Journal of Food Composition and Analysis 86 (2020) 103363

Contents lists available at ScienceDirect

Journal of Food Composition and Analysis

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

Original Research Article

Evaluation of quality, phenolic and carotenoid composition of fresh-cut T

purple Polignano carrots stored in modified atmosphere
B. Pacea,1, I. Capotortoa,1, M. Cefolaa,*,1, P. Minasib, N. Montemurroa, V. Carboneb
a
Institute of Sciences of Food Production, National Research Council (CNR), Via Michele Protano c/o CS-DAT, 71121 Foggia, Italy
b
Proteomic and Biomolecular Mass Spectrometry Center, Institute of Food Sciences, National Research Council (CNR), Via Roma, 64, 83100 Avellino, Italy

A R T I C LE I N FO A B S T R A C T

Chemical compounds studied in this article: Purple Polignano carrot is a traditional Italian landrace cultivated in Apulia region rich in antioxidants and with
3-O-Caffeoylquinic acid (PubChem CID: a high nutritional value. On the other hand, these carrots showed a high perishability. The postharvest behaviour
1794427) of fresh-cut purple Polignano carrots stored in open bags (AIR) or passive modified atmosphere (pMA) was
5-p-Coumaroylquinic acid (PubChem CID: studied, analysing the main qualitative parameters and the polyphenols and carotenoids profile. The storage in
90478782)
pMA allowed to preserve visual quality and respiration rate better than the storage in AIR. Polyphenols (hy-
Caffeoyl-N-tryptophan (PubChem CID:
droxycinnamic acids, their derivatives and anthocyanins) increased during storage of about 249 % in samples
15228042)
Cyanidin 3-xylosyl(sinapoylglucosyl) stored in pMA and of about 306 % in those stored in AIR at 4 °C during the first 4 days of storage, respect to fresh
galactoside (PubChem CID: 71587512) carrots. Instead, carotenoids mean content in carrots just after harvest (6.28 ± 0.48 mg kg-1 fw) did not change
Cyanidin 3-xylosyl(feruloylglucosyl) significantly during storage at 4 °C (both in AIR or pMA). According to our results, pMA resulted a valid solution
galactoside (PubChem CID: 56776253) to cold store fresh-cut purple Polignano carrots until two weeks, preserving quality and nutritional value.
Cyanidin 3-xylosyl(coumaroylglucosyl)
galactoside (PubChem CID: 70698351)
Lutein (PubChem CID: 5281243)
α-carotene (PubChem CID: 6419725)
β-carotene (PubChem CID: 5280489)

Keywords:
Fresh-cut purple carrots
Cold storage
HPLC-ITMSn
DPPH assay
Polyphenols
Carotenoids
Food analysis
Food composition
Nutritional value

1. Introduction including antioxidant, anti-inflammatory and antitumor activities (Hou

Carrots are an important source of phytochemicals, including phe-
nolics and carotenoids, whose chemical composition is responsible for
the peel colour; indeed, purple carrots are richer in antioxidants than
orange typology (Akhtar et al., 2017). In particular, the colour of purple
carrots is due to anthocyanins, a family of red, blue, and purple water-
soluble pigments that have been linked to a variety of biological activity
et al., 2004; Wang and Stoner, 2008). In addition, purple carrots also
contain a valuable amount of carotenoids and phenolics (Kammerer
et al., 2004; Kramer et al., 2012; Mizgier et al., 2016; Nicolle et al.,
2004). Purple Polignano carrots are a traditional landrace cultivated in
Apulia region, rich in antioxidants, with a mean content about 4-fold
higher than the commercial orange carrots (Cefola et al., 2012).
Moreover, compared to commercial orange carrots, purple Polignano

