Innovative Food Science and Emerging Technologies 12 (2011) 6266

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Innovative Food Science and Emerging Technologies

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / i f s e t

Extending shelf-life of Fresh-cut Fuji apples with chitosan-coatings

Haiping Qi a, Wenzhong Hu a,, Aili Jiang a, Mixia Tian a, Yingqiu Li b
a College of Life Science, Dalian Nationalities University, 18 Liaohe Road West, Dalian Economic and Technological Development zone, Education Department and national nationality Key Lab Dalian 116600, China b College of Food and Bioengineering, Shandong Institute of Light Industry, Jinan 250353, China

a r t i c l e

i n f o

a b s t r a c t
The effect of coatings in combination with anti-browning agents (1%chitosan; 2%ascorbic acid + 0.5%CaCl2 and 2% ascorbic acid + 0.5%CaCl2 + 1% chitosan) on minimally processed apple slices was studied during storage. Chitosan-coating treatments effectively retarded enzymatic browning on minimally processed apples during storage and they effectively retarded or avoided tissue softening, apple slices underwent a little loss of rmness. Chitosan-coating did not perform very well as water vapor barriers in apple slices. To control initial respiration rate of apple slices, edible coatings were applied to cut apples as semi-permeable barriers against air. Initial respiration rate showed a decrease in 2% ascorbic acid + 0.5%CaCl2 + 1% chitosan apple slices at 5 C. Industrial relevance: Recently, there has been an increasing market demand for minimally processed fruits and vegetables due to their fresh-like character, convenience, and human heath benets. Minimal processing includes grading, washing, sorting, peeling, slicing, chopping, and then packaging. Since minimal processing results in quality deterioration associated with water loss, softening, microbial contamination, increased respiration and ethylene, and cut-surface browning, minimally processed products become more perishable. Edible coating was used for cut apples to reduced respiration and control physiological changes. The color of products, such as apple slices, is an important quality index. Brown apple slices are aesthetically unattractive. Chitosan has been reported to maintain the quality of fruit and vegetables (EI Ghaouth et al.,1991). The objective of this study was to develop procedures for the use of edible coatings in combination with antibrowning agent and to extend the shelf-life of minimally processed apple slices. 2010 Elsevier Ltd. All rights reserved.

Article history: Received 31 August 2009 Accepted 3 November 2010 Editor Proof Receive Date 20 December 2010 Keywords: Minimally processed apples Edible coatings Initial respiration rate Antibrowning agents Color

1. Introduction Recently, there has been an increasing market demand for minimally processed fruits and vegetables due to their fresh-like character, convenience, and human health benets. Minimal processing includes grading, washing, sorting, peeling, slicing, chopping, and then packaging. Since minimal processing results in quality deterioration associated with water loss, softening, microbial contamination, increased respiration and ethylene, and cut-surface browning, minimally processed products become more perishable (Rolle & Chism, 1987). The respiration activity of minimally processed products increases 1.27.0 fold, or even higher than unprocessed, depending on the products, cutting grade, and temperature (Ahvenainen, 1996). Because the rate of respiration indicates how quickly a product may deteriorate, increased respiration by tissue injury results in a greatly reduced shelf-life compared to whole fruits and vegetables. The use of edible coatings can reduce respiration, thus prolonging product shelflife (Baldwin, Nisperos-Carriedo, & Baker, 1995a, b). Edible coatings provide a semi-permeable barrier against oxygen, carbon dioxide

Corresponding author. Tel.: +86 41187656213. E-mail address: hwz@dlnu.edu.cn (W. Hu). 1466-8564/$ see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.ifset.2010.11.001

