LWT - Food Science and Technology 51 (2013) 9e15

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LWT - Food Science and Technology

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Improved multilayered antimicrobial alginate-based edible coating extends

the shelf life of fresh-cut watermelon (Citrullus lanatus)
R.E. Sipahi a, M.E. Castell-Perez a, *, R.G. Moreira a, C. Gomes a, A. Castillo b
a
Biological and Agricultural Engineering Department, Texas A&M University, College Station, 303G Scoates Hall, TX 77843-2117, USA
b
Animal Science Department, Texas A&M University, College Station, TX 77843-2471, USA

a r t i c l e i n f o a b s t r a c t

Article history: This study evaluated the effectiveness of an improved multilayered antimicrobial alginate-based edible
Received 17 August 2012 coating in increasing the shelf life of fresh-cut watermelon without affecting its quality attributes. A set of
Received in revised form solutions containing sodium alginate (0.5, 1, 2 g/100 g), beta-cyclodextrin and microencapsulated trans-
5 November 2012
cinnamaldehyde (natural antimicrobial agent), pectin, and calcium lactate were used as coating systems
Accepted 9 November 2012
and made into a coating using the layer-by-layer (LbL) technique. The samples were coated using the
layer-by-layer dipping technique and stored at 4
C for 15 days. Texture, color, weight loss, oBrix, pH, and
Keywords:
growth of total coliforms, yeasts and molds, aerobics, and psychrotrophs were monitored every 3 days
Layer-by-layer
Microencapsulated trans-cinnamaldehyde
throughout storage. Controls were uncoated washed fruits. A consumer acceptance test showed high
Fresh-cut fruit acceptance (P < 0.05) of the coated samples except for the 2 g/100 g alginate coating. Texture (firmness)
Safety was particularly enhanced (P < 0.05) by application of the coating. Both the 1 and 2 g/100 g alginate
Quality coatings extended the shelf life of fresh-cut watermelon from 7 (control) to 12e15 days. The application
of the multilayered edible coating with 1 g/100 g alginate will maintain the quality and sensory
acceptance of fresh-cut watermelon while extending its shelf life.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction
Fresh-cut fruits are becoming more attractive to consumers who
are aware of the importance of healthy eating habits, but have less
time available for food preparation. Watermelon (Citrullus lanatus)
is rich in lycopene and other bioactive compounds (Perkins-Veazie,
Collins, Pair, & Roberts, 2001; Tarazona-Diaz, Viegas, Moldao-
Martins, & Aguayo, 2011). Therefore, the economic importance of
the fresh-cut fruit industry is becoming progressively more
significant (Barret & Garcia, 2004; Tapia et al., 2008). According to
the National Watermelon Promotion Board, over the last decade,
fresh-cut watermelon accounts for more than 10% of all water-
melon sales; however, there are some challenges in processing of
fresh-cut fruits such as watermelon and maintaining their fresh-
ness to increase their economic value.
Fresh-cut processing of fruits promotes faster deterioration,
since fruits are living tissues, they could be wounded and undergo
enzymatic browning, off-flavor development, texture breakdown,
and microbial contamination. Hence, the shelf life of fresh-cut
products tends to be very short if they are wounded. Especially,
microbial and enzymatic deterioration in fresh-cut products mainly
* Corresponding author. Tel.: þ1 979 862 7645; fax: þ1 979 845 3932.
E-mail address: ecastell@tamu.edu (M.E. Castell-Perez).
occurs on their wounded surfaces, while the inner part is generally
considered sterile. For this reason, a surface treatment, even if not
effective on the whole product, could be sufficient to extend the
product shelf life (Artes-Hernandez, Robles, Gomez, Tomas-
Callejas, & Artes, 2010; Manzocco, Da Pieve, & Maifreni, 2011;
Saftner, Luo, McEvoy, Abbott, & Vinyard, 2007; Watada, 1990).
Similar to other fresh-cut fruits, quality loss of fresh-cut water-
melon can be described as loss of texture, color, and sweetness
(Mao, Jeong, Que, & Huber, 2006; Perkins-Veazie & Collins, 2004;
Rushing, Fonseca, & Keinath, 2001).
The enhancement of shelf life of these products is very important
as even a few days extension of shelf life could represent a significant
economic advantage for food companies. By providing barrier to
moisture, edible coatings are potential systems for the improvement
of quality and shelf life of fresh-cut commodities (Vargas, Pastor,
Chiralt, McClements, & Gonzalez-Martinez, 2008). One major
advantage of coatings is that they can be used as a means for including
natural or chemical antimicrobial agents, antioxidants, enzymes or
probiotics, minerals and vitamins. Many types of polysaccharide,
protein, lipid-based and composite coatings have been tested in
a wide variety of items (de Azevedo, 2012; Olivas & Barbosa-Canovas,
2009; Valencia-Chamorro, Palou, del Río, & Pérez-Gago, 2011).
The multilayer coating technique has been used as a good
alternative to overcome challenges when using the dipping

