See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/235731681

Bioactive compounds and antioxidant activities during fruit ripening of

watermelon cultivars

Article  in  Journal of Food Composition and Analysis · April 2011

DOI: 10.1016/j.jfca.2011.03.016

CITATIONS READS

73 4,159

6 authors, including:

Tlili Imen Chafik Hdider

National Institute of Agronomic Research of Tunisia National Institute of Agronomic Research of Tunisia
44 PUBLICATIONS   600 CITATIONS    48 PUBLICATIONS   957 CITATIONS   

SEE PROFILE SEE PROFILE

Marcello Lenucci Riadh Ilahy

Università del Salento University of Carthage
101 PUBLICATIONS   1,939 CITATIONS    61 PUBLICATIONS   580 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Amélioration de la qualité de la tomate View project

micropropagation View project

All content following this page was uploaded by Chafik Hdider on 27 September 2019.

The user has requested enhancement of the downloaded file.

This article appeared in a journal published by Elsevier. The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/copyright
 Author's personal copy

Journal of Food Composition and Analysis 24 (2011) 923–928

Contents lists available at ScienceDirect

Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca

Original Article

Bioactive compounds and antioxidant activities during fruit ripening of

watermelon cultivars
Imen Tlili a,b, Chafik Hdider b, Marcello Salvatore Lenucci c,*, Riadh Ilahy a,b, Hager Jebari b,
Giuseppe Dalessandro c
a
Department of Biology, Faculty of Sciences of Bizerte, Zarzouna, 7021 Bizerte, Tunisia
b
Laboratory of Biotechnology and Plant Physiology, National Agricultural Research Institute of Tunisia, Rue Hédi Karray, 2049 Ariana, Tunisia
c
Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Via Prov.le Lecce-Monteroni, 73100 Lecce, Italy

A R T I C L E I N F O A B S T R A C T

Article history: Changes in bioactive compounds and antioxidant activities of three commercial selections (Crimson
Received 29 July 2010 sweet, Dumara and Giza) and two new selections (P503 and P403) of watermelon cultivars were
Received in revised form 21 March 2011 investigated at four different fruit ripening stages (white, white-pink, pink and red-ripe). Lipophilic and
Accepted 29 March 2011
hydrophilic antioxidant activities (LAA and HAA, respectively) were determined, and their correlations
Available online 15 April 2011
with total vitamin C, phenol, flavonoid, lycopene and b-carotene contents were studied. Ripening stage
significantly influenced lycopene and b-carotene contents, as well as the LAA of all investigated
Keywords:
watermelon cultivars. Good correlations between LAA and lycopene and b-carotene contents were
Antioxidant activity
found using the TEAC assay. At the red-ripe stage of ripeness, P503 cultivar showed the highest amount
b-Carotene
Citrullus lanatus of lycopene (64.5 mg kg1 fw), whereas Dumara cultivar showed the highest level of b-carotene
Cultivar difference (2.1 mg kg1 fw). Giza cultivar scored first for total phenol (260.1 mg GAE kg1 fw), flavonoid
Flavonoids (260.0 mg RE kg1 fw) and total vitamin C (204.0 mg kg1 fw) contents. Although, the HAA of the
Food composition studied watermelon cultivars was significantly influenced by the ripening stages, it only correlated to the
Lycopene amount of total phenols and flavonoids. These data confirm the important role played by genetic
Phenols background and ripening stage in determining antioxidant potential of watermelon fruits. They also give
Ripening stages
valuable insights into the synthesis and accumulation of bioactive compounds in such fruits, and
Total vitamin C
furthermore move us closer to identifying the harvesting period with the highest antioxidant potential.
Food analysis
ß 2011 Elsevier Inc. All rights reserved.

1. Introduction the risk of certain types of cancers, cardiovascular diseases and

European consumers are becoming very health conscious, and
this attitude is also supported by governments which invest high
levels of resources in promoting the consumption of fresh fruits
and vegetables. Watermelon (Citrullus lanatus Thunb. Mansfeld) is
largely consumed as refreshing summer fruit throughout the
Mediterranean basin. It is also imported in northern Europe, where
its consumption is seasonal and occurs mainly between May and
September. Watermelon provides a wide variety of dietary
antioxidants such as carotenoids (lycopene and b-carotene),
phenols, vitamins (A, B, C and E) and specific amino acids
(citrulline and arginine) (Perkins-Veazie, 2002; Perkins-Veazie et
al., 2007), which are thought to exert a protective role in reducing
Abbreviations: AsA, ascorbic acid; DHA, dehydroascorbic acid; GAE, gallic acid
equivalents; HAA, hydrophilic antioxidant activities; LAA, lipophilic antioxidant
activities; RE, rutin equivalents.
* Corresponding author. Tel.: +39 0832 298612; fax: +39 0832 298858.
E-mail address: marcello.lenucci@unisalento.it (M.S. Lenucci).
age-related degenerative pathologies (Rice-Evans et al., 1996;
Giovannucci, 1999; Rao, 2006). The health benefits of eating
watermelon, as well as its low caloric value, make it a very
attractive fruit.
Identification and quantification of bioactive compounds and
antioxidant properties of many fruits and vegetables are well
defined; however, studies on the characterization and quantification
of the phytochemical and antioxidant properties of watermelon are
very limited. It has been reported that the levels of the health-
promoting bioactive compounds and the antioxidant activity of
fruits and vegetables are strongly influenced by genotype differ-
ences and external factors such as agro-technical processes,
environmental conditions, ripening stage, and harvest and post-
harvest manipulations (Waterman and Mole, 1994; Abushita et al.,
2000; Dumas et al., 2003; Lenucci et al., 2009). Recently, Perkins-
Veazie et al. (2006, 2007) and Leskovar et al. (2004) emphasized the
importance of genotype when assessing the lycopene, soluble solids
and ascorbic acid contents in watermelons. Previous research (Tlili
et al., 2011) also highlighted this unexploited genotype variability in