(CO2), moisture, and solute movement; thereby reducing respiration, water loss, and oxidation reaction rates (Park, 1999). They are also useful as a carrier of food additives such as antibrowning agents. Edible coating was used for cut apples to reduced respiration and control physiological changes. The color of products, such as apple slices, is an important quality index. Brown apple slices are aesthetically unattractive. The change in color is due to oxidative reactions of phenolic compounds by polyphenol oxidase and the reaction products, oquinones, to various polymerized products. Ascorbic acid (AA), cysteine, citric acid (CA), and some sulfur-containing amino acids have been used as substitutes for sulte to prevent enzymatic browning, alone or in combination with rming or antimicrobial agent (Pizzocaro, Toregiani, & Gilardi, 1993a,b; Dudley & Hotchkiss, 1989; Baldwin, Nisperos, Chen, & Hagenmaier, 1996). Son, Moon, and Lee (2001) investigated the relative anti-browning activity of 36 commonly known anti-browning agents under same conditions. Edible coating can improve the quality of life fruits and vegetables (Wong, Camirand, & Pavlath, 1994; Wong, Gregorski, Hudson, & Pavlath, 1996; Li & Barth, 1998; Baldwin et al., 1996). Until recently, there has been little research that investigated the effect of the combination of Chitosan-based edible coatings and antibrowning agents for minimally processed products to reduce respiration rate and inhibit enzymatic browning. Chitosan has been reported to

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maintain the quality of fruit and vegetables (El Ghaouth, Arul, Ponnampalam, & Boulet, 1991). The objective of this study was to (1) develop procedures for the use of edible coatings in combination with anti-browning agent and to extend the shelf-life of minimally processed apple slices. (2) Evaluate the effects of chitosan coatings on color loss, moisture, rm, respiration and PPO in fresh-cut apple pieces.

2.6. Measurement of respiration rate The effect of coating on CO2 was measured by analyzing the headspace gas composition. Typically, apple slices (100 g), coated or noncoated, were stored in a 100 mL tight-sealed glass container at 25 C for 24 h. Headspace samples were withdrawn at various time intervals and analyzed for CO2 by gas chromatography (Hitachi Model 163, USA), equipped with a thermal conductivity detector and a CTR 1 column (Alltech Associates, Inc., USA). Helium was the carrier gas at 35 mL/min. The injector was 50 C, detector 100 C, and the column was 50 C. All respiration rate measurements were done in three replicates. 2.7. Assay for polyphenol oxidase (PPO) activity

2. Materials and methods 2.1. Preparation of the lm forming solutions Chitosan, with a deacetylation degree of 97%, derived from shrimp shells were purchased from Fluka Biochemica. The chitosan solution (1%) was prepared as follows: weigh 1 g chitosan and put into a 200-mL beaker, then slowly added 100 mL 1% citric acid solution and meanwhile stirred with a magnetic stirrer until the mixture solution pH 3.54.0 became clear and coating formed. The solution was vacuumed for 2 h to remove trapped air bubbles introduced during the mixing. The lm forming solution was used for apple slices fruit coatings.

2.2. Preparation of apple slices Apples (Fuji) were purchased from a local wholesale distributor at commercial maturity. Apples were selected for uniform size and appearance. These fruits were rinsed gently with tap water by hand and dried naturally. Then apples were peeled, cored and cut into 1 cm thick cubes (average weight at 10.3 g) with a sharp stainless steel knife at 10 C. The knife and cutting board were washed with deionized water and rinsed with 1000 l/L sodium hypochlorite solution prior to use.

