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Postharvest Biology and Technology 46 (2007) 201–211

Impact of combined postharvest treatments (UV-C light, gaseous O3,

superatmospheric O2 and high CO2) on health promoting
compounds and shelf-life of strawberries
Ana Allende, Alicia Marı́n, Begoña Buendı́a, Francisco Tomás-Barberán, Maria I. Gil ∗
Research Group on Quality, Safety and Bioactivity of Plant Foods, CEBAS-CSIC, P.O. Box 164, Espinardo, Murcia 30100, Spain
Received 11 January 2007; accepted 21 May 2007

Abstract
Modified atmosphere packaging (MAP), ozone (O3 ) and ultraviolet-C (UV-C) light, have been suggested as postharvest treatments to control
decay of strawberries. However, the influence of these treatments on strawberry phytochemicals has not been thoroughly evaluated. Thus, the
impact of individual and combined UV-C light (1 kJ m−2 ), gaseous O3 (5000 mg L−1 ) and two active MAP conditions (superatmospheric O2 and
CO2 -enriched atmospheres) on the polyphenols, vitamin C content and shelf-life of strawberries was studied. Samples were taken initially, and after
5, 9 and 12 days of storage and microbial, nutritional and organoleptical qualities were evaluated. None of the evaluated samples showed visual
signs of fungal growth after 12 days of storage, including non-treated samples stored in air. However, phenolic contents of UV-C and O3 treated
strawberries were significantly reduced after treatments, mainly due to a significant decrease in procyanidins. Ozonated samples showed the lowest
vitamin C contents at the end of storage. On the other hand, when compared with storage in air, strawberries stored under superatmospheric O2 and
CO2 -enriched concentrations showed lower total phenolic contents after 5 days and a vitamin C reduction after 12 days of storage, accompanied
by a more pronounced conversion from reduced to oxidized forms under superatmospheric O2 . In general, overall quality was good in all samples
throughout the shelf-life except for flavour scores of MAP strawberries, which were clearly lower than air-stored samples after 9 and 12 days of
storage. No additional effect was observed when combining the postharvest treatments compared with the effect of individual treatments.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Fragaria ananassa; Microbial growth; Vitamin C; Procyanidins; Polyphenols; Anthocyanins; Modified atmosphere packaging; Quality

1. Introduction etables, since they have a growth inhibitory effect on bacteria,

yeast and moulds and prevent undesired anoxic fermentation
The short postharvest shelf-life of strawberries encourages (Allende et al., 2002; Van der Steen et al., 2002).
the use of decay-control techniques. The most commonly used Many researchers have reported the effects of MAP on the
postharvest treatments to control fungal growth and reduce res- antioxidant constituents and microbial quality of strawberries
piration rate of strawberries are low temperature and modified (Gil et al., 1997; Pérez and Sanz, 2001; Pelayo et al., 2003; Siro
atmosphere packaging (MAP) at 15–20 kPa CO2 (Mitcham, et al., 2006; Zheng et al., 2007). Among the different postharvest
2004). Carbon dioxide (CO2 )-enriched atmospheres are used technologies that can be used to maintain quality and increase
to reduce the incidence and severity of decay and therefore safety of strawberry fruit, ultraviolet-C (UV-C) light and gaseous
extend the postharvest life of strawberries (Li and Kader, 1989). ozone (O3 ) are two promising preservation techniques. Numer-
However, in some cases, adverse effects on colour and flavour ous studies have already been focused on the bactericidal and
after exposure to very low O2 and very high CO2 concentra- fungicidal capacity of O3 and UV-C treatments (Kim et al., 1999;
tions have been reported (Ke et al., 1991; Shamaila et al., 1992). Marquenie et al., 2002a; Palou et al., 2002; Pan et al., 2004).
Superatmospheric oxygen (O2 ) conditions (i.e. >70%) have been Additionally, UV-C light and gaseous O3 applications have been
suggested as an alternative to conventional MAP of fruit and veg- proposed to promote synthesis of health promoting compounds
(Cantos et al., 2000; Artés-Hernández et al., 2003; González-
Barrio et al., 2006). Nevertheless, little information is available
∗ Corresponding author. Tel.: +34 968 396 315; fax: +34 968 396 213. on the effect of these treatments on the phytochemicals of straw-
E-mail address: migil@cebas.csic.es (M.I. Gil). berries when individually applied or in combination with other

