Research Article

Received: 4 February 2015 Revised: 20 June 2015 Accepted article published: 25 June 2015 Published online in Wiley Online Library:

(wileyonlinelibrary.com) DOI 10.1002/jsfa.7319

Potential use of microwave treatment on

fresh-cut carrots: physical, chemical and
microbiological aspects
Ginés Benito Martínez-Hernández, Maria Luisa Amodio
and Giancarlo Colelli*

Abstract
BACKGROUND: The effect of microwave treatments (900 and 750 W for 45 and 60 s) on the microbial, physicochemical and
sensory properties of fresh-cut carrot slices and the contents of several bioactive compounds was studied. Carrot samples were
stored for 7 days at 5 ∘ C.

RESULTS: The microwaving of fresh-cut carrots reduced the initial respiration rate (8.6 CO2 mL kg−1 h−1 ) by 55–74% compared
with untreated samples, although the rates then increased during storage. The initial pH (6.7), titratable acidity (0.036%), soluble
solid content (8.2 ∘ Brix) and shelf-life of the samples did not differ greatly from those of the untreated samples. Microwaving
prevented the incipient whitening and surface dryness during storage. In general, no significant changes in phenylalanine
ammonia lyase activity (5.5 𝛍mol t-cinnamic acid kg−1 h−1 ), total phenolics (TP, 81.3 mg chlorogenic acid equivalent kg−1 fresh
weight (FW)) or total antioxidant capacity (TAC, 74.2 𝛍mol Trolox equivalent kg−1 FW) were observed on the processing day or
over storage. However, the mildest treatment (750 W for 45 s) caused TP and TAC enhancements of 118 and 394% respectively
after 7 days of shelf-life. Microwave treatments reduced the initial microbial loads of the samples by up to 1.8 log units, although
their microbial growth was greater than that of the untreated samples throughout storage.

CONCLUSION: Mild microwave treatments such as 750 W/45 s and 750 W/60 s are a good sustainable alternative to the use
of NaOCl; however, combining them with other sanitizing techniques is needed to control microbial growth throughout the
shelf-life of fresh-cut carrot slices.
© 2015 Society of Chemical Industry

Keywords: minimally fresh processed; dielectric heating; whiteness; phenylalanine ammonia lyase; phenolics; antioxidant

INTRODUCTION FC carrots represent an important segment of the FC mar-

Mild heat treatments have been used to improve the shelf-life
quality of fresh-cut (FC) products by reducing the microbial load
and decreasing enzyme activities.1 The application of microwave
energy has many advantages over conventional hot water treat-
ments, including (1) in-depth heating in the absence of a temper-
ature gradient; (2) higher heating rates and reduced heating times;
(3) avoidance of the leaching of vitamins, flavours, pigments, car-
bohydrates and other water-soluble components and (4) inacti-
vation of enzyme complexes.2 Very little information is available
about the use of microwave energy as a postharvest treatment
to reduce bacterial populations on fruits or vegetables, with pre-
vious studies being limited to whole fruits.3 – 6 These scarce stud-
ies showed that microwave irradiation reduced the incidence of
postharvest diseases (related to moulds) for whole peaches, nec-
tarines and pears. It has been reported that microwave irradiation
is a very effective approach for inactivating the enzymes respon-
sible for browning. The microwave blanching of carrots resulted
in drastic peroxidase inactivation after 30 s,7 resulting in better
flavour, colour, texture and nutritional value than water-blanched
carrots.8 However, no information is available about the use of
microwave treatment for FC fruits or vegetables.
ket owing to their versatility of use, pleasant flavour and nutri-
tional and health-promoting benefits, which are mainly derived
from phenolic and carotenoid compounds. However, minimal pro-
cessing of FC produce induces tissue damage, which triggers
non-microbial and microbial spoilage during storage and a sub-
sequent negative impact on sensory quality. The most prominent
quality changes in FC carrots during their shelf-life are loss of
orange intensity and/or whitening and a reduction in the flavour,
aroma and texture properties associated with freshness.9
Washing with 50–150 mg L−1 chlorinated water (NaClO) is a
widely used method of sanitizing FC fruits and vegetables, thus
reducing their initial microbial loads and ensuring the food safety
of these products. However, the formation of carcinogenic chlo-
rinated by-products such as chloramines and trihalomethanes
has led to safety concerns regarding chemical hazards for both
humans and the environment.10 These apprehensions have
∗ Correspondence to: Giancarlo Colelli, Dipto SAFE, Università degli Studi di
Foggia, Via Napoli 25, I-71122 Foggia, Italy. E-mail: giancarlo.colelli@unifg.it
Dipto SAFE, Università degli Studi di Foggia, Via Napoli 25, I-71122, Foggia, Italy

