Modified Atmosphere Packaging of Date Fruit (Phoenix dactylifera L.

) Cultivar ‘Barhee’ in Khalal Stage

Abstract

Considering the date fruit (Phoenix dactylifera L.) ‘Barhee’ is mainly harvested at the khalal stage,
the quality changes of fresh fruits were studied under modified atmosphere packaging (MAP). Fruit
was packed in barrier bags and exposed to the ambient air (passive MAP) and different
concentrations of CO2 (5, 15 and 30%) within the packaging (active MAP) by using microperforated
films as control. Fruits were stored at 4°C and physiochemical changes were studied at 7 days
intervals during 28 days of storage. Modified atmosphere containing 30% CO2 caused almost half of
the fruits to turn into low quality rutab, while the best quality and longest shelf-life of khalal fruits
was gained with 5% CO2 concentration. Also, passive MAP compared with control samples, showed
acceptable results by extending the shelf-life of khalal date fruits.

INTRODUCTION Date (Phoenix dactylifera L.) is a berry fruit whose development divides into five
stages namely hababook, kimri, khalal, rutab and tamar. At the khalal stage, fruits are physiologically
mature, hard and crisp with over 50% moisture content, bright yellow or red in color and very
perishable. In general, fruits at khalal stage are ready for commercial trade as “fresh” fruit but this
applies only to those cultivars which are sweet, with a low amount of tannin and low astringency
(Barreveld, 1993). Some date cultivars are suitable for marketing at the khalal stage including
‘Barhee’, ‘Bereim’, ‘Hayany’ and ‘Khalas’ among them ‘Barhee’ is the most popular cultivar
worldwide (Mortazavi et al., 2007). The khalal fruits that are usually harvested at the end of July are
highly perishable and must be transported to the market as soon as possible (Glasner, 2002). Any
delay in transport or improper storage conditions result in quick appearing of rutab spots and surface
wrinkling accompanied by a loss of flavor and taste (Mortazavi et al., 2007). The strategy for
exporting khalal dates would make it necessary to develop new methods to delay fruit ripening
during handling and storage. Modified atmosphere packaging (MAP) is now being used for extending
the shelf-life and reducing the waste of a wide range of fruits and vegetables. There are few studies
about use of vacuum and modified atmosphere packaging for date fruits at the khalal stage. Al-
Redhaiman (2004) stored full mature ‘Barhee’ date fruits under three CO2 concentrations (5, 10 or
20%) at 0°C and reported that fruits under 20% CO2 have a statically longer storage period, lasting
for 26 weeks. Also, Achour et al. (2003) determined that dehydrating of ‘Deglet Nour’ dates at tamar
stage decreased at MA condition with 20% CO2 and 80% N2 during the storage period. A previous
study by authors has demonstrated the potential interest of using passive MAP to reduce the weight
loss and appearance of low quality rutab spots of ‘Barhee’ date picked at khalal stage. However, in
the vacuum packaging, a large part of fruits turned to low quality rutab and fruit firmness reduced
considerably (Mortazavi et al., 2007). The objective of this work was to study the influence of some
active and passive MAP conditions accompanied by fruit to stalk junction status on the physiological
properties, quality attributes and storability of ‘Barhee’ date fruits at the khalal stage.
MATERIALS AND METHODS

‘Barhee’ date fruits were harvested at khalal stage from a commercial orchard in Ahvaz, Khuzestan
province, Iran according to yellow skin color and about 30% soluble solids concentration. The fruit
were precooled immediately and transported to the laboratory. They were selected for no visual
defects and basis of uniform size, color and without rutab spots, then washed with sodium
hypochlorite solution 0.5% for 2 min, rinsed with tap water and dried prior to packaging.

