Pectin-based edible coatings formulated with pomegranate

peel extracts and other antibrowning agents to extend shelf

life of fresh-cut ‘Rojo Brillante’ persimmon
V. Taberner1, E. Sanchı́s1, M. Mateos2, L. Palou1 and M.B. Pérez-Gago1,a
1Centro de Tecnologı́a Poscosecha (CTP), Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada
(Valencia), Spain; 2Universidad CEU-Cardenal Herrera, 46113 Moncada (Valencia), Spain.

t (PG) and (4) 15 g kg-1 PG plus 15 g kg-1 CaAsc. Persimmon slices were dipped in the pectin-based edible coatings or in water (contro
atments and fruit were considered neither soft nor firm. Overall, only persimmon slices treated with the coating containing PG and C

Keywords: minimally processed persimmon, antioxidant edible coatings, colour, sensory

quality, natural extracts, Punica granatum

INTRODUCTION
‘Rojo Brillante’ persimmon is the most important cultivar in the Ribera del Xú quer
area (Valencia, Spain) and its cultivation is experiencing an important increase in the last
years. When harvested, it is an astringent cultivar, but the application of high levels of CO 2
allows the removal of astringency without affecting fruit firmness, which enables the fruit to
be commercialized as a fresh-cut commodity. However, physical damage during processing
leads to major physicochemical changes that reduce the product shelf life, enzymatic
browning and softening being the main causes of quality loss (Sanchı́s et al., 2015).
The use of anti-browning agents based on citric acid (CA) or ascorbic acid, combined
with low temperature storage is probably one of the main approaches to extend the shelf life
of fresh-cut fruits. Nowadays, edible coatings are gaining importance as they provide a semi-
permeable barrier to gases and water vapour and their protective function may be enhanced
with the addition of antioxidants and other active ingredients. In recent previous studies by

a
E-mail: perez_mbe@gva.es

Acta Hortic. 1194. ISHS 2018. DOI 10.17660/ActaHortic.2018.1194.125

887
Proc. VIII International Postharvest Symposium:
Enhancing Supply Chain and Consumer Benefits – Ethical and Technological Issues
Eds.: F. Artés-Hernández et al.
our group, a pectin-based edible coating amended with citric acid and CaCl 2, as antioxidant
and firming agent, and nisin as antimicrobial agent prevented enzymatic browning of fresh-
cut persimmon ‘Rojo Brillante’ and extended the commercial shelf life up to 8 days of
storage (Sanchı́s et al., 2016). However, the combination did not improve the firmness
of coated persimmon slices compared to control samples. Thus, further studies with other
calcium salts, such as calcium ascorbate (CaAsc), calcium lactate, calcium propionate, etc.,
would be appropriate. Among different calcium salts, calcium ascorbate (CaAsc) has been
reported to reduce browning and softening of minimally processed eggplants (Barbagallo et
al., 2012) and apples (Aguayo et al., 2010).
Recent works devoted to find natural alternative compounds with antioxidant
properties have explored a wide range of plant extracts as potential inhibitors of enzymatic
browning (Saisung and Theerakulkait, 2011; Wessels et al., 2014). These extracts are
characterized by high phenolic contents that act as competitive and non-competitive
inhibitors of the enzyme polyphenol oxidase (PPO). Pomegranate peel, which is the major
waste from pomegranate processing industries, has been reported as an abundant source of
phenolic compounds (Fischer et al., 2011; Tehranifar et al., 2011). However, as far as we
know, pomegranate peel extracts have not been investigated for enzymatic browning
inhibition of fresh-cut fruits. Therefore, the aim of this work was to study the antioxidant
activity and the effects on the physicochemical and sensory quality of fresh-cut ‘Rojo
Brillante’ persimmon of an aqueous pomegranate peel extract incorporated into a pectin-
based edible coating and compare these effects with those of other antibrowning agents.

MATERIALS AND METHODS

Pomegranate peel extract preparation

Pomegranate peel extract (PG) was obtained from pomegranates (Punica granatum L.
‘Mollar de Elche’) cultivated in Elche (Alicante, Spain) and harvested at commercial maturity.
Fruit were washed and peeled by hand. The peel was dried in a hot air oven at 55°C until
constant weight (ca. 48 h). After drying, the peel was ground into a fine powder with an
impact mill (Fritsch Pulverisette 14.702, Germany) and stored at -80°C until use. For the
extraction, 25 g of powder were mixed with 100 mL of water and shaken in an orbital
shaker at 200 rpm for 1 h at room temperature. The solution was centrifuged at 18,514.08 G
for 20 min at 4°C and vacuum-filtered through Whatman nº41 filter paper in a Buchner
funnel. A second extraction for 1.5 h shaking time was done and filtered extracts were mixed
and frozen at -80°C until use.