J Sci Food Agric (2015) www.soci.org © 2015 Society of Chemical Industry

 www.soci.org
caused some European countries (e.g. Germany, The Netherlands,
Switzerland and Belgium) to ban the use of NaClO in FC products.
Accordingly, there is a need to find new sustainable and effective
alternatives to NaClO.
Phenylalanine ammonia lyase (PAL, EC 4.3.1.5) is the key enzyme
in phenolic biosynthesis. Accordingly, the enhancement of this
enzyme’s activity may be of great interest for increasing the bioac-
tive properties of FC carrots and other products; some authors
have proposed using different abiotic stresses such as wound-
ing, mild heat treatments and UV-C.11 However, the activities of
other enzymes (polyphenol oxidase and peroxidase) responsible
for the oxidative degradation of phenolic compounds to brown
polymers (melanins) must be diminished or even eliminated to
extend the shelf-life of FC products. However, in the case of FC car-
rots, post-cutting browning is not a major problem.
The main objective of this study was to investigate the poten-
tial use of microwave irradiation during fresh-cut processing by
different operative parameter combinations (power and time of
treatment) to reduce microbial loads and extend the shelf-life of
FC carrots and ultimately design a fresh-cut processing plant. Fur-
thermore, the effect of microwave treatment on the bioactive com-
pounds of carrots was also studied. To the best of our knowledge,
the data provided by this study about the use of microwave treat-
ment for FC products are novel and innovative.
EXPERIMENTAL
Plant material
Carrots (Daucus carota) were obtained from a local market (Fog-
gia, Italy) and stored at 5 ∘ C and 90–95% relative humidity (RH)
until the next day, when they were processed. Minimal process-
ing was accomplished in a disinfected cold room at 10 ∘ C. Plant
material was inspected; carrots free from defects and with similar
visual appearance were selected. Next, the carrots were washed
(2 min) with NaClO (100 mg L−1 , 5 ∘ C, pH 6.5 ± 0.1) at a ratio of 300 g
of plant material to 5 L of disinfected water, rinsed (1 min) with tap
water (5 ∘ C) and drained in a perforated basket. The carrots were
peeled using a manual peeler and cut into slices of 8 mm thick-
ness using a manual slicer. The peeler and slicer were regularly
disinfected with 700 g L−1 ethanol during preparation. Slices cor-
responding to the central part of the carrots ranging from 14 to
18 mm in diameter were selected for the treatments.
Microwave treatment
Samples (150 g) were placed in 1 L rigid polypropylene baskets
(12 cm × 17 cm × 5 cm) (Carton Pack, Rutigliano, Italy). Three
baskets (replicates) per treatment were prepared. Sample bas-
kets were treated in a household microwave oven with a glass
turntable (Samsung CE116KT, Seul, Korea). The power generator
for this unit operated at a frequency of 2450 MHz. According
to preliminary sensory tests performed to prevent excessive
tissue damage, the treatments applied to the carrot slices were
750 W for 45 s (MW1), 750 W for 60 s (MW2), 900 W for 45 s
(MW3) and 900 W for 60 s (MW4). A non-microwave treatment
was used as a control (MW0). A separate (not included in the
experiment) set of five baskets with carrots was subjected to the
various treatments while monitoring the surface temperature
of the samples every 5 s with an infrared thermometer (ScanT-
emp 410, TFA Dostmann GmbH & Co., Wertheim-Reicholzheim,
Germany). Owing to the low thickness of the samples, the
temperature differences between surface and centre were
always <2 ∘ C. After the microwave treatments, the samples
wileyonlinelibrary.com/jsfa © 2015 Society of Chemical Industry
G Martínez-Hernández, ML Amodio, G Colelli
were stored at 5 ∘ C (90–95% RH) under air conditions using
individual polyethylene bags to reduce water loss. The weight
loss of the carrots after treatment and during storage was also
monitored.
Analyses were conducted on the processing day and after 3 and
7 days of shelf-life. After obtaining microbiological tissue aliquots,
carrot samples were stored at −80 ∘ C until further analysis for total
phenolics (TP), total antioxidant capacity (TAC) and PAL.
Analyses
Respiration rate
The respiration rate (RR, mL CO2 kg−1 h−1 ) of carrot slices was
measured using a dynamic system,12 in which 2 L gas-tight jars
were filled with 150 g of slices per jar and stored at 5 ∘ C. Samples
of headspace gas (0.1 mL) were taken from the inlet and outlet
flows of each jar and injected into a gas chromatograph (Shimadzu
17A, Kyoto, Japan) equipped with a thermal conductivity detector
(200 ∘ C). Separation of CO2 was achieved on a Carboxen 1006 PLOT
capillary GC column (30 m × 0.53 mm; Supelco, Bellefonte, PA, USA)
with a column flow of 7 mL min−1 and an oven temperature of
180 ∘ . The difference in concentration was then referred to the
sample weight and the airflow rate. Gas samples from three jars
per treatment were monitored every day for up to 7 days.
pH, titratable acidity and soluble solid content
The samples were analysed for soluble solid content (SSC), pH and
titratable acidity (TA). Sample juice was prepared from five carrot
slices by grinding with an Ultra-Turrax T-25 (IKA, Staufen, Germany)
instrument and filtering through four layers of cheesecloth. A pH
meter was used to determine the pH. The SSC of the juice was
determined by a digital hand-held refractometer (Atago N1, Tokyo,
Japan) at 20 ∘ C and expressed as ∘ Brix. TA was determined by
the titration of 7 mL of juice plus 33 mL of distilled water with
0.1 mol L−1 NaOH to pH 8.1 (T50, Mettler Toledo, Milan, Italy) and
expressed as % (g malic acid per 100 mL). Three replicates per
treatment were analysed.
Colour and image analysis
Colour was determined using a colorimeter (Minolta CM-2600d,
Osaka, Japan). The standard tristimulus parameters (L*, a*, b*) of
the CIELab system were recorded on three equidistant points of
each replicate. Three colour readings were recorded for each piece,
and the mean measurement of five pieces for each replicate was
recorded. The whiteness index (WI) was calculated according to a
previously reported equation:13
[ ]1∕2
WI (%) = 100 – (100 –L∗ )2 + a∗2 + b∗2
The white portions inside the carrot slices were studied by image
analysis as described by Amodio et al.14 A digital colour camera
(Canon EOS 400D, Melville, NY, USA) located inside a dark box con-
taining four fluorescent lamps was used to capture images. All
treatments were captured in the same image (two carrot slices
per treatment) with the following camera settings: manual expo-
sure mode with a lens aperture of f = 8.0, speed 1/10, resolution of
3888 × 2592 pixels and storage in TIFF format. To partition the RGB
image of carrot quarters from the background, the Red image was
thresholded using the Otsu method.15 A second thresholding was
then applied in the pixel corresponding to the carrot slices using
the Blue image. All algorithms for image analysis were performed
using the MATLAB® Version 7.0 image-processing toolbox (Math-
Works, Natick, MA, USA).
J Sci Food Agric (2015)
Potential use of microwave treatment on fresh-cut carrots
Firmness and damage to cell membranes
The firmness of the slices, being defined as the force (N) required