Fruits were divided into eight lots each of 60 fruit, then each lot was divided into three replicates of
20 (200±5 g). Each set of three replicates was put into a dish tray and placed in a barrier
nylon/polyethylene plastic bag (25×35 cm) given one of the eight treatments: (5%-J) joint fruits+5%
O2: 5% CO2; (5%-D) detached fruits+5% O2: 5% CO2; (15%-J) joint fruits+5% O2: 15% CO2; (15%-D)
detached fruits+5% O2: 15% CO2; (30%- J) joint fruits+5% O2: 30% CO2; (30%-D) detached fruits+5%
O2: 30% CO2; (PM) detached fruits in passive MAP; and (C) control (detached fruits, perforated film
providing ambient air atmosphere within the packages). All active MAP treatments (T1 to T6) were
performed by creating a vacuum in a Henkelman vacuum pack instrument (200A) followed by
flushing the gas mixture 1 bar pressure before heat sealing and N2 used as a balance gas. All samples
were stored at 4°C for up to 28 days. The experiment was conducted based on a completely
randomized design (CRD). The data were analyzed with MSTAT-C (version 1.42) statistical package,
and means compared by Duncan’s Multiple Range Test (DMRT) at 0.01 and 0.05 probability levels.
Visual examinations and other quality attributes were evaluated initially and periodically at seven day
intervals. The gas atmosphere in the head space of the bags was analyzed using an O2/CO2 gas
analyzer. The fruit subjected to all treatments were weighed before and after storage and data were
expressed as percentage of weight loss. Fruit firmness was measured by a Wagner Penetrometer (FT
011). Titratable acidity was calculated as percentage of malic acid by titrating 10 g/100 ml of the date
extract with a solution of NaOH (0.01 N) till pH 8.1. The pH was measured by a Metrohm pH meter.
The level of sugars was measured as °Brix by an Atago refractometer. Water activity (aw) values of
fruits were measured at 30°C with a hygrometer. To determine the wrinkled area or rutab spots, to
have more accuracy of data, an image processing procedure was used (Rocculi et al., 2005). Images
were obtained, using a color plane scanner (true color-24 bit, resolution of 600×600 dpi), and by
positioning the fruit halves on a scanner held on a black box, to exclude the surrounding light. The
saved images were opened by a program that had been written by authors in Matlab® software to
calculate the percentage of selected area fraction of fruit and averaged data were considered as RSA
or WAP for each unit.

RESULTS AND DISCUSSION

During the storage period of 28 days, the fruit respiration resulted in a modification of the internal
atmospheres and as expected, all treatments showed a reduced O2 and an increased CO2 level (Fig.
1). The oxygen concentration decreased sharply in the first seven days, from 5 to less than 2.7% and
continued with a little decreasing amplitude in the following days. The final O2 concentration was
less than 0.07% for all treatments after 28 days storage. However the CO2 production rate was
higher at the first week for pouches containing 5% CO2 (5%-J and 5%-D) but the final concentration
of CO2 was in keeping with its concentration at the start of the experiment. Clear correlation was
observed between final and initial concentration of CO2 and its concentration reached to about 60%
for pouches containing 30% at the start of experiment. According to the O2 curves, differences
between passive and active MAP were seen during the transient period, then, drawing near and
steady state was obtained after approximately 14 days of storage. These were in agreement with the
results of Charles et al. (2008) for fresh endives.

It was evident from this study that the rutab spots area (RSA) that appeared during storage was
correlated significantly with CO2 concentration within pouches. The higher CO2 concentration
promoted RSA and image processing of fruit surfaces in 30%-J and 30%-D treatments (30% CO2)
showed 92.3 and 82.5% RSA respectively at the end of 28 days storage. Fruits in pouches filled with
15% CO2, showed 49.5 and 58.8% rutab spots for 15%-J and 15%-D respectively and samples in 5%
CO2 condition (5%-J and 5%-D) undergoing a smaller turning to rutab spots. Also when the storage
period prolonged, greater part of fruits changed to rutab in all treatments, particularly in fruits
detached from stalk (5%-D, 15%-D and 30%-D). Fruits in the perforated bags exposed to the air (C),
exerted highest WAP (24.25%) after 28 days of storage. WAP was negligible for other treatments and
the lowest value was recorded in fruits that received 30% CO2 (30%-J and 30%-D) at which a large
part of fruits turned to rutab. Fruit to stalk junction had no major effectiveness on the wrinkled area
percentage. Appearing rutab spots and wrinkled surface are the two main disorders restricting
marketing, storage and exports of khalal dates after harvest (Fig. 2a,b). However, turning fruit surface
color to brown considered as rutab spot, comparing obtained rutab fruits in this study, with those
ripened naturally on the tree, showed some major differences. Rutab spots in the fruits coming out
of storage can be described as CO2 injury similar to that reported by Serrano et al. (2005), for sweet
cherry at high CO2 concentrations. Elevated CO2 can prove to be fruit damage, often inducing
fermentation, particularly when fruit is sealed in packaging film of insufficient permeability (Betts,
1996).

