ARTICLE IN PRESS

JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 19 (2006) 11–19
www.elsevier.com/locate/jfca

Original Article

Seasonal variations in antioxidant components of cherry tomatoes

(Lycopersicon esculentum cv. Naomi F1)
Antonio Raffoa,, Giuseppe La Malfab, Vincenzo Foglianoc,
Giuseppe Maiania, Giovanni Quagliaa
a
Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, Via Ardeatina 546, 00178 Roma, Italy
b
Dipartimento di Orto-Floro-Arboricoltura e Tecnologie Agroalimentari, Università di Catania, Via Valdisavoia 5, 95123 Catania, Italy
c
Dipartimento di Scienza degli Alimenti, Università di Napoli ‘‘Federico II’’, P.co Gussone, 80055 Portici, Napoli, Italy
Received 21 April 2004; received in revised form 11 January 2005; accepted 7 February 2005

Abstract

To evaluate seasonal variations in antioxidant components of greenhouse cherry tomatoes, the compositional profile of fruits
(‘‘Pomodoro di Pachino’’, cv. Naomi F1) harvested at six different times of the year was compared. Among tomato antioxidants,
phenolic compounds (naringenin content ranged from 1.84 to 9.04 mg/100 g, rutin from 1.79 to 6.61 mg/100 g) and a-tocopherol
(40–1160 mg/100 g) showed the greatest variability. Ascorbic acid (31–71 mg/100 g), carotenoids (8350–15119 mg/100 g), phenolics
and a-tocopherol concentration did not show definite seasonal trends, nor correlation with solar radiation or average temperature.
Nevertheless, tomatoes harvested in mid-summer were characterized by lowered lycopene levels. Greenhouse growing conditions
induced the accumulation of relatively high level of antioxidants for most of the year: one serving of raw tomatoes (100 g) could
provide from 50% to 120% of the recommended daily intake of vitamin C, from 13% to 27% of that of vitamin A, from 0.4% to
12% of that of vitamin E, and from 15% to 35% of the flavonoid daily intake estimated for an Italian diet.
r 2005 Elsevier Inc. All rights reserved.

Keywords: Cherry tomato; Season; Carotenoids; Ascorbic acid; Phenolic compounds; a-tocopherol; Antioxidant activity

1. Introduction et al., 2004). This protective action is typically attributed

Several epidemiologic studies suggest that consump-
tion of tomatoes and tomato-based products reduces the
risk of chronic diseases such as cardiovascular disease
and cancer (Sesso et al., 2003; Weisburger, 2002; Willcox
et al., 2003). In particular, intake of tomato and tomato-
based products has been relatively consistently asso-
ciated with a lower risk of cancers of the prostate, lung
and stomach (Giovannucci, 1999). Recent intervention
studies demonstrate that regular intake of small
amounts of tomato products can increase cell protection
from DNA damage induced by oxidant species (Riso
Corresponding author. Tel.: +39 06 5149 4409;
fax: +39 06 5149 4550.
E-mail address: raffo@inran.it (A. Raffo).
to antioxidant components like lycopene and other
carotenoids, ascorbic acid, flavonoids and vitamin E.
Due to its relatively high average consumption
tomato is an important source of these dietary
antioxidants. Tomatoes are the most highly consumed
vegetable in Italy, with the highest average consumption
among European countries (NETTOX, 1998). In a
study on the Italian food consumption patterns in the
1990s, a consumption of tomatoes (both for salad and
ripe) of 75.5 g/day/capita has been estimated (Turrini
et al., 2001). As a matter of fact, tomatoes have been
assessed to be the second most important source of
vitamin C in the Italian diet (La Vecchia, 1998).
Moreover, in a recent study on dietary sources of
vitamin C, vitamin E and specific carotenoid in Spain
(Garcı́a-Closas et al., 2004), tomatoes ranked first as a