to compress each slice by 3 mm, was determined between two
parallel plates using an Instron Universal Testing Machine (Model
3343 Instron, Norwood, MA, USA) at a speed of 50 mm min−1 . The
peak force was reported as the maximum force and recorded as the
indicator of firmness. Ten slices were tested for each of the three
replicates corresponding to each treatment.
Relative electrolyte leakage (REL) was measured according to
the method described by Martínez-Hernández et al.,16 but with
modifications. Mannitol isotonic solution was used instead of
water to avoid the osmotic shock that water imparts to the tissue.17
An optimal mannitol concentration of 0.2 mol L−1 was determined
according to Peng et al.18 One carrot slice (approximately 3–4 g)
was placed in a Falcon tube (NEST Biotech, Rahway, NJ, USA), and
25 mL of 0.2 mol L−1 mannitol (Sigma Aldrich, Steinheim, Germany)
was added. The electrical conductivity of the bathing mannitol
solution was measured with a conductivity meter (CM35, Crison,
Carpi, Italy) after 1 min (C1) and 60 min (C60) of incubation with
orbital shaking (DAS12500, Intercontinental Equipment, Rome,
Italy) at a speed of 60 cycles min−1 . The samples were then heated
at 95 ∘ C for 2 h in a water bath, and the conductivity (CT) of the
bathing mannitol solution was measured after cooling in an ice
bath for 4 min. The REL was calculated using the equation REL
(%) = [(C60 − C1)/CT] × 100. Three replicates per treatment were
analysed.
Malondialdehyde (MDA) content was measured by the thiobar-
bituric acid (TBA) method described by Bi et al.,19 but with slight
modifications. Briefly, 4 g of tissue was added to 5 mL of 100 g L−1
trichloroacetic acid (Sigma Aldrich), ground to homogenate using
an Ultra-Turrax T-25 (IKA), transferred to a 50 mL tube (NEST
Biotech) with another 15 mL of trichloroacetic acid and cen-
trifuged (PK 121R, ALC International, Cologno Monzese, Italy) at
2200 × g for 15 min. Next, 2 mL of supernatant was mixed with
2 mL of 6 g L−1 TBA (Sigma Aldrich). The mixture was heated at
95 ∘ C for 20 min, then quickly cooled in an ice bath. After centrifu-
gation at 5260 × g for 15 min, the absorbances of the supernatant
at 450, 532 and 600 nm were recorded and calculated according
to Bi et al.19 The concentration of MDA was expressed as nmol g−1
fresh weight (FW). Three replicates per treatment were analysed.
PAL activity
PAL was extracted and determined according to Heredia and
Cisneros-Zevallos.20 Briefly, carrot tissue samples (7 g) were sup-
plemented with polyvinylpolypyrrolidone (Sigma Aldrich) (0.7 g)
and homogenized in ice-cold 50 mmol L−1 borate (Sigma Aldrich)
buffer, pH 8.5 (18 mL), containing 400 μL L−1 2-mercaptoethanol
(Sigma Aldrich). Homogenates were filtered through four layers
of cheesecloth and then centrifuged at 12 500 × g for 20 min at
2 ∘ C. The supernatants were collected and used as PAL extracts.
Two sets of glass tubes (Fisher Scientific, Padova, Italy) contain-
ing 5 mL of PAL extract were prepared for every sample and
pre-incubated at 40 ∘ C for 5 min. Afterwards, 0.55 mL of either
water (blank) or 100 mmol L−1 L-phenylalanine (Sigma Aldrich)
substrate solution (freshly prepared before assay) was added to
each of the tubes for every sample set. The absorbances of the
sample sets at 290 nm were measured (Shimadzu UV1700 spec-
trophotometer) at time 0 and after 1 h of incubation at 40 ∘ C.
The PAL activity was calculated as μmol t-cinnamic acid (Sigma
Aldrich) synthesized kg−1 FW h−1 using a t-cinnamic acid stan-
dard curve (0.02–0.3 μmol mL−1 ). All measurements were made in
duplicate.
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TP and TAC
TP and TAC were determined according to Amodio et al.,21 but
with slight modifications. Briefly, carrot samples (5 g) were homog-
enized (Ultra-Turrax T-25, IKA) at 18 000 rpm with 5 mL of methanol
(Sigma Aldrich). The TP and TAC extracts were centrifuged at
13 500 × g for 15 min at 4 ∘ C.
The amount of TP compounds was determined according to
Amodio et al.14 The TP content was expressed as mg chlorogenic
acid equivalent (CAE) kg−1 FW. All extracts were analysed in dupli-
cate.
The TAC was determined according to Martínez-Hernández
et al.,16 but with slight modifications. A solution of 100 μmol L−1
2,2-diphenyl-1-picrylhydrazyl radical (DPPH• , Sigma Aldrich)
in methanol (Sigma Aldrich) was prepared and adjusted to
1.1 ± 0.02 nm. A 50 μL aliquot of the previously described extract
was added to 950 μL of this solution and incubated for 24 h at room
temperature in darkness. The decrease in absorbance at 515 nm
was measured using a spectrophotometer. The results were
expressed as μmol Trolox equivalent kg−1 FW. All measurements
were made in duplicate.
Microbial analysis
Standard enumeration methods were used to determine micro-
bial growth according to Rinaldi et al.,22 but with slight modifi-
cations. A 10 g sample of carrots was homogenized in 90 mL of
sterile saline solution (8.5 g NaCl L−1 , Sigma Aldrich) for 1 min
with a stomacher (Colwort Stomacher 400 Lab, Seward Medical,
London, UK). A 1 mL aliquot of the appropriate sample dilution
was pour-plated on (1) plate count agar (Oxoid, Basingstoke, UK)
and incubated at 30 ∘ C/24–48 h and 5 ∘ C/7 days for total aerobic
mesophilic and psychrophilic bacteria respectively, (2) violet red
bile dextrose agar (Oxoid) and incubated at 37 ∘ C/48 h for enter-
obacteria and (3) de Man–Rogosa–Sharpe agar (Sigma Chemical
Co., St Louis, MO, USA) overlaid with the same medium and incu-
bated aerobically at 37 ∘ C/48 h for lactic acid bacteria (LAB). For
yeast and moulds (Y&M), 100 μL of the appropriate sample dilu-
tion was spread-plated on potato dextrose agar base (Oxoid) with
100 mg L−1 chloramphenicol (Sigma Chemical Co.) at 25 ∘ C/4–6
days. All microbial counts were reported as log colony-forming
units (CFU) g−1 . Each of the three replicates per treatment was anal-
ysed in duplicate.
Sensory evaluation
Sensory analyses were performed according to international
standards.23 The panel consisted of eight assessors (four women
and four men aged 24–40 years) screened for sensory ability
(colour, odour detection, firmness and basic taste). A five-point
scale of damage incidence and severity was scored for surface
dryness, off-odours and bitter aftertaste (5 = none; 4 = slight;
3 = moderate, limit of usability; 2 = severe; 1 = extreme), orange
colour intensity (5 = strong bright orange colour, freshly cut;
4 = bright; 3 = slightly pale, limit of usability; 2 = pale; 1 = very
pale) and fibrous aftertaste (5 = bite clean break; 4 = moderately
clean; 3 = fair, limit of usability; 2 = slightly woody; 1 = woody).
General appearance, fresh flavour and overall quality were
assessed using another five-point hedonic scale of acceptabil-
ity (5 = excellent; 4 = good; 3 = fair, limit of usability; 2 = poor;
1 = extremely bad).
Statistical analysis
The experiment was a 2 × 2 × 3 three-factor (microwave
power × treatment time × storage time) design subjected to
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Table 1. Results of three-way ANOVA for microwave power (750 and 900 W), treatment time (45 and 60 s), storage time (0, 3 and 7 days) and their
interactions on quality attributes of fresh-cut carrot slices