Abbreviations: pMA, passive modified atmosphere; fw, fresh weight; HPLC, high performance liquid chromatography; UV, ultraviolet; Vis, visible; HPLC-ITMSn,
High Performance Liquid Chromatography-Multistage Ion Trap Mass Spectrometry; PP, polypropylene; PE, polyethylene
⁎
Corresponding author.
E-mail address: maria.cefola@ispa.cnr.it (M. Cefola).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.jfca.2019.103363
Received 30 July 2019; Received in revised form 8 November 2019; Accepted 8 November 2019
Available online 10 November 2019
0889-1575/ © 2019 Elsevier Inc. All rights reserved.
B. Pace, et al.
carrots showed a lower nitrate content (50 %), a lower content in su-
crose and a higher content in fructose (Cefola et al., 2012). Starting
from these first results, purple Polignano carrots can be considered a
vegetable with a high nutritional value. However, from a physiological
point of view, these carrots showed a high perishability, with a Q10
value 6-fold higher than the commercial orange type (Renna et al.,
2013). Processing carrots as fresh-cut vegetables could increase their
diffusion and availability towards new categories of consumers (chil-
dren, single, working women), who are more and more inclined to
purchase ready to use products. The use of modified atmosphere is a
technology generally applied to storage of perishable fruits and vege-
tables in order to preserve the compositional quality, since it reduce the
respiration rate of commodity extending their shelf-life; and it is widely
used to package fresh-cut vegetables, as cut causes the increase in
biochemical and physiological reactions, shortening shelf-life (Kader,
2002).
To the best of our knowledge, there are no research papers on the
characterization of fresh-cut purple Polignano carrots. Therefore, this
paper aimed to study the postharvest performance of fresh-cut purple
Polignano carrots during cold storage in AIR or passive modified at-
mosphere (pMA) packaging. In particular, for the first time, a com-
prehensive study of the phenolic and carotenoid profiles of purple
Polignano carrots, both just after harvest and during cold storage in AIR
or pMA, was carried out by high-performance liquid chromatography
coupled with multistage ion trap mass spectrometry (HPLC/ITMSn).
2. Materials and methods
2.1. Plant materials and storage conditions
Purple Polignano carrots (Daucus carota L. var. sativus) were pur-
chased from a local market in Polignano a Mare (Bari, Italy), and
transported in refrigerated conditions within two hours to the
Postharvest Laboratory c/o CS-DAT to be processed. In details, 5 Kg of
carrots were brushed, washed in tap water, dried and sliced using a
cutter (Robot coupe CL52, Vinvennes-Cedex, France) with a thickness
of approximately 5 mm. Carrots slices were then immediately dipped in
tap water for 5 min, dried using tissue paper, and closed in passive
modified atmosphere (pMA) using 13 × 18 cm microperforated poly-
propylene bags (PP MF1 D100, Carton Pack Srl, Rutigliano, Italy) or
kept in air as control (AIR) using the same bags unsealed (about 100 g
each bag). A total of 24 bags (3 replicates x 2 packages x 4 storage
times, after 4, 7, 10 and 14 days) were prepared.
Just after harvest (fresh sample) and at each storage time, samples
were subjected to sensory, physical and chemical analyses as detailed
below. Headspace gas composition (O2 and CO2) within each package
was monitored daily using a gas analyzer (CheckPoint, PBI Dansensor,
Ringsted, Denmark).
2.1.1. Selection of the proper packaging material
The proper packaging material, used in the trial, was selected by
comparing the following three typologies in a specific test: poly-
propylene (PP) MF1 D50 (40 holes m-2, 30 μm thickness), PP MF1 D100
(80 holes m-2, 30 μm thickness) (Carton Pack Srl, Rutigliano, Italy) and
polyethylene (PE) 40 μm thickness (Corapack, Brenna, Italy). Thus,
carrots slices (about 100 g each bag) were prepared and packaged as
above reported, using five replicates for each packaging material. All
samples were stored for about 2 weeks at 4 °C and headspace gas
analysis was performed during storage. The proper packaging material
used in the trial was chosen on the base of O2 and CO2 measured inside
bags.
2.2. Chemicals and reagents
For chromatographic and mass spectrometric analyses, methanol,
acetonitrile, acetone, ethyl acetate (all HPLC grade) and formic acid
2
Journal of Food Composition and Analysis 86 (2020) 103363
were obtained from Merck (Darmstadt, Germany). Caffeic acid, ferulic
acid and cumaric acid were purchased from Sigma-Aldrich Co. 3-O-
Caffeoylquinic acid (chlorogenic acid), cyanidin-3-O-glucoside
chloride, lutein and β‑carotene were obtained from Extrasynthese
(Genay, France). HPLC grade water (18.2 MΩ) was prepared by using a
Millipore Milli-Q purification system (Merck Millipore, Darmstadt,
Germany).
2.3. Sensory visual quality
Carrots slices were examined by a group of 8 trained researchers to
assess the sensory visual quality, based on a colour photographic scale
associated with brief description of freshness, colour uniformity, and
brightness. Coded (3 digits) samples were presented to the judges in-
dividually, to enable them to make independent evaluations. Visual
quality was evaluated on a 5-point rating scale, where 5 = excellent,
fresh appearance, full marketable, 4 = good, product marketable, slight
loss of visual quality, 3 = lack of visual quality, limit of marketability
(5–10% unmarketable slices), 2 = product has notable visual defects
(10–30% unmarketable slices), 1 = severe visual defects (> 50 % un-
marketable slices). Slices scored below 3 were considered unmarketable
for the loss of the overall visual quality (loss of turgor and brightness).
2.4. Respiration rate
The respiration rate was measured using a closed system as reported
by Kader (2002). About 100 g of carrot slices for each replicate were put
into 6 L sealed plastic jars (one jar per replicate) where CO2 was al-
lowed to accumulate up to 0.1 %. The time taken to reach this value
was calculated, by taking CO2 measurements at regular time intervals.
For CO2 analysis, 1 mL gas sample was taken from the head space of the
plastic jars through a rubber septum and injected into a gas chroma-
tograph (p200 micro GC, Agilent, Santa Clara, CA, USA) equipped with
dual columns and a thermal conductivity detector. CO2 was analyzed
with a retention time of 16 s and a total run time of 120 s on a 10 m
porous polymer (PPU) column at a constant temperature of 70 °C. Re-
spiration rate was expressed as mL CO2 kg-1 h-1.
2.5. Firmness, electrolyte leakage and dry weight
The firmness of carrot slices was measured using a firmness tester
(ZwickLine Z0.5, Zwick/Roell, Ulm, Germany) equipped with a 3-mm
probe of diameter, measuring the relative deformation (%) of the slices
up to a 30 N load (Cefola and Pace, 2016). To determine the electrolyte
leakage, the procedure described by Pace et al. (2015) was used with
slight modification. Disks of 16 mm of sliced carrots (5 g for each re-
plicate), obtained using a cork borer, were placed in tubes and im-
mersed in 20 mL of distilled water. After 30 min of storage at 4 °C,
conductivity of the solution was measured using a conductivity meter
(Model Cond.61, XS Instruments, Carpi, Italy). The tubes were then
frozen at −20 °C and, after 24 h, samples were thawed and the con-
ductivity was measured and considered as total conductivity. Electro-
lyte leakage was calculated as the percentage ratio of initial over total
conductivity. To measure dry weight, chopped carrots were maintained
in a forced-draft oven at 65 °C until constant weight was reached.
2.6. Extractions of phenolic and carotenoid compounds for profiles analyses
Samples of fresh-cut slices of purple Polignano carrots, both just
after harvest and at each sampling times (samples in AIR or pMA) were
finely chopped. Aliquots of 5 g were treated with 20 mL of 2 % formic
acid in methanol/water (60/40 v/v) to extract phenolic compounds.
Extraction was carried out for 1 h on a horizontal shaker in the dark.
