Impaginato


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Adv. Hort. Sci., 2023 37(1): 15­23                                                                                                        DOI: 10.36253/ahsc­13850

Effects of cold storage on quality para­
meters and nutraceutical compounds 

of pomegranate fruits (cv. Acco) 
 
 
A. Cirillo 1 (*), A. Magri 2, 3, M. Petriccione 2, C. Di Vaio 1 
1     Department of Agricultural Sciences, University of Naples Federico II, 

Via Università, 100, 80055 Portici (NA), Italy. 
2     Research Centre for Olive, Fruit and Citrus Crops, Council for 

Agricultural Research and Economics (CREA), Via Torrino, 3, 81100 
Caserta, Italy. 

3     Department of Environmental, Biological and Pharmaceutical Sciences 
and Technologies, University of Campania Luigi Vanvitelli, Via Vivaldi, 
43, 81100 Caserta, Italy. 

 
Key words: Bioactive compounds, cold storage, Punica granatum L., quality fruit. 
 
Abstract: Punica granatum L. contains several bioactive compounds with 
antioxidant activity that have a positive effect on human health. This study 
aims to investigate the changes in the chemical­physical and qualitative para­
meters of pomegranate fruits cv. Acco from harvest up to +90 days of cold stor­
age (+4°C and 95% RH). Morphological parameters, juice yield, weight loss, 
total soluble solids content (TSS), pH, titratable acidity, the color of the epicarp 
(L*, a*, b*), content of polyphenols, anthocyanins, flavonoids, and antioxidant 
activity were analyzed. The results showed an increase (about 29%) in the juice 
content (%) at +60 days of cold storage. Cold storage has also shown positive 
effects on some bioactive compounds. Flavonoids and anthocyanins content 
increased from 287.98 mg CE/100 ml of juice to 389.23 mg of CE/100 ml of juice 
and from 8.32 to mg/100 ml of juice to 11.13 mg/100 ml of juice at + 90 days of 
cold storage, respectively. On the basis of our results that confirmed the litera­
ture data, the pomegranate fruit is rich in bioactive compounds that exert ben­
eficial actions on human health, and it has also been demonstrated that such 
nutraceutical compounds increased during cold storage, allowing the fruit to be 
preserved a long term. 
 
 
1. Introduction 
 
      The pomegranate (Punica granatum L.) is generally cultivated in the 
Mediterranean Basin, in the regions of Southern Asia, in India and in North 
and South America, where long and hot summer favor an optimal fruits 
ripening (Erkan and Dogan, 2018). Its adaptation to the Mediterranean cli­
mate has favored its spread in several countries giving rise, over the cen­
turies, to numerous local genotypes. The fruits are generally harvested when 
fully ripe and displayed a smooth shining leathery skin with a color varies 
from green, to pink, reddish, or dark red (Love et al., 2014). The pomegran­

(*) Corresponding author:  
aurora.cirillo@unina.it 
 
 
Citation: 
CIRILLO A., MAGRI A., PETRICCIONE M., DI VAIO 
C., 2023 ­ Effects of cold storage on quality para‐
meters and nutraceutical compounds of pome‐
granate fruits (cv. Acco). ­ Adv. Hort. Sci., 37(1): 
15­23. 
 
 
Copyright: 
© 2023 Cirillo A., Magri A., Petriccione M., Di 
Vaio C. This is an open access, peer reviewed arti­
cle published by Firenze University Press 
(http://www.fupress.net/index.php/ahs/) and 
distributed under the terms of the Creative 
Commons Attribution License, which permits 
unrestricted use, distribution, and reproduction 
in any medium, provided the original author and 
source are credited. 
 
 
Data Availability Statement: 
All relevant data are within the paper and its 
Supporting Information files. 
 
 
Competing Interests: 
The authors declare no competing interests. 
 

 
Received for publication 11 October 2022 
Accepted for publication 11 January 2023

AHS 
Advances in Horticultural Science

https://doi.org/10.36253/ahsc-13850
http://www.fupress.net/index.php/ahs/
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Adv. Hort. Sci., 2023 37(1): 15­23

16

ate fruits have a low respiration rate and a non­climac­
teric respiratory pattern (Ben­Arie et al., 1984). The 
edible part of the fruit is called arils and constitutes 
about 52% of total fruit (w/w), comprising 78% juice 
and 22% seeds. The fresh juice contains 85.4% mois­
ture and considerable amounts of total soluble solids 
(TSS), total sugars, reducing sugars, anthocyanins, phe­
nolics, ascorbic acid and proteins (El­Nemr et al., 1990) 
and has also been reported to be a rich source of 
antioxidants (Gil et al., 2000; Kulkarni et al., 2004). The 
pomegranate is highly valued for health­promoting 
benefits of its fruit and processed product as demon­
strated by numerous in vivo and in vitro studies (de 
Nigris et al., 2007; Bell and Hawthorne, 2008; Davidson 
et al., 2009). The consumers pose more attention to 
the nutraceutical value of fruits, in fact several studies 
aim to valorize this aspect, as reported by Graziani et 
al. (2020, 2021). The recent growing awareness of con­
sumers to health aspects of fresh fruits and processed 
products greatly increased the interest in consumption 
of this fruit and its processed products; consequently, 
worldwide pomegranate production expanded consid­
erably. Apart from its demand as fresh fruit and juice, 
the processed products such as carbonated drinks, 
syrup, wine, and candy are also gaining importance in 
world trade (Dak and Pareek, 2014). The fruit juice of 
pomegranate was found to have an exceptionally high 
antioxidative capacity (Reddy et al., 2007; Cirillo et al., 
2022). The shelf­life of pomegranate is about 12­14 
days at ambient conditions (Naveena et al., 2008); but 
cold storage with recommended temperature from 0 
to 10°C can be used from 2 weeks to 5 months influ­
encing in different manner cultivars storability 
(Ehteshami et al., 2019). Storage method is very impor­
tant because physiological and enzymatic processes 
cause the loss of quality with browning of the skin, 
necrotic pitting, pallor of the arils that depreciate the 
product during storage (Fawole and Opara, 2013 a; 
Dorostkar and Moradinezhad, 2022). In the present 
study, cv. Acco pomegranate storability, one of the 
main cultivars marketed in Italy, was analyzed, evaluat­
ing physical­chemical and biochemical changes during 
refrigerated storage up to 90 days from harvest to 
define the maximum storage time so that the fruits are 
still appreciated by consumers both organoleptic and 
nutritional point of view. 