0023-6438/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.lwt.2012.11.013
10 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15
technique when the coating may not stick to the hydrophilic
surface of the cut fruit (Soliva-Fortuny, 2010; Soliva-Fortuny &
Martin-Belloso, 2003). Hence, more than two layers of film mate-
rial are used in the layer-by-layer (LbL) technique to bond physi-
cally and chemically to each other (Skurtys et al., 2010; Weiss,
Takhistov, & McClements, 2006). In this method, fruits and vege-
tables are dipped into different solutions that contain opposite
charged polyelectrolytes, and the excess of coating material from
the fruits’ surface is allowed to be removed by a drying step
between each dipping step. Coatings manufactured with this
technique have been proven to be successful in papaya (Brasil,
Gomes, Puerta-Gomez, Castell-Perez, & Moreira, 2012), pineapple
(Mantilla, 2012), and cantaloupe (Martinon, 2011). However,
further optimization of the coating composition is still necessary
since these studies used calcium chloride which imparted some
bitterness to the coated fruits.
The main objective of this study was to assess the effectiveness
of a multilayered edible coating manufactured using the layer-by-
layer technique and containing a natural antimicrobial (trans-cin-
namaldehyde) in extending the shelf life of fresh-cut watermelon
with acceptable physical, chemical, and sensory quality attributes.
Optimum concentration of sodium alginate was also assessed.
2. Materials and methods
2.1. Fruits
Watermelons (C. lanatus) were purchased at a local store and
kept at 10
C (50% relative humidity) until processing. On average,
fruits had average total soluble solids (Brix) values of 7.50  0.18
(triplicates from six watermelons) measured using a refractometer
(Reichert Analytical Instruments, High Precision Brix15 HP, NY).
Average pH and titratable acidity were 5.23  0.35 and 0.11  0.05,
respectively. Fruits were sanitized by immersion in chlorine solu-
tion (300 mg/kg for 1 min), rinsed with distilled water, and dried at
room temperature (Lee, Park, Lee, & Choi, 2003; Saxena, Saxena,
Raju, & Bawa, 2011; Tapia et al., 2008) All the utensils and
surfaces used during cutting were also sanitized with chlorine
solution. Although fresh-cut watermelons are marketed as quar-
ters, halves with rind, or as cubes without rind, in this study, the
cleaned fruits were cut into small cylinders with 3 cm in diameter
and 2.54 cm length. This decision was made to ensure uniformity in
sample size and dimensions for quality testing.
2.2. Antimicrobial inclusion complex
trans-cinnamaldehyde (2.0 g, 99þ%, C80687, SigmaeAldrich,
MO) and beta-cyclodextrin (18.16 g, hydrate Alfa Aesar,
AAA14529-22, Lancashire, UK) were dispersed in 1 L of aqueous
solution to have a molecular ratio 1:1 and mixed in a laboratory
stirrer (Fisher stirrer 60 Hz, Fisher Scientific, PA) for 24 h at room
temperature. The suspension was filtered through a 0.45 mm nylon
filter (VWR International, PA), the filtrate frozen at 20
C and then
freeze-dried at 50
C under 6.67  103 kPa vacuum for 48 h in
a Labconco Freeze Dry-5 dryer (Labconco, MO). The freeze-dried
material was stored in a desiccator placed inside a freezer
(20
C) until further use (Gomes, Moreira, & Castell-Perez, 2011).
This encapsulation process was selected to improve aqueous solu-
bility of trans-cinnamaldehyde (lipophilic nature) and conse-
quently enhance its antimicrobial activity.
2.3. Materials for coating preparation
The edible coating was prepared with three different solutions:
sodium-alginate, pectin, and calcium lactate, as a replacement for
calcium chloride (Brasil et al., 2012). These materials and their
order were chosen based on the polyetrolyte interaction among
opposite charges to obtain a stable and uniform coating. Glycerol
used as a plasticizer (Glycerin USP, J T Baker, NJ) at 2 g/100 g was
weighted and dissolved in sterile distilled water. Upon that,
sodium-alginate (food grade, SAFC, MO) was added to the solution
in three different concentrations (0.5, 1 and 2 g/100 g) while
heating on a stirring hot plate (Corning, model PC-220, MA) at 45
C
until total dissolution of the components was reached. The natural
antimicrobial agent (trans-cinnamaldehyde encapsulate powder at
2 g/100 g) was then added to the solution while being stirred
(600 rpm) until the solution reached total homogeneity. Pectin