0889-1575/$ - see front matter r 2005 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2005.02.003
 ARTICLE IN PRESS
12 A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19
source of lycopene (71.6%), second as a source of
vitamin C (12.0%), pro-vitamin A carotenoids (14.6%)
and b-carotene (17.2%), and third as a source of vitamin
E (6.0%).
In Italy, cherry tomatoes are largely used for fresh
consumption (more than 25% of the market), and their
commercial importance is continuously increasing (Leo-
nardi et al., 2000a, b). In Sicily, as well as in other
regions of the Mediterranean basin, cherry tomatoes are
grown year round in unheated greenhouses, which have
no climate control systems and are covered with plastic
film; consequently, development and ripening of fruits
occur under varying climatic conditions. Temperature
and light intensity exert a direct influence on the quality
attributes of tomato fruit, such as appearance, firmness,
texture, dry matter and sensory properties (Dorais et al.,
2001). On the other hand, environmental factors can
also affect the antioxidant content of tomatoes: Dumas
et al. (2003) have recently reviewed the available
knowledge about the effects of climatic and other pre-
harvest factors on the antioxidant contents of tomatoes.
At present, a thorough understanding of the influence of
environmental factors and their interactions with
agronomic practices on the accumulation of antiox-
idants during the fruiting period is still lacking, although
some relationships have been observed in general. Light
exposure is favourable to vitamin C accumulation
(Dumas et al., 2003; Lee and Kader, 2000). Several
studies have reported on the effect of shading (by leaves
or artificial covers) in decreasing the ascorbic acid
content (El-Gizawi et al., 1993; Venter, 1977), whereas
greenhouse-grown tomatoes were usually found to have
lower vitamin C levels than those grown outdoors,
chiefly because of the lower light intensity (Lopez-
Andreu et al., 1986). On the contrary, lycopene synthesis
is severely inhibited by exposure to intense solar
radiation, and it has been suggested that radiation
injury to tomato fruit might be due to the general effects
of overheating on irradiated tissues (Adegoroye and
Jolliffe, 1987; Dumas et al., 2003). In fact, the formation
of lycopene depends on the temperature range and
seems to occur between 12 and 32 1C, whereas higher
temperatures specifically inhibit its accumulation (Ha-
mauzu et al., 1998; Leoni, 1992; Robertson et al., 1995).
As regards phenolic compounds, although genetic
control represents the main factor in determining their
accumulation in vegetable foods, external factors may
also have a significant effect on this (Macheix et al.,
1990). In many plant species the flavonol content may be
enhanced in response to elevated light levels, in
particular to increased UV-B radiation (Brandt et al.,
1995). As a matter of fact, it has been reported that
cherry tomato plants grown in greenhouse under high
light accumulated an approximately two-fold greater
soluble phenols content (rutin and chlorogenic acid)
than low-light plants (Wilkens et al., 1996).
On the other hand, it has been only infrequently been
investigated whether these effects might produce definite
seasonal trends in tomato antioxidant compositional
profile. Only a few studies have investigated seasonal
fluctuations of nutrients, and in particular the phytonu-
trient content of tomatoes. Vanderslice reported on
seasonal differences in ascorbic acid content of tomato,
with higher levels in summer than in spring (Vanderslice
et al., 1990). Seasonal variations in vitamin C content
were observed in greenhouse-grown tomatoes at the
mature-green stage, and were directly correlated with
temperature variations (Liptay et al., 1986). As regards
phytonutrients, marked variations have been observed
in the level of quercetin in cherry tomatoes grown at
different times of the year, but no definite seasonal
trends have been brought into evidence (Crozier et al.,
1997; Stewart et al., 2000). Only very little information is
available on the seasonal variations of carotenoid
content in the tomato fruit (Heinonen et al., 1989).
On the basis of the above observations, variations in
climatic conditions between different seasons can be
expected to significantly influence the compositional
profiles of tomato typologies produced year round.
Thus to better evaluate the real contribution of tomato
consumption to total daily intake of antioxidants, it is
interesting to know not only the average content of
antioxidants, but also the extent of their seasonal
fluctuations.
To develop information on seasonal variations in
cherry tomato antioxidant properties, we compared the
detailed compositional profile (ascorbic acid, phenolic
compounds, carotenoids, a-tocopherol) of greenhouse
cherry tomatoes (cv. Naomi F1), grown in Sicily within
the same geographical area (‘‘Pomodoro di Pachino’’),
and harvested at six different times of the year. The aim
of this work was to estimate the extent of variation in
cherry tomato antioxidant content and antioxidant
activity for 1 year of production, and to show possible
relationships between antioxidant concentrations and
climate factors such as temperature and solar radiation.
2. Material and methods
2.1. Fruit sampling
Cherry tomatoes (‘‘Pomodoro di Pachino’’, cv.
Naomi F1) were sampled on plants grown in cold
greenhouses, within farms belonging to the Association
for the protection of the typical products of Pachino
(Province of Siracusa, Sicily). ‘‘Pomodoro di Pachino’’
tomatoes, which have recently achieved the PGI
(Protected Geographical Indication) European recogni-
tion (Official Journal of European Communities, 2002),
are produced in a protected environment (glasshouses
and/or tunnels) covered with polyethylene netting or
 ARTICLE IN PRESS
A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19
other plastic material; planting out takes place through-
out the year, at a density of 2–6 plants/m2. The plants
are grown upright and have one or more branches;
irrigation is carried out using groundwater taken from
wells sunk with a salinity level ranging from 1500 to
10 000 mS.
For our experiment, harvests were carried out at six
different times of the year: April, June, July, December
(1999), January and March (2000). Tomatoes were
harvested at full ripeness, as usually occurs for market-
ing. To mediate the effects of technical growing
conditions, within each sampling, fruits were harvested
from ten different representative farms, selected for their
uniformity, and pooled in one sample. Then, antiox-
idant content and antioxidant activity were determined
on a group of 30 fruits taken at random from each
pooled sample.
On a separate group of fruits taken from the same
pooled samples, dry matter and skin colour were
determined. Dry matter content was determined by
gravimetry; about 5 g of homogenized sample was dried
in a thermoventilated oven at 70 1C until constant
weight was reached. Chromatic coordinates (L*, a*, and
b*) were determined on the equatorial part of the fruit as
described by McGuire (1992), using a tristimulus
Minolta Chroma meter (model CR-200, Minolta Corp.).
Colour was described by the ratio a*/b*.
2.2. Biochemical analyses
Whole tomatoes were homogenized in a Waring
blender for 1 min. The homogenate was then frozen at