 
2. Materials and Methods 
 
Experimental orchard and sampling 
     The experiment was conducted in 2019 in Eboli 

(Salerno, Italy) (40° 33’ 29” N; 14° 58’ 28” E a 15 m 
a.s.l.) at “Improsta” Regional Experimental Farm, 
where were cultivated the pomegranate trees of cv. 
Acco. The trees were trained to sapling system and 
spaced 3.5 m on the rows and 4 m between the rows.  
Pomegranate fruits were hand­harvested 3th 
September 2019 according to a randomized block 
design from homogeneous trees, for grown and pro­
duction load. Fruits (n=120) were collected, and 12 
lots (10 fruits each) were prepared. Three lots were 
analyzed at harvest while the others were stored in 
the cold chamber at a temperature of 4±1°C and 
95±1% relative humidity, and analyzed after 30, 60 
and 90 days of storage. In this work, 0 indicates the 
harvest time and +30, +60 and +90 the days of refrig­
erated storage. 

Pomological and physico‐chemical traits of fruits 
     Pomological and physico­chemical traits of the 
fruits were carried out at the Pomology Laboratory of 
the Department of Agriculture of the University of 
Naples “Federico II”. Fruit and seed weight (g) was 
determined by an electronic digital balance (Precisa 
Instruments AG, model XB220A, Dietikon, 
Switzerland) to monitor the weight loss of fruits dur­
ing cold storage. Equatorial diameter (mm) and fruit 
length without calyx (mm) were measured by electric 
digital caliper with ±0.01 mm accuracy (Mitutoyo, 
Kawasaki, Japan). Color of epicarp was determined 
with a colorimeter (Minolta, model CR­400, Tokyo, 
Japan) that was capable of quantifying colors accord­
ing to international standards and expressed in 
defined color spaces. The instrument was calibrated 
with “white” managed by the light source on a white 
tile, before each measurement. The L* a* b* (CIELAB) 
color space is the most common method for measur­
ing the color of an object or materials of different ori­
gins and it is widely used in all sectors. In this color 
space, L* indicates brightness, while a* and b* the 
chromaticity coordinates: +a* is the direction of red, 
­ a* is the direction of green, +b* is the direction of 
yellow, and ­ b* is the direction of blue. The measur­
ing was repeated four times in different points of the 
fruit. The pomegranate seeds were hand­separated 
from the epicarp and carpellary membranes, count­
ed, and squeezed using a small press. A juice yield of 
300 g of pomegranate seeds was measured and 
expressed as a percentage (w:v). Juice (200 mL) was 
stored at ­20°C and afterwards used for the evalua­
tion of physico­chemical and nutraceutical traits. The 
pH, titratable acidity (TA) and total soluble solids 
(TSS) were assessed on the arils juice. The TSS con­

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Cirillo et al. ‐ Quality parameters of pomegranate fruits during cold storage

17

tent was determined with a HI 96.814 digital refrac­
tometer of Hanna instruments and results were 
expressed as °Brix. The pH was determined with a pH 
meter by the Hanna Instruments laboratory and total 
acidity was evaluate by acid­base titration, with 0.1N 
sodium hydroxide standard solution and the results 
were expressed as g citric acid 100 mL­1. 

Determination of total phenolic content 
     Total phenolics content in juice was determined 
according to a Folin­Ciocalteu procedure (Singleton 
and Rossi, 1965). The assay was carried out in dupli­
cate for each sample using 5 µL of extract, 100 µL of 
Folin­Ciocalteu reagent and 300 µL of 7.5% (w/v) 
Na2CO3 solution; the mixture assay was left in the dark 
for 2 hours at room temperature, then the absorbance 
was determined at the wavelength of 765 nm The 
results were expressed as mg of gallic acid equiva­
lent/100 mL of juice (mg of GAE/100 mL of juice). 
 
Determination of flavonoids content 
     The assay was carried out according to Zhishen et 
al. (1999) by the aluminum chloride colorimetric 
method using 20 µL of juice and the absorbance was 
determined at 510 nm. Results were expressed as mg 
of catechin equivalent/100 mL of juice (mg CE/100 
mL of juice). 

Determination of anthocyanins content 
     The assay was carried out according to Magri et 
al. (2020) by a pH­differential method using 50 µL of 
juice in KCl pH 1 and CH3COONa pH 4 buffer. The 
absorbances at 510 and 700 nm were determined. 
Results were expressed as mg of cyanidin­3­glucoside 
equivalent/100 mL of juice (mg C3G/100 mL of juice). 