0889-1575/$ – see front matter ß 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2011.03.016
 Author's personal copy

924 I. Tlili et al. / Journal of Food Composition and Analysis 24 (2011) 923–928

the antioxidant content of different cultivars of watermelon grown

in Tunisia, focusing on the importance of sampling areas when such
compounds are determined.
In this study we investigated changes in major antioxidant
compounds and antioxidant activities of five watermelon cultivars
grown simultaneously in an open-field of the south of Italy and
harvested at four different stages of ripening. The hydrophilic and
lipophilic antioxidant activities (HAA and LAA, respectively) and
their correlation to different classes of antioxidants were also
examined.

2. Materials and methods

2.1. Plant material

Five watermelon cultivars, including three of the most important

commercial selections worldwide (Crimson sweet [Clause], Dumara
[Nunhems] and Giza [Egyptian cultivar selected and improved by
the National Agricultural Research Institute of Tunisia – INRAT]) and
two new cultivars (P503 and P403) selected by the INRAT, were used
for all the experimental work. Sowing was carried out in alveolar
boxes at the beginning of April 2008. One-month-old watermelon
seedlings were transplanted with a spacing of approximately
200 cm within the row and 250 cm between rows matching a
density of about 2000 plants/ha into a sandy soil of an open-field in
the province of Lecce in southern Italy (latitude 408230 1600 80N,
longitude 178570 4100 40E; decimal degrees 40.3881; 17.9615).
All cultivars under investigation were subjected to identical
cultural and agronomical practices as described by Tlili et al.
(2011) and, of course, environmental conditions in order to Fig. 1. Longitudinal sections of the fruit of the five analysed watermelon cultivars at
the four different stages of ripening.
minimize the influence of pre- and post-harvest factors.

2.2. Watermelon harvest and sampling
Watermelon fruits were harvested from the rows at different
ripening stages. Three independent samples of at least 3 injury-free
watermelon fruits for each cultivar were hand harvested randomly at
four ripening stages indicated as white (60 days after sowing): low
fruit size and white flesh; white-pink (66 days after sowing): not yet
mature fruit with white-pink flesh; pink (73 days after sowing): large
fruit size with pink flesh and green tendril; red-ripe (80–90 days after
sowing): mature fruit with red flesh, brown tendril and yellow
ground spot (Fig. 1). Watermelon fruits were quickly delivered to the
laboratory and immediately cut longitudinally from the stem-end to
the blossom-end through the ground spot. Flesh samples were taken
from the heart area (between locular and the fruit centre) of each
watermelon. Soluble solids content (8Brix) was measured by cutting
a wedge of flesh from the heart area and squeezing the juice into a
digital refractometer (Atago PR-100, NSG Precision Cells, Inc,
Farming dale, NY, USA) calibrated with a 10% sucrose solution.
Since soluble solid content increases during watermelon ripening the
measured values were used to analytically identify the four ripening
stages as follows: white stage 2.0 Brix, white-pink stage 4 Brix,
pink stage 6 Brix and red-ripe stage 8 Brix.
Approximately 1 kg of the obtained samples was homogenized in
a mixer (Waring Laboratory & Science, Torrington, CT, USA) for 5 min.
The homogenates were frozen at 80 8C and used to determine
the carotenoid, phenol, flavonoid and total vitamin C contents, as
well as the HAA and LAA within less than one week to minimize the
depletion of nutrients.
2.3. Analytical determinations
Carotenoids were extracted from triplicate aliquots of the
homogeneous suspension (0.5 g) according to the method of Sadler
et al. (1990) as modified by Perkins-Veazie et al. (2001), and their
content was determined by HPLC as previously described (Lenucci
et al., 2006). Briefly, carotenoids were extracted with 0.05% (w/v)
butylated hydroxytoluene (BHT – 99%, W218405 – Sigma–
Aldrich, Milan, Italy) in acetone and 95% ethanol (1:1, v/v),
separated by partition into hexane and directly assayed by a
Dionex HPLC (Dionex s.r.l., Milan, Italy) with an AD 25 UV–Vis
detector. The separation was performed at 31 8C on an Acclaim
HPLC column C18 (5 mm, 250 mm  4.6 mm) by using a linear
gradient of acetonitrile (A), hexane (B) and methanol (C) as follow:
from 70% A, 7% B, 23% C to 70% A, 4% B, 26% C within 35 min, with a
flow rate of 1.5 mL/min. Peaks were detected at 503 nm. Sigma–
Aldrich HPLC grade solvents were used.
Total phenols were extracted as described by Martinez-
Valverde et al. (2002) on triplicate aliquots of homogeneous
suspension (0.3 g). The total phenol assay was performed by using
the Folin-Ciocalteu reagent (F9252, Sigma–Aldrich) as described
by Spanos and Wrolstad (1990) on triplicate 50 mL aliquots of the
supernatant. The absorbance was read at 750 nm using a
spectrophotometer (Beckman DU 650, Beckman Coulter, Inc., CA,
USA). Results were expressed in mg gallic acid equivalent (GAE)/kg
fresh weight (fw). Even if Folin-Ciocalteu assay usually over-
estimates the content of phenolic compounds due to the
interference from other reducing agents present in food (i.e.
ascorbic acid), it was chosen over HPLC separation methods since it
meant the large number of samples was processed immediately
after harvesting thus minimizing nutrient depletion which can
occur during long term storage of samples even at low
temperatures.
The flavonoid content was determined as described by Zhishen
et al. (1999) on triplicate aliquots of the homogeneous suspension
(0.3 g). The absorbance was read at 510 nm using a Bechman DU
650 spectrophotometer. The linear reading of the standard curve
was from 0 to 250 mg rutin mL1 and flavonoid content was
expressed as mg of rutin equivalent per kg of fw (mg RE/kg fw).
 Author's personal copy