20 1C and stored until analysed. All data reported in
tables were expressed on fresh weight basis.
2.2.1. Ascorbic acid
Ascorbic acid, reduced and total (ascorbic plus
dehydroascorbic acid), was extracted from freshly
homogenized tomatoes and quantified by HPLC ac-
cording to the method described by Margolis and
Schapira (1997). HPLC separation was carried out
using an ESA HPLC system with an eight-channel
coulometric electrode array detector (ESA), on a
Capcell Pak NH2 column (250  4.4 mm) (Shiseido) at
a flow rate of 1 mL/min, at 40 1C; the mobile phase was
as described in the above-mentioned paper. Pure
standards (Sigma Chemicals) were used for identifica-
tion of peaks and for quantification.
2.2.2. Phenolic compounds
Phenolics were either hydrolysed or not, in order to
obtain free (naringenin, caffeic, p-coumaric and ferulic
acid) and conjugated (chlorogenic acid and rutin) forms,
and extracted as described by Hertog et al. (1992).
Quantitative analysis was performed using the ESA
HPLC system previously mentioned. HPLC separation
13
was carried out at a flow rate of 1 mL/min, at 30 1C,
using a Supelcosil LC-18 column (250  4.6 mm) with a
Perisorb Supelguard LC-18 (Supelco); the mobile phase
and the elution programme were as described by Hertog
et al. (1992). The calibration curve for quantification
was obtained using authentic standards (Sigma Chemi-
cals).
2.2.3. Carotenoids
The procedure described by Tonucci et al. (1995) was
followed with slight modification. Homogenized toma-
toes were extracted in THF in the presence of butylated
hydrohytoluene (BHT) (from Sigma Chemicals), and
resuspended in 5 mL of CHCl3. A further 1:10 dilution
of the extracted material in 40% CH3CN, 20%
methanol, 20% hexane, and 20% CH2Cl2 was per-
formed before the chromatographic analysis. HPLC
separation was carried out at a flow rate of 0.8 mL/min
and a temperature of 30 1C using a Shimadzu HPLC
with diode array detection and a Supelcosil C18 column
(250  4.6 mm). Carotenoid elution was achieved using
the following linear gradient: starting condition, 82% A,
18% B; 20 min, 76% A, 24% B; 30 min, 58% A, 42% B;
40 min, 39% A, 61% B. ‘‘A’’ was CH3CN and ‘‘B’’ was
methanol/hexane/CH2Cl2 1:1:1 v/v. Quantification of
carotenoids was achieved by calibration curve obtained
with authentic standard (b-carotene from Fluka) or
HPLC-purified compounds (lycopene and other deter-
mined compounds). The concentration of the standards
was calculated using the extinction coefficient.
2.2.4. a-tocopherol
A-tocopherol from homogenized tomatoes was mea-
sured according to Baldini et al. (1996). Briefly, the
extraction consists of base hydrolysis of the sample
followed by C2H4Cl2 addition. The organic phase was
dried, and the residue resuspended in 1 mL of CH3OH
and run on HPLC column (Phenomenex Prodigy C18,
5 mm; 4.6  250 mm). The mobile phase was isocratic
CH3OH: H2O (98:2) with a flow rate of 1 mL/min. A-
tocopherol was identified by comparison with a pure
standard (Sigma Chemicals) and quantified by measur-
ing absorbance at 290 nm.
2.2.5. Antioxidant activity
A quantity of 10 mL of deionized water was added to
5 g of freshly homogenized tomatoes, and the suspension
was centrifuged at 15 000 rpm for 15 min (4 1C). The
supernatant (water-soluble fraction) was recovered,
filtered, and after suitable dilution (1:5) with phosphate
buffer (10 mM, pH 7.0), used for the crocin bleaching
inhibition test (Tubaro et al., 1996). Antioxidant activity
was expressed as equivalent millimolar Trolox (6-
hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid)
for 100 g of tissue fresh weight. The pulp resulting from
centrifugation of the homogenate was extracted with
 ARTICLE IN PRESS
14 A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19

10 mL of CH2Cl2, centrifuged at 15 000 rpm for 5 min 3. Results and discussion

(4 1C), filtered, and the supernatant was recovered;
this extraction step was repeated three times, and
supernatant fractions were pooled. The extract (water-
insoluble fraction) was used for the 2,20 -azinobis(3-
ethylbenzothiazoline-6-sulphonic acid) (ABTS) test
(Pellegrini et al., 1999), and antioxidant activity was
expressed as equivalent millimolar Trolox for 100 g of
tissue fresh weight.
2.3. Statistical evaluation of data
We performed analysis of variance (ANOVA) in
order to test the significance of the observed differences.
When the effects of harvesting date were significant
(Pp0:05), the mean values for each parameter were
compared by a multiple comparison Duncan test to look
for grouping (at P ¼ 0:05).
The content of some tomato antioxidants and the
antioxidant activity of tomato extracts are greatly
affected by ripening stage (Cano et al., 2003; Dumas
et al., 2003; Raffo et al., 2002). In our experiment,
tomato fruits were harvested at full ripeness, the stage at
which they are usually consumed, and chromaticity
values determined on the skin showed only slight
differences between different sampling times, denoting
quite similar stage of ripeness. The a*/b* ratio, a simple
and appropriate ripening index (Giovanelli et al., 1999),
did not show significant differences at P ¼ 0:05 (a*/b*
mean values were: 1.31 (April), 1.14 (June), 1.42 (July),
1.33 (December), 1.35 (January), 1.35 (March)). More
marked variations between sampling times were ob-
served in dry matter content (mean values were,
respectively, 6.97%, 8.93%, 8.14%, 7.37%, 9.10%,
8.67%).

2.4. Climatic data

3.1. Ascorbic acid
Climatic data in the area of production were recorded
daily at the weather station in Santo Pietro. Average Fruits harvested at different times were characterized
temperature and mean solar radiation detected during by markedly variable ascorbic acid levels (CV 34.5%
the month preceding the harvest are presented in and 24.4%, for reduced and total ascorbic acid);
Table 1. reduced ascorbic acid ranged from 16 to 44 mg/100 g
(230–493 mg/100 g of dry weight), total ascorbic acid
(the sum of ascorbic and dehydroascorbic acid, repre-
Table 1
senting vitamin C content) from 31 to 71 mg/100 g
Average climate data detected during the month preceding the harvest
(445–795 mg/100 g d.w.) (Table 2). A range of seasonal
Date of harvest Average temperature Mean solar radiation variation from 7 to 23 mg/100 g was observed by Liptay
(1C) (MJ m
2 day
1) et al. (1986) on greenhouse grown fruits of cv. Jumbo at
April 6 11.5 17.2
the mature-green stage. In the samples of our experi-
June 8 22.4 24.5 ment, the level of total ascorbic acid was relatively high
July 26 23.6 26.1 when compared with published data on different
December 21 11.0 6.8 cultivars for fresh consumption grown in several
January 25 7.1 7.7 countries. Davies and Hobson (1981) reported the range
March 20 10.5 14.0
from 10 to 45 mg/100 g f.w. for reduced ascorbic acid

Table 2
Ascorbic acid, reduced and total, and phenolic compounds content (mg/100 g) in tomatoes harvested at different times of the year

Compound Apr Jun Jul Dec Jan Mar CV

Ascorbic acid
Reduced 1673 aa 4471 e 2575 bc 2871 c 2271 b 3372 d 34.5
Total 3177 a 7179 c 6473 bc 5573 b 5571 b 6175 bc 24.4