Antioxidant activity 
     The assay was conducted as described by 
Petriccione et al. (2015) using 20 µL of juice and 1480 

µL of 1,1­diphenyl­2­picril­hydrazyl (DPPH). The 
change in absorbance was observed at 515 nm and 
the results were expressed as mg of Trolox equiva­
lent/100 mL of juice (mg Teq/100 mL of juice). 
 
Statistical analysis 
     Analysis of variance (ANOVA) on the complete 
randomized block design on the data and mean sepa­
ration by Duncan’s multiple range test (p<0.05) and 
Principal component analysis (PCA) were carried out 
using XLSTAT, version 2013, statistical software pack­
age (New York, NY, USA). 
 
 
3. Results 
 
Fruit pomological characterization 
     The changes of the pomological parameters in the 
cv. Acco pomegranate fruits during the refrigerated 
storage at + 30, + 60 and + 90 days are shown in table 
1. There is a significant reduction in the fruit weight 
loss about 10.5% already after 30 days of cold stor­
age, up to 23.7% at +90 days, while no significant dif­
ferences are highlighted for the equatorial diameter, 
the fruit length, and the seeds weight. One of the 
most important commercial parameters is the juice 
content in the seeds, our study showed a significant 
increase in the percentage of juice from collection to 
storage in the fridge, after +60 days about 63.5% 
compared to harvest (Table 1). 

Juice quality parameters and fruit color characteriza‐
tion during cold storage 
     The results of the physico­chemical properties of 
cv. Acco during cold storage are shown in table 2. A 
significant reduction in total soluble solids (TSS) was 
highlighted, from 14.25°Brix at harvest to 11.85°Brix 
already at +60 days of refrigerated storage, up to a 

Table 1 ­ Pomological parameters (fruit weight, equatorial diameter, fruit length, seed weight (n=100), % weight loss, % juice) at harvest 
and during the refrigerated storage (+30, +60, +90 days) of pomegranate fruits (cv. Acco). 0 days is the time of harvest

All the data are expressed as mean ± SE (standard error). The same letter indicates not significant differences according to Duncan’s mul­
tiple range test (p<0.05). Level of significance at the ANOVA are indicated as NS (not significant), * (0.01<P <0.05), ** (0.01> P> 0.001), 
and *** (P <0.001).

Days Fruit weight 
(g)

Equatorial  
diameter  

(mm)

Fruit length 
(mm)

Seed weight 
(g)

Weight loss 
(%)

Juice 
(%)

0 216.38 ± 9.86 a 80.38 ± 1.53 a 63.5 ± 4.12 a 113.88 ± 5.76 b 0 ± 0.00 a 47.60 ± 0.47 c

30 187.18 ± 6.68 b 78.63 ± 0.92 a 65.5 ± 0.79 a 111.44 ± 4.86 b 10.5 ± 0.85 b 54.43 ± 1.55 b

60 171.27 ± 8.84 b 80.31 ± 1.35 a 67.0 ± 1.32 a 113.38 ± 6.70 b 19.6 ± 1.76 bc 63.50 ± 1.69 a

90 167.13 ± 7.48 b 80.88 ± 1.15 a 69.56 ± 0.73 a 119.87 ± 4.70 b 23.7 ± 2.10 c 61.35 ± 1.63 a

Significance *** NS NS NS ** *** 



Adv. Hort. Sci., 2023 37(1): 15­23

18

final reduction of about 25.75% at +90 days while the 
pH showed an increase at +60 days of refrigerated 
storage equal to 17.66%. Color coordinates (L*, a*, 
b*) of pomegranate peel during the refrigerated stor­
age are shown in Table 2. L* and fruit peel redness 
(a*) showed a reduction with progressed refrigerated 
storage. The highest a * index was found at harvest 
(47.14), while after + 90 days of refrigerated storage 
this parameter was reduced to 40.03. b* was 26.98 
at harvest while after + 30 days of refrigerated stor­
age a reduction of about 23.42% was observed, with­
out significant differences up to 90 days of storage. 
 
Content of bioactive compounds and antioxidant 
activity during cold storage 
     The current use of pomegranate fruit regards 
especially the nutritional and, still potential, health 
benefits that come out from the various parts com­
posing this one (carpellary membranes, arils, seeds 
and bark). Indeed, the phytochemical composition of 

the fruit abounds in compounds (flavonoids, ellagi­
tannins, proanthocyanins, mineral salts, vitamins, 
lipids, organic acids) presenting a significant biologi­
cal and nutraceutical value. Table 3 shows the bioac­
tive compounds of the pomegranate fruits cv. Acco 
during the refrigerated storage. The polyphenol con­
tent did not display significant differences up to 60 
days of storage, with average values of 272 mg GAE/ 
100 ml of juice, while at 90 days an increase in the 
polyphenol content was shown up to a value of 
389.23 mg GAE/100 mL juice; the flavonoid content 
showed a small increase at 30 days of storage 
(124.58 mg EC/100 mL juice), without statistical sig­
nificance, for the other two periods considered with 
an average content of 103 mg CE/100 mL juice; the 
anthocyanin content displayed a slight decrease up 
to 60 days of refrigerated storage with average val­
ues of about 3 mg C3G/100 mL of juice, followed by 
an increase after 90 days, up to 11.13 mg C3G/100 
mL of juice. The antioxidant activity of pomegranate 

Table 2 ­ Total soluble solids (TSS), titratable acidity (TA), pH and color attributes (L*, a* and b*) at harvest and during cold storage (+30, 
+60 and +90 days) of pomegranate fruits (cv. Acco). 0 Days is the time of harvest

Table 3 ­ Total polyphenol, anthocyanins, flavonoids content and antioxidant activity (D) at harvest and during cold storage (+30, +60 
and +90 days) of pomegranate fruits (cv. Acco). 0 Days is the time of harvest

All the data are expressed as mean ± SE (standard error). The same letter indicates not significant differences according to Duncan’s mul­
tiple range test (p<0.05). Level of significance at the ANOVA are indicated as NS (not significant), * (0.01<P <0.05), ** (0.01> P> 0.001), 
and *** (P <0.001).

