPO activity is observed at pH 6.0–6.5, while little activity isetected below pH 4.5 (Whitaker, 1995). Jiang et al. (2004) reportedhat citric acid at concentrations lower than 0.02 M stimulated PPOctivity of fresh-cut Chinese water chestnut, but at concentrationsbove 0.1 M the enzyme activity was markedly inhibited. Acidu-ants are not often used alone because it is difficult to achievefficient browning inhibition, and combination with a chemicaleductant may have a major effect. Pizzocaro et al. (1993) reportedore than a 90% inhibition of PPO activity in apple cubes by using aixture of 1% AA + 0.2% citric acid or 1% AA + 0.5% sodium chloride.

itric acid and/or AA dips were not effective in controlling brown-ng of pear slices. In contrast, color was substantially improvedhroughout refrigerated storage when dipping into a 1% CaCl2 solu-ion (Rosen and Kader, 1989). Other carboxylic acids such as oxaliccid and oxalacetic acid showed higher antibrowning activity thanitric acid on fresh-cut apples (Son et al., 2001). Immersion ofanana and apple slices in oxalic acid solutions was effective againstrowning (Son et al., 2001; Yoruk et al., 2004). Oxalic acid is aatural component of a large number of plants, such as aspara-us, broccoli, Brussels sprouts, carrot, garlic, lettuce, onion, parsley,ea, potato, radish, spinach, tomato and turnip (Yoruk et al., 2004).lthough the mechanism of browning inhibition is unknown, oxaliccid seems to inhibit PPO per se, by chelating copper from the activeite of the enzyme, since oxalic acid has a high affinity to form metalomplexes with copper ion (Tong et al., 1995). The extent of inhibi-ion is influenced not only by oxalic acid concentration, but also byH (Altunkaya and Gökmen, 2008). In addition, PPO enzymes fromifferent sources exhibit different types of inhibition mechanismsSon et al., 2001; Aydemir and Akkanli, 2006).

.1.4. Phenolic acidsCertain phenolic acids inhibit PPO activity by binding to

he active site of the enzyme, whereas others can promotehe enzymatic browning reaction (Janovitz-klapp et al., 1990).ome researchers have proposed that kojic acid [5-hydroxy--(hydroxymethyl)-�-pyrone], a fungal metabolite produced byany species of Aspergillus and Penicillium acts as a reducing agent

s well as an inhibitor to the PPO enzyme per se (Chen et al., 1991).ojic acid is thought to inhibit PPO activity by interfering with

he uptake of O2 required for the enzyme reaction, by reducing-quinones to diphenols to prevent melanin formation via poly-erization and/or by combination of the two previous processes

Chen et al., 1991). The minimal concentration of kojic acid for effec-ive antibrowning activity on fresh-cut apples was 0.05%, which issimilar concentration to that reported for oxalic acid and cysteine

Son et al., 2001).

.1.5. ResorcinolsAmong several resorcinol derivatives, 4-hexylresorcinol has

een proved to be effective in controlling browning on fresh-cutruit such as apples and pears (Monsalve-González et al., 1993;ong et al., 2000; Son et al., 2001; Oms-Oliu et al., 2006; Rojas-raü et al., 2006). 4-Hexylresorcinol has a structural resemblance

o phenol substrates and could have a competitive inhibitory effectn PPO activity (McEvily et al., 1992). Hence, 4-hexylresorcinol maypecifically interact with PPO, and render it unable to catalyze thenzymatic reaction (Kahn and Andrawis, 1985). Its applicability onresh-cut fruit has been proven, especially when used in combi-ation with reducing agents (Monsalve-González et al., 1993; Luond Barbosa-Cánovas, 1997; Dong et al., 2000; Arias et al., 2008).ome combinations have been proven to extend the storage life of

resh-cut produce. A mixture of 0.001 M 4-hexylresorcinol + 0.5 Msoascorbic acid + 0.05 M calcium propionate + 0.025 M homocys-eine maintained freshness of apple slices for 4 weeks at 5 ◦C (Butat al., 1999). Other combinations have been suggested to preventresh-cut pears from browning. For instance, 4-hexylresorcinol in

nd Technology 57 (2010) 139–148

combination with sodium erythorbate had a significant effect onmaintaining the color of fresh-cut Anjou pears (Sapers and Miller,1998). A dip in 0.01% 4-hexylresorcinol + 0.5% AA + 1% calcium lac-tate solutions also provided color stability on fresh-cut pears for30 d (Dong et al., 2000).