Regardless of gas composition within the package, fruit weight loss ranged between 1.13-1.82 g/100
g under MAP conditions, while control fruits in perforated bags (C) lost 10.48 g/100 g. The data
revealed the pattern of reduction in fruit weight in all MAP packages was almost the same after 7, 14,
21 and 28 days of storage and packaging was effective in limiting weight loss of fruits (Table 1). As
expected, the wrinkled area percentage (WAP) followed by a similar pattern with fruit weight loss
(Weight loss = 0.525 WAP + 0.499). Weight loss is a physiological event caused by loss of water from
the fruit surface to the surrounding atmosphere and loss of carbon on formation of CO2 during
respiration (Rizzo and Muratore, 2009). It can be controlled by temperature, humidity, and using
proper packaging. In this work, for using barrier films, and minimum package condensation observed,
it can be assumed that a gentle increase in weight loss of all treatments (except control), is caused
mainly by the biochemical reactions of fruit cell components.

The initial firmness measured at the start of experiment was 3.4 kg. The following decrease in
firmness during 28 days cold storage to 0.66 kg (average of all treatments) showed a clear response
to the different MA conditions applied (Fig. 2c,d). Prolonging the storage duration, decreased the
firmness of all samples gradually. The lowest flesh firmness was obtained in fruits stored under MA
conditions with 30% CO2 (30%-J and 30%-D). Date fruit at khalal stage had a hard and crisp texture
and physico-chemical changes that caused arising rutab spots, decreased fruit firmness. Fruits in the
pouches with 5% O2 (5%-J and 5%-D) and control (C) showed the highest firmness (2.1, 1.9 and 2.2 kg
respectively).
 Significant differences between the treatments were found in terms of titratable acidity. TA level of
date fruit in passive MAP (PM) and control (C) treatments were consistently lower than those of the
other treatments. At harvest, the levels of total acidity (TA) calculated as malic acid was 22.48
mg/100 g fresh weight and TA increased significantly throughout the evaluation period for all
samples (Fig. 2e,f). At the end of the experiment, pouches containing 30% CO2 recorded nearly 1.5-
1.7 fold higher acidity of the initial value. Also the results showed the fruits in pouches filled with 5%
CO2 had consistently lower acidity than those of treatments with 15 and 30% CO2. However,
detaching fruit from stalk, increased TA in 5 and 15% CO2 treatments, but it did not show any
significant effect on acidity levels. Similarly to rutab spots area, an increase in acidity level was
apparent during storage time and direct correlation was observed among these studied parameters.
Reversely, a gradual decrease in pH from 7.5 to 6.3 was seen during the 28 days of storage (Fig. 2e,f).
Pouches filled with 30% CO2 had maximum reduction in pH.

The pattern of changes in flesh softening, reducing pH and rising acidity level in fruits with high rutab
spot area exhibit correlation with a high CO2 level and it can be 1066 concluded that the highest CO2
level (30%) is not appropriate for storage of date fruits at khalal stage. This was nearly in contrast
with the results of Al-Redhaiman (2004) that reported khalal date fruits have significantly longer
storage period under higher CO2 concentration.

Khalal fruits that lost their hard and crisp texture had lower quality and price. Softening of fruit
texture is related to activation of pectin decomposing enzymes. The role of high CO2 level in slowing
texture softening and other enzymatic reactions has been underlined in different reports. Our results
showed that CO2 at 30% concentration had a negative effect on firmness due to stimulated
fermentation, during that degradation of hemicellulosic polysaccharides and cellulose, leads to
disorganization of cell wall, decrease in cellular turgidity and loss of texture firmness (Kader et al.,
1989).

Increasing the acidity level upon storage and more specifically at the onset of deterioration of fruit
texture postulated the second generation of organic acids formation (Barreveld, 1993). The
accompanying increment of acidity level and brown surface discoloration of fruits also suggested that
organic acids can react with reducing sugars to produce brown pigments (Lozano, 2006). Organic
acids are a useful index of authenticity in fruits and have an important influence on the sensory
properties of fruits especially in combination with sugars. Major organic acids that have been isolated
from date flesh are citric, malic and oxalic acid (Barreveld, 1993). Fruit juice pH is affected by alkaline
and acidic compounds of cells and any change in concentration of these compounds will change the
pH quickly (Wills, 1998). Barreveld (1993) reported the most common pH values for ‘Deglet Noor’
date range from 5.3 to 6.3 and definite correlation was observed between increasing the pH and the
commercial quality for this cultivar.