Edible coatings formulation

The edible coating was prepared from a base solution of apple pectin (Sigma-Aldrich,
St. Louis, MO, USA) at 10 g kg-1 for 60 min. The pectin was emulsified with 2.5 g kg -1 oleic
acid (Panreac Quı́mica, S.A., Barcelona, Spain) and 2.5 g kg -1 Tween 80 (Sigma-Aldrich).
Glycerol (Panreac Quı́mica) was added as plasticizer at 10 g kg-1. Treatments consisted of
the pectin-based coating amended with the following antibrowning agents: (1) 15 g kg -1
calcium ascorbate (CaAsc); (2) 10 g kg -1 citric acid (CA) plus 15 g kg-1 CaAsc; (3) 15 g kg-1
aqueous pomegranate peel extract (PG); and (4) 15 g kg -1 PG plus 15 g kg-1 CaAsc. Citric acid
(CA), was supplied by Quimivita (Barcelona, Spain) and CaAsc was acquired from Sigma-
Aldrich. The coating emulsions were kept at 5°C until application.

Plant material and samples preparation

‘Rojo Brillante’ persimmons were provided by the ‘Denominació n de Origen Kaki
Ribera del Xuquer’ (Valencia, Spain). Astringency was eliminated by placing fruit in closed
chambers at 20°C with 95% CO2 for 24 h. After removal from the chambers, fruit were
stored in air at 5°C for 1 day until processing. Persimmons were sanitized in 150-mg L -1
NaClO solution for 2 min, rinsed with tap water, and dried prior to cutting operations.
Persimmons were peeled, cut into wedges with a sharp stainless-steel knife to reduce
mechanical bruising and dipped in the pectin-based coatings or in water (control) for 3 min.
After

888
dipping, persimmon pieces were removed and left to dry at 5°C. Four persimmon pieces
(115±10 g) were placed into polypropylene trays and sealed with perforated
polypropylene- polyethylene terephthalate film (64-μm thickness) (P12-2050PXNP ILPRA
systems, Españ a
S.L. Mataró, Spain). To study only the effect of the edible coating and ensure that the
atmosphere within the trays was not modified, the film was perforated with a needle (four
perforations, 1 mm in diameter). Three trays per treatment and sampling day were stored
up to 9 days at 5°C.

Quality and sensorial evaluation

Colour measurements were made periodically with a Minolta chroma meter (Model
CR-300, Ramsey, NY, USA) on 12 persimmon pieces per treatment using the CIE L*, a*, b*
coordinates. Firmness was measured on 12 persimmon slices as the force (N) required for
an 8-mm diameter rod to penetrate the sample to a depth of 2 mm at a speed of 5 mm s -1
(Instron Universal Machine, Mod. 3343). Polyphenol oxidase (PPO) activity was determined
by adding 3 mL of 0.05 M 4-methylcatechol to 100 μL of the enzyme extract (Sanchı́s et al.,
2016). The extract was obtained by mixing 15 g of fresh persimmon with a McIlvaine buffer
solution (1:1) at pH 6.5, containing 1 M sodium chloride and 5% polyvinylpolypyrrolidone.
Changes in absorbance were determined at 420 nm every 5 s for up to 2 min in a
spectrophotometer (UV-1, Thermo Electron Corporation, UK). Each treatment was
measured in triplicate. One unit of PPO activity was defined as a change in absorbance of
0.001 min-1 mL-1 enzymatic extract immediately after extract addition. The sensory
evaluation included visual and taste assessment. General visual quality was evaluated based
on the following scale: 9 = excellent, just sliced; 7 = very good; 5 = good, limit of
marketability; 3 = fair, limit of usability; and 1 = poor, inedible (Gorny et al., 2002). The effect
of the coatings on taste included the assessment of overall quality (a 9-point scale, where 1
represented a poor quality and 9 an excellent quality) and off-flavour development (from 1
= absence to 5 = marked presence). Sensory firmness was evaluated according to the
following scale: 5 = very firm; 3 = neither firm nor soft; and 1 = very soft. Samples for taste
analysis were presented to the judges in a random order, labelled with 3-digit codes and
served at room temperature (20±1°C) in separate cabins with colour lights to avoid
discrimination of samples due to colour differences. Visual evaluation was performed under
white lights at 5°C. The sensory panel consisted of 15 trained members.

Statistical analysis
Statistical analysis was performed using Statgraphics Plus v. 5.1 (Statpoint
Technologies Inc., Warrenton, VA, USA). Specific differences between means were
determined for each evaluation date by the least significant difference (LSD) test at P≤0.05
applied after a one-way analysis of variance (ANOVA). Results are presented as
mean±standard deviation.