0925-5214/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.postharvbio.2007.05.007
202 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211

postharvest techniques. Therefore, the aim of this study was 2.3. Respiration rate and gas analysis
to determine the effect of UV-C light, gaseous O3 , superatmo-
spheric O2 and CO2 -enriched atmospheres applied individually Strawberries (250 g) were placed in a 1 L glass jar at 2 ◦ C and
and in combination on the health promoting compounds and 95% RH. A continuous humidified air flow was pumped into the
shelf-life of strawberries. jars to avoid dehydration and excessive CO2 accumulation. The
respiration rate as CO2 production was monitored every day for
2. Materials and methods up to 6 days. On each sampling date, triplicate jars per treatment
were evaluated. Samples of 1 mL of headspace gas were taken
2.1. Plant material from each glass jar with a calibrated syringe and monitored by
using a gas analyser (Horiba Via 510, Irving, USA).
Strawberry fruit (Fragaria × ananassa D. cv. Camarosa) at
commercial maturity were obtained from commercial grow- 2.4. Superatmospheric O2 and CO2 -enriched atmospheres
ers in Huelva (Spain) between April and June 2004. Fruit
were transported in refrigerated conditions to CEBAS-CSIC Strawberry samples of 250 g each, were packaged and sealed
(Murcia, Spain) and stored in darkness at 2 ◦ C and 95% rel- in polypropylene (PP) trays (185 mm × 138 mm × 32 mm)
ative humidity (RH). The day after, strawberries were graded using perforated film (control air) as well as in an active
for uniformity of colour and size, and damaged berries were modified atmosphere by using an O2 barrier film with an
removed. OTR of 0 pmol s−1 m−2 Pa−1 (Tecknopack S.L., Barcelona,
Spain). The perforated packages were prepared using the same
2.2. UV-C and O3 treatments barrier film with 50 perforations made with a syringe (Ø
0.6 mm, 979 perforations m−2 ). Active modified atmospheres
The UV-C equipment consisted of a bank of 17 unfiltered ger- were generated by flushing the packages with the desired gas
micidal emitting lamps (λ = 254 nm) (Sylvania, G30T8, Philips, composition: (1) superatmospheric O2 of 80 kPa O2 (balanced
The Netherlands). A wooden box covered with aluminium foil with N2 ) and (2) CO2 -enriched concentrations of 10 kPa CO2
and supported by a metal framework enclosed the UV-C lamps, (10 kPa O2 , balanced with N2 ). A gas exchange device with a
reflectors, and treatment area, providing UV protection for the vacuum packaging machine (Zermat, Carburos Metálicos S.A.,
operator. The lamp bank was horizontally suspended over the Madrid, Spain) and a mixing station (Witt-Gasetechnik, model
illumination vessel at a distance of 60 cm. Strawberries were KM 100-3 M, Carburos Metálicos S.A.) were used. All prepa-
placed in a single layer on the illumination vessel for the treat- ration steps were performed at 10 ◦ C under sanitary conditions.
ment, and different UV-C doses were applied by altering the Subsequent evaluation of atmospheric composition, microbial
exposure time at the fixed distance. Twenty-five strawberries growth, vitamin C and polyphenols of strawberry samples were
were treated for each illumination batch and each fruit was carried out at day 0 and after 5, 9 and 12 days of storage at 2 ◦ C
rotated manually once to ensure even surface exposure to UV and 95% RH.
light. The fluence rate of the lamps at the level of the samples
was 2.82 ± 0.44 mW cm−2 , measured with a Blak-Ray J-225 2.5. Microbial analysis
photometer (Ultra-Violet Products, Inc., San Gabriel, CA). To
produce O3 , extra-dry compressed air (0.7 Pa) was let through a Strawberry samples of 30 g each were homogenized in a 1:10
water-cooled corona discharge generator (model 1A, Steriline, dilution of 1% sterile peptone buffered water (AES Labora-
Ozono Electónica Ibérica, Granada, Spain) (González-Barrio et toire, Combourg, France) by using a stomacher (IUL Instrument,
al., 2006). Gaseous O3 concentration was measured with an O3 Barcelona, Spain) for 90 s. Filter stomacher bags (Seward Lim-
gas analyser (model H1-SPT, IN USA Inc., Needham, MA). ited, London, UK) were used to eliminate solid particles from the
Ozone gas was supplied in air constant flow of 0.16 N m3 h−1 , strawberry homogenates. Ten-fold dilution series were made in
with productions of 1.7, 3.5, 5.2 and 6.9 g h−1 produced by the peptone buffered water as needed for pour plating. The following
O3 generator in a 50 L methacrylate chamber at 22 ◦ C and 95% culture media and conditions were used to evaluate the microbial
relative humidity (RH). The chamber temperature and RH were growth: 1 mL of the appropriate sample dilution was pour-plated
determined with a data logger (model Tinytag TGU-1500, Gem- on (1) plate count agar (PCA, Scharlau Chemie S.A., Barcelona,
ini Dataloggers Ltd., Sussex, UK). The values of O3 obtained Spain) incubated at 30 ◦ C for 24–48 h for aerobic mesophilic
were constant throughout the experiments, and the gas mix- bacteria and at 4 ± 1 ◦ C for 7 days for aerobic psychrophilic
ture expelled from the chamber was destroyed by a catalytic bacteria; (2) Chromocult coliform agar (Merck, Darmstadt, Ger-
destroyer (OIE, Granada, Spain). The experiments with O3 were many) incubated at 37 ◦ C for 24–36 h for coliform counts; (3)
made in the pilot plant of CEBAS-CSIC in accordance with strict Lactobacilli MRS broth (Difco Lab, Sparks, MD, USA) with
safety and protection rules. Preliminary assays were carried out addition of BactoTM agar (10 g L−1 ) (Difco Lab, Sparks) and
to select the optimum UV-C light and gaseous O3 treatment overlaid with the same medium, incubated aerobically at 30 ◦ C
doses. Four different UV-C light doses (1, 5, 10 and 15 kJ m−2 ) for 72 h for lactic acid bacteria, and (4) 100 ␮L of the appropriate
and O3 concentrations in the gas carrier (air) of 5000, 10,000, sample dilution was spread-plated on Rose Bengal agar (Schar-
15,000 and 20,000 mg L−1 were tested. All the treatments were lau Chemie S.A.) and incubated at 30 ◦ C for 48–72 h for moulds
applied in a cold room at 10 ◦ C. and yeasts. Microbial counts were expressed as log10 cfu g−1 .
 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211 203