The water activity value in ‘Barhee’ date fruits at the beginning of the experiment was 0.97 (Fig.
2g,h). Evidently, after 28 days of storage, a slight decrease was observed in this parameter in all
treatments but with different rates. The highest rate of aw decline during the storage period
occurred in fruits stored under MA with 30% CO2 (0.94 in T5 and T6). On the contrary, the highest aw
value was observed in fruits stored under MA with 5% CO2. A clear correlation was identified
between decrease in water activity and rutab spots area, the less water activity, the higher Rutab
spots (Aw = -0.0004 RSA + 0.9687). Water activity is the ratio of the partial vapor pressure of water in
food to the partial saturation vapor pressure of water vapor in the air at the same temperature and
describes the energy state of water in the food as an important quality factor for dates during
storage (Fontana, 2000). Decreasing the level of aw by storage duration revealed bonding more H2O
molecules to the solutes released from the fruit texture degradation.

As expected, significant changes in SSC were seen between different MAP conditions applied. Fruits
exposed to 30% CO2, displayed maximum amounts of SSC (33.2 and 32.9 for 30%-J and 30%-D
respectively). Generally, most of the full mature khalal fruits gradually turned to low quality rutab
fruits during storage. In all treatments by increasing the storage period, a greater part of fruits turned
to rutab, and SSC increased from 27.7 to 34.3%. A direct correlation (SSC = 0.1039 RSA + 27.945) was
observed between SSC and RSA. The lowest amounts of increase in the SSC and RSA were seen in
pouches filled with 5% CO2, passive MAP and control (31.4, 32.8, 34.1 and 33.6% respectively). SSC is
one of the most important maturity and quality indices in various fruits and in date fruits at khalal
stage, should be more than 28% (Barreveld, 1993). In contrast with produced rutab fruits in this work
from khalal dates, the SSC content of rutab date fruits naturally ripened on the tree is about 70% and
those have an acceptable quality. These findings are similar to those reported earlier by other
workers on various date cultivars (Barreveld, 1993). It is postulated that the immobile form of the
ripening enzymes existed in fruits harvested at khalal stage, is in contrast with dates left on the palm
till they turn naturally to rutab. Formation of some metabolites including soluble contents arises by
releasing and activating enzymes such as amylase, invertase, polygalacturonase and polyphenol
oxidases (Saleem et al., 2005).

CONCLUSIONS The date fruit of cultivar ‘Barhee’ lose their astringency at khalal stage and are sold as
fresh fruit. This stage will last for a couple of weeks, terminating the supply for this purpose, leaving
other fruits to mature further on the palm (Glasner et al., 2002). Fruit at khalal stage has a limited
shelf-life and generally is sold shortly after harvest and mostly in the local markets. Based on our
results we can conclude the significant improvement in postharvest storage of khalal ‘Barhee’ date
fruits that can be achieved, under modified atmosphere conditions. Detrimental color changes of
khalal dates were related to packaging conditions and, CO2 injury was the direct result of the high
CO2 levels. A conservative recommendation to minimize quality losses would be to keep the khalal
dates under 5% CO2 level during storage. However, the best beneficial effects were obtained in
active MAP, significant but not major differences were seen in passive MAP conditions and due to
lower costs, it can be recommended when needed. Detaching fruit from stalk, showed negligible
positive effects on maintaining the studied quality factors. Evident correlations were displayed
between RSA and firmness, acidity, water activity and SSC postulated the biochemical changes of
fruit issued from texture degradation by means of different enzymes. To understand better these
biochemical changes, research dealing with and focused on the understanding of the processes
involved might be proposed. ACKNOWLEDGEMENTS We would like to thank the Departments of
Horticultural Science and Food Science & Technology, Tarbiat Modares University (TMU) for
providing technical and laboratory facilities, also the Date Palm and Tropical Fruits Institute of Iran in
Ahwaz, for providing fruit samples