After centrifugation at 4000 rpm for 10 min, the supernatant was re-
moved, the pellet was suspended in 20 mL of 2 % formic acid in me-
thanol/water 60/40 (v/v). The extraction was repeated twice. The
B. Pace, et al.
supernatants were pooled and dried in a rotary evaporator (LaboRota
4000/HB Efficient, Heidolph, Schwabach, Germany) and stored at
−20 °C until the use.
Extraction of carotenoids from aliquots of 2.5 g of chopped fresh-cut
carrot samples was carried out with acetone until discoloration.
Extracts were filtered through a filter paper and filtrates were evapo-
rated under a stream of nitrogen and analyzed by HPLC.
2.7. High Performance Liquid Chromatography-Ultraviolet/Visible
(HPLC–UV/Vis) analyses
Both the polyphenolic and carotenoid extracts were analyzed by
HPLC-UV/Vis on a HP 1110 series HPLC (Agilent, Palo Alto, CA, USA)
equipped with a binary pump (G-1312A) and an UV detector (G-
1314A). For polyphenol analyses, extracts were reconstituted in me-
thanol/water (60/40 v/v) and individual phenols were separated on a
XBridge BEH C18 column (130 Å, 5 mm, 4.6 mm × 150 mm, Waters) at
a flow rate of 1 mL min-1; solvent A was water/acetonitrile/formic acid
(95/4/1 v/v/v) and solvent B was water/acetonitrile/formic acid (44/
55/1 v/v/v). After a 2 min hold at 6 % solvent B, elution was performed
by a linear gradient from 6 to 20 % solvent B in 20 min, from 20 to 40 %
solvent B in 15 min, from 40 to 60 % solvent B in 5 min and from 60 to
95 % solvent B in 5 min, followed by 20 min of maintenance.
Hydroxycinnamic acid derivatives were detected at 280 nm and an-
thocyanins at 520 nm.
Quantification of hydroxycinnamic acids was performed with ex-
ternal calibration curves generated by repeated injections of each hy-
droxycinnamic acid standard prepared over a concentration range of
1 − 300 μg mL-1 with six different concentration levels and duplicate
injections at each level.
As to anthocyanins, quantification was performed with external
calibration curves generated by repeated injections of standard solu-
tions of cyanidin 3-O-glucoside over a concentration range of
0.1 − 200 μg mL-1 with six different concentration levels and duplicate
injections at each level. All samples were prepared and analyzed in
duplicate. Results were expressed as mg Kg-1 fw. For carotenoids ana-
lyses, extracts were reconstituted in mobile phase and aliquots were
injected on the C18 chromatographic column already described for
polyphenol analyses. Carotenoids separation was carried out by a
60 min’ isocratic elution with acetonitrile/methanol/ethyl acetate (80/
17/3 v/v/v) at a flow rate of 1 mL min-1. The chromatogram was
monitored at 490 nm. Quantification was carried out using external
analytical curves of lutein and β‑carotene over a concentration range of
1 − 50 μg mL-1 with six concentration levels and duplicate injections at
each level. All samples were prepared and analyzed in duplicate.
Results were expressed as mg Kg-1 fw.
2.8. High Performance Liquid Chromatography-Multistage Ion Trap Mass
Spectrometry (HPLC-ITMSn) analysis
Identification of phenolic compounds in the different extracts of
purple Polignano carrots was carried out by HPLC coupled to multistage
ion trap mass spectrometry with electrospray ionization (HPLC/ESI-
ITMSn) on a Surveyor MS micro HPLC coupled with a Finnigan LCQ
DECA XP Max ion trap mass spectrometer, equipped with Xcalibur®
system manager data acquisition software (Thermo Finnigan, San Jose,
CA, USA). Individual phenols were separated on a XBridge BEH C18
column (130 Å, 5 mm, 2.1 mm × 150 mm, Waters) at a flow rate of
200 μL min-1; solvent A was water/acetonitrile/formic acid (96:4:0.1 v/
v/v) and solvent B was water/acetonitrile/formic acid (45:55:0.1 v/v/
v). The HPLC conditions were as described in paragraph 2.7. Mass
spectra were recorded from mass-to-charge ratio (m/z) 80–1500 both in
negative and positive ionization mode. The capillary voltage was set at
−16 V, the spray voltage was -3.5 kV, and the tube lens offset was
−10 V in negative ion mode, while, in positive ion mode, the capillary
voltage was set at 36 V, the spray voltage was 3 kV, and the tube lens
3
Journal of Food Composition and Analysis 86 (2020) 103363
offset was 45 V. The capillary temperature was 275 °C. Data were ac-
quired in MS, MS/MS, and MSn scanning modes.
The same HPLC-IT-MSn system, but equipped with atmospheric
pressure chemical ionization (APCI) source, was used for carotenoids
characterization in the different extracts of purple Polignano carrots.
Carotenoids separation was carried out using the C18 chromatographic
column already described by a 60 min’ isocratic elution with acetoni-
trile/methanol/ethyl acetate (80/17/3 v/v/v) at a flow rate of 200 μL
min-1. The APCI vaporizer temperature was set at 450 °C, the capillary
voltage at 13 V, the discharge current at 5 μA, and the tube lens offset at
−15 V. The capillary temperature was 250 °C. Data were acquired in
MS, MS/MS, and MSn scanning modes and recorded in the
400–2000 m/z range in positive ionization mode.
2.9. Statistical analysis
A multifactor ANOVA for P ≤ 0.05, was performed with the aim of
evaluating the effects of packaging condition (AIR or pMA), storage
time (4, 7, 10 and 14 days) and their interaction on all parameters
measured. When the interaction was significant, data were presented as
graphs with mean values ± standard deviation. When interaction be-
tween factors resulted not significant and data were affected only by
main factors, mean values were separated applying Least Significant
Difference (LSD) multiple range test with significant difference when
P ≤ 0.05.
3. Results and discussion
3.1. Selection of the proper packaging material
Results of the test, carried out to select the proper packaging ma-
terial for the experimental trial, showed that in PP MF1 D50 bags the
gas composition remained similar to air, while in PE anaerobiosis oc-
curred after 5 days of storage (data not shown). Instead, in PP MF1
D100 bags the equilibrium-modified atmosphere (EMA) of about 15 %
O2 and 5 % CO2 was reached after roughly 2 days of storage. Thus, this
last packaging material was used for the trial. During the experimental
trail, the same EMA was reached in pMA bags (Fig. 1).
3.2. Sensory visual quality, respiration rate, dry weight, firmness, and
electrolyte leakage in fresh-cut purple Polignano carrots stored in AIR or
pMA
Sensory visual quality was affected by packaging condition and
storage time, while only packaging condition significantly influenced
respiration rate and, on the opposite, dry weight was affected only by
storage time. Finally, electrolyte leakage and texture were not affected
by factors and their interactions (Table S1). Sensory visual quality was
scored significantly higher for sample stored in pMA than for each one
stored in AIR, while a significant decrease in visual quality was reported
during storage in both packaging conditions (Table 1). As for dry
weight, it was affected only by storage time, slight significantly in-
creasing at the end of the storage (Table 1). The positive effect of
modified atmosphere was observed on respiration rate data too. In
particular, samples stored in pMA showed a mean respiration rate about
60 % lower than samples stored in AIR, which remained almost con-
stant during storage (Table 1). Thus, while samples stored in AIR
showed a significant increase in respiration rate compared to the fresh
sample (11.2 ± 0.12 mL CO2 kg-1 h-1), sliced carrots in pMA main-
tained the value of fresh sample during the entire storage. Modified
atmosphere of 15 % O2 and 5 % CO2 minimized the stress caused by the
cutting process, that was evident in AIR samples (Simões et al., 2011).
Just after harvest, the electrolyte leakage measured on carrots was
7.0 ± 0.3 %, then it slight increased after 4 days at 4 °C in storage in
both packaging condition (mean value of about 8.4 %); then no sig-
nificant difference due to treatments and storage time were measured.
B. Pace, et al. Journal of Food Composition and Analysis 86 (2020) 103363