3.2. Edible coatings

Enzymatic browning is often inhibited by a direct immer-sion of the fruit pieces into an aqueous solution of antibrowningagents. However, the potential use of edible coatings with fresh-cut fruit as carriers of antibrowning agents has been investigated(Baldwin et al., 1996; Lee et al., 2003; Perez-Gago et al., 2006;Rojas-Graü et al., 2007b, 2008b; Oms-Oliu et al., 2008a,b). Baldwinet al. (1996) demonstrated very effective browning inhibition onfresh-cut apples by AA, when incorporated into an edible coatingformulation; this was better than when dipping fruit pieces into anaqueous solution containing this compound. The incorporation ofantioxidant agents such as N-acetylcysteine and glutathione intoalginate- and gellan-based coatings also helped to prevent fresh-cut apples, pears and papayas from browning (Tapia et al., 2005;Rojas-Graü et al., 2007b, 2008b; Oms-Oliu et al., 2008b). Olivas etal. (2003) also reported the positive effect of the incorporation ofsome additives (ascorbic acid, calcium chloride and sorbic acid) intomethylcellulose and methylcellulose-stearic acid coatings on thebrowning control of fresh-cut pears. Polysaccharide-based coatingsare generally applied at a first step and the antibrowning agentsare incorporated afterwards in the dipping solution containing cal-cium for crosslinking and instant gelation of the coating (Wong etal., 1994; Lee et al., 2003). On the other hand, the effect of incor-poration of AA, cysteine or 4-hexylresorcinol into protein-basededible coatings was investigated on fresh-cut apples by dipping thefruit pieces into the coating emulsion containing the antioxidants(Perez-Gago et al., 2006). The results showed that the inclusion ofantioxidants into the coating matrix reduced browning comparedto the use of the antioxidants alone, treatments with 1% AA or 0.5%cysteine being the most effective.

The incorporation of N-acetylcysteine and/or glutathione intopolysaccharide-based coatings not only prevented fresh-cut applesand pears from browning but also reduced microbial growth com-pared to samples without antioxidants (Rojas-Graü et al., 2008b;Oms-Oliu et al., 2008b). In addition, the addition of AA as an antiox-idant in alginate- and gellan-based coatings helped to preserve thenatural AA content of fresh-cut papaya, contributing to maintain-ing its nutritional quality (Tapia et al., 2008). Other authors havealso observed increased vitamin C and total phenolic contents inpear wedges coated with alginate, gellan and pectin including N-acetylcysteine and glutathione compared with control samples,thus contributing to maintenance of the antioxidant potential ofthe fruit (Oms-Oliu et al., 2008b).

4. Maintaining texture of fresh-cut fruit

Eating quality of fresh-cut fruit products is not only influencedby the stage of ripeness at cutting (Gorny et al., 1998b) but also ishighly dependent on the postharvest history of fruit before process-ing. Some studies have revealed that prolonged storage of apples,even in controlled conditions, causes changes in the fruit that leadto a shorter sensory shelf-life (Varela et al., 2005, 2008). In pro-longed storage of pears, a decrease in acceptability was related to

losses in texture (softer and less crisp fruit), lower acid/sugar ratio,and a loss in cell wall integrity (Salvador et al., 2007).

During mechanical operations, cut surfaces are damaged, releas-ing enzymes which spread through the tissue and come into contactwith their substrates. The softening of fresh-cut fruit is mainly due

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o the enzymatic degradation of the cell wall, which is mainly com-osed of cellulose, hemicellulose and pectins. Enzymes such asectinmethylesterase (PME) and polygalacturonase (PG) generallylay an important role in the fruit softening. PME demethylatesectin, resulting in the production of methanol and a pectinolecule with a lower degree of methylation. This allows depoly-erization by PG, which breaks down �-1,4 glycosidic bonds,

eading to cell wall degradation (Alandes et al., 2006). However,alcium can interact with the free carboxyl groups liberated byhe de-esterification of pectin by PME to form insoluble calciumectates, which strengthen the structure of the cell wall. There-ore, in order to avoid any loss of texture and to preserve thetructure, fresh-cut fruit can be treated with calcium salts such asalcium chloride, calcium lactate, calcium ascorbate and calciumropionate.