(USP, Citrus, galacturonic acid 74.0% (anhydrous basis), P8471,
Spectrum Chemical Mfg. Corp., CA) at 2 g/100 g was added to sterile
distilled water previously heated at 45
C on a stirring hot plate
(Corning, model PC-220, MA) until it was completely dissolved.
Calcium lactate (USP/NF, SigmaeAldrich, MO) was used to cross-
link the polymers and 2 g/100 g was dissolved in sterile distilled
water at room temperature.
2.4. Manufacture of coating using the layer-by-layer technique
The layer-by-layer deposition process consisted of five steps that
ensure proper coating of the fruit pieces. First, watermelon pieces
previously stored at 10
C were dipped into each coating solution
for 2 min, with the excess coating allowed to drip off for 2 min
before submerging the samples into the calcium lactate solution.
Then, the samples were dipped into several solutions in the
following order: (1) alginate and antimicrobial solution, (2) calcium
lactate, (3) pectin, and (4) a third dipping onto calcium lactate
(Fig. 1). Controls were only dipped into sterile distilled water for
2 min and then allowed to drip off for two more min. After 8 min of
drying at room temperature, eight pieces from each treatment
(coated samples and controls) were placed into plastic containers
(ZiplocÒ Smart SnapÔ Seal, 591-ml) with polyethylene lid, and
stored at 4
C for 15 days.
2.5. Shelf life study
Changes on physical, chemical, microbiological, and sensory
quality attributes of coated and uncoated samples were monitored
Fig. 1. Layer-by-layer coating manufacturing process.
 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15
on days 1, 3, 7, 12, and 15 of storage. All tests were performed at
room temperature and at least in triplicate for each treatment
(coated samples and controls) throughout the shelf life study. These
results were used to determine the feasibility of the coating to
increase the shelf life of fresh-cut watermelon.
2.5.1. Physical and chemical properties
The juice of watermelon samples was squeezed out to avoid any
solid particles from the samples, and the pH was measured using
a digital pH meter (Cole Parmer, Ph 500 series, model #59003-20,
Singapore) (AOAC 981.12, 1990). To measure moisture content (in
wet basis), a sample’s weight was recorded before and after drying
in a vacuum oven at 60
C for 10 h and 30 kPa (AOAC method
920.151). Fruit samples were sliced into thin pieces and placed in
the small chamber of a HygroLab 3 m (Rotronic Co.) for measure-
ments of water activity.
Weight loss was determined by recording the weight of three
samples per treatment using a Mettler Toledo Laboratory Balance
(PG5002-S, Switzerland). New samples were tested each day to
avoid cross-contamination. Results were reported as ((initial
sample weight e final sample weight)/initial sample weight)  100.
The attribute of texture (firmness) of watermelon pieces was
identified as the maximum force to compress the sample down to
50% strain before the sample failed. Ten samples (coated and
controls) were compressed with a cylindrical probe (TA3/100,
diameter 5.2 cm) traveling at 0.5 mm/s using a CT3 Brookfield
Texture Analyzer (Brookfield Engineering Laboratories, Middle-
boro, MA). The effect of the multilayered antimicrobial coating on
the color of the fruits was evaluated using a Labscan XE colorimeter
(Hunter Lab, Inc., Reston, VA, USA) and the CIELAB system cali-
brated with a standard plate (Y ¼ 94.00, x ¼ 0.3578, y ¼ 0.4567)
with illuminant/viewing geometry of D65/10
and a measuring
aperture diameter of 36 mm. Readings of L* (lightness), a* (green-
red chromaticity) and b* (yellow-blue chromaticity) were taken
from five samples per treatment.
The headspace composition was measured to determine
whether application of the coating helped delay the onset of decay.
Glass jars (0.473 L, Mason Jars, USA) with hermetic metal lids were
used instead of the ZiplocÒ trays to prevent gas leakages. The lids
were perforated and then sealed with septum. Four pieces of
watermelon fruit per treatment (coated samples and controls) were
placed inside the glass containers and the gas composition (CO2
and O2) in the jars recorded using a MOCON headspace analyzer
(Model 650, Dual Head Space Analyzer, MN). On each sampling day,
a new container was opened to avoid loss of gases.
2.5.2. Consumer acceptance test
Color, odor, flavor, texture, and overall quality sensory attributes
were evaluated by 40 untrained panelists for days 1, 3, 7, 13, and 15