Phenolic compounds
Chlorogenic acid 5.4470.70 c 3.0870.71 ab 3.1270.08 ab 3.7670.67 ab 4.3570.13 bc 2.6770.08 a 28.0
Caffeic acid 0.2770.01 a 0.2870.09 a 0.2470.10 a 0.1770.01 a 0.2970.01 a 0.5670.24 b 44.3
p-Coumaric acid 0.6870.04 c 0.4570.04 b 0.3670.02 a 0.4870.05 b 0.7970.05 d 0.6370.08 c 28.6
Ferulic acid 0.1670.01 a 0.1570.02 a 0.0970.02 a 0.0970.02 a 0.2970.07 b 0.0470.01 a 63.6
Rutin 1.7970.07 a 2.0370.17 a 2.7670.14 b 2.9270.20 b 3.8770.17 c 6.6170.07 d 53.0
Naringenin 9.0470.55 d 2.6770.41 b 5.4070.35 c 2.1870.21 ab 1.9270.10 a 1.8470.13 a 74.8

Values are means (7 standard deviations) of three samples; CV represents coefficient of variation of mean values.
a
In this and in the following tables, different letters indicate significant differences according to the Duncan test (P ¼ 0:05).
 ARTICLE IN PRESS
A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19
content in commercial cultivars from different parts of
the world, whereas Souci et al. (1994) reported the range
from 20.0 to 28.8 mg/100 g for vitamin C. Several factors
could have contributed to the relatively high accumula-
tion of ascorbic acid in the tomatoes of our experiment.
The first factor is the cultivar, since it is known that
tomatoes from cherry cultivars contain higher ascorbic
acid levels, as well as dry matter and soluble solids, than
normal-sized fruits (Kader et al., 1977). The second
factor is the relatively high salinity of the irrigation
water used in the considered production area (Leonardi
et al., 2000a, b); it has been observed, in fact, that
irrigation water of high salinity may enhance the
ascorbic acid content, as well as dry matter, soluble
solids and titratable acidity (Petersen et al., 1998; De
Pascale et al., 2001). The third factor is the particularly
sunny climate of the production area; it has been
reported that the region where ‘‘Pomodoro di Pachino’’
are grown has the highest temperatures and receives the
greatest amount of solar radiation, averaged out
over the year, in mainland Europe (Official Journal
of European Communities, 2002). As expected from
its antioxidant-based functions in plant metabolism, the
level of ascorbic acid is responsive to a wide variety
of environmental stress factors, including light and
temperature; in fact, several works have reported
that plants increase their ascorbic acid levels in response
to light (Davey et al., 2000) and that high light intensity
is associated with a high vitamin C content in
tomato fruit (Dumas et al., 2003; Lee and Kader,
2000). In our study, tomato samples harvested at
different times did not show any significant correlation
between ascorbic acid content and climatic parameters.
Nevertheless, the effect of relatively high light and
temperature (Table 1) could have contributed, at least
partly, to the relatively high level of total ascorbic acid
observed in the fruits harvested in June and July. The
effects induced by climatic variations were probably
superimposed upon other factors, such as sanitary and
nutritional status of the plant, and changes in water
availability.
We found, moreover, a relatively high amount of
dehydroascorbic acid (40–60% of total ascorbic acid),
whereas lower proportions have been previously re-
ported for ripe tomatoes: 20–30% according to Van-
derslice et al. (1990) and Davies and Hobson (1981),
whereas even lower proportions have been observed by
Cano et al. (2003) and Jimenez et al. (2002). On the
contrary, over 90% of the vitamin C was found as
dehydroascorbic acid in fresh tomatoes grown in warm
climates (De Serrano et al., 1993). In any case, it is
evident that, in the tomatoes we considered, the simple
ascorbic acid quantification would lead to a substantial
underestimation of vitamin C content. From a nutri-
tional point of view, one serving of the considered raw
cherry tomatoes (100 g) may provide from 50% to 120%
15
of the recommended daily intake of vitamin C for the
Italian people (Società Italiana di Nutrizione Umana,
1996).
3.2. Phenolic compounds
Among main tomato antioxidants, phenolic com-
pounds showed the greatest variations between different
sampling times (Table 2). Naringenin content varied
from 1.84 to 9.04 mg/100 g (21–130 mg/100 g d.w.) with
a CV of 74.8%, and chlorogenic acid from 2.67 to
5.44 mg/100 g (31–78 mg/100 g d.w.) with a CV of
28.0%. Rutin level ranged from 1.79 to 6.61 mg/100 g
(0.99–3.66 mg/100 g when expressed as conjugated quer-
cetin), with a range of variation (CV of 53.0%)
similar to that found by Stewart et al. (2000) in Spanish
field-grown cherry tomatoes (cv. Paloma) sampled over
a period of 13 months. Rutin concentration relative to
dry weight ranged from 27 to 76 g/100 g. Hertog et al.
(1992) observed variations in flavonol levels associated
with seasonal influences in some leafy vegetables
(lettuce, endive and leek), with the highest levels in
summer. On the other hand, in our experiment, as well
as in that on Spanish cherry tomatoes, no definite
seasonal trends were evidenced in the main phenolic
level; moreover, we did not observe any correlation
between rutin, naringenin and chlorogenic acid content
and climatic parameters. Limited amounts (less than
1 mg/100 g f.w.) of other hydroxycinnamic acids (p-
coumaric, caffeic and ferulic acid) were also found in all
samples.
As observed in the case of ascorbic acid, it is possible
that phenolic concentration was affected by a wide
range of cultural and environmental factors. In plants,
moreover, phenolic biosynthesis is particularly sensitive
to induction by various biotic and abiotic stresses
(Dixon and Paiva, 1995): for instance, flavonols
like quercetin and kaempferol can be synthesized in
response to a pathogen attack or to wounding, whereas
wound-induced chlorogenic acid and phenolic esters
may act directly as defence compounds or may serve
as a precursor for the synthesis of polyphenolic barriers.
Consequently, a limited number of samples are not
adequate to evidence significant relationships with
single climatic factors. Interestingly, the phenolics
pattern showed noticeable variations between different
sampling times, suggesting that each compound was
affected in a different way by the same environmental
changes.
From our data, it resulted that one serving of cherry
tomatoes provided from 3.7 to 10.3 mg of flavonoids
(conjugated quercetin plus naringenin), which was equal
to approximately 15–35% of the flavonoid daily intake
estimated for the Italian diet (23–34 mg/diet) (Hertog
et al., 1995), confirming the relevance of tomatoes as
dietary sources of flavonoids.
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16 A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19