Fig. 1. Changes in gas composition inside pMA bags of fresh-cut purple Polignano carrots stored at 4 °C for 14 days. Mean values ± standard deviation.

As for firmness, the relative deformation measured just after harvest
was 31.9 ± 1.35 %, it slightly decreased after 4 days of storage at 4 °C
(mean value of about 26 % whether in AIR or pMA). Then, no sig-
nificant changes during storage in both packaging conditions were
measured. Probably, the storage temperature (4 ± 1 °C) applied during
the experiment was sufficient to preserve the firmness and the tissue
integrity measured by electrolyte leakage (Watada and Qi, 1999).
3.3. Identification of polyphenols in fresh-cut purple Polignano carrots
Identification of polyphenols present in samples of fresh-cut slices of
purple Polignano carrots, just after harvest and during storage in AIR or
pMA at different sampling times, was achieved through the analysis and
characterization of extracts by HPLC-UV/Vis and HPLC-ESI-ITMSn. The
HPLC-UV/Vis chromatograms of polyphenolic extracts from fresh
sample of purple Polignano carrots are shown in Fig. 2 (Panels A and B).
These carrots contained mainly hydroxycinnamic acids and their deri-
vatives, and anthocyanins, in accordance with previous studies on dif-
ferently coloured carrot cultivars (Kammerer et al., 2004; Kramer et al.,
2012; Mizgier et al., 2016). In particular, we confirmed the presence of
caffeic acid hexoside (peak 1), 3-O-caffeoylquinic acid (chlorogenic
acid, peak 2), a caffeoylquinic acid isomer (peak 3), ferulic acid hexo-
side (peak 4), 5-p-coumaroylquinic acid (peak 5) and feruloylquinic
acid (peak 6) (Fig. 2A). Compounds present in peaks 7, 8 and 9 (Fig. 2A)
displayed pseudo molecular ions at m/z values of 365, 527 and 541,
respectively. Molecules with these m/z values have already been found
in extracts from different carrot cultivars (Kammerer et al., 2004;
Kramer et al., 2012; Mizgier et al., 2016), but they have been gener-
ically identified as caffeic or ferulic acid derivatives. Interestingly, the
study of their fragmentation spectra allowed us to identify these mo-
lecules as cinnamoyl-amino acid conjugates. As a matter of fact, in the
MS2 spectra, all of them presented a fragment ion at m/z 203 that could
be attributed to the presence of the amino acid tryptophan, while peaks
8 and 9 showed prominent fragment ions at m/z 365 and m/z 379 re-
spectively, that could be originated from the loss of a hexose moiety
(162 Da). Comparing these data with those previously reported in the
literature, for conjugated cinnamoyl-amino acid identified in green
coffee beans and in Coriandrum sativum L. (Alonso-Salces et al., 2009;
Barros et al., 2012; Asamenew et al., 2019), compounds eluted in peaks
7–9 were tentatively identified as caffeoyl N-tryptophan (peak 7), caf-
feoyl N-tryptophan hexoside (peak 8) and feruloyl N-tryptophan
hexoside (peak 9). Their occurrence well paralleled with the presence of
the aromatic amino acid tryptophan in differently coloured carrot cul-
tivars (Kramer et al., 2012). To the best of our knowledge, the presence
of cinnamoyl-amino acid conjugates in carrots is here reported for the
first time.
Additionally, six anthocyanins were identified in purple Polignano
carrots. Five of them are cyanidin-based anthocyanins containing dif-
ferent sugar moieties non-acylated (peaks 10 and 11), or acylated with
sinapic acid (peak 12), ferulic acid (peak 13) or coumaric acid (peak 14)
(Fig. 2B). Instead, the sixth anthocyanin (peak 15) was detected in very
small amount and identified as pelargonidin 3-xylosylglucosylgalacto-
side acylated with ferulic acid. These anthocyanins have been pre-
viously reported in black and purple carrots by other authors (Montilla
et al., 2011; Algarra et al., 2014; Mizgier et al., 2016). The list of
compounds identified in fresh sample of purple Polignano carrot is
reported in Table 2.
3.4. Polyphenols composition in fresh-cut purple Polignano carrots stored in
AIR or pMA
Total and individual polyphenol contents in fresh-cut samples of
purple Polignano carrots just after harvest determined by HPLC-UV/Vis
is showed in Table 3. Hydroxycinnamic acids and their derivatives were
the most predominant class of phenolic compounds, accounting for
about 95 % of total phenolic content in fresh sample. In particular, 3-O-

Table 1
Main effects of packaging condition (pMA or AIR) and storage time (4, 7, 10 and 14 days at 4 °C) on visual quality, dry weight and respiration rate of fresh-cut purple
Polignano carrots.
Packaging Storage time

pMA AIR 4 7 10 14

Visual quality score (5-1) 3.0 a 2.8 b 3.7 a 2.8 b 2.9 b 2.3 c
Dry weight (%) 9.4 ns 9.7 ns 9.3 b 9.4 b 9.8 ab 9.9 a
Respiration rate (mL CO2 kg-1 h-1) 8.7 b 22.6 a 17.7 ns 16.7 ns 15.9 ns 12.3 ns

For packaging condition, data are mean values of 12 samples (3 replicates × 4 storage time); for storage, data are mean values of 6 samples (3 replicates × 2
packaging condition). Different letters indicate statistical differences within storage conditions and storage time, according to LSD test (P ≤ 0.05).

4
B. Pace, et al. Journal of Food Composition and Analysis 86 (2020) 103363

Fig. 2. HPLC-UV/Vis chromatograms of polyphenol compounds recorded at 280 (A) and at 520 (B) nm and carotenoids recorded at 490 (C) present in purple
Polignano carrots (just after harvest). For chromatographic conditions, see text. For peak assignments, see Table 2.

caffeoylquinic acid (chlorogenic acid) was a major hydroxycinnamic
acid in fresh sample of purple Polignano carrots, representing the 61 %
of total phenolic compounds. As to the second class of phenolic com-
pounds detected in these samples, among the six anthocyanins identi-
fied in the purple Polignano carrot extracts, cyanidin 3-O-xylosyl (fer-
uoylglucosyl) galactoside was the dominant compound. It constituted
70.6 % of the total anthocyanins and 3.6 % of total phenolic content.
Furthermore, changes in polyphenol content of purple Polignano
carrots slices were monitored during storage. Fresh-cut purple
Polignano carrots just after harvest showed an initial mean content of
total polyphenols of 116.14 ± 8.86 mg kg-1 fw (Table 3) that drasti-
cally increased during cold storage in both packaging conditions. In
fact, the mean content of total polyphenols became
405.45 ± 183.35 mg kg-1 fw (pMA condition) and 471.86 ± 78.20 mg
kg-1 fw (AIR condition) after only four days of cold storage, and
670.69 ± 130.86 mg kg-1 fw and 705.07 ± 122.20 mg kg-1 fw after 14
days of cold storage in pMA and AIR, respectively (Table S2). The ef-
fects of wounding on the accumulation of phenolic compounds in fruits
and vegetables has been evaluated in several studies, demonstrating
that stresses such as light, temperature and water-stress and cutting