of storage. Panelists were students, staff and faculty from our
institution who liked watermelon. Four samples (control and three
coated samples) were offered to the panelists in white plastic cups
labeled with 3 random digits. The panelists then scored the samples
on a sensory evaluation sheet using a nine-point hedonic scale
(1 ¼ dislike extremely; 9 ¼ like extremely). Samples were consid-
ered acceptable when they received scores higher or equal to 5.
Scores below 5 indicated that the samples were disliked by the
consumers (Lawless & Heymann, 1998).
2.5.3. Microbiology of fruits
Total aerobic plates, psychrotrophic, coliforms, and yeast and
molds counts were determined on days 0, 1, 3, 7, 12 and 15 of
storage at 4
C. Under sterile conditions, two watermelon pieces
from each treatment were stomached inside a sterile stomacher
bag. A 10 g aliquot of the blended material was transfer to another
11
stomacher bag, mixed with 90 ml of 0.1% buffered peptone water,
and homogenized for 1 min; subsequently, 10-fold dilutions were
made g/100 ml in these diluents. All counts were performed using
petrifilms (3M aerobic plate count, 3M coliform count, and 3M
yeast and mold count plates, 3M microbiology, St. Paul, MN). All
inoculated 3M aerobic plate count plates (APC) were incubated at
37
C for 48 h (AOAC official method 990.12); for the psychro-
trophics count the APC plates were incubated at 4
C for 7 days
(Brasil et al., 2012), all coliform count plates were incubated at 35
C
for 24 h (AOAC official method 991.14), and all 3M yeast and mold
count plates were incubated at 20
C for 7 days (AOAC official
method 997.02). After incubation, colonies were enumerated and
results reported as log CFU/g of sample.
2.6. Coating microscopic examination
Microscopic observations helped evaluate coating uniformity
and adherence to the watermelon’s surface. Samples from each
treatment (0.5, 1 and 2 g/100 g of sodium alginate) and uncoated
controls were analyzed each sampling interval at room tempera-
ture. Coating thickness was measured based on green dye color
illumination (Assorted food color & egg dye, McCormick & Co. Inc.,
MD) that was first introduced in the pectin component of the edible
coating formulation by adding 7.25 ml (175 drops) of blue dye in
one litter of pectin (2 g/100 g) solution. The coating thickness was
imaged using an Inverted Microscope (Nikon Eclipse TS100, Nikon
Instruments Inc., NY) illuminated with a pre-centered 6Ve30W
halogen lamp. Small pieces (cylinders) of the surface of the
coated watermelon fruit were excised with a cork borer (#7, 4-mm
diameter) and a transversal cut made using a stainless steel blade.
Sample surfaces of coated watermelon pieces were observed with
10 magnification lens (numerical aperture 0.25) and the images
were analyzed and recorded with software NIS-Elements BR 3.2
(Nikon Instruments Inc., NY).
2.7. Statistical analysis
For all analyses, determinations were made in triplicate as
independent experiments. Data analysis was performed using SASÒ
software (SAS 9.2. for Windows, 2010). The effect of the antimi-
crobial coating on watermelon quality and shelf life was evaluated.
Differences between variables were tested for significance by one-
way analysis of variance (ANOVA) with a randomized block
experimental design with four treatments for all the quality and
sensory analysis, and three repetitions, respectively. Significantly
different means were separated using Tukey’s test. The statistical
significance was expressed at the 95% significance level (P < 0.05).
3. Results and discussion
3.1. Effect of coating on quality and shelf life of fresh-cut
watermelon
3.1.1. Physical and chemical properties
Application of the antimicrobial edible coating did not (P > 0.05)
affect the pH (5.2  0.3), oBrix (7.57e7.88  0.23), and water
activity of the fruits (0.99  0.01) throughout storage (data not
shown). On the other hand, moisture content of uncoated samples
was lower (P < 0.05) by Day 15 of storage compared to the coated
samples (Table 1). This result correlates with the effect (P < 0.05) of
concentration of alginate in the percentage of weight loss by Day
15, with the coating with 1.0 and 2.0 g/100 g sodium alginate being
very effective (Table 1). This finding is evidence of the benefits of
applying the edible coating to fresh-cut watermelon pieces, mainly
because of the formation of a polymeric barrier, which reduced
12 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15