All the data are expressed as mean ± SE (standard error). The same letter indicates not significant differences according to Duncan’s mul­
tiple range test (p<0.05). Level of significance at the ANOVA are indicated as NS (not significant), * (0.01<P <0.05), ** (0.01> P> 0.001), 
and *** (P <0.001).

Days TSS 
(Brix°)

TA 
(g citric acid 100 

mL­1)
pH L* a* b*

0 14.25 ± 0.25 a 8.75 ± 0.25 a 3.00 ± 0.00 b 47.08 ± 0.66 a 47.14 ± 0.88 a 26.98 ± 0.49 a
30 13.78 ± 0.09 a 8.10 ± 0.11 a 3.10 ± 0.09 b 40.51 ± 1.68 b 43.47 ± 1.32 b 20.66 ± 0.62 b
60 11.85 ± 0.72 b 6.60 ± 0.14 b 3.53 ± 0.07 a 39.09 ± 0.87 b 44.14 ± 0.55 b 20.63 ± 0.74 b
90 10.58 ± 0.21 c 5.63 ± 0.31 c 3.63 ± 0.07 a 41.21 ± 1.08 b 40.03 ± 0.56 c 20.68 ± 0.75 b
Significance *** *** *** *** *** ***

Days Polyphenols 
(mg GAE/100 mL juice)

Anthocyanins  
(mg C3G/100 mL juice)

Flavonoids 
(mg CE/100 mL juice)

Antioxidant activity 
(µmol TE/ 100 mL juice)

0 287.98 ± 8.86 b 8.32 ± 1.67 a 99.49 ± 3.25 a 271.63 ± 4.13 a
30 285.25 ± 26.07 b 3.53 ± 2.67 b 124.58 ± 9.27 a 268.11 ± 5.56 a
60 244.15 ± 23.92 b 2.72 ± 3.36 b 102.99 ± 1.90 a 267.67 ± 3.49 a
90 389.23 ± 29.01 a 11.13 ± 4.71 a 104.21 ± 10.46 a 267.97 ± 3.21 a
Significance * ** NS NS



Cirillo et al. ‐ Quality parameters of pomegranate fruits during cold storage

19

juice showed no significant differences during the 
refrigerated storage with average values of 268 µmol 
TE/100 mL juice. 
 
Principal component analysis (PCA)  
     To obtain a broad overview of all parameters eval­
uated in cv. Acco fruit following the refrigerated stor­
age a principal component analyses (PCA) was con­
ducted. Figure 1 shows the PCA of the changes of 
nutraceutical compounds and qualitative parame­
ters, during refrigerated storage. The first two princi­
pal components (PCs) disclosed 88.39% of the cumu­
lative variance with PC1 detailing for 55.55% and PC2 
for 32.83%. At harvest this cultivar showed a higher 
antioxidant activity, while a higher flavonoids content 
is shown between +30 and +60 days of refrigerated 
storage, at +90 days of refrigerated storage a higher 
polyphenols and anthocyanins content was highlight­
ed. The qualitative parameters showed higher values 
at +30 days of refrigerated storage for TSS and TA, 
while at + 60 days there was a higher juice yield. 
 

 
 
4. Discussions and Conclusions 
 
     Both fresh market and processing industry drive 
pomegranate consumption, and it is crucial to 
acknowledge all fruit characteristics to not only clas­
sify varieties from a botanical point of view, but also 
to meet the current market demand for quality fruits 
(Martìnez et al., 2006). In the recent years, the 
demand and the consumption of pomegranate fruit 
have shown an extensive growth in several countries 

worldwide becoming a valuable commodity due to 
high levels of nutraceutical compounds and greater 
awareness of health­promoting benefits (Asgary et 
al., 2022). The major postharvest losses can occur 
among harvest and consumption affecting either fruit 
quantity or quality at any stage in the postharvest 
chain. Freshly harvested agricultural products are a 
living thing that breathes and undergoes changes 
during postharvest handling (Kiaya, 2014; Erkan and 
Dogan, 2018; Siddiqui et al., 2022). Postharvest dis­
eases caused by bacteria, fungi, and other microor­
ganisms are compromised factors and a consistent 
problem for long term storage of pomegranate fruits 
(Munhuweyi et al., 2016; Akhila et al., 2022; Gurtler 
and Garner, 2022). These spoilages greatly affect the 
appearance, aroma and taste of fruits and the control 
of postharvest diseases represents the most signifi­
cant economic challenges in agriculture. High water 
and sugars content combined with soft and delicate 
texture make pomegranate fruits susceptible to 
weight loss, mechanical damage, and attack of 
pathogens (Sayarri et al., 2012). Physiological and 
enzymatic disorders are responsible to qualitative 
decay and storage life in pomegranate like fruit wrin­
kling, browning, and drying of skin and seeds, seeds 
paleness and the pathogens often that cause damage 
to the tissues, thereby making the fruit unsaleable 
(Caleb et al., 2012 a; Nazoori et al., 2022). As previ­
ously mentioned, an important aspect is the juice 
content. Our results reported an increase in the juice 
content during refrigerated storage most likely due 
to the “softening” of the tissues of the arils increasing 
the extraction of the juice. At harvest, pomological 
and physico­chemical traits in cv. Acco fruit showed a 
slight difference compared to what is reported by 
Ferrara et al. (2014); these differences may be 
ascribed to different cultural practice, climatic and 
soil conditions (Fadavi et al., 2005; Ferrara et al., 
2014). Weight loss is one of the major problems asso­
ciated with stored pomegranate fruit which cause 
hardening of the skin and browning of the rind and 
seeds (Artés et al., 2000 a; Caleb et al., 2012 b; 
Pareek et al., 2015). Several studies have demon­
strate that weight loss increased with increasing tem­
perature and prolonged storage in different pome­
granates cultivars and it is due to water being lost 
through natural porosity of the skin (Al­Mughrabi et 
al., 1995; Al­Yahyai et al., 2009; Wasker, 2011; 
Fawole and Opara, 2013 b). Our results, on the physi­
co­chemical parameters of pomegranate fruits during 
cold storage, are in agree with those reported in the 