The PPO activity apple slices was measured according to the method proposed by Galeazzi, Sgarbieri, and Constantinides (1981) (4), with slight modications, as follow: apple slices were pulped with deionized water, homogenized, and centrifugated. The supernatant was carried out in the presence of 0.2 M phosphate buffer (50:50 w/w) at pH 6.4 containing 5% (w/w) polyvinyl pyrrolidone. Polyvinyl pyrrolidone was used to eliminate the polyphenolic fraction of apples. The PPO activities were assayed at 398 nm by means of a Beckman DU 640 spectrophotometer (Beckman, Fullerton, CA) in 3 mL of incubation medium containing 0.5 mL of enzymatic extract. The rate of enzymatic reaction was expressed as OD398 nm/ming FW. 3. Results and discussion 3.1. Color changes for storage In general, the enzymatic browning of apple pieces during storage was accompanied by a decrease in lightness L* and an increase in colorimetric a*. Figs. 1 and 2 show effects of chitosan-coating on L* and a* of apple pieces. In comparison with control sample, chitosan sample almost remained stable L* and a* values at 20 C for 24 h. L* values of all sample decreased rapidly after 24 h for 20 C (no show) and decreased slightly after 24 h for 5 C (Fig. 1b). But a* values of all samples increased rapidly after 24 h for 20 C (no show) and increased slightly after 24 h for 5 C (Fig. 2b). This indicated that low temperature storage was better than high temperature storage to retard browning of apple pieces. L* values of control, 1%chitosan, 2% AA+0.5% CaCl2, 2% AA+0.5% CaCl2 +1% chitosan samples were 59.78, 68 .84, 68.73, and 70.88 on 8 d at 4 C, respectively, which were 75.15%, 91.11%, 91.00%, and 93.84% of L* values of control sample on 0 d at 4 C. a* values of control, 1%chitosan, 2% AA+0.5% CaCl2, 2% AA+0.5% CaCl2 +1% chitosan samples were 0.88, 1.94, 2.96, and 3.14 on 8 d at 4 C, respectively, which were 23.04%, 50.79%, 75.96%, and 82.20% of a* values of control sample on 0 d at 4 C. These results showed that chitosan inhibited effectively browning of apple pieces and ascorbic acid and citric acid increased inhibitory efciency of browning. 3.2. Firmness An undesirable consequence of cutting is softening; tissue stress results in a loss of rmness, principally owing to enzymatic hydrolysis of cell wall pectic substances and the action of pectinolytic enzymes, the decrease in cellulose crystallinity and the thinning of cell walls. Ascorbic acid has long been applied in combination with organic acids and calcium salts, particularly calcium chloride, to prevent enzymatic browning and maintain fruit rmness. The rmness of the control did not signicantly decrease over the 8 days of storage. Coating with chitosan effectively retarded or avoided tissue softening in the samples. This is in agreement with many previous research papers, which have reported that CaCl2 treatment rms apple tissue by strengthening the cell wall and middle lamella. The results (Fig. 3) showed that 0.2%AA + 0.5%CaCl2 +1%chitosan treated apples had the highest rmness, since calcium chloride, a known

2.3. Dipping and storage conditions of apple slices Dipping solutions of AA (Ascorbic Acid), CA (Citric Acid), CaCl2, and 1% chitosan were designed as follows: (1) control sample: deionized water, (2) 1% chitosan, (3) 2% AA + 0.5% CaCl2, (4) 2% AA + 0.5% CaCl2 + 1% chitosan. The apple pieces were rst dipped into the before four solutions pH 3.54.0 fully immersed by using forceps for 2 min. Residual solutions of slices were allowed to drip off for 1 min, The samples were kept at 5 C until the excess of water was drained. Then these apple slices were placed on plastic-coated wire racks inside plastic containers in the conditions of 8 d at 5 C and 24 h at 20 C before testing.

2.4. Measurement of color The color of apple slices was measured with a Minolta Chroma Meter Model CR-300 (Minolta. Tokyo, Japan). The degree of browning was expressed as L-value and a value. The parameter L is a measure of brightness/whiteness that ranges from 0 to 100 (white if L = 100, black if L = 0). The parameter a is an indicator of redness that varies from a* to + a* (a = green, a = red). The results were expressed as a mean value from three replications of the 10 measured samples.

2.5. Water loss To determine the effectiveness of chitosan coatings as moisturebarriers, the weight of eight apples slices in each treatment was monitored during storage. It was assumed that weight loss corresponded entirely with water loss. The weight loss percent relative to initial weight was calculated by weighing the samples every 2 days in triplicate.