2.6. Extraction and analysis of vitamin C ples were injected by means of an autosampler (model L-7200)
and compound separations were achieved on a Mediterranean
Ascorbic acid (AA) and dehydroascorbic acid (DHA) con- sea C18 column (250 mm × 4.6 mm i.d., 5 ␮m, Teknokroma,
tents were determined as described by Zapata and Dufour Barcelona, Spain) using H2 O/formic acid (95/5, v/v) (A) and
(1992) with some modifications (Gil et al., 1999). The extrac- acetonitrile (B) as mobile phases. Elution was performed using
tion analysis was carried out with fresh samples. Ten grams of a gradient ranging from 5% B in A to reach 60% B after 60 min,
fresh strawberries were added to 10 mL of extraction medium 90% B at 61 min and 90% B at 66 min. The flow rate was
(MeOH/H2 O (95/5, v/v), 0.1 M citric acid, 0.05% (w/v) EDTA 1 mL min−1 and chromatograms were recorded at 280 nm for
disodium salt and 4 mM NaF). Homogenates were then fil- procyanidins and ellagitanins, 320 nm for hydroxycinnamics
tered through cheesecloth, centrifuged at 10,500 × g for 5 min at acids, 360 nm for flavonols, ellagic acid and its derivatives and
2–5 ◦ C and filtered into a C18 Sep-Pak cartridges (Waters, Mil- 510 nm for anthocyanins.
ford, MA) and finally through a 0.45 ␮m filter. Then, 250 ␮L of The HPLC–MS system was equipped with a DAD detector
1,2-phenylenediamine dihydrochloride (OPDA) solution (35 mg and mass detector in series consisted of a G1312A binary pump,
100 mL−1 ) was added to 750 ␮L extract for derivatisation a G1313A autosampler, a G1322A degasser, a G1315B photodi-
of DHA into the fluorophore 3-(1,2-dihydroxyethyl)furo[3,4- ode array detector and an ion trap mass spectrometer equipped
b]quinoxaline-1-one (DFQ). After 37 min in darkness, AA and with electrospray ionization (ESI) and operated in the negative
DHA analyses were performed by using a high-performance ion mode controlled by software (v. 4.0.25) from Agilent Tech-
liquid chromatograph (HPLC) (Merck Hitachi, Tokyo, Japan), nologies (Waldbronn, Germany). The heated capillary and the
equipped with a L-6000 pump and coupled to a D-2500 voltage were maintained at 350 ◦ C and 4 kV, respectively. Mass
variable-wave-length UV detector. Twenty microlitre samples scan (MS) and daughter (MS–MS) spectra were measured from
were injected on a reversed-phased Kromasil 100 C18 col- m/z 100 to ∼1500. Collision-induced fragmentation experiments
umn (250 mm × 4 mm i.d. 5 ␮m particle size; Tecnokroma, were performed in the ion trap using helium as the collision gas,
Barcelona, Spain) with a ODS guard C18 precolumn (Tec- and the collision energy was set at 50%. Mass spectrometry data
nokroma). The mobile phase was MeOH/H2 O (5/95, v/v) were acquired in the negative mode for all phenolic compounds.
containing 5 mM cetrimide and 50 mM potassium dihydrogen Procyanidins were quantified as catechin, hydroxycinnam-
phosphate at pH 4.5. The flow rate was kept at 0.9 mL min−1 . ics acids as chlorogenic acid (5-caffeoylquinic acid), flavonoids
The detector wavelength was initially set at 348 nm and after as quercetin 3-rutinoside, ellagic acid and derivatives as ellagic
elution of DFQ, it was manually shifted to 261 nm for AA detec- acid and anthocyanins as cyanidin 3-glucoside. Standards were
tion. The variation coefficient was less than 8%. A standard of acquired from Sigma (St. Louis, MO). The phenolic identifica-
AA was supplied by Sigma Chemical Co. Vitamin C content tion was carried out by means of their UV spectra, molecular
was calculated by adding AA and DHA contents and the results weights and their MS–MS fragments. Total phenolics were cal-
expressed as mg per 100 g fresh weight. culated as the addition of the individual phenolics quantified in
the HPLC analyses. The results were expressed as mg per 100 g
2.7. Extraction and analysis of phenolic compounds fresh weight.