3.3. Carotenoids skin) do not necessarily contain similar amounts of total

Carotenoid content was also characterized by marked
variations between different harvesting times, ranging
from 8350 to 15 119 mg/100 g (101.9–175.5 mg/100 g
d.w.) (Table 3). In all samples, lycopene was by far the
main component of carotenoid fraction (76–85%),
ranging from 7061 to 11 969 mg/100g (86.7–138.1 mg/
100 g d.w.). The coefficient of variation was 20.4%,
20.1% and 25.2% for total carotenoids, lycopene and b-
carotene contents, respectively. Our results were similar
to previously published data: lycopene content of 24
tomato varieties grown in southern Italy in 1998 ranged
from 3400 to 15 000 mg/100 g (mean value 8600 mg/
100 g), whereas in 29 varieties grown in 1999 it varied
from 4500 to 16 300 mg/100 g (mean value 8700 mg/100 g)
(Dumas et al., 2003). Few data have been reported in
literature on seasonal fluctuations of carotenoid levels in
tomatoes. Heinonen et al. (1989) observed that lycopene
concentration was relatively high in summer (June to
August) (3800–6600 mg/100g) and low in winter (Octo-
ber to March) (2600–3100 mg/100 g) in tomatoes pur-
chased from retail food stores in Finland; on the
contrary, they found the minimum level of b-carotene
and lutein in summer. Our results did not evidence clear
seasonal trends of carotenoid content or any association
with climatic parameters. Nevertheless, lycopene con-
tent was at the lowest in July (Table 3), similar to that
observed by Martinez-Valverde et al. (2002) on tomato
samples harvested in mid-summer, confirming that the
high radiation level and hot temperatures reached in this
period of year in the Mediterranean area, may exert a
significant inhibition effect on lycopene accumulation.
Moreover, total carotenoid and lycopene content did
not correlate with dry matter content nor with
chromaticity values; as previously observed, tomatoes
showing similar fruit colour (a*/b* determined on the
carotenoids or lycopene (Leonardi et al., 2000a, b;
Giovanelli et al., 1999).
Different from phenolic compounds, carotenoid
profile was similar in all samples, whereas it was
reported to change quite rapidly during the ripening
process (Raffo et al., 2002). It was confirmed the
absence of z-carotene, and the presence of phytoene
and phytofluene, in relevant amounts, and of g-carotene,
lutein, neurosporene and lycopene 1,2-epoxide, in minor
quantities (Table 3).
Our data confirmed that cherry tomatoes are im-
portant source of carotenoids, not only lycopene but
also b-carotene, one serving providing from 13% to
27% of the recommended daily intake of vitamin A
(Società Italiana di Nutrizione Umana, 1996).
3.4. a-tocopherol
a-tocopherol level was rather low at each sampling
time, with a considerable variation from December,
40 mg/100 g (0.54 mg/100 g d.w.), to June, 1160 mg/100 g
(13.0 mg/100 g d.w.) (Table 3). Previous papers on
several tomato varieties cultivated in Hungary reported
a less marked extent of variation, from 90 to 612 mg/
100 g (Abushita et al., 1997; Abushita et al., 2000).
However, our data confirmed that a-tocopherol con-
tribution to the antioxidant properties of tomato is of
minor relevance, while the contribution to vitamin E
intake of one serving of fruits ranged from 0.4% to 12%
of the recommended daily intake (Società Italiana di
Nutrizione Umana, 1996). Moreover, in tomatoes,
vitamin E is present mainly within seeds, which are
not readily digested by humans, so the nutritional
relevance of the vitamin E content of cherry tomatoes
could be lower than the whole fruit content would
suggest.

Table 3
Carotenoid and a-tocopherol content (mg/100 g) in tomatoes harvested at different times of the year

Compound Apr Jun Jul Dec Jan Mar CV

Carotenoids
Lycopene 81007140 b 10818778 d 70617271 a 101927233 c 82027427 b 119697218 e 20.1
b-Carotene 950724 c 1063733 d 519730 a 8437127 bc 623735 a 820719 b 25.2
Phytoene 733710 c 608713 b 36777 a 1215719 d 1231756 de 1284741 e 42.8
Phytofluene 386717 b 41274 b 30078 a 52575 c 563725 d 846723 e 38.1
Lutein 1772 a —* 2574 b 1473 a 1676 a 2072 ab 55.0
Neurosporene 2971 a 3675 a —* 3272 a 3578 a 4573 b 52.4
g-Carotene 3572 c 3874 c 1172 a 2374 b 1573 a 2472 b 43.9
Lycopene 1,2-epoxide 134715 c 13379 c 7178 a 91718 ab 91721 ab 11079 bc 24.1
Total carotenoids 103847190 b 131087116 d 83537315 a 129357400 c 107757550 b 151197340 e 20.4
a-Tocopherol 1100720 e 1160710 f 800710 c 40710 a 1000720 d 100710 b 71.9

*Below the detection limit.