Table 1 Table 2
Effect of multilayered antimicrobial edible coating formulation on physico-chemical Effect of multilayered antimicrobial edible coating formulation on color parameters
properties of fresh-cut watermelon samples. a* and b* of fresh-cut watermelon samples.

Time (days) Control* Sodium alginate (g/100 g) Time (days) Control* Sodium alginate (g/100 g)

0.5 1.0 2.0 0.5 1.0 2.0

Moisture content (g/100 g w.b.) a*
a a a a a
0 x0.93  0.01 x0.92  0.01 x0.94  0.01 x0.93  0.02 0 x19.50  2.19 x,y16.62a  1.79 x,y16.32
a
 1.40 y14.58a  2.40
a a a a a
1 x0.92  0.01 x0.92  0.01 x0.94  0.01 x0.93  0.02 1 x19.49  2.01 x16.54a  1.82 x,y16.11
a
 1.38 y14.38a  2.32
a a ab
3 x0.92  0.01 x0.93  0.01 x0.93  0.01 ,x0.93a  0.02 3 x19.33
a,b
 2.48 y14.89a,b  1.60 a
y14.80  0.53 y12.22a,b  1.54
b a a,b a ab
7 x0.89  0.02 xy0.92  0.02 y0.93  0.01 y0.92  0.02 7 x16.42  1.44 y,z11.26b  2.22 y13.28
a,b
 1.29 z9.61
b
 0.26
b a a a b
12 x,z0.88  0.01 x,y0.92  0.01 y0.93  0.01 y0.92  0.02 12 x15.83  1.49 y11.50b  1.92 y13.34
a,b
 2.51 y9.84
b
 2.33
b a,b a a a,b
15 x0.88  0.01 x,y0.90  0.01 y0.91  0.01 y0.92  0.01 15 x18.10  1.71 xy12.76b  2.20 b
y,z11.28  1.78 y9.29
b
 1.55
Weight loss (g/100 g) b*
a a a a
0 e e e e 0 x10.55  0.71 y7.15  1.20 y7.22  0.40 y7.08  0.64
a a a a a a a
1 x4.12  2.40 x3.01  1.39 x2.20  0.74 x2.79a  0.59 1 x10.41  0.61 y7.10  1.16 y7.15  0.39 y7.02  0.84
a,b,c
3 x8.42  2.29 x,y5.60a  0.41 y3.76
a
 0.51 y2.49
a
 0.94 3 a
x10.51  0.71 y6.36
a,b
 0.78 y6.21
a
 0.91 y5.04
a,b
 1.11
a,b
7 x7.08  1.48 x,y3.75a  1.19 x,y3.15
a
 2.16 y1.92
a
 0.94 7 x9.65
ab
 0.61 y3.77 
c
1.71 y5.68a,b  0.62 b
y3.95  1.07
b,c a a a c
12 x12.31  4.09 y3.48a  0.31 w3.95  1.86 y3.36  0.98 12 x8.16  1.57 y4.15  2.22 x,y6.34a,b  1.80 b
y4.33  1.71
c b a a a b,c b b
15 x14.29  1.06 w10.00  1.14 z5.53  1.04 z3.37  1.95 15 x9.49  2.03 xy4.68  0.84 y4.30  0.48 y3.47  0.24
Texture (maximum force in N)
a a a a All samples were stored at 4
C for 15 days.
0 x,y31.30  7.03 x31.10  5.23 x41.83  3.05 x42.20  4.02 a,b
a a a a Values within a column followed by a common superscript letter indicate that
1 x,y31.28  7.52 x31.09  4.87 y,x41.90  4.52 z42.32  4.88
a a a a mean values are not significantly different (P < 0.05).
3 x27.74  3.62 x28.46  0.01 y41.52  2.49 ,y41.55  6.67
a,b x,y Values within a row followed by a common subscript letter indicates that mean
7 x23.51  3.00 x26.19a  2.28 y37.24
a
 4.55 y36.00
a
 3.89
a,b values are not significantly different (P < 0.05).
12 x23.18  4.70 x27.44a  6.53 y35.88
a
 1.23 y35.97
a
 3.24
b a a a *Controls were uncoated fruits.
15 x16.65  1.66 x22.23  4.48 y34.92  5.09 y36.03  1.72

All samples were stored at 4
C for 15 days.

a,b
Values within a column followed by a common superscript letter indicate that
mean values are not significantly different (P < 0.05).
x,y Values within a row followed by a common subscript letter indicates that mean
values are not significantly different (P < 0.05).
*Controls were uncoated fruits.
water loss from fresh-cut samples as found with other fruits (Brasil
et al., 2012; Brecht, 1995; Mantilla, Castell-Perez, Gomes, & Moreira,
2012; Martinon, 2011).
As expected, in general, coated samples were firmer (P < 0.05)
than the uncoated controls by Day 15 of storage (Table 1). On Day 1,
the average firmness of fruits coated with the 1 and 2 g/100 g
sodium alginate formulations was preserved (P < 0.05) in contrast
with the controls, due to the firming action of the calcium lactate.
Studies on pears and other fruits showed that calcium interacts
with cell wall components esuch as pectins ethus maintaining the
structure of the cell wall longer (Alandes, Perez-Munuera, Llorca,
Quiles, & Hernando, 2009; Dong, Wrolstad, & Sugar, 2000) This
finding confirms the benefits of using this multilayered antimi-
crobial edible coating to preserve fresh-cut watermelon firmness.
Application of the coating affected (P < 0.05) the color of fresh-
cut watermelon pieces. The uncoated controls were more reddish
softening of fresh-cut watermelon, as previously supported by the
firmness results.
3.1.2. Consumer acceptance test
Sensory results are shown in Table 4. Panelists gave lower
(P < 0.05) color scores to all samples throughout storage time. By
Day 13, there was a significant difference between controls and the
coated samples with those coated with 2 g/100 g sodium alginate
receiving scores lower than 5.0, mostly because this coating was
whitish in color. In contrast, the panelists ranked the samples with
0.5 g/100 g and 1 g/100 g sodium alginate coating higher than the
controls. This finding supports the observations made in this study
based on the color measurements (Table 2).
In terms of odor, panelists clearly differentiated (P < 0.05)
between the control and the coated samples and showed a prefer-
ence for the uncoated samples and those with the 1 g/100 g sodium
alginate coating until Day 7 while the samples with the 2 g/100 g
Table 3
Effect of multilayered antimicrobial edible coating formulation on headspace CO2
and O2 concentration (%) during storage at 4
C for 15 days.