Table 2
List of phenolic compounds and carotenoids identified in purple Polignano carrots (just after harvest and during storage) by HPLC-UV/Vis and HPLC-ITMSn. Specific
quasi-molecular ions and fragment ions are reported for each compound.
Peak [M-H]- m/z MSn ions m/z Proposed structure

2
1 341 MS [341]: 179 Caffeic acid hexoside
2 353 MS2 [353]: 179, 191 3-O-Caffeoylquinic acid (Chlorogenic acid)
MS3 [353→191]: 85, 93, 127, 173
3 353 MS2 [353]: 179, 191 Caffeoylquinic acid isomer
MS3 [353→191]: 85, 93, 127, 173
4 355 MS2 [355]:193, 175 Ferulic acid hexoside
5 337 MS2 [337]: 163, 191 5-p-Coumaroylquinic acid
MS3 [337→191]: 85, 93, 127, 173
6 367 MS2 [367]:191 Feruloylquinic acid
7 365 MS2 [365]: 179, 185, 203 Caffeoyl N- tryptophan
MS3 [365→203]: 69, 97, 115, 141, 185
8 527 MS2 [527]: 203, 365 Caffeoyl N-tryptophan hexoside
MS3 [527→365]: 141, 179, 185, 203
MS4 [527→365→203]: 69, 97, 115, 123, 141, 159, 185
9 541 MS2 [541]: 379 Feruloyl N-tryptophan hexoside
MS3 [541→379]: 203, 185, 141
MS4 [541→379→185]: 141
Peak [M]+ m/z MSn ions m/z Proposed structure
10 743 MS2 [743]: 449, 287 Cyanidin 3-xylosyl(glucosyl)galactoside
11 581 MS2 [581]: 449, 287 Cyanidin 3-xylosylgalactoside
12 949 MS2 [949]: 449, 287 Cyanidin 3-xylosyl(sinapoylglucosyl)
galactoside
13 919 MS2 [919]: 449, 287 Cyanidin 3-xylosyl(feruloylglucosyl) galactoside
14 889 MS2 [889]: 449, 287 Cyanidin 3-xylosyl(coumaroylglucosyl) galactoside
15 903 MS2 [903]: 271 Pelargonidin 3-xylosyl(feruloylglucosyl) galactoside
Peak [M+H]+ m/z MSn ions m/z Proposed structure
16 569 MS2 [569]: 551, 533,459 Lutein
17 537 MS2 [537]: 481, 413, 401, 399 α-carotene
18 537 MS2 [537]: 481, 413, 401, 399 β-carotene

5
B. Pace, et al. Journal of Food Composition and Analysis 86 (2020) 103363

Table 3 Table 4
Concentration of individual and total polyphenols and carotenoids of purple Effect of packaging condition (AIR or pMA) (A), storage time (4, 7, 10 and 14
Polignano carrots determined by HPLC-UV/Vis in the fresh samples. Data are days at 4 °C) (B), and their interaction (A x B) on phenolic compounds and
mean of 3 replicates ± standard deviation. carotenoids identified in fresh-cut purple Polignano carrots.
Phenolic Compounds mg kg-1 fw A B AxB