Attributea Microwave power (A) Treatment time (B) Storage time (C) A×B A×C B×C A×B×C

RR *** *** *** *** *** *** ***

pH NS *** *** NS NS NS NS
TA NS ** *** ** NS NS NS
SSC ** NS ** *** *** ** NS
Firmness NS NS NS NS NS NS NS
REL *** ** *** * *** ** *

MDA *** *** *** NS *** *** ***

L* NS *** *** *** NS NS NS

WI NS *** *** *** NS NS NS
PAL *** *** *** *** *** *** ***

TP *** *** *** *** *** *** **

TAC NS *** *** NS *** *** NS

Mesophiles ** *** *** NS NS NS *

Psychrophiles * *** *** NS NS NS *

Enterobacteria * *** *** NS NS NS *

LAB * *** *** NS NS * NS

Yeasts and moulds NS *** *** NS NS NS NS

NS, not significant;

* P ≤ 0.05;
** P ≤ 0.01;
*** P ≤ 0.001.
a RR, respiration rate; TA, titratable acidity; SSC, soluble solid content; REL, relative electrolyte leakage; WI, whitening index; MDA, malondialdehyde;
PAL, phenylalanine ammonia lyase; TP, total phenolics; TAC, total antioxidant capacity; LAB, lactic acid bacteria.

analysis of variance (ANOVA) using Statgraphics Plus Version 5.1

software (Statpoint Technologies, Warrenton, VA, USA). When
treatment effect was compared with the control samples, a
one-way ANOVA for the treatment effect was run at each time of
storage. Statistical significance was assessed at the level P = 0.05,
and Tukey’s multiple range test was used to separate means.

RESULTS AND DISCUSSION

The ANOVA results for the effects of microwave power (750 and
950 W), treatment time (45 and 60 s) and storage time (0, 3 and
7 days) and their interactions on quality attributes and bioactive
compounds of FC carrots are reported in Table 1. Microwave power
affected RR, SSC, REL, MDA, PAL, TP and microbial counts, except
Y&M, whereas time of treatment and time of storage affected most
of the quality parameters. For some of the studied parameters,
the effect of the three factors was not independent, showing
second-order interactions and sometimes also the interaction of Figure 1. Respiration rate of fresh-cut carrot slices subjected to different
third order (for RR, REL, MDA, PAL, TP, mesophiles, psychrophiles microwave treatments and stored for up to 7 days at 5 ∘ C (n = 3 ± SD).
and enterobacteria). Data were than further analysed according Different letters denote significant differences (P ≤ 0.05) among treatments
for the same sampling day.
to order of significant interactions as discussed in the following
subsections.
temperatures after treatment of 68–70 ∘ C (Fig. 2), as previously
Respiration rate reported for microwaved carrots.25 These temperatures may lead
For RR, all interactions were significant, so in Fig. 1 are shown the to the thermal inactivation of enzymes involved in pathways that
values of treated and untreated samples at each time of storage. lead to quality losses of FC products during their shelf-life, such
The initial (day 1) RR of the MW0 samples was 8.6 mL CO2 kg−1 h−1 . as ethylene production, which has been reported to be inhibited
Microwaved samples showed lower initial RR than MW0 samples, at temperatures near or above 35 ∘ C in many forms of produce.26
with values ranging between 2.2 and 3.9 mL CO2 kg−1 h−1 without Accordingly, the higher RR of MW0 compared with microwaved
significant differences among them. The wounding of carrots samples may be due to the increase in the wound-induced RR (for
induces an initial increase in RR, as previously modelled by Sur- MW0 samples) and the inactivation of respiration pathway-related
jadinata and Ci.24 The microwaved samples achieved their final enzymes (for microwaved samples). Furthermore, although no