in color with higher (P < 0.05) a* values compared to the coated Time Control* 0.5 g/100 g 1 g/100 g 2 g/100 g
samples (Table 2), most likely because the coating had a whitish (days) Na-alginate Na-alginate Na-alginate
color. The average b* values were significantly higher for controls CO2 (%)
while the values decreased (P < 0.05) for all the coated samples 0 e e e e
a d c d
1 x1.40  0.28 x1.80  0.14 x1.20  0.01 x1.35  0.07
throughout storage. No significant differences in lightness (L* d d,c b,c c
3 x2.80  0.14 x2.85  0.07 y1.95  0.07 x2.60  0.14
values) were observed for all samples by Day 15 (data not shown). 7 x5.15
c
 0.49 x4.60
b,c
 0.57 x3.90
a,b
 0.14 x4.00
b
 0.14
Coating formulation did not (P > 0.05) affect headspace 12 x8.40
b
 0.28 y5.95
a,b
 0.49 y5.40 
a
0.99 y5.55
a
 0.35
a a a a
composition (Table 3). However, the application of the multilayered 15 x12.5  1.06 y7.80  0.99 y5.70  0.99 y6.25  0.07
antimicrobial edible coating to fresh-cut watermelon helped to O2 (%)
0 e e e e
inhibit the production of CO2 during the respiration process since 1 a
 a
 a
 a

x20.2 1.13 x19.4 0.01 x19.6 0.07 x19.5 0.01
the jars containing uncoated controls had higher (P < 0.05) CO2 3 x18.2
a,b
 0.57 x18.3
a
 0.35 x18.7
a
 0.57 x18.0
a,b
 0.57
concentrations (and lower O2 concentrations) by Day 15. This is due 7 b
x16.4  0.35 x17.7
a
 0.21 x17.1
a,b
 0.78 x17.1
a,b
 1.13
c b a,b a,b
to the coating acting as a barrier to the gases needed for the 12 y11.6  0.85 x,y15.3  0.92 y16.6  0.85 x15.9  1.06
c b b b
15 y8.95  0.23 x13.8  0.85 x15.2  1.20 x15.6  1.27
respiration process. Similar results were obtained with studies on
a,d
cantaloupe (Martinon, 2011), papaya (Brasil et al., 2012) and pine- Values within a column followed by a common superscript letter indicate that
apple (Mantilla et al., 2012), among others. Low O2 levels and high mean values are not significantly different (P < 0.05).
x,y Values within a row followed by a common subscript letter indicates that mean
CO2 accumulation in the internal atmosphere enhances softening of values are not significantly different (P < 0.05).
fruits (Rojas-Grau, Soliva-Fortuny, & Martin-Belloso, 2009). Hence, *Controls were uncoated fruits stored at 4
C up to 15 days. Values are means of two
the multilayered antimicrobial edible coating aided in delaying the replications.
 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15 13

Table 4
Effect of multilayered antimicrobial coating formulation (g/100 g sodium alginate)
on selected sensory attributes of fresh-cut watermelon stored at 4
C for 15 days.