Values are means (7 standard deviations) of three samples; CV represents coefficient of variation of mean values.
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A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19
Table 4
Antioxidant activity (equivalent mM Trolox/100 g of fresh weight of food) in tomatoes harvested at different times of the year
Fraction Apr Jun
Water-soluble fraction 0.19170.009 a 0.26370.010 c
Water-insoluble fraction 0.02970.001 b 0.02770.001 a
Values are means (7 standard deviations) of three samples; CV represents coefficient of variation of mean values.
3.5. Antioxidant activity
Several studies have investigated the effects of
ripening stage, genotype and processing on hydrophilic
and lipophilic antioxidant activity of tomato (Cano
et al., 2003; George et al., 2004; Lavelli et al., 2000;
Leonardi et al., 2000a, b; Raffo et al., 2002) by using
different test reactions for antioxidant activity measure-
ment. Generally, hydrophilic antioxidant activity was
significantly affected by genotype and processing, and
almost independent of the ripening stage; on the other
hand, lipophilic antioxidant activity was more influ-
enced by ripening stage than by genotype. In our
experiment, total antioxidant activity (the sum of
activity of both water-soluble and water-insoluble
fractions, Table 4) varied from 0.199 to 0.454 equivalent
mM Trolox/100 g, and in all samples the water-soluble
fraction represented by far the main percentage con-
tribution (83–92%) to total activity. Similar ratios
between hydrophilic and lipophilic tomato antioxidant
activity were previously observed in full ripened
tomatoes (Cano et al., 2003; Raffo et al., 2002). The
water-soluble fraction showed more variable antioxi-
dant activity between different samples than the water-
insoluble fraction (CV of 40% and 11.5%, respectively);
moreover, the antioxidant activity of the two fractions
was not correlated with climatic data. Tomato sample
harvested in March showed a markedly higher total
activity than other samples, due to a higher activity of
the water-soluble fraction. This high value could be, at
least partly, due to a relatively high content of rutin and
a medium–high level of reduced ascorbic acid. In fact,
rutin is characterized by the highest antioxidant activity
among main phenolics of tomato (rutin 2.4 TEAC,
naringenin 1.53 TEAC, chlorogenic acid 1.24 TEAC, as
determined using in vitro test by Rice-Evans et al.
(1996)). Nevertheless, it is hard to explain changes in
antioxidant activity of water-soluble fractions on the
basis of variations in ascorbate and phenolics content.
First, it has been observed that the antioxidant activity
of food extracts depends on both concentrations and
synergistic effects (Diplock et al., 1998). Secondly, test
reactions for antioxidant activity measurement might be
affected by other components (glutathione and enzy-
matic components) that are involved in complex
antioxidant systems of tomato fruits (Jimenez et al.,
17
Jul Dec Jan Mar CV
0.21070.006 b 0.17170.005 a 0.17070.013 a 0.42070.020 d 40.3
0.02670.001 a 0.03470.001 c 0.02970.001 b 0.03470.001 c 11.5
2002). Already in a previous study, we did not observe
significant correlation between the antioxidant activity
of water-soluble fraction and any hydrophilic antiox-
idant compounds (Raffo et al., 2002). On the other
hand, lipophilic antioxidant activity is, generally, well
correlated to total carotenoid content, or to lycopene
content, in tomato samples (Cano et al., 2003, George
et al., 2004, Raffo et al., 2002). In our data, the sample
with the lowest carotenoid content (July) showed the
lowest water-insoluble antioxidant activity, and that one
with the highest content (March) had the highest
activity, even though the correlation between the two
parameters was lower (R ¼ 0:68, Po0:1) than that
observed on tomato sampled at different ripening stages
(Cano et al., 2003; Raffo et al., 2002).
4. Conclusions
In summary, cherry tomatoes of the same cultivar,
greenhouse-grown under similar conditions in the same
geographical area, and harvested at similar stage of
ripeness but at different times of the year showed
marked differences in the antioxidant content. In spite
of wide variations in antioxidant content according to
the season having been observed, neither a clear
seasonal trend, nor a correlation between antioxidant
content and mean solar radiation or average tempera-
ture, was found. In practice, for an accurate assessment
of effects of climate factors on phytonutrient content,
large-scale field trials over several years and in several
locations would be necessary. Nevertheless, our results
confirmed that the hot temperatures of mid-summer in
the Mediterranean basin may produce a significantly
negative effect on lycopene accumulation.
Despite the marked variability showed by antiox-
idants content, greenhouse-growing conditions in Sicily
induced the accumulation of relatively high levels of
ascorbic acid, phenolic compounds and carotenoids in
cherry tomatoes for most of the year.
Acknowledgements
We are indebted to Elena Azzini and Aldo Bertone
for technical assistance.
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References
Abushita, A.A., Hebshi, E.A., Daood, H.G., Biacs, P.A., 1997.
Determination of antioxidant vitamins in tomatoes. Food Chem-
istry 60, 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.
Adegoroye, A.S., Jolliffe, P.A., 1987. Some inhibitory effects of