I. Tlili et al. / Journal of Food Composition and Analysis 24 (2011) 923–928 925

Total vitamin C content was determined as reported by
Kampfenkel et al. (1995) on triplicate samples of the homoge-
neous suspension (0.1 g). Briefly, ascorbic acid (AsA) and dehy-
droascorbic acid (DHA), which both contributes to total vitamin C
content, were extracted with 6% metaphosphoric acid. DHA, was
reduced to AsA by pre-incubation of the sample with dithiothreitol
(DTT). Exceeding DTT was removed with N-ethylmaleimide (NEM)
and total Vitamin C (corresponding to total AsA) was determined
by the 2,20 -dipyridyl method. This method is based on the
reduction of Fe3+ to Fe2+ by AsA and detection of Fe2+ complexed
with 2,20 -dipyridyl at 525 nm in a Beckman DU 650 spectropho-
tometer. Total vitamin C was expressed in mg/kg fw. The linear
reading of the standard curve was from 0 to 700 mmol AsA.
The HAA and LAA were measured using the trolox equivalent
antioxidant capacity (TEAC) assay (Miller and Rice-Evans, 1997).
Hydrophilic and lipophilic antioxidants were extracted from 0.3 g
homogeneous suspension (three replicates) with 50% methanol or
50% acetone, respectively at 4 8C under constant shaking (300 rpm)
for 12 h. Samples were centrifuged at 10,000  g for 7 min and
each supernatant was recovered and used for antioxidant activity
measurements. The antioxidant activity was measured at 734 nm
in a Beckman DU 650 spectrophotometer. Two different calibration
curves were constructed using freshly prepared trolox solutions for
HAA and LAA determinations. The linear reading of the standard
curve was from 0 to 16 mM Trolox for HAA and LAA. Values were
obtained from three replicates as mM Trolox/100 g fw.
The inter- and intra-day variability of each analytical method
was measured repeating the analyses three times on the same day
and three times on three consecutive days using the red-ripe stage
homogenate of the Crimson sweet cultivar as sample. The
coefficients of variation of the intra-day and the inter-day
variability were calculated to be below 3% and 4%, respectively,
for all the analytical methods.
2.4. Statistical analysis
Effects of genotype and ripening stage on the nutritional
properties of watermelon cultivars were assessed by analysis of
variance (ANOVA). When a significant difference was detected,
means were compared using the least significant difference (LSD)
test (P < 0.05). All statistical comparisons were performed using
SAS Version 6.1 software (SAS Institute, Cary, NC, USA). Correla-
tions were done using Person’s correlation coefficient (r).
3. Results and discussion
The amount of lycopene, b-carotene, total phenols, flavonoids
and total vitamin C in watermelon cultivars grown in the south of
Italy and harvested at four successive stages of fruit development
and ripening corrensponding to the white, white-pink, pink and
red-ripe colour of the fruit flesh, are reported in Table 1.
For all the investigated watermelon cultivars, the amount of
lycopene and b-carotene, expressed on a fresh weight (fw) basis,
markedly increased during fruit ripening. In particular, at the white
stage, lycopene was not detected in Giza, Dumara and P403
cultivars, whereas it was detected, at a very low level, in Crimson
sweet (0.3 mg/kg fw) and P503 (0.2 mg/kg fw) cultivars.
The highest rate of synthesis and accumulation of lycopene in
chromoplasts was detected in the transitional phase between the
white-pink and the pink stage. This may possibly be due to a
progressive activation of the molecular mechanisms involved in
carotenogenesis regulation during the transition between the
white and pink ripening stages, followed by a feedback inhibition
of the pathway by end-products towards the end of fruit ripening.
These mechanisms may include regulation at the transcriptional
and/or post-transcriptional level, metabolite flux into the carot-
enoid pathway and carotenoid sequestration, as has been found in
tomatoes (Bramley, 2002). In Crimson sweet cultivar, most of
lycopene (78%) present in the red-ripe fruit had already been
synthesized and stored by the pink stage, whereas only a lower
percentage, ranging from 31% to 53%, had already been synthesized
and stored in P503, Dumara, P403 and Giza cultivars.
Differences among cultivars were also evidenced at the red-ripe
stage, where the highest amount of lycopene was detected in P503
cultivar (64.5 mg/kg fw) followed by Giza (62.6 mg/kg fw), Dumara
(47.1 mg/kg fw), P403 (44.8 mg/kg fw) and, finally, Crimson sweet

Table 1
Lycopene, b-carotene, total phenols, flavonoids and total vitamin C contents in watermelon cultivars harvested at four different ripening stages.a

Cultivar Lycopene (mg/kg fw) b-Carotene (mg/kg fw) Total phenols (mg GAE/kg fw) Flavonoids (mg RE/kg fw) Total vitamin C (mg/kg fw)