Fresh, frozen, frozen and stored and freeze-dried strawber- 2.8. Sensory quality
ries were compared to select the sample preparation procedure,
avoiding underestimation of the phenolic constituents. Thus, The sensory analysis was used to determine the organolep-
lyophilised strawberry fractions (1 g each) were homogenized tical quality of the strawberries. The end of the shelf-life was
by using an Ultra Turrax (Ika, Staufen, Germany) in 20 mL of reached when the average value of the samples was judged unac-
extraction solution (acetone/H2 O/acetic acid; 70/29.5/0.5, v/v/v) ceptable for consumption for the sensory panel. Sensory quality
for 1 min at 4 ◦ C. Homogenates were sonicated in a bath-type was evaluated by a six-member expert panel, trained to score the
ultrasonic cleaner (5510E-MTH, Branson Ultrasonic Corpora- quality attributes of strawberries prior to the test. Samples were
tion, USA) at 37 ◦ C for 10 min and centrifuged at 4000 × g scored for overall visual quality by using an interval hedonic
for 15 min (Centromix centrifuge, Selecta, Barcelona, Spain). scale, where the extremes and centre of the interval were repre-
Supernatants were concentrated under reduced pressure at 35 ◦ C sented as follows: 0 ‘dislike extremely, no characteristic of the
to remove acetone. The aqueous fraction was dissolved in 10 mL product’, 5 ‘neither like nor dislike, limit of acceptance from the
of H2 O and filtered through a C18 Sep-Pak cartridge (Waters consumers point of view’, and 10 ‘like extremely, very charac-
Corp.), which was previously activated with MeOH followed teristic of the product’. The remaining attributes such as flavour,
by H2 O and air. Phenolic compounds were absorbed onto the colour, firmness and freshness were evaluated on a 5 point scale,
column, while sugars, acids and other water-soluble compounds where 5 = full characteristic of the product, 2.5 = moderate and
were eluted with 10 mL of H2 O. Phenolic compounds were then 0 = no characteristic. Defects of the product such as off-odours,
recovered with 1.5 mL of MeOH and passed through a 0.45 ␮m decay and calyx damage were evaluated as follows: 5 = severe,
pore filter (Millex HV13, Millipore, Bedford, MA). 2.5 = moderate and 0 = absence. The samples were coded with
Samples of 50 ␮l of extracts were analysed using a HPLC random three-digit numbers to mask the treatment identity to
system (Merck Hitachi, Tokyo, Japan) equipped with a L-7100 minimize subjectivity and to ensure test accuracy. All qual-
pump and a L-7455 photodiode array UV/VIS detector. Sam- ity evaluations were performed in a sensory room, equipped
204 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211