Fig. 1 ­   Principal component analysis (PCA) based on quality 
parameters and bioactive compounds at harvest and 
during cold storage (+30 +60 and +90 days) of 
pomegranate fruits (cv. Acco). 0 is the time of harvest. 



20

Adv. Hort. Sci., 2023 37(1): 15­23

literature where a reduction in TSS and TA is shown. 
According to a similar study on ‘Mollar’ pomegranate 
after only 7 days of storage at 4°C, the TSS content is 
reduced about 11% (Gil et al., 1996), while Artes et 
al. (2000 b) showed a significant reduction in the 
acidity of pomegranate fruit juice in cv. Molla de 
Elche stored at 5°C for 90 days and subsequently, 
held at 20°C for six days. The decrease in TSS content 
could be a result of the degradation of sugars with 
prolong storage period and the changes in TA levels 
are strong indications of the ongoing metabolism in 
the fruit during storage since pomegranate is a non­
climacteric fruit (Fawole and Opera, 2013 a). The 
decrease in fruit acidity can be attributed to the 
cumulative effects of the increase in juice content 
and the use of organic acids which act as a substrate 
for cellular respiration that occurs during fruit ripen­
ing (Diakou et al., 2000). Organic acids present in 
pomegranate include citric, malic, acetic, fumaric, 
tartaric and lactic acid, but citric acid is the main one 
which represents the titratable acidity of pomegran­
ate fruits (Melgarejo et al., 2000). The results 
obtained on the peel color changes during refrigerat­
ed storage are in agreement with the results 
obtained by other studies, where similar storage con­
ditions on ‘Ganesh’ pomegranate fruit induced the 
slight change in stored fruit color over a 12­week 
duration (Nanda et al., 2001). Furthermore, during 
cold storage our findings highlighted a reduction of 
fruit lightness and a darker and saturated red color of 
the skin. In addition to being appreciated for its quali­
tative aspects the pomegranate is known for the high 
nutritional value of its fruits and for its beneficial 
effects on health, and the medicinal properties of the 
different parts of the tree are also well known. The 
main phytochemicals responsible for these beneficial 
health effects are polyphenols which include ellagic 
acid, ellagitannins (eg punicalagin), punicic acid, 
anthocyanins, flavonols, flavan­3­oils and flavones. 
Ellagic acid can be free or condensed with different 
sugars (glucose, rhamnose and arabinose), with dif­
ferent concentrations between various cultivars 
(Zaouay et al., 2012). Although ellagitannins are the 
main polyphenols in pomegranates, punicalagin and 
punicalin are the compounds most characterized for 
their antiatherogenic properties (Seeram et al., 
2005). Anthocyanins are the pigments responsible for 
the typical pomegranate fruit color and include del­
phinidin, cyanidin and pelargonidin 3­glucoside and 
3,5­diglucosides. The changes in the composition and 

concentration of anthocyanins have been shown in 
the different cultivars, the main anthocyanin in the 
Spanish sweet pomegranate cultivar “Mollar de 
Elche” is cyanidin 3­glucoside, while cyanidin 3,5­
diglucoside has been found as the main compound in 
sour pomegranate cultivars (Sayyari et al., 2011 a, b). 
Several studies highlighted the changes in the bioac­
tive compounds content during the refrigerated stor­
age of the pomegranate (Zaouay et al.,  2012; 
Ehteshami et al., 2020). Fawole and Opara (2013 a) 
have shown that the polyphenol content did not vary 
during the first 4 weeks of storage in the ‘Bagwa’ 
fruit but increased after 8 weeks due to an accumula­
tion of anthocyanins; variations of bioactive com­
pounds during cold storage are also shown in apples 
fruits (Graziani et al., 2020). In our study, no 
decrease in polyphenol content was observed during 
refrigerated storage in agreement to Baltacioglu et 
al. (2011) and this is attributed to the action of oxida­
tive enzymatic activities following low temperature 
stress (Ehteshami et al., 2020). The anthocyanin con­
tent in the pomegranate showed different trends 
related to the cultivars and storage temperatures 
(Fawole and Opara, 2013 a), which suggests that 
varietal differences represent a key factor in the 
post­harvest biosynthesis of anthocyanins (Turfan et 
al., 2011). Furthermore, several studies have shown 
that the antioxidant activity is linked to the bioactive 
compounds content and that a decrease in this activ­
ity is observed in the pomegranate during refrigerat­
ed storage after 4 weeks compared to harvest 
(Fawole and Opara, 2013 a). Our study showed dur­
ing refrigerated storage a significant weight loss, 
while the juice percentage increased, with maximum 
values at +60 days. Acidity (TA) and TSS decreased 
during refrigerated storage, but the reduction in 
acidity was more significant than that of TSS, there­
fore overall good organoleptic characteristics were 
showed. The polyphenols and anthocyanins 
increased during refrigerated storage, while the 
flavonoids and antioxidant activity were constant. On 
basis of our results suggest that pomegranate fruits 
cv. Acco have a maximum storage time in the fridge 
of 60 days for fresh consumption while they can 
exceed 60 days to produce the juice, as the chemical­
nutraceutical characteristics remain optimal. During 
the cold storage the pomegranate fruits of the cv. 
Acco cultivar did not show the appearance of plant 
diseases (molds or rot fungi), thus allowing a good 
conservation of the product. 
 