Crimson sweet
White 0.3  0.0d nd 42.4  0.2c 136.1  6.5b 118.6  0.7b
White-pink 6.4  0.0c nd 61.8  0.4b 138.6  3.6c 144.9  6.1a
Pink 34.8  0.1b 0.1  0.0b 54.8  0.3b 145.6  2.9c 121.8  4.2b
Red-ripe 44.5  1.0a 1.2  0.1a 137.2  0.2a 234.4  5.9a 152.7  6.2a
Giza
White nd nd 126.8  0.4b 156.1  8.1c 304.5  3.8a
White-pink 2.9  0.0c nd 118.9  0.3b 195.0  3.4b 155.8  11.0c
Pink 29.5  0.1b 0.3  0.0b 96.0  0.3 c 155.6  3.1c 217.9  10.8b
Red-ripe 62.6  1.5a 1.4  0.2a 260.2  0.6a 260.0  7.2a 204.0  1.9b
Dumara
White nd nd 181.5  0.1b 162.2  5.3b 233.4  5.1a
White-pink 4.3  0.1c nd 161.6  0.2c 199.2  4.8a 169.9  1.2b
Pink 25.1  0.0b 0.2  0.0b 151.7  0.4d 88.1  3.9c 108.4  3.8d
Red-ripe 47.1  0.7a 2.1  0.1a 246.4  0.5a 193.1  4.8a 130.8  3.7c
P403
White nd nd 132.9  0.5 c 212.5  6.5a 206.3  37.5a
White-pink 3.5  0.0c nd 173.2  0.3b 51.4  2.4c 76.1  4.0b
Pink 23.8  0.1b 0.4  0.2b 144.0  0.1c 178.9  4.6b 76.5  4.8b
Red-ripe 44.8  0.9a 1.0  0.1a 200.8  0.6a 196.7  4.3ab 119.7  6.7b
P503
White 0.2  0.1d nd 66.1  0.3d 121.1  4.6c 134.2  2.3b
White-pink 2.5  0.1c nd 169.0  0.2b 137.8  3.1b 124.6  2.7b
Pink 20.3  0.6b 0.2  0.1b 120.7  0.1c 86.7  6.3d 149.2  4.7a
Red-ripe 64.5  0.2a 1.6  0.2a 183.4  0.5a 230.6  2.7a 137.7  5.0ab
a
Values are given as mean  SEM (n = 3). Letters indicate mean separation within ripening stages of each cultivar; values with different letters are significantly different at
P < 0.05 by LSD test; nd: not detected. LOD was 0.1 mg/kg for lycopene and 0.05 mg/kg for b-carotene. GAE = gallic acid equivalent; RE = rutin equivalent.
 Author's personal copy
926 I. Tlili et al. / Journal of Food Composition and Analysis 24 (2011) 923–928
(44.5 mg/kg fw) cultivars. These values were from 17% to 37%
lower than those we found in the same cultivars grown in Tunisia
during the season 2008, with the exception of Dumara and P403
cultivars in which lycopene content was almost identical (Tlili
et al., 2011).
b-Carotene was not detected at the white and white-pink
stages of ripening in all watermelon cultivars investigated. At the
pink stage, the amount of b-carotene was generally very low and
ranged between 0.1 mg/kg fw and 0.4 mg/kg fw for Crimson sweet
and P403 cultivars, respectively. Intermediary values were found
for the other cultivars under investigation. The amount of b-
carotene increased markedly from 4- to 9-fold during the
transition from the pink to the red-ripe stage of ripening reaching
the highest value (2.1 mg/kg fw) in Dumara cultivar and the lowest
(1.0 mg/kg fw) in P403 cultivar. The lack of b-carotene during the
white and the white-pink stages of ripening indicates that the
enzyme involved in cycling lycopene to b-carotene (lycopene b-
cyclase) is not-expressed, or inactive, at the very early stage of
watermelon fruit development, but it starts to be active during the
transition from the white-pink to the pink and red-ripe stages.
Other carotenoids such as a-carotene and lutein were not
detectable in all analysed watermelon cultivars at any stage of
ripening. This indicates the absence or a low expression of the
lycopene a- and b-cyclase enzymes involved in cyclization of
lycopene to a-carotene, which is converted into lutein by the
action of b-ring hydroxylase. However, a small amount of lutein
(1–3 mg/kg fw) has been reported in watermelon by other authors
(Agarwal and Rao, 2000; Perkins-Veazie et al., 2006).
The amounts of lycopene and b-carotene measured in red-ripe
watermelons concur and fall within the ranges (35–112 mg/kg fw
and 0.9–10.2 mg/kg fw, respectively) reported by Perkins-Veazie
et al. (2006) for ripe red-fleshed watermelon cultivars grown in
Oklahoma. Although it has been reported that the cultivars
characterized by high lycopene have, in parallel, higher b-carotene
content (Perkins-Veazie et al., 2006), the data reported in this
study do not seem to corroborate this assumption, since no
correlation was observed between lycopene and b-carotene
amounts in red-ripe watermelons.
Taken together these sets of data indicate that the general
pattern of lycopene and b-carotene synthesis and accumulation is
genotype dependent and highly variable. Differences in agro-
technical and environmental conditions may amplify this variabil-
ity contributing to the substantial discrepancy observed in
watermelons grown in different geographical areas.
As for carotenoids, genetic control is the primary factor in
determining the amount of phenols in fruits and vegetables,
however, variations could also depend on ripening stage at the
time of harvesting, environnemental factors (mainly light and
temperature) (Macheix et al., 1990; Dumas et al., 2003) and
analytical methodology. In most fruits the degree of ripeness
considerably affects the quality and quantity of the various
phenols. In general, it is known that phenolic acid concentrations
decrease during ripening, whereas flavonoid concentrations