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Potential use of microwave treatment on fresh-cut carrots
Figure 2. Surface temperature of fresh-cut carrot slices during microwave
treatments at 750 and 900 W for up to 60 s (n = 5 ± SD). Vertical bars
represents standard errors.
significant differences were observed between the initial RR
of microwaved samples, the treatment with higher power and
longer time (MW4) registered up to 3 days the lowest RR, while
the treatment with lower power and shorter time (MW1) showed
the highest RR among the microwave treatments. Following the
initial wound-induced maximum (days 1–3) RR of MW0 samples,
a typical decrease was observed on day 4, followed by a steady
state, as previously found.24 However, the RR of microwaved
samples, which remained quite stable until day 3 or 4, increased
dramatically at the end of storage, except for treatment MW4,
and this trend can explain the significant interactions of factors
A and B (power and time of treatment) with factor C (time of
storage). Moreover, while up to 3 days the high-energy treatments
(MW3 and MW4) showed lower RR than the treatments with low
energy (MW1 and MW2), at the end of storage a clear effect of
power and time of treatment could not be found (significant inter-
actions A × B, A × C and B × C), with MW3 showing the highest
RR, followed by MW1, MW2 and MW4. It has been reported that
postharvest heat treatments cause a transitory inhibition of gene
expression or a decrease in enzymatic activities, which recover
when the tissue is returned to non-stressed temperatures.27 In this
way, during storage, the activity of those enzymes was reactivated
and even enhanced owing to the abiotic stress applied by the
thermal treatments. Furthermore, the enhanced microbial growth
of the microwaved samples (as will be shown later) could have
significantly contributed to these RR increments.
pH, TA and SSC
Carrot pH was significantly affected by treatment time and storage
but not by treatment power (Table 1). Untreated carrots showed an
initial pH and TA of 6.7 and 0.036% (data not shown) respectively,
as previously reported for FC carrots.28,29 The pH decreased by
0.4–0.5 pH units after microwave treatments (Table 2). The pH
of microwave-treated carrots was 0.4–0.5 pH units lower than
the initial value and 0.5–0.8 pH units lower compared to control
samples after 7 days (Table 2), with a higher reduction for longer
time of treatment (60 s) at both treatment powers.
In accordance with pH data, microwave treatments increased TA
levels of carrots by 0.010–0.02 TA units (data not shown). Among
microwave treatments, treatment time increases from 45 to 60 s
led to higher TA increments (from 0.01 to 0.02 pH units), but only
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Table 2. Simple effects of microwave power (750 and 900 W)
and treatment time (45 and 60 s) on data averaged throughout
storage time for pH, titratable acidity (TA), L* and whitening
index (WI) of fresh-cut carrot slices subjected to different
microwave treatments and stored for up to 7 days at 5 ∘ C
Parameter Power (W) MW0 45 s 60 s
pH 750 6.35 ± 0.32 6.23 ± 0.01Aa 6.04 ± 0.02Bb
900 6.17 ± 0.11Aa 6.06 ± 0.03Ab
TA (%) 750 0.039 ± 0.004 0.052 ± 0.001Aa 0.051 ± 0.003Aa
900 0.046 ± 0.04Aa 0.053 ± 0.001Ba
L* 750 55.6 ± 2.2 49.0 ± 1.1Aa 48.8 ± 0.8Aa
900 50.5 ± 1.1Ab 47.2 ± 0.8Bb
WI (%) 750 46.6 ± 2.3 43.0 ± 1.2Aa 42.7 ± 0.3Aa
900 43.0 ± 0.8Aa 41.0 ± 0.7Bb
Different capital letters denote significant differences (P ≤ 0.05) between treat-
ment times for the same microwave power. Different lowercase letters denote
significant differences (P ≤ 0.05) between microwave powers for the same treat-
ment time. A column with the average value for control samples (MW0) has been
included as reference.
at 900 W power (as a consequence of the A × B interaction). During
storage, there were no differences between the two microwave
powers for the same treatment times. A similar TA increase and
corresponding pH reduction have been reported in untreated
shredded carrots stored for up to 10 days at 2 or 10 ∘ C under air
conditions.13,29 As Rocha et al.29 stated, the observed pH decrease
could be due to the production of CO2 , which, by reacting with the
water of the tissues, might induce a release of H+ .
The initial SSC (8.2 ∘ Brix) increased slightly after microwaving
(Fig. 3). In particular, this increase at 750 W power was higher
for longer treatment (0.8 ∘ Brix increment), while, on the con-
trary, as the effect of the A × B interaction, at 900 W power the
increase was more accentuated for the shorter time of treatment,
although it was only significant on days 3 and 7. Similarly, Zhang
et al.6 observed an increase in SSC and TA (not significant) after
microwaving (450 W for 2 min) of pears. However, Alegria et al.28
found that topped carrots registered an SSC reduction of 11% after
heating (boiling in water for 45 s), probably due to a leaching of
soluble solids into the boiling water. The TA and SSC increments
found herein may be attributed to a better extractability of acids
and other soluble solids respectively, due to the plant cell disrup-
tion during microwaving.
Colour and image analysis
The white discoloration of the surface of FC carrots is considered
an important form of quality reduction and is regarded by con-
sumers as a loss of freshness.30 In general, microwave treatment
delayed the whitening of carrot slices, as can be observed by a
reduction in sample WI (Table 2 and Figs 4A and 4C) and by lower
L* values, which showed an increase over time in control samples
(Table 2). Treated samples showed a slightly more intense orange
colour, although no significant colour differences were found by
the panellists (data not shown). Among microwave treatments, L*
and WI were significantly affected by treatment and storage times.
The microwave power × treatment time interaction (A × B) was sig-
nificant (P ≤ 0.001) for both L* and WI. Among samples treated for
60 s, those treated at 900 W showed lower L* and WI, while no dif-
ferences were observed for those treated for 45 s. Regarding the
time of treatment at 900 W, a lower whitening (lower WI and L*)
was observed for higher time of exposition (Table 2), therefore the
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Figure 3. SSC of fresh-cut carrot slices subjected to microwave treatments
and stored for up to 7 days at 5 ∘ C (n = 3 ± SD). Different capital letters
denote significant differences (P ≤ 0.05) among microwave treatments for
the same sampling day. Different lowercase letters denote significant dif-
ferences (P ≤ 0.05) among sampling days for the same microwave treat-
ment.
whitening was better controlled as the power and time of treat-
ments increased (Table 2 and Figs 4B and 4D). Accordingly, the
colour and surface dryness of MW0 samples were scored by panel-
lists below the limit of usability after 7 days. However, those param-
eters of microwaved samples were scored high (3–4), although no
significant differences among them were found (data not shown).
The superior bright orange colour and lower surface dryness of the
microwaved samples compared with the non-microwaved sam-
ples after 7 days can also be observed in Fig. 4. However, the
MW4 samples showed an incident pale orange colour (Fig. 4). The
observed discoloration of non-heat-treated FC carrots throughout
storage is a well-documented defect of these products that has
been attributed to surface dehydration and/or lignin synthesis.31
Furthermore, the control of whitening by heat treatment has pre-
viously correlated with the partial inhibition of enzymes related
to discoloration.32 In the case of our study, enzymatic inhibition
was assumed to play a greater role than surface dehydration,
because the microwaved samples registered higher dehydration
levels (3–7% after 7 days, being higher for treatments with higher
power and longer time as a consequence of the cell disruption
induced by heat treatment) than the MW0 samples (approximately
1% after 7 days, data not shown). Accordingly, the microwave treat-
ment of FC carrot slices reduced the usual unpleasant whitening of
FC carrots during conservation, with the treated samples exhibit-
ing better overall quality than the untreated samples.
Firmness and damage to cell membranes
Together with colour, firmness is one of the most important
quality attributes of FC carrots, strongly influencing their accep-
tance by consumers. Microwave treatments caused membrane
disruption of the carrots (Fig. 5) related to water loss and firm-
ness reduction (data not shown) and therefore to softening.
Accordingly, softening of carrot slices ranged from 38 to 51%
after microwaving (data not shown). However, initial water losses
(2.1–4.4%, data not shown) were not detected by panellists, as no
significant differences in dehydration scores among treatments
were found on the processing day (data not shown). Similarly,
no significant differences in sample fibrosity (data not shown)
were observed among treatments in the sensory analyses on the
wileyonlinelibrary.com/jsfa © 2015 Society of Chemical Industry
G Martínez-Hernández, ML Amodio, G Colelli
processing day. Previous studies reported sudden firmness loss
(>50%) in carrots from the initial state within a few minutes from
non-microwave heating.33,34 Microwave treatments avoided large
textural changes, maintaining acceptable firmness and ensur-
ing the acceptance of the FC carrot slices by the consumer. No
influence of power and treatment time on firmness was observed
among microwave treatments throughout storage (Table 1).
Carrot REL was significantly affected by the factors microwave
power, treatment time and storage time as well as their interac-
tions (Table 1), therefore the REL changes over time for each single
treatment are reported in Fig. 5. The membrane permeability of
FC carrot slices was expressed by the change in REL. FC carrots
registered an initial REL of 1.1% (Fig. 5) before microwave treat-
ments (MW0), being slightly reduced to 0.7% after 7 days. On the
other hand, REL increased greatly after microwave treatments (up
to 30–40%) as a consequence of the membrane, particularly for
the treatments with higher power and longer time. Accordingly,
MW1 samples registered a REL of 30%, while MW4 samples showed
a REL of 44%. These microwave treatments induced similar REL val-
ues to those achieved after the water blanching of carrots slices for
5 s.35 MW1 showed the lowest REL on the processing day and after
3 days of storage, and both MW1 and MW2 showed values con-
siderably lower than those of MW3 and MW4 at 7 days of storage.
Moreover, all treated samples showed a decrease in REL compared
with the processing day (except for MW3), but the samples cor-
responding to the treatments with lower power (MW1 and MW2)
registered a more pronounced decrease, being almost 50% lower
after 7 days than on the processing day (Fig. 5).
Malondialdehyde (MDA), a secondary end product of oxidative
lipid degradation, is often adopted as a suitable index to reflect
tissue damage. Accordingly, Hodges et al.36 reported a substantial
MDA increase of 25-fold in carrot slices after UV-C treatment for
7 days. MW0 registered an initial MDA content of 0.76 nmol g−1
(data not shown). No significant MDA changes were observed after
microwave treatments on the processing day (data not shown).
Similarly, Bi et al.19 did not find significant differences in the MDA
content after high-pressure carbon dioxide (HPCD) treatment (up
to 5 MPa for 15 min) of FC carrot slices. During cold storage, no
significant MDA changes occurred except for the MW1-treated
samples (data not shown), which registered a higher MDA content
(approximately twofold higher after 7 days). This was likely due to
an activation of lipid peroxidation-related enzymes after the mild
microwave treatment.
According to our data, and as Bi et al.19 found, REL is a better
cell membrane indicator than MDA content in carrot slices after
microwave treatments, as shown by the correspondence between
firmness and REL.
PAL activity
The PAL activity of FC carrot slices subjected to different microwave
treatments and stored at 5 ∘ C for up to 7 days is shown in Fig. 6.
As reported in Table 1, all factors and their interactions were sig-
nificant for this parameter. The MW0 carrot slices showed an ini-
tial PAL activity of 5.5 μmol t-cinnamic acid kg−1 h−1 , similar to that
reported by other authors.20,37 No significant differences in PAL
activity were observed after microwave treatments on the pro-
cessing day. Among microwave treatments on the processing day,
samples treated at 750 W for 60 s showed lower PAL activity than
those treated at 900 W, whereas carrots treated at 750 W for 45 s
showed intermediate values. A large PAL increase of approximately
tenfold was registered in MW0 carrots after 3 days and maintained
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Potential use of microwave treatment on fresh-cut carrots www.soci.org