-1
Caffeic acid hexoside 2.35 ± 0.45 Phenolic compounds (mg kg fw)
3-O-Caffeoylquinic acid (Chlorogenic acid) 70.94 ± 8.33 Caffeic acid hexoside ns ns ns
Caffeoylquinic acid isomer 4.64 ± 1.31 3-O-Caffeoylquinic acid (Chlorogenic acid) ns ns ns
Ferulic acid hexoside 3.60 ± 0.74 Caffeoylquinic acid isomer ns ns ns
5-p-Coumaroylquinic acid 10.52 ± 0.40 Ferulic acid hexoside ns *** ns
Feruloylquinic acid 6.05 ± 0.35 5-p-Coumaroylquinic acid ns ns ns
Caffeoyl N- tryptophan 4.81 ± 0.54 Feruloylquinic acid ns * ns
Caffeoyl N-tryptophan hexoside 6.60 ± 0.81 Caffeoyl N- tryptophan ns ns ns
Feruloyl N-tryptophan hexoside 0.74 ± 0.05 Caffeoyl N-tryptophan hexoside ** ** ns
Total hydroxycinnamic acids and derivatives 110.3 ± 8.63 Feruloyl N-tryptophan hexoside ** ** ns
Cyanidin 3-xylosyl(glucosyl)galactoside 0.31 ± 0.05 Cyanidin 3-xylosyl(glucosyl)galactoside ns * ns
Cyanidin 3-xylosylgalactoside 0.61 ± 0.06 Cyanidin 3-xylosylgalactoside ns ns ns
Cyanidin 3-xylosyl(sinapoylglucosyl) galactoside 0.39 ± 0.06 Cyanidin 3-xylosyl(sinapoylglucosyl) galactoside ns ns ns
Cyanidin 3-xylosyl(feruloylglucosyl) galactoside 4.16 ± 0.65 Cyanidin 3-xylosyl(feruloylglucosyl) galactoside ns * ns
Cyanidin 3-xylosyl(coumaroylglucosyl) galactoside 0.37 ± 0.01 Cyanidin 3-xylosyl(coumaroylglucosyl) galactoside ns * ns
Pelargonidin 3-xylosyl(feruloylglucosyl) galactoside 0.05 ± 0.01 Pelargonidin 3-xylosyl(feruloylglucosyl) galactoside ns * ns
Total anthocyanins 5.89 ± 0.83 Total Phenolic Contents ns * ns
Total Phenolic Contents 116.14 ± 8.86 Carotenoids (mg kg-1 fw)
Carotenoids Lutein *** ns *
Lutein 1.45 ± 0.32 α-carotene * ns ns
α-carotene 0.91 ± 0.08 β-carotene ns ns ns
β-carotene 3.92 ± 0.43 Total carotenoid contents ns ns ns
Total Carotenoid Contents 6.28 ± 0.48
Asterisks indicate the significance level for each factor of the ANOVA test (ns,
not significant; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001). For packaging con-
affect the physiology of fresh products by triggering responses that dition mean values of 12 samples (3 replication x 4 storage duration) were used;
could induce the accumulation of phenolic compounds or other sec- for storage duration mean values of 6 samples (3 replication x 2 packaging
ondary metabolites (Reyes et al., 2007). In particular, wounding could condition) were used.
induce the production of phenolic compounds in a large variety of
horticultural crops including carrots (Surjadinata and Cisneros- condition (Table 4). The content of all the other compounds was not
Zevallos, 2012); in fact, a 10 % increase of phenolic content was de- affected by these factors. In particular, mean values of the content of
tected in slices of carrots (Daucus carota, cv Choctaw) stored 6 days at total polyphenols, ferulic acid hexoside, feruloylquinic acid, cyanidin 3-
15 °C (Heredia and Cisneros-Zevallos, 2009). It is worth to stress that xylosyl(glucosyl)galactoside, cyanidin 3-xylosyl(feruloylglucosyl) ga-
the increase in polyphenol content in purple Polignano carrots reached lactoside, cyanidin 3-xylosyl(coumaroylglucosyl) galactoside and pe-
about 249 % in samples stored in pMA and about 306 % in those store largonidin 3-xylosyl(feruloylglucosyl) galactoside showed an increase
in AIR at 4 °C during the first 4 days of storage respect to fresh carrots. throughout storage time (both in AIR or pMA) (Table 5). Also the
The changes in phenolic content during storage of fresh-cut carrots content of caffeoyl N-tryptophan hexoside and feruloyl N-tryptophan
could be associated with wound-induced stimulation of the enzyme hexoside increased during the storage period, but samples stored in AIR
phenylalanine ammonia lyase (PAL) (Hager and Howard, 2006). Babic showed mean values of these two compounds significantly higher than
et al. (1993) reported a significant increase in this enzyme activity in fresh-cut carrots stored in pMA throughout storage time (Table 5). Han
stored shredded carrot roots and, in turn, in the phenolic compounds et al. (2017) demonstrated that wounding also stimulated the pathways
synthesis. Moreover, increased PAL activity during storage has been of “biosynthesis of amino acids”, “phenylalanine, tyrosine, and tryp-
observed in carrots stored in AIR (Howard et al., 1994; Leja et al., tophan”, and “phenylalanine metabolism”, which are involved in the
1997). PAL catalyzes the conversion of L-phenylalanine to trans-cin- biosynthesis and degradation of some specific amino acids.
namic acid by a non-oxidative deamination and this reaction is the first
step in the phenylpropanoid pathway that leads to various phenylpro-
3.5. Identification of carotenoids in fresh-cut purple Polignano carrots
panoids and their derivatives, such as flavonoids, isoflavonoids, re-
stored in AIR or pMA
sveratrol, coumarins, stilbenes, anthocyanins, lignin, and other phe-
nolic compounds (Kong, 2015). Other authors reported that carrots
Identification and quantification of carotenoids present in samples
responded to wounding stress producing and accumulating caffeoyl-
of fresh-cut purple Polignano carrots, just after harvest and during
quinic acids (Heredia and Cisneros-Zevallos, 2009; Jacobo-Velázquez
storage in AIR or pMA at different sampling times, was achieved
et al., 2011). As a matter of fact, in this study on purple Polignano
through the analysis and characterization of extracts by HPLC-UV/Vis
carrots both hydroxycinnamic acids and their derivatives, and antho-
and HPLC-APCI-IT-MSn. The HPLC-Vis chromatograms of carotenoids
cyanins increased their content during storage.
extracted from fresh sample of purple Polignano carrots are shown in
Results of the multifactor ANOVA performed on data obtained by
Fig. 2 (Panel C). APCI-IT-MSn based structural characterization con-
quantitative analysis of total and individual polyphenols in extracts of
firmed the presence of lutein, α-carotene and β-carotene (Table 2), in
purple Polignano carrots stored in AIR or in pMA at 4 °C for 14 days are
agreement with previous studies (Alasalvar et al., 2001; Nicolle et al.,
reported in Tables 4 and 5. These results showed that total polyphenols
2004). Table 3 shows the carotenoid content in fresh sample of purple
content, ferulic acid hexoside, feruloylquinic acid, cyanidin 3-xylosyl
Polignano carrots determined by HPLC-UV/Vis. β-carotene was the
(glucosyl)galactoside, cyanidin 3-xylosyl(feruloylglucosyl) galactoside,
major carotenoid (3.92 ± 0.43 mg kg-1 fw), followed by lutein
cyanidin 3-xylosyl(coumaroylglucosyl) galactoside and pelargonidin 3-
(1.45 ± 0.32 mg kg-1 fw) and α-carotene (0.91 ± 0.08 mg kg-1 fw).
xylosyl(feruloylglucosyl) galactoside were significantly affected only by
Total carotenoids level was 6.28 ± 0.48 mg kg-1 fw. These data were
storage time, while caffeoyl N-tryptophan hexoside and feruloyl N-
similar to those reported by Nicolle et al. (2004) in a previous study on
tryptophan hexoside were affected by both storage time and packaging
two different purple carrots (Violette jordanienne and Violette turque),

6
B. Pace, et al. Journal of Food Composition and Analysis 86 (2020) 103363

Table 5
Main effect of packaging condition (AIR or pMA) (A) and storage time (4, 7, 10 and 14 days at 4 °C) (B) on phenolic compounds and carotenoids identified in fresh-cut
purple Polignano carrots.
Packaging Storage time

pMA AIR 4 7 10 14

Phenolic compounds (mg kg-1 fw)

Ferulic acid hexoside 6.0 – 7.3 – 5.0 bc 4.4 c 7.2 b 10.0 a
Feruloylquinic acid 14.5 – 16.6 – 13.7 b 10.8 b 14.2 b 23.5 a
Caffeoyl N-tryptophan hexoside 25.7 B 48.1 A 23.4 b 25.2 b 48.0 a 51.0 a
Feruloyl N-tryptophan hexoside 1.4 B 2.0 A 1.3 b 1.4 b 1.8 ab 2.2 a
Cyanidin 3-xylosyl(glucosyl)galactoside 4.1 – 4.5 – 3.5 b 2.7 b 4.2 ab 6.8 a
Cyanidin 3-xylosyl(feruloylglucosyl) galactoside 40.3 – 45.0 – 35.3 b 27.9 b 43.1 ab 64.2 a
Cyanidin 3-xylosyl(coumaroylglucosyl) galactoside 6.3 – 7.0 – 5.9 b 3.5 b 5.8 b 11.5 a
Pelargonidin 3-xylosyl(feruloylglucosyl) galactoside 0.9 – 1.1 – 0.8 b 0.6 b 1.0 ab 1.7 a
Total Phenolic Contents 469.8 – 548.3 – 438.7 b 381.4 b 528.2 ab 687.9 a
Carotenoids (mg kg-1 fw)
α-carotene 0.8 B 1.3 A 1.1 – 1.0 – 0.9 – 1.2 –

Different uppercase and lowercase letters indicate statistical differences within packaging condition and storage time, respectively, according to LSD test (P ≤ 0.05).
For packaging condition mean values of 12 samples (3 replication x 4 storage duration) were used; for storage duration mean values of 6 samples (3 replication x 2
packaging condition) were used.