increase (Macheix et al., 1990; Manach et al., 2004).
When total phenol content was assayed in ripening tomatoes
results were often contradictory, since it changed differentially
depending on the cultivars (Ilahy et al., in press) or it remained
pretty stable (Helyes and Lugasi, 2006). Our results (Table 1) show
that, in watermelon, the amounts of total phenols and flavonoids
significantly change during ripening (P < 0.05). The highest
amount of phenols and flavonoids was usually detected at the
red-ripe stage in all the studied cultivars, with the exception for
Giza and P403 cultivars.
An increase of flavonoids during ripening was only observed in
Crimson sweet cultivar, whereas a more complex pattern of change
was observed in the other analysed cultivars. The lowest flavonoid
content was detected at the pink stage in Dumara, Giza and P503
cultivars, but at the white-pink stage in P403 cultivar. This lack of a
clear trend in total phenol and flavonoid contents may be
attributed to differential variations in the different compounds
that fall within phenols and flavonoids during ripening, as was
reported by Buta and Spaulding (1997) and Raffo et al. (2002) for
tomato fruits.
At the red-ripe stage, Giza ranked first for total phenol content
(260.2 mg GAE/kg fw), followed by Dumara (246.4 mg GAE/kg fw),
P403 (200.8 mg GAE/kg fw), P503 (183.4 mg GAE/kg fw) and
Crimson sweet (137.2 mg GAE/kg fw) cultivars. A different ranking
was observed for flavonoids which content ranged from 260.0 mg
RE/kg fw in Giza cultivar to less than 200 mg RE/kg fw in P403 and
Dumara cultivars. The amount of flavonoids was in the range of the
values reported for cherry and high-pigment tomato cultivars
(134–622 mg RE/kg fw) by Lenucci et al. (2006), confirming that
watermelon constitute a significant source of flavonoids.
In most cultivars, phenol and flavonoid contents were 77–121%
and 33–74%, respectively higher than those previously reported for
the same cultivars grown in Tunisia (Tlili et al., 2011). An exception
was represented by Crimson sweet cultivar, in which the amount
of total phenols remained almost identical. These differing results
are mainly due to environmental and soil conditions, since the
applied agronomical techniques were identical. A much higher
content of phenols, ranging between 870 and 910 mg GAE kg1 fw,
was reported by Perkins-Veazie (2002) in red fleshed watermelon
cultivars grown in Oklahoma.
Although, the phenol and flavonoid contents in watermelons
are lower than in other fruits and vegetables such as cranberry and
onion, the large consumption of this fruit in the Mediterranean
diet, may considerably contributes to the daily intake of such
compounds. Vinson et al. (2001) reported that watermelon is
classified forth among the eight fruits providing 80% and 50% of the
daily phenols intake in the American and Spanish diet, respective-
ly.
Watermelon fruits have been also identified as a good source of
vitamin C (Vanderslice et al., 1990). Total vitamin C levels were
significantly different between the studied ripening stages
(P < 0.01) (Table 1). Such variability was genotype dependent, in
fact different patterns of change were evidenced for the studied
watermelon cultivars. The vitamin C content of many fruits (apple,
mango, citrus, etc.) is higher when they are immature and decline
as the fruits reach peak ripeness. For other fruits, such as apricot,
peach, papaya, jujube and peppers, the vitamin C content does the
opposite, it rises during ripening (Lee and Kader, 2000; Osuna-
Garcı́a et al., 1998). In muskmelon, vitamin C content is correlated
with the refractive index and with sucrose content of the juice,
hence it increases during fruit ripening (Wagner et al., 1940).
The highest amount of total vitamin C was recorded at the red-
ripe and pink stages in Crimson sweet and P503 cultivars,
respectively, although these cultivars showed a moderate vari-
ability in total vitamin C content during ripening. Contrastingly,
the highest values were measured at the white stage for Giza,
Dumara and P403 cultivars. Similar conflicting results were
reported by Raffo et al. (2002) and Abushita et al. (1997) on
the variability of total vitamin C content during tomato ripening. In
the flesh of red-ripe fruits, total vitamin C values ranged from
119.7 mg/kg fw in P403 cultivar to 204.0 mg/kg fw in Giza cultivar.
If we compare our results with those regarding the same cultivars
grown in Tunisia (Tlili et al., 2011), a reduction in total vitamin C in
the Italian grown Giza, Dumara and P403 is evident. However, the
amount of total vitamin C in the Crimson Sweet and P503 cultivars
does not appear to be influenced by the growing region. These data
are among the first reports on vitamin C content in watermelon
cultivars and confirm that of Leskovar et al. (2004) who reported
that watermelon is considered a good source of vitamin C (10% of
 Author's personal copy