A B

MW0 MW1 MW2 MW3 MW4 MW0 MW1 MW2 MW3 MW4

C D

MW0 MW1 MW2 MW3 MW4 MW0 MW1 MW2 MW3 MW4

Figure 4. Original RGB image (A, processing day; B, 7 days at 5 ∘ C) and binary image (C, processing day; D, 7 days at 5 ∘ C), where ‘0’ (black) is the background
and ‘1’ (white) is the whiteness region.

Figure 6. PAL activity of fresh-cut carrot slices subjected to microwave

Figure 5. REL of fresh-cut carrot slices subjected to microwave treat-
treatments and stored for up to 7 days at 5 ∘ C (n = 3 ± SD). Different capi-
ments and stored for up to 7 days at 5 ∘ C (n = 3 ± SD). Different capital
tal letters denote significant differences (P ≤ 0.05) among microwave treat-
letters denote significant differences (P ≤ 0.05) among microwave treat-
ments for the same sampling day. Different lowercase letters denote signif-
ments for the same sampling day. Different lowercase letters denote signif-
icant differences (P ≤ 0.05) among sampling days for the same microwave
icant differences (P ≤ 0.05) among sampling days for the same microwave
treatment.
treatment.

samples.38,40 These authors hypothesized that heat shock inhibits

up to day 7. It has been widely reported that the physical wound-
ing of plant tissues triggers several important changes in plant
metabolism, such as PAL activation, and thereby the biosynthe-
sis of phenolic compounds.37 – 39 Microwaved samples showed a
delayed PAL activation throughout storage, as observed by the
maximum PAL levels registered on day 7 for these samples. At 3
days of storage, samples microwaved at 900 W showed lower PAL
activity than samples treated at 750 W for 60 s, followed by the
treatment at 750 W for 60 s. Furthermore, samples microwaved at
750 W for 45 s showed the highest PAL activity after 7 days of stor-
age, even compared with the control samples. Accordingly, the
MW1 carrots registered a 64% higher PAL activity than the con-
trol samples on day 7, which may be attributed to an enzymatic
activation with this mild treatment. On the other hand, increas-
ing the time of treatment at both powers delayed the PAL activ-
ity up to 7 days, particularly at 900 W; in fact, MW4 registered a
91% lower PAL activity than the MW0 samples. Similar to our data,
heat-shocked (water at 45 ∘ C for 120 s) lettuce registered a delayed
maximum PAL activity, with 30% lower activity than non-heated
the increase in wound-induced PAL activity by inhibiting the accu-
mulation of PAL proteins (and thereby increasing enzyme activity)
by either preventing the translation or accelerating the turnover
of PAL proteins.38 Furthermore, heat shock treatments of plant tis-
sues have been reported to induce the synthesis of a unique set
of proteins called heat shock proteins, the accumulation of which
correlates with an increased tolerance to chilling injuries.40
TP content
As seen in Table 1, all factors and interactions were significant for
this parameter, therefore the single effects are reported in Fig. 7A,
where also the control samples (MW0) have been included. The
MW0 carrots showed an initial TP content of 81.3 mg CAE kg−1 FW,
which is in agreement with previous TP data on peeled carrots.41
Microwave treatments did not induce significant TP changes in car-
rots on the processing day. However, other authors reported that
microwave treatments for longer time (2.5 min at 900 W) used on
minimally processed kailan-hybrid broccoli induced an increase in