4. Conclusions

A deeper qualitative and biochemical characterization of fresh-cut

Fig. 3. Changes in lutein content in purple fresh-cut Polignano carrots stored at
4 °C in AIR or pMA (EMA = 15 % O2+ 5 % CO2) for 14 days. Mean va-
lues ± standard deviation.
although in those cultivars α-carotene was not detected. Also
Grassmann et al. (2007) found similar levels of α- and β-carotene in a
purple carrot cultivar, but a higher level of lutein. However, it is well
known that growing conditions, growing season, soil, year, variety and
genetic factors affect not only the total carotenoids content but also the
proportions of the different molecules (Hart and Scott, 1995; Rosenfeld
et al., 1997; Desobry et al., 1998).
The total carotenoids content did not change significantly during
storage at 4 °C (both in AIR or pMA). As a matter of fact, results of the
multifactor ANOVA performed on data obtained by quantitative ana-
lysis of total and individual carotenoids in extracts of fresh-cut purple
Polignano carrots stored in AIR or in pMA at 4 °C for 14 days demon-
strated that total carotenoids content was not affected either by storage
time or packaging condition (Table 4). Instead, α-carotene content was
significantly affected only by packaging condition (Table 4) and AIR
samples showed mean values of this compound significantly higher
than fresh-cut carrots stored in pMAP throughout storage time
(Table 5).
Moreover, lutein content was affected by storage time and by the
interaction of both parameters (packaging condition and storage time)
(Table 4). In particular, fresh-cut carrots stored in pMAP showed a
slight increase in lutein content after 7 days, which was followed by a
decrease after 10 days, while the content remained almost constant
during the entire trial in AIR samples (Fig. 3). These results demon-
strated that storage at 4 °C for 14 days, both in AIR and in pMA, had
little or no influence on carotenoids content, and this result was in
agreement with that reported by Lee et al. (2011) in a study on changes
in nutritional content during storage at 2 °C and 4 °C for 4 weeks of
sliced or whole roots of purple and orange carrots.
purple Polignano carrots during cold storage in AIR or pMA was carried
out using PP MF1 D100 bags as proper packaging material for the trial;
in fact, the EMA of about 15 % O2 and 5 % CO2 was reached after
roughly 2 days at 4 °C. The storage in pMA preserved the sensory visual
quality and the respiration rate, while a decrease in the appearance
(loss of turgor and brightness) with an increase in respiration rate was
measured in samples stored in AIR. Fifteen polyphenol compounds
(belonging to the classes of hydroxycinnamic acids and their deriva-
tives, and anthocyanins) and three carotenoids were identified and
their content was monitored during cold storage in both packaging
conditions. Hydroxycinnamic acids, their derivatives and anthocyanins
drastically increased their content during storage, mainly in AIR, as
consequence of wounding. As for carotenoids, β-carotene was the most
represented, followed by lutein and α-carotene, and no significant
changes of these compounds were reported during storage in AIR or
pMA samples.
In conclusion, pMA resulted a valid solution to cold store fresh-cut
purple Polignano carrots until two weeks, and preserve their quality
and nutritional value. This would prompt a spread of this perishable
and high value product from this small area of Southern Italy where it is
grown to the wider national and global markets, thus suitably sup-
porting the local economy.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.jfca.2019.103363.
References
Alasalvar, C., Grigor, J.M., Zhang, D., Quantick, P.C., Shahidi, F., 2001. Comparison of
volatiles, phenolics, sugars, antioxidant vitamins, and sensory quality of different
colored carrot varieties. J. Agric. Food Chem. 49 (3), 1410–1416.
Algarra, M., Fernandes, A., Mateus, N., de Freitas, V., Esteves da Silva, J.C.G., Casado, J.,
2014. Anthocyanin profile and antioxidant capacity of black carrots (Daucus carota L.
ssp. sativus var. atrorubens Alef.) from Cuevas Bajas, Spain. J. Food Compos. Anal. 33
(1), 71–76.
Alonso-Salces, R.M., Guillou, C., Berrueta, L.A., 2009. Liquid chromatography coupled
with ultraviolet absorbance detection, electrospray ionization, collision induced
dissociation and tandem mass spectrometry on a triple quadrupole for the on-line
characterization of polyphenols and methylxanthines in green coffee beans. Rapid
Commun. Mass Spectrom. 23 (3), 363–383.
Akhtar, S., Rauf, A., Imran, M., Qamar, M., Riaz, M., Mubarak, M.S., 2017. Black carrot
(Daucus carota L.), dietary and health promoting perspectives of its polyphenols: a
review. Trends Food Sci. Technol. 66, 36–47.