with individual cabinets with white and red lights. Off-odours, differences were observed among UV-C treatments except for
flavour and firmness were evaluated under red light. Subse- the highest dose (15 kJ m−2 ) (Fig. 1A). This treatment increased
quently, colour, decay, calyx damage and overall visual quality the strawberry RR at the beginning of storage, but after 6 days
were evaluated under white light. at 2 ◦ C, no differences were observed for the different treat-
ment intensities. On the other hand, neither of the different
2.9. Experimental design and statistical analysis doses of O3 affected the RR when compared with untreated fruit
(Fig. 1B).
A preliminary experiment was carried out to adjust the When the overall visual quality of treated strawberries was
optimal dose of UV-C and gaseous O3 treatments. The main evaluated, the colour of the calyx was a decisive parameter of
experiment was repeated twice and results from the second fruit quality, as browning of the calyx leaves was the most rele-
assay were used for the discussion of the study, as the same ten- vant damage. Strawberries subjected to high UV-C and gaseous
dency was observed for both experiments. Experimental units O3 doses showed brown discoloration and shrivelling of sepals
were packages and there were three replications per treatment and, when treatment doses increased, the destructive effect of
and evaluation period. A total of nine treatments were eval- UV-C and O3 was more apparent on the strawberry sepals.
uated: untreated fruit in air (Untreated + air), untreated fruit Therefore, the lowest UV-C (1 kJ m−2 ) and O3 (5000 mg L−1 )
in superatmospheric O2 (Untreated + O2 ), untreated fruit in doses were selected for the two main experiments, since the rest
CO2 -enriched atmosphere (Untreated + CO2 ), O3 -treated fruit of the doses had the negative effect of browning and drying of
in air (O3 + Air), UV-C treated fruit in air (UV-C + Air), O3 - the strawberry calyx (data not shown).
treated fruit in superatmospheric O2 (O3 + O2 ), UV-C treated
fruit in superatmospheric O2 (UV-C + O2 ), O3 -treated fruit in 3.2. Modified atmosphere development
CO2 -enriched atmosphere (O3 + CO2 ), UV-C treated fruit in
CO2 -enriched atmosphere (UV-C + CO2 ). Statistical analysis of Strawberries were exposed to active MAP containing super-
the data was carried out using SPSS 14.0 for Windows (SPSS atmospheric O2 (80 kPa O2 ) (Fig. 2A) and CO2 -enriched
Inc., Chicago, IL, USA). atmospheres (10 kPa O2 + 10 kPa CO2 ) (Fig. 2B). In superatmo-
spheric O2 packages, the target O2 conditions were maintained
3. Results ≥65 kPa during the storage period. The CO2 level accumulated
inside these packages, reached bacteriostatic or bactericidal lev-
3.1. Selection of optimal UV-C light and gaseous O3 els (≥20 kPa) at the end of storage because of respiration. In
treatments this case, a superatmospheric O2 conditions combined with high
CO2 concentrations was established after 5 days of storage. In
The selection of optimal doses was based on their effect on the case of the CO2 -enriched atmosphere (10 kPa O2 + 10 kPa
the respiration rate (RR) and visual quality of the samples stored CO2 ) initially flushed in the packages, a marked modification
in air. A higher metabolic activity of the product is manifested by of the atmosphere was observed during storage (Fig. 2B). Thus,
a higher rate of respiration. Thus, the direct relation between the CO2 concentrations of about 15 kPa were reached after 5 days,
respiration rate and the rate of deterioration of the product sub- going up to 25 kPa at the end of storage. On the other hand,
jected to different UV-C light and O3 treatments was evaluated. partial pressures of O2 declined with time, reaching very low val-
Fig. 1 shows the RR of treated strawberries up to 6 days of storage ues (≈1 kPa) after 5 days of storage. Therefore, an O2 -depleted
at 2 ◦ C. It was observed that strawberry RR was very similar for atmosphere was reached after 9 days. Perforated packages main-
all treatments. A general decrease in the RR was observed dur- tained air atmospheres (21 kPa O2 –0.03 kPa CO2 ) during the
ing storage from 335.0 ± 59.4 to 79.3 ± 10.1 nmol kg−1 h−1 . No storage period (data not shown).

Fig. 1. Respiration rate of strawberries untreated and treated with (A) UV-C light (1, 5, 10 and 15 kJ m−2 ) and (B) gaseous O3 (5000, 10,000, 15,000, 20,000 mg L−1 )
doses and stored at 2 ◦ C for 6 days. Error bars represent the standard deviation.
 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211 205

Fig. 2. Headspace O2 (unfilled symbols) and CO2 (filled symbols) levels in strawberry packages untreated and treated with UV-C light and gaseous O3 stored at
2 ◦ C for 12 days under air and active MAP: (A) superatmospheric O2 and (B) CO2 -enriched atmosphere. Error bars represent the standard deviation.

3.3. Influence of postharvest treatments on microbial spheres maintained the initial counts up to 5 days, although
analysis no differences were observed at the end of storage (data not
shown).
Bacteria growth was not critical in the spoilage of Visible signs of fungal growth were not observed in strawber-
strawberries, as their number remained low during storage ries after 12 days of storage at 2 ◦ C. Microbial analyses indicated
(≈102 –104 cfu g−1 ). Individual application of UV-C promoted that moulds and yeast ranged from 102 to 104 log cfu g−1
psychrophilic growth with a final count of 4.8 log cfu g−1 . (Fig. 3). No differences were observed after application of indi-
However, UV-C combined with active MAP controlled the expo- vidual postharvest treatments. However, the combination of
nential growth of these microorganisms. On the other hand, gaseous O3 with and superatmospheric O2 and CO2 -enriched
samples treated with gaseous O3 stored in air and active MAP atmospheres reduced yeast and mould growth by approximately
did not reduced mesophilic and psychrophilic loads with final 1 log for up to 5 days and throughout the storage period, respec-
counts very similar to untreated samples. CO2 -enriched atmo- tively.

Fig. 3. Yeast and mould counts of strawberries untreated and treated with UV-C light and gaseous O3 stored at 2 ◦ C for 12 days under air and active MAP
(superatmospheric O2 and CO2 -enriched atmospheres). Error bars represent the standard deviation.
206 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211

Fig. 4. Content of ascorbic acid (uncross-hatched bars) and dehydroascorbic acid (cross-hatched bars) of strawberries treated with UV-C light and gaseous O3
stored at 2 ◦ C for 12 days under air and active MAP (superatmospheric O2 and CO2 -enriched atmospheres). Error bars represent the standard deviation. LSD values
(time × treatment × atmosphere) = 0.4.