radiation stress on tomato fruit ripening. Journal of the Science of
Food and Agriculture 39, 297–302.
Baldini, M., Fabietti, F., Giammarioli, S., Onori, R., Orefice, L.,
Stacchini, A., 1996. Metodi di analisi per il controllo chimico
degli alimenti. Report of Health Higher Institute, No. 34, Rome,
p. 265.
Brandt, K., Giannini, A., Lercari, B., 1995. Photomorphogenic
responses to UV radiation III: a comparative study of UVB effects
on anthocyanin and flavonoid accumulation in wild type and aurea
mutant of tomato (Lycopersicon esculentum Mill.). Photochemistry
and Photobiology 62, 1081–1087.
Cano, A., Acosta, M., Arnao, M., 2003. Hydrophilic and lipophilic
antioxidant activity changes during on-vine ripening of tomatoes
(Lycopersicon esculentum Mill.). Postharvest Biology and Technol-
ogy 28, 59–65.
Crozier, A., Lean, M.E., Mc Donald, M.S., Black, C., 1997.
Quantitative analysis of the flavonoid content of commercial
tomatoes, onions, lettuce and celery. Journal of Agricultural and
Food Chemistry 43, 590–595.
Davey, M.W., Van Montagu, M., Inzé, D., Sanmartin, M., Kanellis,
A., Smirnoff, N., Benzie, I.J.J., Strain, J.J., Favell, D., Fletcher, J.,
2000. Plant L-ascorbic acid: chemistry, function, metabolism,
bioavailability and effects of processing. Journal of the Science of
Food and Agriculture 80, 825–860.
Davies, J.N., Hobson, G.E., 1981. The constituents of tomato fruit.
The influence of environment, nutrition, and genotype. Critical
Reviews in Food Science and Nutrition 15, 205–280.
De Pascale, S., Maggio, A., Fogliano, V., Ambrosino, P., Ritieni, A.,
2001. Irrigation with saline water improves carotenoids content
and antioxidant activity of tomato. Journal of Horticultural
Science and Biotechnology 76, 447–453.
De Serrano, J.Q., De Gonzalez, L., Solomons, N.W., 1993. The
partition of ascorbic and dehydroascorbic acid in vitamin C-
containing Guatemalan food. Food Chemistry 47, 87–92.
Diplock, A.T., Charleux, J.L., Crozier-Willi, G., Kok, F.J., Rice-
Evans, C., Roberfroid, M., Stahl, W., Vine-Ribes, J., 1998.
Functional food science and defence against reactive oxygen
species. British Journal of Nutrition 80, S77–S112.
Dixon, R.A., Paiva, N.L., 1995. Stress-induced phenylpropanoid
metabolism. The Plant Cell 7, 1085–1097.
Dorais, M., Papadopoulos, A.P., Gosselin, A., 2001. Greenhouse
tomato fruit quality. Horticultural Reviews 26, 239–319.
Dumas, Y., Dadomo, M., Di Lucca, G., Grolier, P., 2003. Effects of
environmental factors and agricultural techniques on antioxidant
content of tomatoes. Journal of the Science of Food and
Agriculture 83, 369–382.
El-Gizawi, A.M., Abdallah, M.M.F., Gomaa, H.M., Mohamed, S.S.,
1993. Effect of different shading levels on tomato plants. 2. Yield
and fruit quality. Acta Horticulturae 323, 349–354.
Garcı́a-Closas, R., Berenguer, A., Tormo, M.J., Sánchez, M.J.,
Quirós, J.R., et al., 2004. Dietary sources of vitamin C, vitamin
E and specific carotenoids in Spain. British Journal of Nutrition 91,
1005–1011.
George, B., Kaur, C., Khurdiya, D.S., Kapoor, H.C., 2004.
Antioxidants in tomato (Lycopersicon esculentum) as a function
of genotype. Food Chemistry 84, 45–51.
Giovanelli, G., Lavelli, V., Peri, C., Nobili, S., 1999. Variation in
antioxidant components 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.
Hamauzu, Y., Chachin, K., Ueda, Y., 1998. Effect of postharvest
temperature on the conversion of 14C-mevalonic acid to carotenes
in tomato fruits. Journal of the Japanese Society for Horticultural
Science 67, 549–555.
Heinonen, M.I., Ollilainen, V., Linkola, E.K., Varo, P.T., Koivistoi-
nen, P.E., 1989. Carotenoids in Finnish foods. Journal of
Agricultural and Food Chemistry 37, 655–659.
Hertog, M.G.L., Hollman, P.C.H., Katan, M.B., 1992. Content of
potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits
commonly consumed in The Netherlands. Journal of Agricultural
and Food Chemistry 40, 2379–2383.
Hertog, M.G.L., Kromhout, D., Aravanis, C., Blackburn, H., Buzina,
R., Fidanza, F., Giampaoli, S., Jansen, A., Menotti, A.,
Nedeljkovic, S., et al., 1995. Flavonoid intake and long term risk
of coronary heart disease and cancer in the Seven Countries Study.
Archives of Internal Medicine 155, 381–386.
Jimenez, A., Creissen, G., Kular, B., Firmin, J., Robinson, S.,
Verhoeyen, M., Mullineaux, P., 2002. Changes in oxidative
processes and components of the antioxidant system during tomato
fruit ripening. Planta 214, 751–758.
Kader, A.A., Stevens, M.A., Albright-Holton, M., Morris, L.L.,
Algazi, M., 1977. Effect of fruit ripeness when picked on flavor and
composition in fresh market tomatoes. Journal of the American
Society for Horticultural Science 102, 724–731.
La Vecchia, C., 1998. Mediterranean epidemiological evidence on
tomatoes and the prevention of digestive tract cancers. Proceedings
of the Society for Experimental Biology and Medicine 218,
125–128.
Lavelli, V., Peri, C., Rizzolo, A., 2000. Antioxidant activity of tomato
products as studied by model reactions using xanthine oxidase,
myeloperoxidase, and copper-induced lipid peroxidation. Journal
of Agricultural and Food Chemistry 48, 1442–1448.
Lee, S.K., Kader, A.A., 2000. Preharvest and postharvest factors
influencing vitamin C content of horticultural crops. Postharvest
Biology and Technology 20, 207–220.
Leonardi, C., Ambrosino, P., Esposito, F., Fogliano, V., 2000a.
Antioxidative activity and carotenoid and tomatine contents in