Time (days) Control* 0.5 g/100 g 1 g/100 g 2 g/100 g

Color
a a a a
1 y7.28  1.25 y6.45  1.55 y6.63  1.67 x5.08  1.79
a a a a
3 y 7.10  1.71 y6.20  1.71 y6.63  1.66 x4.90  1.77
a a a a,b
7 y,z6.70  1.49 y6.03  1.07 z7.00  1.11 x4.57  1.51
b a a b
13 y5.09  1.29 z6.34  1.51 z6.28  1.73 x3.84  1.50
b a a ab
15 x4.77  1.37 y6.35  1.37 y6.16  1.16 x4.26  1.48
Odor
a b a a
1 y7.00  1.24 x6.23  1.74 x5.90  1.52 x5.55  1.78
a b a a
3 y6.95  1.50 x5.67  1.71 xy6.20  1.56 x5.35  1.64
a b a b
7 z 6.23  1.25 y5.63  1.76 yz5.93  1.48 x 4.33  1.51
b b a a,b
13 x,y5.06  1.28 y6.70  1.09 y5.69  1.79 x4.75  1.36
b b a a,b
15 x4.84  1.02 z4.77  1.37 y,z5.55  0.81 x,y5.03  1.33
Flavor
a a a a
1 y8.15  0.70 x6.13  1.64 x6.08  1.49 x5.75  1.43
a a a a
3 z8.13  0.99 x,y5.70  1.76 y6.25  1.51 x5.30  1.98
b a a a,b
7 y7.27  0.99 x5.60  1.27 x5.90  1.71 x5.27  1.65
c a a b
13 x,y4.88  1.33 y5.84  1.63 y5.25  2.03 x4.22  1.74
c a a a,b
15 x4.65  1.39 y5.71  1.56 y5.71  1.02 x,y4.90  1.64
Texture Fig. 2. Effect of multilayered edible coating (containing 0 (control) C, 0.5 -, 1 :, or 2
a a a a
1 y7.80  0.99 x6.80  1.30 x6.18  1.39 x6.13  2.11 A g/100 g sodium alginate coating) on the growth of aerobics on fresh-cut water-
a b a a
3 y7.95  0.88 x6.23  1.78 x6.03  1.54 x5.83  1.63 melon samples stored at 4
C for 15 days.
b a a a
7 x6.27  1.18 x6.13  1.48 x6.63  1.25 x5.87  1.35
c a a a
13 x4.94  1.71 y6.00  1.71 y5.97  1.83 x,y5.75  1.28
c a a a
15 x 4.42  1.37 y6.16  1.42 y6.29  1.20 y5.68  1.65
Overall quality microorganisms increased (P < 0.05) with time for both controls
a a a,b a
1 y7.85  0.80 x6.35  1.55 x6.10  1.32 x5.68  1.42
a a a,b a
and coated samples, mainly due to the high moisture content of
3 y7.78  0.95 x5.95  1.54 x6.20  1.57 x5.35  1 72
7 b
 a
 a
 a,b

watermelon, though the coating (1 and 2 g/100 g sodium alginate
z6.97 0.97 y6.17 1.13 y,z6.37 1.37 x5.20 1.44
13 x,y4.88
c
 1.37 z5.94
a
 1.60 y,z,5.34
b
 1.93 b
x4.28  1.62 formulations) effectively inhibited their growth by Day 15 (Fig. 2). It
c a a,b a,b
15 x4.45  1.28 y5.77  1.30 y5.81  0.97 x4.77  1.33 is worth noticing that other types of microorganisms such as
a,b
Values within a column followed by a common superscript letter indicate that Pseudomonas must have also grown in the samples, because the pH
mean values are not significantly different (P < 0.05). remained constant throughout the shelf life study (Raybaudi-
x,y Values within a row followed by a common subscript letter indicates that mean Massilia, Mosqueda-Melgar, Soliva-Fortuny, & Martin-Belloso,
values are not significantly different (P < 0.05).
2009).
*Controls were uncoated fruits stored at 4
C. Nine-point hedonic scale: 1 ¼ dislike
extremely; 9 ¼ like extremely. n ¼ 40.
Although not significantly different, the samples coated with
2 g/100 g sodium alginate formulation had the lowest counts of
psychrotrophic microorganisms, with an approximate decrease of
sodium alginate coating received scores lower than 5.0. This result 1.30 logs. Furthermore, the growth of these microorganisms
is explained by the fact that the coated the samples had a particular was significantly inhibited by Day 7 in samples coated with
cinnamon odor imparted by the trans-cinnamaldehyde. In contrast, the 1 g/100 g (3.4 log CFU/g) and 2 g/100 g (3.04 log CFU/g)
the panelists found both the samples with 0.5 and 1 g/100 g sodium sodium alginate formulations compared to the control (4.62 log
alginate coating acceptable (scores > 5.0) by Day 15 of storage. CFU/g).
Similar results were obtained for flavor. Controls received the
highest values for flavor until Day 7 of storage. However, there was
no significant difference in the scores for the control and all coated
samples by Day 13 of storage. By Day 15, samples with 0.5 and 1 g/
100 g sodium alginate coating received scores significantly higher
than uncoated controls.
The panelists preferred the firmness of the control samples until
Day 7 of storage when these samples had significantly lower scores
(<5.0). On the other hand, panelists found all the coated samples
acceptable (scores > 5.0) until Day 15 of storage. As shown by the
firmness measurements (Table 1), the controls became softer
(P < 0.05) throughout storage and the panelists noticed that. In
terms of overall quality, the samples coated with 0.5 and 1 g/100 g
sodium alginate formulations were acceptable to the consumers by
Day 15 while the control and samples with the 2 g/100 g coating
were unacceptable (<5.0). These lower scores were due to the
problems with color and odor described above. In summary, the
consumer acceptance test highlights the benefits (texture and fla-
vor) of applying the coating with 0.5 and 1 g/100 g sodium alginate
formulations.