3.4. Influence of postharvest treatments on vitamin C were only reduced by gaseous O3 and no effect was observed
in ellagitanin content (Fig. 6). Thus, procyanidin content was
Vitamin C content was maintained well during storage reduced to 64% in O3 treated samples and 74% in UV-C treated
in untreated and treated samples, although reductions were strawberries compared to untreated samples at day 0. It was also
observed at the end of storage in UV-C and O3 treated samples observed that throughout storage, oxidizing treatments such as
stored in air and active MAP, as well as in untreated strawberries UV-C and gaseous O3 , decreased procyanidins and, in general,
stored in active MAP (Fig. 4). AA was the main form of vitamin slightly reduced flavonols, ellagic acid and derivatives and p-
C in all untreated and treated samples during storage (87% AA coumaroyl glucose (data not shown). Active MAP also affected
and 13% DHA), except for strawberries exposed to superatmo- the total phenolic content, as it was reduced during the first
spheric O2 , which increased the proportions of oxidized forms 5 days of storage (Fig. 5). In fact, superatmospheric O2 and
by approximately 13 to 41% after 12 days of storage. CO2 -enriched atmospheres decreased the ellagitanin content to
70 and 60%, respectively, after 5 days of storage, when com-
3.5. Influence of postharvest treatments on phenolic pared to untreated samples (data not shown). Additionally, a
contents marked reduction of the procyanidin content was also observed
in strawberries stored under superatmospheric O2 conditions
The HPLC analysis showed that the phenolic profile mostly (Fig. 6). The combination of different postharvest treatments
comprised ellagitanins, anthocyanins and procyanidins, and to also influenced the total phenolic content of strawberries. The
a lower extent flavonols, ellagic acid and its derivatives, and initial reduction of phenolic compounds observed after O3 and
p-coumaroyl glucose. Initially, total phenolic contents were in UV-C treatments, seemed to be emphasized after 5 days of stor-
the range of 48.5–53.0 mg 100 g−1 fw and they decreased during age, when superatmospheric O2 was applied (Fig. 5). However,
storage at 2 ◦ C in all the samples. This decrease was substantial at the end of storage, total phenolic reductions were very similar
at day 5 in strawberries treated with individual and combined in all the strawberry samples.
postharvest treatments, mainly after gaseous O3 and UV-C light
(Fig. 5). However, no difference in total phenolic contents was 3.6. Influence of postharvest treatments on sensory quality
observed in untreated and treated samples after 12 days of stor-
age. When the individual phenolic compounds were evaluated, The visual quality of all strawberry samples decreased dur-
it was found that procyanidins were reduced after the appli- ing storage with a similar trend for all the treatments (Fig. 7).
cation of both, O3 and UV-C treatments, while anthocyanins After 9 days of storage, this reduction was more noticeable in
 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211 207

Fig. 5. Total phenolic content of strawberries treated with UV-C light and gaseous O3 stored at 2 ◦ C for 12 days under air and active MAP (superatmospheric O2
and CO2 -enriched atmospheres). Error bars represent the standard deviation. LSD values (time × treatment × atmosphere) = 0.6.