RESULTS AND DISCUSSION

Colour change of fresh-cut persimmon

Table 1 shows the effect of the pectin-based edible coatings on lightness (L*) and a*
values of fresh-cut ‘Rojo Brillante’ persimmon. Coated samples presented lower a* values
than control samples, which indicated an effective browning inhibition on fresh-cut
persimmon during storage. No significant differences were observed among coatings.
However, only the coatings amended with PG, alone or in combination with CaAsc, showed
higher L* values than the control throughout the 9 days of storage. Barbagallo et al. (2012)
also reported lower L* values on fresh-cut eggplants dipped in calcium salts (CaAsc or
calcium citrate) than on untreated samples. Whereas, Gomes et al. (2010) showed that dips
in calcium lactate or calcium proprionate enhanced browning of fresh-cut pears under acidic
conditions. Similarly, Ghidelli et al. (2013) reported a decrease in L* on fresh-cut persimmon
as ascorbic acid concentration was increased, suggesting an oxidative damage of the tissue
with cell disruption and subsequent decompartmentalization of enzymes.
Table 1. Colour L* and a* values of fresh-cut ‘Rojo Brillante’ persimmons dipped in water
(control) or pectin-based coatings (PC) amended with antibrowning agents and
stored at 5°C for up to 9 days.
Storage time (days)
Treatments
0 2 5 7 9
L* PC_15 g kg-1 CaAsc 68±2z 64±2z 60±3y 58±3y 57±3x
PC_10 g kg-1 CA + 15 g kg-1 CaAsc 66±2z 60±2y 57±4x 54±1x 53±4w
PC_15 g kg-1 PG 67±2z 64±4z 63±4z 62±4z 60±2zy
PC_15 g kg-1 PG + 15 g kg-1 CaAsc 68±2z 65±2z 64±2z 62±2z 61±3z
Control 66±3z 62±2y 58±3yx 57±2y 58±2yx
a* PC_15 g kg-1 CaAsc 7±2zy 7±1y 6±2y 7±1yx 6±1x
PC_10 g kg-1 CA + 15 g kg-1 CaAsc 8±2z 7±1y 7±2y 9±2y 7±2y
PC_15 g kg-1 PG 7±2zy 7±2y 7±1y 7±2x 8±1y
PC_15 g kg-1 PG + 15 g kg-1 CaAsc 6±2y 7±1y 7±2y 6±1x 7±2y
Control 8±1z 9±1z 10±2z 11±2z 9±1z
CaAsc: calcium ascorbate; CA: citric acid; PG: aqueous pomegranate peel extract.
In each column, different letters indicate significant differences among treatments by LSD test ( P≤0.05).
Shown values are mean±standard deviation (n=12).
Several research works describe the antibrowning effect of different plant extracts in
both in vitro and in vivo studies (Saisung and Theerakulkait, 2011; Zocca et al., 2011;
Wessels et al., 2014). This has been attributed to the antioxidant capacity of plant secondary
metabolites acting as reducing agents and/or to phenolic compounds acting as competitive
or non-competitive PPO inhibitors (Wessels et al., 2014). Thus, extracts from pomegranate
arils reduced browning of stem artichoke slices and this was related to the inhibition of PPO
activity (Zocca et al., 2011). However, Wessels et al. (2014) reported that despite the high
antioxidant capacity of a ‘superberry’ fruit extract obtained from seven different berries,
with grapes and pomegranates as main ingredient sources, no PPO inhibition was observed
on apple slices. Hence, the browning inhibition capacity of this extract was explained by the
presence of proanthocyanidins acting as reducing agents, whereas catechin, gallic acid,
chlorogenic acid and ellagic acid, all present at high contents, could have acted as PPO
substrates rather than as competitors. In our work, only the coatings containing 15 g kg -1
CaAsc significantly reduced PPO activity compared to uncoated samples during 5 days of
storage at 5°C, while PG amended coatings had no significant effect (data not shown). These
results would confirm that the antibrowning effect of PG from fruit peel is based on the
reduction of o-quinones to their uncoloured precursors diphenols. The differences between
the lack of effectiveness of PG from peel to inhibit persimmon PPO activity and the
effectiveness of a PG from arils to inhibit artichoke PPO activity reported by Zocca et al.
(2011) could be related to differences in ellagic and gallic acid content and/or differences in
PPO substrate. The quantification of individual phenolics in pomegranate peel and juices
showed values two orders of magnitude higher in peel than in juice (Fischer et al., 2011).