.1. Calcium treatments

Calcium chloride (CaCl2) has always been one of the most fre-uently used calcium salts when treating minimally processed fruitTable 3). Applying CaCl2 to cantaloupe melons made the fruitrmer (Luna-Guzmán et al., 1999); the higher the concentrationf calcium chloride used, the firmer the fruit. The combination ofCaCl2 treatment and packaging with a low O2 concentration wasore effective than the use of CaCl2 alone to maintain firmness of

resh-cut ‘Piel de Sapo’ melons (Oms-Oliu et al., 2007), ‘Conference’ears (Soliva-Fortuny et al., 2002b) and ‘Golden Delicious’ applesSoliva-Fortuny et al., 2003) over several weeks of storage. By thend of storage of the apples and pears, microstructural observa-ions showed that the original cellular structure of the fruit was notubstantially altered. Quiles et al. (2004) observed that CaCl2 pre-erved the structure of ‘Granny Smith’ apples during the processf osmotic dehydration. The drawback to this calcium salt is thatt contributes to a bitter taste in the product (Luna-Guzmán andarrett, 2000; Saftner et al., 2003; Lamikanra and Watson, 2003;ernández-Munoz et al., 2006). An integrated sensory approach,

nvolving instrumental/sensory correlations, has been published byarela et al. (2007) in order to evaluate the use of different calciumalts in fresh-cut apples. Their results showed that a dipping of 1%aCl2 for 3 min for fresh-cut ‘Fuji’ apples maintained the overallcceptability of the samples for at least 8 d of storage. However, anstringent aftertaste in treated samples detected by a trained panelid not affect the apple taste liking score in a consumer test.

In recent years, calcium lactate has been used as an alternativeource of calcium, since it does not leave a residual taste in theroduct, it prevents browning and it acts as an acidity regulatorManganaris et al., 2005). In this way, calcium lactate stabilises theood’s pH when acids or bases are added. This effect is importantor the food industry as pH of foods can affect the effectiveness ofther additives such as preservatives and flavourings, which onlyct in a narrow pH range. Dong et al. (2000) and Gorny et al. (2002)xtended shelf-life of fresh-cut pears treated with calcium lactateue to a reduced softening during storage. Alandes et al. (2006)reserved the texture of fresh-cut ‘Fuji’ apples by treating themith calcium lactate. Peaches treated with calcium lactate were

lso found to be firmer than untreated samples (Manganaris et al.,007).

Other calcium salts that are used as firming agents for thetructure of fresh-cut fruit are calcium propionate and calciumscorbate. A Cryo-SEM and LM study of fresh-cut ‘Fuji’ applesreated with calcium propionate showed that calcium consolidated

he structure and preserved the integrity of the apple parenchymaor at least 2 weeks of storage at 4 ◦C (Quiles et al., 2007). A dip withalcium ascorbate reduced firmness loss of fresh-cut ‘Gala’ applesy approximately 13% after 3 weeks at 10 ◦C (Fan et al., 2005). Treat-

ng fresh-cut ‘Golden Delicious’ apples with calcium ascorbate and

nd Technology 57 (2010) 139–148 145

electrolysed water reduced softening for 21 d at 4 ◦C (Wang et al.,2007).