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 www.soci.org
Figure 7. (A) TP and (B) TAC of fresh-cut carrot slices subjected to
microwave treatments and stored for up to 7 days at 5 ∘ C (n = 3 ± SD).
Different capital letters denote significant differences (P ≤ 0.05) among
microwave treatments for the same sampling day. Different lowercase let-
ters denote significant differences (P ≤ 0.05) among sampling days for the
same microwave treatment.
TP of approximately twofold due to the cellular tissue disruption
and subsequently higher phenolic extractability.42 The lower phe-
nolic extractability of carrot tissue compared with kailan-hybrid
broccoli could be explained by the higher thermal resistance of the
carrot tissue compared with broccoli as well as the longer treat-
ment time.
The TP contents during storage are in accord with the PAL activ-
ity. Hence only the MW0 samples registered an early TP enhance-
ment of 36% after 3 days, and the treatments at 900 W (MW3 and
MW4) showed lower TP content than the control samples. Simi-
larly, large TP enhancements have been reported in FC carrot slices
stored in air conditions at 8 ∘ C for 12 days.43 Among microwaved
samples, those treated for 45 s registered TP increments of 37%
(900 W) and 118% (750 W) after 7 days compared with their respec-
tive initial levels. On the other hand, samples treated for 60 s did
not show great changes after 7 days compared with their respec-
tive initial levels. Conclusively, the microwaving of FC carrot slices
for 60 s did not alter the initial TP content. Furthermore, the TP
enhancement observed for samples treated for 45 s was not cor-
related with sample browning.
TAC
The TAC of samples after treatment and during low-temperature
storage showed a similar behaviour to their TP content, which can
wileyonlinelibrary.com/jsfa © 2015 Society of Chemical Industry
G Martínez-Hernández, ML Amodio, G Colelli
be explained by the strong TAC–TP correlation (R2 = 0.93). An ini-
tial TAC of 74.2 μmol Trolox equivalent kg−1 FW was registered by
the MW0 samples (Fig. 7B). Regarding the microwave treatments,
only the treatment with higher power and longer time (MW4)
induced a significant (P ≤ 0.05) TAC increment (49%) on the pro-
cessing day. A similar trend (not significant) was observed for TP
content. The lack of significance for this trend in the TP data may
be explained by the contribution of other compounds with antiox-
idant capacity, such as carotenoids and vitamin C, which showed
the same behaviour as phenolics. Similarly, TAC enhancements
have been found in several vegetables after microwaving,44,45
which could be explained in different ways: (1) the liberation of
high amounts of antioxidant components due to the thermal
destruction of cell walls and sub-cellular compartments, (2) the
production of strong radical-scavenging antioxidants by thermal
chemical reactions, (3) the suppression of the oxidation capacity of
antioxidants by thermal inactivation of oxidative enzymes and/or
(4) the production of new non-nutrient antioxidants or the forma-
tion of novel compounds with antioxidant activity, such as Maillard
reaction products.
Among microwave treatments, no power × treatment time inter-
action was found. However, the interactions power × storage time
and treatment time × storage time and the third-order interaction
were significant (Table 1). According to TP contents, the TAC of the
MW0 and samples microwaved for 45 s increased by 193% (MW0),
277% (900 W) and 394% (750 W) after 7 days. These increments
corresponded to specific antioxidant activities (defined as the ratio
between the antioxidant capacity and phenolic content of tis-
sue) of 823, 667 and 951 μg Trolox mg−1 phenolics, which were
lower than the value of 1056 μg Trolox mg−1 phenolics reported by
Cisneros-Zevallos11 for wounded unpeeled carrots stored for 48 h
at higher storage temperature (20 ∘ C).
The recommended daily intake of antioxidants for humans
to obtain benefits is an estimated equivalent of approximately
3000–3600 μmol Trolox day−1 .46 Therefore the microwaving of FC
carrot slices at 750 W for 45 s could provide an opportunity to
obtain products with higher antioxidant capacity to meet the daily
requirement.
Microbial analysis
Microbial loads of FC carrot slices were significantly (P ≤ 0.05)
affected by microwave power (except for Y&M), treatment time
and storage time, except for LAB and Y&M (Table 1). Triple interac-
tions were significant (P ≤ 0.05) for mesophiles, psychrophiles and
enterobacteria, therefore data were analysed for single treatment
and storage time (Table 3). The initial mesophilic, psychrophilic,
enterobacteria, LAB and Y&M counts of control samples were 4.6,
4.6, 4.4, 2.4 and 4.1 log CFU g−1 respectively, similar to those found
by Alegria et al.9,28 in FC carrots treated with 200 ppm NaOCl for
1 min. In general, microwave treatments induced significant reduc-
tions in the initial counts for all microbial groups. As expected,
MW4 achieved the greatest microbial reduction, corresponding
to 1.5, 1.8 and 1.6 log CFU g−1 for mesophiles, psychrophiles and
enterobacteria respectively, while the other treatments achieved
reductions of 0.6–0.9 log CFU g−1 without significant differ-
ences between them. The longest treatments (MW2 and MW4)
reduced Y&M counts to detectable levels (<10 CFU g−1 ), while
the shorter treatments (MW1 and MW3) induced Y&M reductions
of 0.7–0.8 log CFU g−1 without significant differences between
them. All microwave treatments reduced LAB to undetectable
levels (<10 CFU g−1 ). Longer microwave treatments (2.5 min
at 900 W) used for minimally processed kailan-hybrid broccoli
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Potential use of microwave treatment on fresh-cut carrots www.soci.org

Table 3. Microbial loads (log CFU g−1 ) of fresh-cut carrot slices subjected to different microwave treatments and stored for up to 7 days at 5 ∘ C
(n = 3 ± SD)

Storage time(days) Treatment Mesophilic Psychrophilic Enterobacteria LAB Yeasts/moulds