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80 75

L-Value

L-Value
0 6 12 18
o

75

70 65 60 55

70

65

24

50

Time(h) 20 C

Time(d) 5oC

Fig. 1. L-value of minimally processed Fuji apples during storage at 5 and 20 C (control sample 1% chitosan,2%AA + 0.5%CaCl2, 2%AA + 0.5%CaCl2 + 1%chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

0 -1

0 -1

a-Value

-2 -3 -4 -5 0 6 12 18 24

a-Value

-2 -3 -4 -5 0 2 4 6 8

Time(h) 20oC

Time(d) 5oC

Fig. 2. a-value of minimally processed Fuji apples during storage at 5 and 20 C (control sample 1% chitosan,2%AA + 0.5%CaCl2, 2%AA + 0.5%CaCl2 + 1%chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

rming agent, helped apple slices maintain rmness. Calcium chloride treatments rm fruit tissue by reacting with pectic acid in the cell wall to form calcium pectate which strengthens molecular bonding between constituents of cell wall. Chitosan at 1% content treated apples also showed constant rmness throughout 8 d of storage due to low level of Ca2+ contained in aqueous chitosan solution. In contrast with chitosan coatings, non-coated apples showed reduced rmness. This softening of apple slices maybe due to the pectic acid undergoing acid hydrolysis (Ponting, Jackson, & Watters, 1972). Therefore, the addition of calcium chloride to an acidic dipping solution could minimize the softening of apple slices. 3.3. Water loss Few articles have reported the effect of coatings on weight loss of fresh-cut produce. McHugh and Senesi (2000) reported no effect of

apple puree coatings, containing ascorbic or citric acid, on moisture loss from either unpacked or packed apple samples. Slicing of apples exposes the skinless tissue to an environment with lower relative humidity, and causes substantial weight loss. A large increase in water loss occurred with uncoated control apples after slicing, as shown in Fig. 4. After 2 days of storage, control apples lost around 19% of their weight, while coated apples lost 15% of their weight (p b 0.05). Chitosan coatings on apple slices did not prove to work effectively as water vapor barriers during the entire storage period (Fig. 4). The three formulations, prevented water loss by producing high relative humidity at the surface of sliced apples, thus reducing the gradient to the exterior. The best coating to prevent water loss was 2%AA+ 0.5%CaCl2 +1%chitosan (p b 0.05), which allowed a 14.3% loss compared to 19% for control apples at day 2 (pb 0.05). No performance differences were found between 1% chitosan and 2%AA+ 0.5%CaCl2,which allowed about 15% less weight loss than the control apple slices (p b 0.05) at day

30

Weight loss(%)
0 2 4
o

Firmness(N)

20

10

Time(d) 5 C
Fig. 3. Mean values and standard deviations of rmness, variation with storage time at 5 C (control sample 1% chitosan,2%AA+ 0.5%CaCl2, 2%AA+ 0.5%CaCl2 +1%chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

Time(d) 5oC
Fig. 4. Weight loss from Fuji apple slices stored 8 days at 5 C and 85%RH (control sample 1% chitosan,2%AA + 0.5%CaCl2, 2%AA + 0.5%CaCl2 + 1%chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

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Respiratory rate CO2mg/kg.h

100 80 60 40 20 0 0 2 4 6 8

aqueous chitosan solution. Calcium ion is known to inhibit respiratory activity and ethylene production. Wong, Tillin, Hudson, and Pavlath (1994) investigated that polysaccharide/lipid bilayer formulation reduced respiration and ethylene production of cut apples due to the diffusion barrier properties of the lipid layer and the inhibitory effect of the ascorbate buffer containing calcium. However, considering the 1 g/ 100 mL concentration of CaCl2 in the 1% chitosan solution, the observed large reduction of initial respiration rate seems to be due to oxygen barrier properties of chitosan coating rather than the effect of calcium ion.