different typologies of fresh consumption tomatoes. Journal of
Agricultural and Food Chemistry 48, 4723–4727.
Leonardi, C., Gennaro, L., Raffo, A., 2000b. Pomodoro ciliegia, in
Prodotti Ortofrutticoli Tipici del Meridione Italiano: Caratterizza-
zione Agronomica e Nutrizionale. Agricoltura Ricerca, No. 185.
ISMEA, Roma, pp. 34–40.
Leoni, C., 1992. Industrial quality as influenced by crop management.
Acta Horticulturae 301, 177–184.
Liptay, A., Papadopoulos, A.P., Bryan, H.H., Gull, D., 1986. Ascorbic
acid levels in tomato (Lycopersicon esculentum Mill) at low
temperatures. Agricultural and Biological Chemistry 50, 3185–3187.
Lopez-Andreu, F.J., Lamela, A., Esteban, R.M., Collado, J.G., 1986.
Evolution of quality parameters in the maturation stage of tomato
fruit. Acta Horticulturae 191, 387–394.
Macheix, J.J., Fleuriet, A., Billot, J., 1990. Changes and metabolism of
phenolic compounds in fruits. In: Fruit Phenolics. CRC Press,
Boca Raton, FL, pp. 149–221.
Margolis, S.A., Schapira, R.M., 1997. Liquid chromatographic
measurement of L-ascorbic acid and D-ascorbic acid in biological
samples. Journal of Chromatography B 690, 25–33.
Martinez-Valverde, I., Periago, M.J., Provan, G., Chesson, A., 2002.
Phenolic compounds, lycopene and antioxidant activity in
 ARTICLE IN PRESS
A. Raffo et al. / Journal of Food Composition and Analysis 19 (2006) 11–19
commercial varieties of tomato (Lycopersicum esculentum). Journal
of the Science of Food and Agriculture 82, 323–330.
McGuire, R.G., 1992. Reporting objective color measurement.
Horticultural Science 27, 1254–1255.
NETTOX, 1998. Compilation of Consumption Data, EU Report No.
4. Danish Veterinary and Food Administration.
Official Journal of the European Communities, 2002. Publication of an
application for registration pursuant to Article 6(2) of Council
Regulation (EEC) No. 2081/92 on the protection of geographical
indications and designations of origin, C 168, vol. 45, 13 July 2002,
pp. 7–10 (2002/C 168/04).
Pellegrini, N., Re, R., Yang, M., Rice-Evans, C., 1999. Screening of
dietary carotenoids and carotenoid-rich fruit extracts for antiox-
idant activities applying 2,20 -azinobis(3-ethylbenzothiazoline-6-
sulphonic acid) radical cation decolorization assay. Methods in
Enzymology 299, 379–389.
Petersen, K.K., Willumsen, J., Kaack, K., 1998. Composition and taste
of tomatoes as affected by increased salinity and different salinity
sources. Journal of Horticultural Science and Biotechnology 73,
205–215.
Raffo, A., Leonardi, C., Fogliano, V., Ambrosino, P., Salucci, M.,
Gennaro, L., Bugianesi, R., Giuffrida, F., Quaglia, G., 2002.
Nutritional value of cherry tomatoes (Lycopersicon esculentum Cv.
Naomi F1) harvested at different ripening stages. Journal of
Agricultural and Food Chemistry 50, 6550–6556.
Rice-Evans, C.A., Miller, N.J., Paganga, G., 1996. Structure-
antioxidant activity relationships of flavonoids and phenolic acids.
Free Radical Biology and Medicine 20, 933–956.
Riso, P., Visioli, F., Testolin, G., Porrini, M., 2004. Lycopene and
vitamin C concentrations increase in plasma and lymphocites after
tomato intake. Effects on cellular antioxidant protection. Eur-
opean Journal of Clinical Nutrition 58, 1350–1358.
Robertson, G.H., Mahoney, N.E., Goodman, N., Pavlath, A.E., 1995.
Regulation of lycopene formation in cell suspension culture of
VFNT tomato (Lycopersicon esculentum) by CPTA, growth
regulators, sucrose, and temperature. Journal of Experimental
Botany 46, 667–673.
19
Sesso, H.D., Liu, S., Gaziano, J.M., Buring, J.E., 2003. Dietary
lycopene, tomato-based food products and cardiovascular disease
in women. Journal of Nutrition 133, 2336–2341.
Società Italiana di Nutrizione Umana, 1996. Livelli di Assunzione
Raccomandata di Energia e Nutrienti per la Popolazione Italiana.
Revisione 1996. SINU, Roma.
Souci, S.W., Fachmann, W., Kraut, H., 1994. Food Composition and
Nutrition Tables, fifth ed. Medpharm Scientific Publishers,
Stuttgart.
Stewart, A., Bozonnet, S., Mullen, W., Jenkins, G.I., Lean, M.E.J.,
Crozier, A., 2000. Occurrence of flavonols in tomatoes and tomato-
based products. Journal of Agricultural and Food Chemistry 48,
2663–2669.
Tonucci, L.H., Holden, J.M., Beecher, G.R., Khachik, F., Davis, C.S.,
Mulokozi, G., 1995. Carotenoid content of thermally processed
tomato-based food products. Journal of Agricultural and Food
Chemistry 43, 579–586.
Tubaro, F., Micossi, E., Ursini, F., 1996. The antioxidant capacity
of complex mixtures by kinetic analysis of crocin bleaching
inhibition. Journal of the American Oil Chemists’ Society 73,
173–179.
Turrini, A., Saba, A., Perrone, D., Cialfa, E., D’Amicis, A., 2001.
Food consumption patterns in Italy: the INN-CA study
1994–1996. European Journal of Clinical Nutrition 55, 571–588.
Vanderslice, J.T., Higgs, D.J., Hayes, J.M., Block, G., 1990. Ascorbic
acid and dehydroascorbic acid content of foods-as-eaten. Journal
of Food Composition and Analysis 3, 105–118.
Venter, F., 1977. Solar radiation and vitamin C content of tomato
fruits. Acta Horticulturae 58, 121–127.
Weisburger, J.H., 2002. Lycopene and tomato products in health
promotion. Experiments in Biology and Medicine 227, 924–927.
Wilkens, R.T., Spoerke, J.M., Stamp, N.E., 1996. Differential
responses of growth and two soluble phenolics of tomato to
resource availability. Ecology 77, 247–258.
Willcox, J.K., Catignani, G.L., Lazarus, S., 2003. Tomatoes and
cardiovascular health. Critical Reviews in Food Science and
Nutrition 43, 1–18.