Cirillo et al. ‐ Quality parameters of pomegranate fruits during cold storage

21

Acknowledgements 
 
     The authors are grateful to Doctor Mariangela 
Scarano for her technical support in the laboratory 
and in field. 
 
 
References 

AKHILA P.P., SUNOOJ K.V., NAVAF M., AALIYA B., 
SUDHEESH C., SASIDHARAN A., SABU S., MIR S.A., 
GEORGE J., KHANEGHAH A.M., 2022 ­ Application of 
innovative packaging technologies to manage fungi 
and mycotoxin contamination in agricultural products: 
Current status, challenges, and perspectives. ­ Toxicon, 
214: 18­29. 

AL­MUGHRABI M.A., BACHA M.A., ABDELRAHMAN A.O., 
1995 ‐ Effects of storage temperature and duration on 
fruit quality of three pomegranate cultivars. ­ J. King 
Saud Univ., 7: 239­248. 

AL­YAHYAI R., AL­SAID F., OPARA L., 2009 ­ Fruit growth 
characteristics of four pomegranate cultivars from 
northern Oman. ­ Fruits, 64(6): 335­341. 

ARTES F., TUDELA J.A., VILLAESCUSA R., 2000 a ­ Thermal 
postharvest treatments for improving pomegranate 
quality and shelf life. ­ Postharvest Biol. Technol., 18: 
245­251. 

ARTÉS F., VILLAESCUSA R., TUDELA J.A., 2000 b ­ Modified 
atmosphere packaging of pomegranate. ­ J. Food Sci., 
65(7): 1112­1116. 

ASGARY S., KARIMI R., POUR P.M., HEYDARPOUR F., 
MOSTAFAEI S., FARZAEI M.H., MORADI S., ANEVA, I.Y., 
2022 ­ Is consumption of pomegranate supplementa‐
tion effective on oxidative stress biomarkers including 
MDA, ox‐LDL, POX 1, GPX, TAC, and TBRAS? A system‐
atic review and meta‐analysis of randomized controlled 
trials. ­ Curr. Probl. Cardiol., 101198. 

BALTACIOĞLU M.K., BAŞAR M.T., ARAT H.T., ERDOĞAN Y., 
2021 ­ Experimental and theoretical fuel property com‐
parison of four different waste biomass with a commer‐
cial counterpart. ­ Research Square, pp. 1­15. 

BELL C., HAWTHORNE S., 2008 ­ Ellagic acid, pomegranate 
and prostate cancer ‐ a mini review. ­ J. Pharm. 
Pharmacol., 60(2): 139­144. 

BEN­ARIE R., SEGAL N., GUELFAT­REICH S., 1984 ­ The mat‐
uration and ripening of the ‘Wonderful’ pomegranate. ­ 
J. Amer. Soc. Hort. Sci., 109(6): 898­902. 

CALEB O.J., MAHAJAN P.V., OPARA U.L., WITTHUHN C.R.,  
2012 a ­ Modelling the respiration rates of pomegran‐
ate fruit and arils. ­ Postharvest Biol. Technol., 64(1): 
49­54. 

CALEB O.J., OPARA U.L., WITTHUHN C.R., 2012 b ­ Modified 
atmosphere packaging of pomegranate fruit and arils: 
a review. ­ Food Bioprocess Technol., 5: 15­30. 

CIRILLO A., MAGRI A., SCOGNAMIGLIO M., D’ABROSCA B., 

FIORENTINO A., PETRICCIONE M., DI VAIO C., 2022 ­ 
Evaluation of morphological, qualitative, and metabo‐
lomic traits during fruit ripening in pomegranate 
(Punica granatum L.). ­ Horticulturae, 8(5): 384. 

DAK M., PAREEK N.K., 2014 ­ Effective moisture diffusivity 
of pomegranate arils undergoing microwave‐vacuum 
drying. ­ J. Food Engineering, 122: 117­121. 

DAVIDSON M.H., MAKI K.C., DICKLIN M.R., FEINSTEIN S.B., 
WITCHGER M., BELL M., MCGUIRE D.K., PROVOST J.­C., 
LIKER H., AVIRAM M., 2009 ­ Effects of consumption of 
pomegranate juice on carotid intima‐media thickness in 
men and women at moderate risk for coronary heart 
disease. ­ Amer. J. Cardiol., 104(7): 936­942. 

DE NIGRIS F., BALESTRIERI M.L., WILLIAMS­IGNARRO S., 
D’ARMIENTO F.P., FIORITO C., IGNARRO L.J., NAPOLI C., 
2007 ­ The influence of pomegranate fruit extract in 
comparison to regular pomegranate juice and seed oil 
on nitric oxide and arterial function in obese Zucker 
rats. ­ Nitric Oxide, 17(1): 50­54. 

DIAKOU P., SVANELLA L., RAYMOND P., GAUDILLERE J.­P., 
MOING A., 2000 ­ Phosphoenolpyruvate carboxylase 
during grape berry development: protein level, enzyme 
activity and regulation. ­ Aust. J. Plant Physiol., 27: 221­
229. 