I. Tlili et al. / Journal of Food Composition and Analysis 24 (2011) 923–928 927

recommended daily intake). Nevertheless, lower values ranging Table 3

Correlation coefficient and related significance between antioxidant content and
from 38.2 to 69.8 mg/kg fw were reported for different watermelon
antioxidant activity. n (sample size) = 72.
cultivars grown under 100% evapotranspiration irrigation (Lesko-
var et al., 2004). Higher values attaining 576.2 mg/kg fw were Compounds TEAC assay
reported by Mélo et al. (2006). The variations are probably R P
ascribed to differences in genotypic and cultural practices as was Hydrophilic fraction
reported by Leskovar et al. (2004). Total vitamin C 0.149 nsa
Due to the complexity of the composition of foods, their Total phenols 0.318 <0.01
antioxidant power depends on the synergistic effects and redox Flavonoids 0.398 <0.01
Lipophilic fraction
interactions between the different nutrient and ‘‘non nutrient’’
Lycopene 0.649 <0.01
molecules, which together contribute to the possible health b-carotene 0.403 <0.01
benefits. Therefore, recently, attention has been given to the a
No significant correlation.
antioxidant activity of fruits and vegetables, a parameter that
allows a real evaluation of the nutritional value of foods (Lenucci
et al., 2006; Pellegrini et al., 2007). very narrow variability ranging between 218.0 and 293.5 mM
The HAA and LAA determined by the TEAC assay (Table 2) were Trolox/100 g fw. In most cultivars, the obtained values were in
significantly different between the studied ripening stages in most agreement with those previously reported for Tunisian grown
of the investigated watermelon cultivars (P < 0.01), with the watermelons ranging between 203 and 291 mM Trolox/100 g fw
exception of P403 cultivar in which LAA values resulted unaffected (Tlili et al., 2011). Only Crimson sweet cultivar showed a consistent
by ripening. In Crimson sweet, Giza, P403 and P503 cultivars the increase (>80%) in both HAA and LAA values when produced in
highest HAA was detected at the red-ripe stage, whereas it was Italy with respect to the same cultivar grown in Tunisia; an even
found at the white stage in Dumara, notwithstanding this cultivar greater increase in HAA value (125%) was recorded in P403 when
presents the highest amount of total phenolics and flavonoids at grown in Italy.
the red-ripe stage. A peak in LAA value was reached at the red-ripe Many authors have studied correlations between bioactive
stage of maturity in Crimson sweet, Giza and P503 cultivars. compounds and antioxidant activities in numerous fruits and
During the early ripening stages, the LAA was the lowest at the vegetables particularly tomatoes (Lenucci et al., 2006; Raffo et al.,
white stage in Crimson sweet and P503 cultivars, whereas it was at 2002; Giovanelli et al., 1999). However, little information is known
the white-pink in Giza cultivar. To our knowledge, this is the first concerning these types of correlations in watermelons. In a
report on the variations of antioxidant activities in watermelon previous study we found that TEAC measured HAA well correlated
cultivars during ripening. with phenols but not with total vitamin C. A negative correlation
In the red-ripe fruits, HAA ranged from 265 mM Trolox/100 g with flavonoids was also observed (Tlili et al., 2011). In this study,
fw in Dumara, Giza and P503 cultivars to 535 mM Trolox/100 g fw after considering data from all watermelon cultivars, we confirm
in Crimson sweet and P403 cultivars. Crimson sweet proved to be that no significant correlation between HAA values and total
the watermelon cultivar with the highest LAA (467.9 mM Trolox/ vitamin C (R = 0.149; P > 0.05) was obtained, whereas they
100 g fw), whereas the other cultivars showed lower values and a correlated well with both phenols (R = 0.318; P < 0.01) and
flavonoids (R = 0.398; P < 0.01) (Table 3). Although, it is likely
Table 2 that HAA depends upon synergistic effects among all hydrophilic
Hydrophilic and lypophilic antioxidant activities in watermelon cultivars harvested antioxidants and their interaction with other constituents of the
at four different ripening stages.a fraction (Diplock et al., 1998; Lenucci et al., 2006), this differential
Cultivar TEAC assay (mM Trolox/100 g fw) presence/absence of correlation between HAA values obtained
HAA LAA
with the different classes of hydrophilic antioxidants, could be due
to a higher sensibility of the TEAC assays for such classes of
Crimson sweet
compounds. Moreover, the test reaction used for antioxidant
White 148.5  0.2d 123.0  11.1c
White-pink 301.4  1.9b 253.1  27.5b activity measurement might be differentially influenced by other
Pink 195.0  2.1c 301.9  2.0b compounds involved in complex antioxidant system such as
Red-ripe 535.1  3.4a 467.9  22.6a glutathione and other enzymatic components (Jiménez et al.,
Giza 2002).
White 250.8  7.3b 209.2  1.0c
White-pink 165.6  5.1c 173.4  0.4d
The LAA has been mainly attributed to the presence of
Pink 161.2  6.4c 253.1  2.3b carotenoids particularly lycopene in tomato fruits (Martinez-
Red-ripe 270.7  16.0a 293.5  7.8a Valverde et al., 2002; Raffo et al., 2002; Cano et al., 2003; George et
Dumara al., 2004). After considering data from all watermelon cultivars,
White 324.2  5.8a 186.9  2.5b
good and significant correlations between TEAC values and
White-pink 211.9  4.9b 239.6  38.3ab
Pink 265.4  0.8c 259.9  12.3ab lycopene (R = 0.649; P < 0.01) and b-carotene (R = 0.403;