7
B. Pace, et al.
Asamenew, G., Kim, H.W., Lee, M.K., Lee, S.H., Lee, S., Cha, Y.S., Lee, S.H., Yoo, S.M.,
Kim, J.B., 2019. Comprehensive characterization of hydroxycinnamoyl derivatives in
green and roasted coffee beans: a new group of methyl hydroxycinnamoyl quinate.
Food Chem.: X 2, 100033.
Babic, I., Amiot, M.J., Nguyen-The, C., Aubert, S., 1993. Changes in phenolic contents in
fresh ready-to-use shredded carrots during storage. J. Food Sci. 58 (2), 351–356.
Barros, L., Dueñas, M., Dias, M.I., Sousa, M.J., Santos-Buelga, C., Ferreira, I.C., 2012.
Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L. Food Chem. 132
(2), 841–848.
Cefola, M., Pace, B., Renna, M., Santamaria, P., Signore, A., Serio, F., 2012. Compositional
analysis of yellow, orange and purple Polignano carrots. Ital. J. Food Sci. 24 (3),
284–291.
Cefola, M., Pace, B., 2016. High CO2-modified atmosphere to preserve sensory and nu-
tritional quality of organic table grape (cv. ‘Italia’) during storage and shelf-life. Eur.
J. Hortic. Sci. 81 (4), 197–203.
Desobry, S.A., Netto, F.M., Labuza, T.P., 1998. Preservation of β-carotene from carrots.
Crit. Rev. Food Sci. Nutr. 38 (5), 381–396.
Grassmann, J., Schnitzler, W.H., Habegger, R., 2007. Evaluation of different colored
carrot cultivars on antioxidative capacity based on their carotenoid and phenolic
contents. Int. J. Food Sci. Nutr. 58 (8), 603–611.
Hager, T.J., Howard, L.R., 2006. Processing effects on carrot phytonutrients. HortScience
41 (1), 74–79.
Han, C., Jin, P., Li, M., Wang, L., Zheng, Y., 2017. Physiological and transcriptomic
analysis validates previous findings of changes in primary metabolism for the pro-
duction of phenolic antioxidants in wounded carrots. J. Agric. Food Chem. 65 (33),
7159–7167.
Hart, D.J., Scott, K.J., 1995. Development and evaluation of an HPLC method for the
analysis of carotenoids in foods, and the measurement of the carotenoid content of
vegetables and fruits commonly consumed in the UK. Food Chem. 54 (1), 101–111.
Heredia, J.B., Cisneros-Zevallos, L., 2009. The effect of exogenous ethylene and methyl
jasmonate on pal activity, phenolic profiles and antioxidant capacity of carrots
(Daucus carota) under different wounding intensities. Postharvest Biol. Technol. 51
(2), 242–249.
Hou, D.X., Fujii, M., Terahara, N., Yoshimoto, M., 2004. Molecular mechanisms behind
the chemopreventive effects of anthocyanidins. Biomed Res. Int. 2004 (5), 321–325.
Howard, L.R., Griffin, L.E., Lee, Y., 1994. Steam treatment of minimally processed carrot
sticks to control surface discoloration. J. Food Sci. 59 (2), 356–358.
Jacobo-Velázquez, D.A., Martínez-Hernández, G.B., del. C. Rodríguez, S., Cao, C.M.,
Cisneros-Zevallos, L., 2011. Plants as biofactories: physiological role of reactive
oxygen species on the accumulation of phenolic antioxidants in carrot tissue under
wounding and hyperoxia stress. J. Agric. Food Chem. 59 (12), 6583–6593.
Kader, A.A., 2002. Postharvest Technology of Horticultural Crops Vol. 3311 University of
California Agriculture and Natural Resources.
Kammerer, D., Carle, R., Schieber, A., 2004. Characterization of phenolic acids in black
carrots (Daucus carota ssp. sativus var. atrorubensAlef.) by high-performance liquid
Journal of Food Composition and Analysis 86 (2020) 103363
chromatography/electrospray ionization mass spectrometry. Rapid Commun. Mass
Spectrom. 18 (12), 1331–1340.
Kramer, M., Maksylewicz-Kaul, A., Baranski, R., Nothnagel, T., Carle, R., Kammerer, D.,
2012. Effects of cultivation year and growing location on the phenolic profile of
differently coloured carrot cultivars. J. Appl. Bot. Food Qual. 85 (2), 235–247.
Kong, J.Q., 2015. Phenylalanine ammonia-lyase, a key component used for phenylpro-
panoids production by metabolic engineering. RSC Adv. 5 (77), 62587–62603.
Lee, E.J., Yoo, K.S., Patil, B.S., 2011. Total carotenoid, anthocyanin, and sugar contents in
sliced or whole purple (cv. Betasweet) and orange carrots during 4-week cold storage.
Hortic. Environ. Biotechnol. 52 (4), 402–407.
Leja, M., Mareczek, A., Wojciechowska, R., Stanisław, R., 1997. Phenolic metabolism in
root slices of selected carrot cultivars. Acta Physiol. Plant. 19 (3), 319–325.
Mizgier, P., Kucharska, A.Z., Sokół-Łętowska, A., Kolniak-Ostek, J., Kidoń, M., Fecka, I.,
2016. Characterization of phenolic compounds and antioxidant and anti-in-
flammatory properties of red cabbage and purple carrot extracts. J. Funct. Foods 21,
133–146.
Montilla, E.C., Arzaba, M.R., Hillebrand, S., Winterhalter, P., 2011. Anthocyanin com-
position of black carrot (Daucus carota ssp. sativus var. atrorubens Alef.) cultivars
Antonina, Beta Sweet, Deep Purple, and Purple Haze. J. Agric. Food Chem. 59 (7),
3385–3390.
Nicolle, C., Simon, G., Rock, E., Amouroux, P., Rémésy, C., 2004. Genetic variability
influences carotenoid, vitamin, phenolic, and mineral content in white, yellow,
purple, orange, and dark-orange carrot cultivars. J. Am. Soc. Hortic. Sci. 129 (4),
523–529.
Pace, B., Capotorto, I., Ventura, M., Cefola, M., 2015. Evaluation of L-cysteine as anti-
browning agent in fresh-cut lettuce processing. J. Food Process. Preserv. 39 (6),
985–993.
Reyes, L.F., Villarreal, J.E., Cisneros-Zevallos, L., 2007. The increase in antioxidant ca-
pacity after wounding depends on the type of fruit or vegetable tissue. Food Chem.
101 (3), 1254–1262.
Renna, M., Pace, B., Cefola, M., Santamaria, P., Serio, F., Gonnella, M., 2013. Comparison
of two jam making methods to preserve the quality of colored carrots. LWT-Food Sci.
Technol. 53 (2), 547–554.
Rosenfeld, H.J., Baardseth, P., Skrede, G., 1997. Evaluation of carrot varieties for pro-
duction of deep fried carrot chips. IV. The influence of growing environment on
carrot raw material. Food Res. Int. 30 (8), 611–618.
Simões, A.D., Allende, A., Tudela, J.A., Puschmann, R., Gil, M.I., 2011. Optimum con-
trolled atmospheres minimise respiration rate and quality losses while increase
phenolic compounds of baby carrots. LWT-Food Sci. Technol. 44 (1), 277–283.
Surjadinata, B.B., Cisneros-Zevallos, L., 2012. Biosynthesis of phenolic antioxidants in
carrot tissue increases with wounding intensity. Food Chem. 134 (2), 615–624.
Wang, L.S., Stoner, G.D., 2008. Anthocyanins and their role in cancer prevention. Cancer
Lett. 269 (2), 281–290.
Watada, A.E., Qi, L., 1999. Quality of fresh-cut produce. Postharvest Biol. Technol. 15 (3),
201–205.