DOROSTKAR M., MORADINEZHAD F., 2022 ­ Postharvest 
quality responses of pomegranate fruit (cv. Shishe‐kab) 
to ethanol, sodium bicarbonate dips and modified 
atmosphere packaging. ­ Adv. Hort. Sci., 36(2): 107­
117. 

EHTESHAMI S., ABDOLLAHI F., RAMEZANIAN A., DASTJERDI 
A.M., RAHIMZADEH M., 2019 ­ Enhanced chilling toler‐
ance of pomegranate fruit by edible coatings combined 
with malic and oxalic acid treatments. ­ Scientia 
Horticulturae, 250: 388­398. 

EHTESHAMI S., ABDOLLAHI F., RAMEZANIAN A., 
MIRZAALIAN DASTJERDI A., 2020 ­ Maintenance of 
quality and bioactive compounds in pomegranate fruit 
(Punica granatum L.) by combined application of organ‐
ic acids and chitosan edible coating. ­ J. Food Biochem., 
44(9): e13393. 

EL­NEMR S.E., ISMAIL I.A., RAGAB M., 1990 ‐ Chemical 
composition of juice and seeds of pomegranate fruit. ­ 
Food/Nahrung, 34(7): 601­606. 

ERKAN M., DOGAN A., 2018 ­ Pomegranate/Roma‐Punica 
granatum, pp. 355­361. ­ In: RODRIGUES S., E.O. SILVA, 
and E.S. BRITO (eds.) Exotic fruits. Reference guide. 
Academic Press, London, UK, pp. 466. 

FADAVI A., BARZEGAR M., AZIZI M.H., BAYAT M., 2005 ­ 
Note. Physicochemical composition of ten pomegran‐
ate cultivars (Punica granatum L.) grown in Iran. ­ Food 
Sci. Technol. Inter., 11(2): 113­119. 

FAWOLE O.A., OPARA U.L., 2013 a ‐ Effects of storage tem‐
perature and duration on physiological responses of 
pomegranate fruit. ­ Industrial Crops Products, 47: 300­
309. 

FAWOLE O.A., OPARA U.L., 2013 b ‐ Changes in physical 



Adv. Hort. Sci., 2023 37(1): 15­23

22

properties, chemical and elemental composition and 
antioxidant capacity of pomegranate (cv. ‘Ruby’) fruit 
at five maturity stages. ­ Sci. Hortic., 150: 37­46. 

FERRARA G., GIANCASPRO A., MAZZEO A., GIOVE S.L., 
MATARRESE A.M.S., PACUCCI C., PUNZI R., TRANI A., 
GAMBARCORTA G., BLANCO A., GADALETA A., 2014 ­ 
Characterization of pomegranate (Punica granatum L.) 
genotypes collected in Puglia region, Southeastern 
Italy. ­ Sci. Hortic., 178: 70­78. 

GIL M.I., ARTES F., TOMAS­BARBERAN F.A., 1996 ­ Minimal 
processing and modified atmosphere packaging effects 
on pigmentation of pomegranate seeds. ­ J. Food Sci., 
61(1): 161­164. 

GIL M.I.,  TOMÁS­BARBERÁN F.A., HESS­PIERCE B., 
HOLCROFT D.M., KADER A.A., 2000 ­ Antioxidant activi‐
ty of pomegranate juice and its relationship with phe‐
nolic composition and processing. ­ J. Agric. Food 
Chem., 48(10): 4581­4589. 

GRAZIANI G., GASPARI A., DI VAIO C., CIRILLO A., RONCA 
C.L., GROSSO M., RITIENI A., 2021 ­ Assessment of in 
vitro bioaccessibility of polyphenols from annurca, 
limoncella, red delicious, and golden delicious apples 
using a sequential enzymatic digestion model .  ­ 
Antioxidants, 10(4): 541. 

GRAZIANI G., RITIENI A., CIRILLO A., CICE D., DI VAIO C., 
2020 ­ Effects of biostimulants on Annurca fruit quality 
and potential nutraceutical compounds at harvest and 
during storage. ­ Plants, 9(6): 775. 

GURTLER J.B., GARNER C.M., 2022 ­ A review of essential 
oils as antimicrobials in foods with special emphasis on 
fresh produce. ­ J. Food Protection, 85(9): 1300­1319. 

KIAYA V., 2014 ­ Post‐harvest losses and strategies to 
reduce them. ­ Technical Paper on Postharvest Losses, 
Action Contre la Faim (ACF), 25: 1­25. 

KULKARNI A.P., ARADHYA S.M., DIVAKAR S., 2004 ­ 
Isolation and identification of a radical scavenging 
antioxidant‐punicalagin from pith and carpellary mem‐
brane of pomegranate fruit. ­ Food Chem., 87(4): 551­
557. 

LOVE K., CHEN N., PAULL R., 2014 ­ Quick harvest and 
postharvest tips for better quality and longer posthar‐
vest life. ­ Fruit, Nut, Beverage Crops, 36: 1­13. 

MAGRI A., ADILETTA G., PETRICCIONE M., 2020 ­ 
Evaluation of antioxidant systems and ascorbate‐glu‐
tathione cycle in feijoa edible flowers at different flow‐
ering stages. ­ Foods, 9(1): 95. 

MARTINEZ J.J., MELGAREJO P., HERNÁNDEZ F.A., SALAZAR 
D.M., MARTINEZ, R., 2006 ­ Seed characterisation of 
five new pomegranate (Punica granatum L.) varieties. ­ 
Sci. Hortic., 110(3): 241­246. 