Red-ripe 261.5  9.8b 266.9  13.7a P < 0.01) contents were obtained during ripening (Table 3). It
P403 should be underlined that no correspondence was found between
White 267.3  1.9b 188.4  1.7a the approximate increase in carotenoid contents in watermelon
White-pink 97.4  10.8d 163.4  26.8a
Pink 143.7  13.0c 176.9  19.0a
fruits (averaged across cultivars), when red-ripe compared to the
Red-ripe 546.5  15.8a 218.0  7.0a white stage of ripeness, and the increase in LAA. This may probably
P503 be due to the presence of other lipophilic antioxidant compounds
White 141.4  9.0b 126.2  0.8c in the watermelon fruits which could account for most of the LAA
White-pink 112.9  4.9b 190.5  18.6b
at the white stage.
Pink 243.3  4.2a 196.0  6.0b
Red-ripe 264.7  2.1a 281.0  7.6a
a
4. Conclusions
Values are given as mean  SEM (n = 3). Letters indicate mean separation within
ripening stages of each cultivar; values with different letters are significantly different
at P < 0.05 by LSD test. HAA = hydrophilic antioxidant activity; LAA = lypophilic Our data highlight the important role played by genetic
antioxidant activity. background and ripening stage in determining the antioxidant
 Author's personal copy
928 I. Tlili et al. / Journal of Food Composition and Analysis 24 (2011) 923–928
potential of watermelon. The amount of lycopene and b-carotene,
as well as the LAA, markedly increased in all the studied
watermelon cultivars with ripening, whereas the contents of total
phenols, flavonoids, total vitamin C and HAA were affected by fruit
ripening in a cultivar-dependent way. Hence, the increase of
antioxidants during watermelon fruit ripening must not be
generalized; rather, their contents reach levels leading to the
maximum nutritional quality depending on their genotypic
differences.
The study of changes in bioactive compounds and antioxidant
activity during ripening is of great relevance both to human health
and commercial purposes. In fact, it provides valuable information to
understand the synthesis of these compounds and for evaluating the
best harvesting period to reach the highest antioxidant potential.
These data are useful to plant breeders and food scientists working to
maximize the nutritional value and antioxidant contents of fresh
watermelon cultivars. Furthermore, they confirm that this refresh-
ing summer fruit represents a nutritionally balanced source of
dietary antioxidants for the population of the Mediterranean area
because of its availability and high consumption.
Acknowledgements
The authors wish to thank Prof. Gabriela Piro for laboratory
facilities and Dr. Gaetano Carrozzo for technical assistance in
watermelon plant cultivation.
References
Abushita, A.A., Hebshi, E.A., Daood, H.G., Biacs, P.A., 1997. Determination of vitamin
and antioxidant in tomatoes. Food Chemistry 60 (2), 207–212.
Abushita, A.A., Daood, H.G., Biacs, P.A., 2000. Change in carotenoids and antioxidant
vitamins in tomato as a function of varietal and technological factors. Journal of
Agricultural and Food Chemistry 48, 2075–2081.
Agarwal, S., Rao, A.V., 2000. Tomato lycopene and its role in human health and
chronic diseases. Canadian Medical Association Journal 163, 739–744.
Bramley, P.M., 2002. Regulation of carotenoid formation during tomato fruit
ripening and development. Journal of Experimental Botany 53 (377),
2107–2113.
Buta, J.G., Spaulding, D.W., 1997. Endogenous levels of phenolics in tomato fruit
during growth and maturation. Journal of Plant Growth Regulation 16, 43–46.
Cano, A., Acosta, M., Arnao, M.B., 2003. Hydrophilic and lipophilic antioxidant
activity changes during on-vine ripening of tomatoes (Lycopersicon esculentum
Mill.). Postharvest Biology and Technology 28 (1), 59–65.
Diplock, A.T., Charleux, J.L., Crozier-Willi, G., Kok, F.J., Rice-Evans, C., Roberfroid, M.,
Stahl, W., Viña-Ribes, J., 1998. Functional food science and defence against
reactive oxygen species. British Journal of Nutrition 80, S77–S112.
Dumas, Y., Dadomo, M., Lucca, G.D., Grolier, P., Di Lucca, G., 2003. Effects of
environmental factors and agricultural techniques on antioxidant content of
tomatoes. Journal of the Science of Food and Agriculture 83, 369–382.
George, B., Kaur, C., Khurdiya, D.S., Kapoor, H.C., 2004. Antioxidants in tomato
(Lycopersicon esculentum) as a function of genotype. Food Chemistry 84 (1),
45–51.
Giovanelli, G., Lavelli, V., Peri, C., Nobili, S., 1999. Variation in antioxidant compo-
nents of tomato during vine and post-harvest ripening. Journal of the Science of
Food and Agriculture 79, 1583–1588.
Giovannucci, E., 1999. Tomatoes, tomato-based products, lycopene, and cancer:
review of the epidemiologic literature. Journal of the National Cancer Institute
91, 317–331.
Helyes, L., Lugasi, A., 2006. Formation of certain compounds having technological
and nutritional importance in tomato fruit during maturation. Acta Alimentaria
35 (2), 183–193.
Ilahy, R., Hdider, C., Lenucci, M.S., Tlili, I., Dalessandro, G., Antioxidant activity and
bioactive compound changes during fruit ripening of high-lycopene tomato
cultivars. Journal of Food Composition and Analysis in press, doi:10.1016/
j.jfca.2010.11.003.
Jiménez, A., Creissen, G., Kular, B., Firmin, J., Robindon, S., Verhoyen, M., Mullineaux,
P., 2002. Changes in oxidative processes and components of the antioxidant
system during tomato fruit ripening. Planta 214, 751–758.