Time(d) 5oC
Fig. 5. Effect of edible coatings on the initial respiration rate of fresh-cut apple slices stored at 5 C for 8 d (control sample 1% chitosan,2%AA+0.5%CaCl2, 2%AA+0.5%CaCl2 +1% chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

3.5. Effect of edible coatings on polyphenol oxidase activity It is generally assumed that minimal processing operations cause disruption of compartmentalization in apple, allowing substrates and PPO to come into contact to result in browning, that PPO is the enzyme mainly responsible for browning. Tissue browning is usually due to oxidation of phenolic compounds to O-quinone, catalysed by PPO. The present data (Fig. 6) show that PPO activity increased with the increase of stored time at beginning, and reach some maximum, and decreased later in tissue at 5 C. The control PPO activity reached maximum 118.1 OD398 nm/min g FW at 2 days, while the PPO activity of 2% AA+ 0.5% CaCl2 + 1% chitosan samples is to reach maximum 120.6 OD398 nm/ min g FW at 6 days, 1%chitosan, and 2% AA + 0.5% CaCl2 samples are to reach maximum 119.6 and 111.8 OD398 nm/min g FW at 4 days, respectively. At 8 days the PPO activities of control, 1%chitosan, 2% AA + 0.5% CaCl2, 2% AA + 0.5% CaCl2 + 1% chitosan samples were 52.5, 36.3, 40.2,76.5 OD398 nm/min g FW, respectively. This result shows AA, CaCl2, and chitosan retard to the decrease of PPO activity and 2% AA + 0.5% CaCl2 + 1% chitosan is best. 4. Conclusions

10, although it is well known that hydrocolloids do not perform very well as water vapor barriers in high water activity foods due to their high hydroscopic nature. However, in this case , 2%AA +0.5%CaCl2 +1% Chitosan and 2%AA+0.5%CaCl2 performed relatively well in limiting water loss. This is probably due to the ability of calcium to cross-link chitosan, making the coating insoluble, bearing in mind that the capacity of hydrocolloid-based lms to function as water vapor barriers increases as their solubility in water decreases (Kester & Fennema, 1986). These results indicate the greater effect pared with coating the fruit, which could be due to the inability of forming a uniform coating in the wet surface of the broken cellular material. The moisture of the fruit surface affects the degree of adhesion of the hydrophobic and hydrophilic coating materials (Baldwin et al., 1995a, b).

3.4. Effect of edible coatings on initial respiration rate The physical damage or wounding caused by peeling and cutting increases respiration rate within minutes, thus the need to reduce initial respiration rate is critical in extending shelf-life of minimally processed fruits. Fig. 5. shows the effect of chitosan edible coatings on initial respiration rate. All formulations tested reduced initial respiration rate, and 2%AA+0.5%CaCl2 +1% chitosan had a signicantly greater effect than others. Initial respiration rate showed a decrease by about 5% and 20% in the 2%AA +0.5%CaCl2 + 1% chitosan apple slices, Their waterinsolubility and oxygen barrier properties made them suitable as coating to achieve internal modied atmospheres (MA) within a fruit system. Another possible reason for the large reduction of initial respiration rate in chitosan-coated apples is the effect of calcium ion contained in the

Our results demonstrate coating treatments effectively retarded enzymatic browning on minimally processed apples during storage and retarded or avoided tissue softening the apple slices, apple slices with chitosan-coating underwent a little loss of rmness. Chitosancoating did not perform very well as water vapor barriers in apple slices. To control initial respiration rate of apple slices, edible coatings were applied to cut apples as semi-permeable barriers against air. Initial respiration rate showed a decrease. Acknowledgement The work is nancially supported by the National Science Foundation of China (Grant Nos. 30571302, 30671458 and 30972038). References

140

PPO activity OD398 nm /ming FW

120 100 80 60 40 20 0 0 2 4 6 8

Time(d) 5oC
Fig. 6. Effect of chitosan coatings on the polyphenol oxidase activity of apple slices stored at 5 C for 8 d (control sample, 1% chitosan, 2%AA + 0.5%CaCl2, 2%AA + 0.5%CaCl2 + 1%chitosan). Values are means of three replicates. Vertical bars represent standard deviation.

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