those samples stored under air compared to strawberries stored found to be effective in reducing fungal decay in strawber-
in active MAP. When flavour was scored, only air-stored sam- ries (Marquenie et al., 2002b; Pan et al., 2004), while gaseous
ples were still accepted after 12 days of storage. Thus, besides O3 reduced decay incidence, weight loss and strawberry soft-
overall visual quality, strawberry flavour determined the end of ness (Nadas et al., 2003). In this study, the selected treatments
shelf-life (Fig. 7). On the other hand, the sensory panel was (1 kJ m−2 UV-C light and 5000 mg L−1 gaseous O3 ) did not sig-
not able to detect significant differences in strawberry firmness nificantly affect microbial growth of strawberry fruit. In fact,
when subjected to different postharvest treatments. After 5 days neither untreated nor treated strawberries showed fungal decay
of storage, all strawberry samples showed damaged calyxes and throughout storage. It should be taken into account that tissue
this was also observed throughout storage, although it did not used for microbial analysis included exposed surface tissues as
affect the overall visual quality scores, which were still accept- well as flesh (non-exposed tissues). Thus, the impact of these
able. postharvest treatments in reducing microbial load was very low.
Several studies have already focussed on the UV-C light and
4. Discussion O3 impact on nutritional constituents of fruit and vegetables
(Pérez et al., 1999; Pan et al., 2004; González-Barrio et al.,
Strawberries are very susceptible to microbial contami- 2006). In general, it has been shown that due to the high oxida-
nation due to the fact that their skins are soft, and easily tive capacity of O3 and UV-C light and their ability to generate
ruptured with numerous indentation and hair-like protuber- toxic molecular species, they act as potent phytotoxic agents.
ances, which allow most organisms to attach and proliferate They elicit plant defence reactions such as the production of
(Tournas and Katsoudas, 2005). However, in this study, straw- phytoalexins (Maharaj et al., 1999), including AA in strawber-
berry fruit stored up to 12 days at 2 ◦ C showed low bacteria ries (Pérez et al., 1999) and resveratrol in grapes (Langcake
growth and reduced yeasts and mould levels ranging from 2.0 and Pryce, 1977; Cantos et al., 2000; González-Barrio et al.,
to 3.6 log cfu g−1 . These yeast and mould counts are similar to 2006). Vitamin C and phenolic compounds are able to scav-
those reported by Ragaert et al. (2006), but relatively lower than enge oxygen radicals and avoid oxidative stress (Klopotek et
those described by Tournas et al. (2006) for fresh-cut strawberry al., 2005). Thus, changes in AA of O3 -treated strawberries can
(3.2–4.7 log cfu g−1 ). be attributed to the activation of an antioxidative system that
UV-C light and O3 have been extensively used to control promotes the biosynthesis of vitamin C from the carbohydrate
microbial decay in fruit and vegetables (Kim et al., 1999; pool (Pérez et al., 1999). However, as UV-C light and O3 can be
Maharaj et al., 1999; Palou et al., 2002; Allende et al., 2006). involved in photo-oxidation reactions in plants by free radical
Different UV-C doses (e.g. 4.1 and 10.0 kJ m−2 ) have been and superoxide production, they might result in the acceleration
208 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211

Fig. 6. Procyanidin, anthocyanin and ellagitanin content in strawberries untreated and treated with gaseous O3 and UV-C light and stored at 2 ◦ C under air and
active MAP (superatmospheric O2 and CO2 -enriched atmospheres) at days 0 and 12. Error bars represent the standard deviation. LSD values are in brackets
(time × treatment × atmosphere) = procyanidins (0.5), anthocyanins (0.8) and ellagitanins (1.7).

of senescence (Maharaj et al., 1999). In this study, the UV-C In accordance with these results, Pérez et al. (1999) found a sig-
light and gaseous O3 treatments did not have a lasting effect nificantly lower value of total anthocyanin contents in Camarosa
in de novo synthesis of vitamin C or phenolic compounds of strawberries treated with ozone (0.35 mg L−1 ). In addition, Pan
strawberries. What it is more, ozonated strawberries presented et al. (2004) reported that total phenolic contents were lower
the lowest vitamin C content at the end of storage, whereas total in UV-C treated (4.16 kJ m−2 ) strawberries using the Folin Cio-
phenolics were reduced after UV-C and gaseous O3 treatments. calteu determination. They concluded that the applied UV-C
 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211 209

Fig. 7. Sensory quality: (A) visual quality, (B) flavour, (C) firmness and (D) calyx damage of strawberries untreated and treated with UV-C light and gaseous O3
stored at 2 ◦ C for 12 days under air and active MAP (superatmospheric O2 and CO2 -enriched atmospheres). Error bars represent the standard deviation. LSD values
are in brackets (time × treatment × atmosphere) = visual quality (0.9), flavour (0.6), firmness (0.6) and calyx (0.6).
210 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211