Kampfenkel, K., Van Montagu, M., Inzé, D., 1995. Extraction and determination of
ascorbate and dehydroascorbate from plant tissue. Analytical Biochemistry
225, 165–167.
View publication stats
Lee, S.K., Kader, A.A., 2000. Preharvest and postharvest factors influencing vita-
min C content of horticultural crops. Postharvest Biology and Technology 20,
207–220.
Lenucci, M.S., Cadinu, D., Taurino, M., Piro, G., Dalessandro, G., 2006. Antioxidant
composition in cherry and high-pigment tomato cultivars. Journal of Agricul-
tural and Food Chemistry 54, 2606–2613.
Lenucci, M.S., Caccioppola, A., Durante, M., Serrone, L., De Caroli, M., Piro, G.,
Dalessandro, G., 2009. Carotenoid content during tomato (Solanum lycopersicum
L.) fruit ripening in traditional and high-pigment cultivars. Italian Journal of
Food Science 4 (21), 461–472.
Leskovar, D.I., Bang, H.J., Crosby, K., Maness, N., Franco, J.A., Perkins-Veazie, P., 2004.
Lycopene, carbohydrates, ascorbic acid, and yield components of diploid and
triploid watermelon cultivars are affected by deficit irrigation. Journal of
Horticultural Science and Biotechnology 79 (3), 75–81.
Macheix, J.J., Fleurient, A., Billot, J., 1990. Phenolic compounds in fruit processing. In:
Mitra, S. (Ed.), Fruit Phenolic. CRC Press, Boca Raton, FL, USA, pp. 295–342.
Manach, C., Scalbert, A., Morand, C., Rémésy, C., Jiménez, L., 2004. Polyphenols: food
sources and bioavailability. American Journal of Clinical Nutrition 79, 727–747.
Martinez-Valverde, I., Periago, M.J., Provan, G., Chesson, A., 2002. Phenolic com-
pounds, lycopene and antioxidant activity in commercial varieties of tomato
(Lycopersicon esculentum). Journal of the Science of Food and Agriculture 82,
323–330.
Mélo, E.A., Lima, V.L.A.G., Mciel, M.I.S., Caetano, A.C.S., Leal, F.L.L., 2006. Polyphenol,
ascorbic acid and total carotenoid contents in common fruits and vegetables.
Brazilian Journal of Food Technology 9 (2), 89–94.
Miller, N.J., Rice-Evans, C.A., 1997. The relative contribution of ascorbic acid and
phenolic antioxidants to the total antioxidant activity of orange and apples
fruits juices and blackcurrant drinks. Food Chemistry 60 (3), 331–337.
Osuna-Garcı́a, J.A., Wall, M.M., Waddell, C.A., 1998. Endogenous levels of toco-
pherols and ascorbic acid during fruit ripening of new mexican-type chile
(Capsicum annuum L.) cultivars. Journal of Agricultural Food Chemistry 46
(12), 5093–5096.
Pellegrini, N., Colombi, B., Salvatore, S., Brenna, O., Galaverna, G., Del Rio, D., 2007.
Evaluation of antioxidant capacity of some fruit and vegetable foods: efficiency
of extraction of a sequence of solvents. Journal of the Science of Food and
Agriculture 87, 103–111.
Perkins-Veazie, P., 2002. Composition of orange, yellow, and red fleshed watermel-
on. Cucurbitacea 436–440.
Perkins-Veazie, P., Collins, J.K., Pair, S.D., Roberts, W., 2001. Lycopene content differs
among red-fleshed watermelon cultivars. Journal of the Science of Food and
Agriculture 81, 983.
Perkins-Veazie, P., Collins, J.K., Davis, A.R., Roberts, W., 2006. Carotenoid content
of 50 watermelon cultivars. Journal of Agricultural and Food Chemistry 54,
2593–2597.
Perkins-Veazie, P., Collins, J.K., Clevidence, B., 2007. Watermelons and health. Acta
Horticulture (ISHS) 731, 121–128.
Raffo, A., Cherubino, L., Vincenzo, F., Ambrozino, P., Salucci, M., Gennaro, L.,
Bugianesi, R., Giuffrida, F., Quaglia, G., 2002. Nutritional value of cherry toma-
toes (Lycopersicon esculentum cv. Naomi F1) harvested at different ripening
stages. Journal of Agricultural and Food Chemistry 50, 6550–6556.
Rao, A.V., 2006. Tomatoes, Lycopene and Human Health. Preventing Chronic Dis-
eases. Caledonian Science Press, Badalona, Spain.
Rice-Evans, C.A., Miller, N.J., Paganga, G., 1996. Structure antioxidant activity
relationships of flavonoids and phenolic acids. Free Radical Biology and Medi-
cine 20 (7), 933–956.
Sadler, G., Davis, J., Dezman, D., 1990. Rapid extraction of lycopene and b-carotene
from reconstituted tomato paste and pink grapefruit homogenates. Journal of
Food Science 55, 1460.
Spanos, G.A., Wrolstad, R.E., 1990. Influence of processing and storage on the
phenolic composition of Thompson Seedless grape juice. Journal of Agricultural
and Food Chemistry 38, 1565–1571.
Tlili, I., Hdider, C., Lenucci, M.S., Ilahy, R., Jebari, H., Dalessandro, G., 2011. Bioactive
compounds and antioxidant activities of different watermelon (Citrullus lana-
tus (Thunb.) Mansfeld) cultivars as affected by fruit sampling area. Journal of
Food Composition and Analysis 24 (3), 307–314.
Vanderslice, J.T., Higgs, D.J., Hayes, I.M., Block, G., 1990. Ascorbic acid and dehy-
droascorbic acid content of foods-as-eaten. Journal of Food Composition and
Analysis 3, 105–118.
Vinson, J.A., Su, X., Zubic, S.K., Bose, P., 2001. Phenol antioxidant quantity and
quality in foods: fruits. Journal of Agricultural and Food Chemistry 49, 5315–
5321.
Wagner, L.E., Hoffman, J.C., Brown, H.D., 1940. Correlation between the vitamin C
content and refractive indices of muskmelon. Proceedings of American Society
for Horticultural Science 40, 839–840.
Waterman, P.G., Mole, S., 1994. Why are phenolic compounds so important. In:
Waterman, P.G., Mole, S. (Eds.), Analysis of Phenolic Plant Metabolites. Black-
well, Oxford, pp. 61–63.
Zhishen, J., Mengcheng, T., Jianming, W., 1999. The determination of flavonoid
contents in mulberry and their scavenging effects on superoxide radicals. Food
Chemistry 64 (4), 555–559.