Fruit firmness
Table 2 shows the effect of the pectin-based edible coatings on instrumental firmness
of fresh-cut ‘Rojo Brillante’ persimmon. Coatings formulated with PG, alone or in
combination with CaAsc, and with CaAsc alone, but not with CaAsc and CA, significantly
reduced firmness loss of persimmon slices with respect to control samples during 5 days of
storage at 5°C. The role of calcium salts in the reduction of tissue softening is well-known.
Calcium ions maintain the cell wall structure in fruits by interacting with the pectins in the
cell wall to form calcium pectate, which assists molecular bonding between constituents of
the cell wall (Poovaiah, 1986). However, several works have described that the use of
additives that alter the surface pH of fresh-cut products (e.g., citric and ascorbic acid) does
not only affects the PPO activity, but it also modulates cell wall metabolism and texture
(Pinheiro and Almeida, 2008; Gomes et al., 2010). These authors showed that acidification
can be detrimental to texture of fresh-cut products by increasing water solubility of pectins.
For instance, pectin solubilisation was higher in pear slices dipped in solutions at pH 3.0
than in slices treated at pH 7.0 (Gomes et al., 2010), and in tomato pericarp disks dipped in
solutions at pH 4.0 than at pH 7.0 (Pinheiro and Almeida, 2008). Furthermore, the
effectiveness of different calcium salts to prevent softening of fresh-cut pears was highly pH-
dependent and the use of acidic solutions with CaAsc, lactate, or propionate was not
recommended to prevent tissue softening (Gomes et al., 2010).

Table 2. Firmness (N) of fresh-cut ‘Rojo Brillante’ persimmons dipped in water (control) or
pectin-based coatings (PC) amended with antibrowning agents and stored at 5°C
for up to 9 days.
Storage time (days)
Treatments 0 2 5 7 9
PC_15 g kg-1 CaAsc 30±14z 33±8z 31±9z 21±9z 21±6z
PC_10 g kg-1 CA + 15 g kg-1 CaAsc 30±14z 25±6y 22±8y 21±5z 19±4z
PC_15 g kg-1 PG 30±14z 31±9zy 33±9z 24±4z 21±7z
PC_15 g kg-1 PG + 15 g kg-1 CaAsc 30±14z 36±8z 31±10z 23±6z 24±6z
Control 30±14z 26±10y 20±8y 22±5z 20±7z
CaAsc: calcium ascorbate; CA: citric acid; PG: aqueous pomegranate peel extract.
In each column, different letters indicate significant differences among treatments by LSD test (P≤0.05).
Shown values are mean ± standard deviation (n=12).

Sensorial quality
The application of edible coatings did not affect the overall flavour of fresh-cut ‘Rojo
Brillante’ persimmons, which were evaluated at the end of the 9 days of storage at 5°C as of
good quality (scored as 7). Likewise, the coatings did not induce off-flavours in the samples
(scored as 1). Furthermore, treated fruit were considered neither soft nor firm (scored as 3)
(data not shown).
Visual quality of persimmon slices, based on colour and general appearance, was
evaluated by a trained sensory panel. Figure 1 represents the time in which fresh-cut ‘Rojo
Brillante’ persimmon reaches the limit of marketability (scored as 5) for the different
treatments. Uncoated samples were evaluated below the limit of marketability even by day
1 of storage. The coatings containing PG or CaAsc as single antibrowning agents were
evaluated as having a good visual quality during the first 2 days of storage, but the limit of
marketability was not further extended. The combination of antibrowning agents increased
the commercial shelf-life, the coating amended with PG and CaAsc being the most effective,
with a limit of marketability of 7 days of storage at 5°C.

CONCLUSION
The application of the pectin-based coating formulated with the combination of CaAsc
(15 g kg-1) and aqueous PG (15 g kg-1) was the most effective treatment to extend the shelf-
life of fresh-cut ‘Rojo Brillante’ persimmon since it reduced enzymatic browning and
maintained flesh firmness for 7 days of storage at 5°C, without negatively affecting fruit
flavour. These results open a potential use to an important waste by-product in the
pomegranate industry, although further studies should be conducted to improve the
performance of these coatings and also to evaluate their antimicrobial properties.
 Treatments

PC_ CaAsc

PC_ CA + CaAsc

PC_ PG

PC_ PG + CaAsc

Control

0 2 4 6 8

Time (days)

Figure 1. Effect of pectin-based coatings (PC) amended with different antibrowning agents
on the marketable shelf-life of fresh-cut ‘Rojo Brillante’ persimmons stored at 5°C.
Shelf-life was defined as the number of days to reach the limit of marketability
(score of 5). CaAsc = calcium ascorbate (15 g kg-1); CA = citric acid (10 g kg-1); PG
= aqueous pomegranate peel extract (15 g kg-1); control = water.

ACKNOWLEDGEMENTS
The ‘Denominació n de Origen Kaki Ribera del Xú quer’ is gratefully acknowledged for
providing fruit. This work was funded by the Instituto Nacional de Investigació n y
Tecnologia Agraria y Alimentaria (INIA) through Project RTA2012-00061-00-00 and the
European Union through the FEDER Program.

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