dose did not significantly increase PAL activity. Contradictory after prolonged storage. Contrary to this, no significant differ-
information about the effects of UV-C treatments on the sen- ences in the total level of phenolics of strawberries in relation to
sory quality of fruit and vegetables has been reported. Some storage atmospheres were found (Pelayo et al., 2003). Addition-
authors found no negative effect on calyx quality (drying or ally, it was found that the content of quercetin and kaempferol
staining of leaves) in strawberries treated with 4.1 and 5.0 kJ m−2 derivatives of strawberries increased after 10 days at 5 ◦ C in
(Lammertyn et al., 2003; Pan et al., 2004), while others reported air and CO2 -enriched atmospheres (10, 20 and 40 kPa), while
that 1.0 kJ m−2 could damage the fruit (Baka et al., 1999). In ellagic content decreased in CO2 -enriched atmosphere (Gil et
fact, it is not uncommon to find these contradictory conclusions al., 1999). We found that total flavonol contents (quercetin and
since there have not been standard procedures used to determine kaempferol) were lower in strawberries after 9 days of storage
UV-C treatment doses. In an attempt to standardise the field of when active MAP was applied.
research using UV, a report on UV units, UV sources specifi- Even though CO2 -enriched atmospheres reduced decay, they
cations and experimental procedures has been developed by the increased production of fermentative metabolites that impart
European COST 924 action to provide consistent and traceable negative organoleptic properties to strawberries (Wszelaki and
practices with which to evaluate experimental results (Lagunas- Mitcham, 2000; Pérez and Sanz, 2001; Van der Steen et al.,
Solar and Gómez-López, 2007). In this study, the lowest doses 2002; Pelayo et al., 2003). According to this, in our experi-
of UV-C (1 kJ m−2 ) and O3 (5000 mg L−1 ) were selected since ments, flavour scores of active MAP strawberries were clearly
higher treatments caused calyx dehydration. In contrast, there is lower than in air-stored samples after storage. Additionally,
an strong evidence that UV-C light increases firmness of straw- it has been reported that strawberries treated with superatmo-
berries (Baka et al., 1999; Marquenie et al., 2002a,b; Pan et al., spheric O2 and CO2 -enriched atmospheres were firmer than
2004), which could be associated with the effect of the UV-C air-stored fruit (Wszelaki and Mitcham, 2000; Van der Steen
light on the activity of enzymes involved in cell wall degrada- et al., 2002; Pelayo et al., 2003). This phenomenon has been
tion (Pan et al., 2004). However, the sensory panel involved in usually linked to the accumulation of CO2 in the packages, but
this study was not able to detect significant differences among it was not directly related to superatmospheric O2 (Kader and
treatments. Ben-Yehoshua, 2000). As a result of CO2 -enriched treatments,
On the other hand, the beneficial effects of CO2 -enriched it was reported that cell-to-cell adhesion increased by 60% due
atmospheres and superatmospheric O2 on microbial and sensory to changes in the pH of the appoplast, with the subsequent pre-
quality during storage of strawberries have been reported (Siro et cipitation of soluble pectins (Harker et al., 2000). Nevertheless,
al., 2006; Zheng et al., 2007). In the present study, CO2 -enriched the sensory panel involved in the present study did not detect
atmospheres reduced psychrophilic growth while superatmo- differences in strawberry firmness after storage.
spheric O2 did not affect microbial growth. Therefore, according
to Van der Steen et al. (2002), in spite of the ‘in vitro’ inhibitory
effect of high O2 treatments on yeast growth, this reduction was 5. Conclusions
not observed in storage experiments of strawberries. Nutritional
constituents could also be affected by atmospheric conditions In the present study, microbial growth was not a critical
during strawberry storage. It has been shown that storage in parameter of strawberry quality since no visible signs of fungal
superatmospheric O2 and CO2 -enriched atmospheres had ben- development were observed in any of the samples. The posthar-
eficial effects on vitamin C retention in the first days of storage, vest treatments tested under the described conditions affected
but the levels decreased after 10 days of storage, probably due the health promoting constituents of ‘Camarosa’ strawberries.
to O2 -promoted oxidation of the main antioxidants (vitamin C, In general, the use of UV-C light and gaseous O3 significantly
anthocyanins and phenolic compounds) (Pérez and Sanz, 2001; reduced phenolic content of strawberries after treatment, mainly
Zheng et al., 2007). Our results agree with these data as the due to a significant decrease in procyanidins. Gaseous O3 nega-
vitamin C content of strawberries stored under active MAP was tively affect the content of vitamin C after 12 days of storage at
similar to that of air-stored samples at the beginning of the stor- 2 ◦ C when compared to untreated or UV-C treated samples. On
age, although it declined after 12 days. Several authors reported the other hand, our results confirm that superatmospheric O2 and
that phenolic contents, particularly anthocyanins, were lower in CO2 -enriched atmospheres decrease total phenolic contents in
strawberries treated with superatmospheric O2 and conventional the first days of storage, mainly ellagitanins, while reducing vita-
CA containing CO2 -enriched atmospheres when compared to min C content after 12 days of storage. It was found that flavour
air-stored fruit (Gil et al., 1997; Pérez and Sanz, 2001; Zheng scores of MAP strawberries were clearly lower than those of
et al., 2007). This phenomenon has been explained as a delay air-stored samples after 9 and 12 days of storage. However, the
of the fruit ripening process caused by the superatmospheric sensory panel was not able to detect differences in strawberry
O2 and CO2 -enriched atmospheres (Kader, 1995; Wszelaki and firmness among treatments. Finally, the combination of different
Mitcham, 2000). Our results confirmed that superatmospheric postharvest treatments had similar effects than individual treat-
O2 reduced total phenolic contents, mostly ellagitanins and pro- ments for ‘Camarosa’ strawberries. The postharvest treatments
cyanidins during the first 9 days of storage. On the other hand, tested in this study, which are commonly proposed to control
Zheng et al. (2007) reported that high O2 may induce the accu- microbial decay in strawberries, could have detrimental effects
mulation of phenolic compounds during the initial treatment from a nutritional point of view, reducing phenolic and vitamin
period, but it may also promote the oxidation of these compounds C content of ‘Camarosa’ strawberries.
 A. Allende et al. / Postharvest Biology and Technology 46 (2007) 201–211 211

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