MELGAREJO P., SALAZA D.M., ARTES F., 2000 ­ Organic 
acids and sugars composition of harvested pomegran‐
ate fruits. ‐ European Food Res. Technol., 211: 185­190. 

MUNHUWEYI K., LENNOX C.L., MEITZ­HOPKINS J.C., CALEB 
O.J., OPARA U.L., 2016 ­ Major diseases of pomegran‐
ate (Punica granatum L.), their causes and manage‐

ment ­ A review. ­ Sci. Hort., 211: 126­139. 
NANDA S., RAO D.S., KRISHNAMURTHY S., 2001 ­ Effects of 

shrink film wrapping and storage temperature on the 
shelf life and quality of pomegranate fruits cv. Ganesh. 
­ Postharvest Biol. Technol., 22(1): 61­69. 

NAVEENA B.M., SEN A.R., KINGSLY R.P., SINGH D.B., 
KONDAIAH N., 2008 ­ Antioxidant activity of pomegran‐
ate rind powder extract in cooked chicken patties. ­ 
Inter. J. Food Sci. Technol., 43(10): 1807­1812. 

NAZOORI F., ZAMANIBAHRAMABADI E., RAFIE A., 
MIRDEHGHAN S.H., 2022 ­ Combined application of 
gamma‐aminobutyric acid and carnauba wax as edible 
coating on pomegranates in cold storage. ­ J. Agric. Sci. 
Technol., 24(3): 591­602. 

PAREEK S., VALERO D., SERRANO M., 2015 ­ Postharvest 
biology and technology of pomegranate. ­ J. Sci. Food 
Agric., 95(12): 2360­2379. 

PETRICCIONE M., PASQUARIELLO M.S., MASTROBUONI F., 
ZAMPELLA L., DI PATRE D., SCORTICHINI M., 2015 ­ 
Influence of a chitosan coating on the quality and 
nutraceutical traits of loquat fruit during postharvest 
life. ­ Sci. Hortic., 197: 287­296. 

REDDY M.K., GUPTA S.K., JACOB M.R., KHAN S.I., FERREIRA 
D., 2007 ­ Antioxidant, antimalarial and antimicrobial 
activities of tannin‐rich fractions, ellagitannins and phe‐
nolic acids from Punica granatum L. ­ Planta Medica, 
53(05): 461­467. 

SAYYARI M., BABALAR M., KALANTARI S., 2012 ­ 
Enhancement of chilling resistance, antioxidant activity, 
and quality of pomegranate (Punica granatum) cv. 
Rabab during cold storage, through an application of 
salicilyc acid. Iran. ­ J. Hortic. Sci., 42(4): 339­347. 

SAYYARI M., BABALAR M., KALANTARI S., MARTINEZ­
ROMERO D., GUILLEN F., SERRANO M., 2011 a ­  
Vapour treatments with methyl salicylate or methyl jas‐
monate alleviated chilling injury and enhanced antioxi‐
dant potential during postharvest storage of pome‐
granates. ­ Food Chem., 124: 964­970. 

SAYYARI M., CASTILLO S., VALERO D., DIAZ­MULA H.M., 
SERRANO M., 2011 b ­ Acetyl salicylic acid alleviates 
chilling injury and maintains nutritive and bioactive 
compounds and antioxidant activity during postharvest 
storage of pomegranates. ­ Postharv. Biol. Technol., 60: 
136­142. 

SEERAM N.P., ADAMS L.S., HENNING S.M., NIU Y., ZHANG 
Y., NAIR M.G., HEBER D., 2005 ­ In vitro antiprolifera‐
tive, apoptotic and antioxidant activities of punicalagin, 
ellagic acid and a total pomegranate tannin extract are 
enhanced in combination with other polyphenols as 
found in pomegranate juice. ­ J. Nutr. Biochem., 16(6): 
360­367. 

SIDDIQUI S.A., BAHMID N.A., SHEKHAWAT G.K., JAFARI 
S.M., 2022 ­ Introduction to postharvest and post‐
mortem technology, pp. 1­38. ­ In: JAFARI S. (ed.) 
Postharvest and postmortem processing of raw food 
materials. Woodhead Publishing, Sawston, Cambridge, 



Cirillo et al. ‐ Quality parameters of pomegranate fruits during cold storage

23

UK, pp. 406. 
SINGLETON V.L., ROSSI J.A., 1995 ‐ Colorimetric of total 

phenolic with phoshomolybdic‐phosphotungstic acid 
reagents. ­ Am. J. Enol. Viticult., 16: 144­158. 

TURFAN Ö., TÜRKYILMAZ M., YEMIS O., ÖZKAN M., 2011 ­ 
Anthocyanin and colour changes during processing of 
pomegranate (Punica granatum L., cv. Hicaznar) juice 
from sacs and whole fruit. ­ Food Chem., 129: 1644­
1651. 

WASKAR D.P., 2011 ­ Studies on extension of postharvest 

l ife of pomegranate fruits’ Bhagawa’.  ­  Acta 
Horticulturae, 890: 455­460. 

ZAOUAY F., MENA P., GARCIA­VIGUERA C., MARS M., 2012 
‐ Antioxidant activity and physic‐chemical properties of 
Tunisian grown pomegranate (Punica granatum L.) cul‐
tivars. ‐ Ind. Crops Prod., 40: 81­89.  

ZHISHEN J., MENGCHENG T., WU J., 1999 ­ The determina‐
tion of flavonoid contents in mulberry and their scav‐
enging effects on superoxide radicals. ­ Food Chem., 
64: 555­559.