Several authors have analysed the role of pectic enzymes suchas PME and PG in fruit softening process. In fresh-cut Cantaloupemelons, there was an increase in the activity rate of PG at the begin-ning of the storage period, but a gradual decrease in this rate after14 d at 4 ◦C. This was related to enzyme inactivation due to fruitsenescence (Lamikanra et al., 2003). On the other hand, PME activ-ity rate in fresh-cut cantaloupe melons suffered a marked decreaseon the first day of storage at 4 ◦C and 15 ◦C (Lamikanra and Watson,2003). In fresh-cut ‘Fuji’ apples, a treatment of calcium propionateresulted in reduced PME activity throughout storage time (Quiles etal., 2007). Many authors have attributed such a reduced enzymaticactivity to the inhibiting effect of the propionate anions (Nari etal., 1991; Moustacas et al., 1991; Charnay et al., 1992). They sug-gested that there is an activation of PME by metal ions that can beexplained by assuming that they interact with the substrate ratherthan with the enzyme. However, calcium lactate treatment of ‘Fuji’apples showed that PME and PG activities increased just after treat-ment, as a result of the enzymatic activation by Ca2+, which bindsto the enzyme itself as a cofactor rather than a direct calcium effecton the product of the reaction. Release of carboxyl groups throughPME activity would favour the formation of calcium pectates, thusstrengthening the structure of the fresh-cut fruit and making up forthe detrimental activity of the enzyme (Alandes et al., 2006).

4.2. Edible coatings

The main ways to apply calcium salts to fruit are by immer-sion and impregnation, with the former the most commonly used.However, the incorporation of calcium salts such as calcium chlo-ride into coatings seems also to have a beneficial effect on firmnessretention of fresh-cut fruit, especially those commodities thatexhibit a substantial softening of tissues (Lee et al., 2003; Olivaset al., 2007; Rojas-Graü et al., 2008b; Oms-Oliu et al., 2008a).The use of CaCl2 for crosslinking carbohydrate polymers such asalginate, gellan or pectin minimized softening of fresh-cut applesand melon (Rojas-Graü et al., 2008b; Oms-Oliu et al., 2008a). Inaddition, the incorporation of antibrowning agents and 1% CaCl2within a whey protein concentrate coating also helped to main-tain firmness of apple pieces (Lee et al., 2003). As already stated,calcium interacts with pectic acids to form a crosslinked polymernetwork that increases mechanical strength and makes molecularbonding between cell wall constituents firmer (Dong et al., 2000;Soliva-Fortuny et al., 2003). For some fresh-cut fruit, water lossis a big problem that can be controlled by use of edible coatingsas carriers of calcium salts. Edible coatings decrease water vapourtransmission rate by forming a barrier on the fruit surface, pre-venting water loss and texture decay (Olivas and Barbosa-Cánovas,2009). In alginate- or gellan-based coatings, the use of CaCl2 forcrosslinking polymers has been reported to prevent moisture lossand loss of turgor of fresh-cut apples (Rojas-Graü et al., 2007b,2008b). Wong et al. (1994) achieved a reduction of between 12 and14 times water loss of apple slices by coating them with a cellu-lose/lipid bilayer edible film containing CaCl2. Similar results werereported by Montero-Calderón et al. (2008) who observed a reduc-tion of juice leakage of fresh-cut pineapple coated with alginateand CaCl2. However, Olivas et al. (2007) showed that the effectof CaCl2 in maintaining texture of apple slices was more impor-tant than the effect of alginate coatings in preventing water lossof fruit pieces, since softening of apples can be attributed more to

cell wall degradation than to a reduction in turgor pressure. Othersalts such as calcium gluconate have been investigated as alterna-tives to CaCl2 since they impart a more neutral taste. Incorporationof gluconate into chitosan coatings contributed to the extension ofshelf-life of strawberries by maintaining fruit firmness. In addition,

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he addition of calcium gluconate could increase the nutritionalalue of fruit without altering the visual appeal of the strawberriesHernández-Munoz et al., 2006, 2008).

. Conclusions

The use of natural GRAS compounds offers interesting pos-ibilities for extending shelf-life and quality of fresh-cut fruit.haracterization of compounds obtained from plant extracts andther natural sources, such as essential oils, reducing agents andrganic acids, among others, is required in order to further inves-igate the potential applications of these compounds in fresh-cutroduce. In addition, the use of edible coatings is gaining impor-ance in recent years since they may allow incorporation of naturaldditives in lower amounts while increasing their effectiveness,hus contributing to greater consumer acceptance.

cknowledgements

This work was supported by the Ministerio de Ciencia y Tec-ología (AGL2003-09208-C03), the Departament d’Universitats,ecerca i Societat de la Informació of the Generalitat de CatalunyaSpain), the Universidad de Lleida (Spain) and the Universidadolitécnica de Valencia (Spain).

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