0 MW0 4.6 ± 0.1Ac 4.6 ± 0.1Ab 4.4 ± 0.1Ab 2.4 ± 0.2Ac 4.1 ± 0.2Ab
MW1 4.1 ± 0.1Bc 4.0 ± 0.1Bc 3.9 ± 0.12ABc 1.5 ± 0.4Bc 3.4 ± 0.5ABc
MW2 3.9 ± 0.1Bc 3.7 ± 0.2Bc 3.7 ± 0.2BCc 1.0 ± 0.1Bb 2.2 ± 0.3BCc
MW3 4.0 ± 0.2Bc 3.9 ± 0.2Bc 3.8 ± 0.1ABCc 1.4 ± 0.3Bc 3.3 ± 0.8ABCb
MW4 3.1 ± 0.3Cc 2.7 ± 0.4Cc 2.8 ± 0.4Cc 1.3 ± 0.4Bb 2.1 ± 0.1Cc
3 MW0 5.9 ± 0.1ABb 6.0 ± 0.2ABa 5.8 ± 0.1ABa 3.4 ± 0.1Ab 5.9 ± 0.1Aa
MW1 6.3 ± 0.4Ab 6.3 ± 0.4Ab 6.1 ± 0.2Ab 3.6 ± 0.1Ab 6.0 ± 0.1Ab
MW2 5.2 ± 0.1Cb 5.2 ± 0.2Cb 5.1 ± 0.1Cb 2.5 ± 0.9ABab 5.5 ± 0.2Ab
MW3 6.0 ± 0.2ABb 6.1 ± 0.1ABb 5.9 ± 0.2Ab 3.1 ± 0.2ABb 5.8 ± 0.1Aa
MW4 5.5 ± 0.1BCb 5.6 ± 0.1BCb 5.5 ± 0.1BCb 2.1 ± 0.1Bb 5.1 ± 0.3Bb
7 MW0 6.3 ± 0.1Ca 6.1 ± 0.1Ba 5.9 ± 0.1Ba 5.0 ± 0.1Ca 4.6 ± 0.6Bb
MW1 8.6 ± 0.4Aa 8.6 ± 0.5Aa 8.4 ± 0.4Aa 5.7 ± 0.7Aa 8.0 ± 0.2Aa
MW2 8.4 ± 0.3ABa 8.1 ± 0.5Aa 7.8 ± 0.6Aa 4.5 ± 1.3ABa 7.6 ± 0.9Aa
MW3 8.4 ± 0.2ABa 8.4 ± 0.3Aa 8.1 ± 0.2Aa 5.2 ± 0.3Aa 7.8 ± 0.1Aa
MW4 7.7 ± 0.5Ba 7.7 ± 0.5Aa 7.5 ± 0.4Aa 4.1 ± 1.7Ba 7.3 ± 0.8Aa

Different capital letters denote significant differences (P ≤ 0.05) among treatments for the same sampling day. Different lowercase letters denote
significant differences (P ≤ 0.05) among sampling days for the same treatment.

achieved microbial reductions ranging from 2.4 to 2.9 CFU g−1 .42
Hot water treatment of FC carrots has also shown greater microbial
reductions,9 probably due to the water treatment itself. Never-
theless, MW4 and MW2 microwave treatments achieved similar
or greater initial microbial reductions than those achieved after
the NaOCl treatment (200 ppm for 1 min) of FC carrots.9,28 Accord-
ingly, microwave treatment could be considered a sustainable
alternative to conventional NaOCl sanitation to reduce the initial
microbial population of FC carrots.
However, the microbial growth of the microwaved samples was
greater than that of the control samples throughout storage, and
this explained the interaction of third order for mesophiles, psy-
chrophiles and enterobacteria and the interaction of second order
with treatment time and storage time. No significant differences
were registered between the MW0 and microwaved samples after
3 days, with microbial loads of approximately 3 log CFU g−1 for
LAB and 6 log CFU g−1 for the other microbial groups. After 7
days, MW0 achieved microbial loads of 5–6 log CFU g−1 , while
the microwaved samples registered approximately 8 log CFU g−1
counts for all microbial groups (5 log CFU g−1 for LAB). How-
ever, no significant differences between microwave treatments
were found after 7 days, except for the LAB counts of MW2 and
MW4 samples, which were 0.5–1 log units lower than those of
MW0 samples. Similar mesophilic increases in untreated FC car-
rot slices on day 7 of storage at 8 ∘ C have been reported.43 How-
ever, Rocha et al.29 reported microbial increases that were approxi-
mately twofold higher than those found in this study for shredded
carrots stored at an even lower temperature (2 ∘ C) for the same
period of time, but this could be due to the higher wounding of
the tissue compared with the present study, which led to greater
cellular disruption, with more nutrients available for microbial pro-
liferation. The high microbial growth of microwaved samples may
be attributed to the high plant cell disruption caused by the ther-
mal treatment, as previously shown by the REL data, which led to
a strong release of nutrients, favouring microbial growth.
Conclusively, the microwave treatment of FC carrot slices
achieved good initial microbial reductions. That fact corroborates
the highly significant effect of heat shock in controlling microbial
development, as also previously reported by Alegria et al.9,28 How-
ever, other combinations of treatments or techniques to control
microbial growth, including modified atmosphere packaging, are
needed to retain those initially achieved microbial reductions
throughout the storage of microwaved samples.
CONCLUSIONS
Microwave treatments greatly retained the physicochemical and
sensory quality of fresh-cut carrots during shelf-life, with the treat-
ments with lower power (750 W for 45 and 60 s) being found to be
the most appropriate for possible implementation in an industrial
plant. Microwave treatment reduced whitening and avoided large
textural changes of samples during storage. The bioactive com-
pounds of microwaved samples were enhanced throughout the
shelf-life by up to 120 and 200% respectively. Although microwave
treatments achieved initial microbial reductions of approximately
1 log CFU g−1 , the microbial growth of these samples was greater
than that of the untreated samples. Accordingly, microwave treat-
ment is a good sustainable alternative to NaOCl but requires sup-
plementation with other sanitizing techniques to control microbial
growth throughout the shelf-life of fresh-cut carrot slices.
ACKNOWLEDGEMENTS
The assistance of Mulugheta Tesfamichael is appreciated. This
work was realized thanks to funding from the project ‘P.O.N. Ricerca
e Competitività’ PON02_00657_00186_3417392/1 (INFOPACK),
2007–2013.
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