3.1.3. Microbiology Fig. 3. Effect of multilayered edible coating (containing 0 (control) C, 0.5 -, 1 :, or 2
Growth curves for all the microorganisms tested in this study A g/100 g sodium alginate coating) on the growth of pyschrotrophs on fresh-cut
are shown in Figs. 2e5. As expected, the population of aerobic watermelon samples stored at 4
C for 15 days.
14 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15
Fig. 4. Effect of multilayered edible coating (containing 0 (control) C, 0.5 -, 1 :, or 2
A g/100 g sodium alginate coating) on the growth of coliforms on fresh-cut water-
melon samples stored at 4
C for 15 days.
The coatings with 1 and 2 g/100 g sodium alginate were very
effective (P < 0.05) in inhibiting the initial population of coliforms
(Fig. 4). By Day 12, application of 1 g/100 g alginate coatings
significantly inhibited the growth of coliforms by approximately
1.5 log CFU/g compared to the control (5.85 log CFU/g).
Application of the coating had a drastic effect (P < 0.05) in the
growth of yeasts and molds (Fig. 5). The control had significantly
higher counts (5.45 log CFU/g) compared to all the coated samples,
with a reduction of almost 4.0 log CFU/g in the fruits coated with
the 2 g/100 g sodium alginate formulation; however, there was no
significant difference in counts between the samples coated with 1
and 2 g/100 g sodium-alginate formulations.
Furthermore, if a maximum tolerable level of microorganisms is
set as 5 log CFU/g (Gilbert et al., 2000), the uncoated fruit will only
last 7 days at 4
C while all the coated samples will have an
extended shelf life between 12 and 15 days, particularly these
samples coated with the 1 and 2 g/100 g sodium alginate formu-
lations. These results suggest that application of the multilayered
Fig. 5. Effect of multilayered edible coating (containing 0 (control) C, 0.5 -, 1 :, or 2
A g/100 g sodium alginate coating) on the growth of yeast and molds on fresh-cut
watermelon samples stored at 4
C for 15 days.
Fig. 6. Coating thickness of fresh-cut watermelon sample coated with 1 g/100 g
sodium alginate multilayer edible coating (cross-sectional view). Image was obtained
using an inverted microscope Nikon Eclipse TS100, 10 magnifications.
edible coating is an effective means to inhibit the growth of
spoilage microorganisms in fresh-cut watermelon. A previous
study proved that the natural antimicrobial compound used (trans-
cinnamaldehyde) is effective against the growth of pathogens such
as Salmonella and Listeria monocytogenes (Gomes et al., 2011).
Hence, the application of the coating in fresh-cut fruits like
watermelon could further ensure their safety.
3.2. Coating characteristics
Microscopic analysis (Fig. 6) shows that the coating with 0.5 g/
100 g sodium alginate, 2 g/100 g pectin and 2 g/100 g natural
antimicrobial, was homogenous and showed good adherence
throughout the entire surface of the fruit (132 mm  1.0). The
thickness of the coatings with 1 g/100 g and 2 g/100 g sodium
alginate showed more variability (180 mm  of 2.0 and
412 mm  0.9, respectively, figures not shown). Thus, coating
thickness is affected by the alginate concentration as reported by
a previous study (Tapia et al., 2008).
4. Conclusion
The alginate-based multilayered antimicrobial coating did not
affect the pH and oBrix of fresh-cut watermelon throughout
storage, and only a slight difference was observed in the color of
coated samples when compared to the controls. Coated samples
had significantly less weight loss and their texture was well
preserved. This is an important result since texture is one of the
main sensory attributes in fresh-cut fruits. Consumer panelists
accepted the coated fruit as long as it did not influence the color,
odor, and flavor attributes. The flavor of the 1 g/100 g alginate-
based coated samples was acceptable to panelists up to Day 13,
while the control sample showed signs of decay.
Microbiological analyses demonstrated the effectiveness of the
coating as a carrier of natural antimicrobial compounds and
the effectiveness of the compound against microbial growth. The
effectiveness against psychrotrophics, coliforms, and yeast and
molds was particularly significant. The recommended coating
formulation to maintain the watermelon’s original quality attri-
butes while extending its shelf life is 1 g/100 g of alginate, 2 g/
100 g of natural antimicrobial complex (trans-cinnamaldehyde)
and 2 g/100 g of pectin.
 R.E. Sipahi et al. / LWT - Food Science and Technology 51 (2013) 9e15
Acknowledgments
We thank Mr. Alex Puerta-Gomez, Research Assistant, depart-
ment of Biological and Agricultural Engineering, for his assistance
with the microscopic analysis of the coatings.
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