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Postharvest and Refrigeration Group, Department of Food Engineering, Technical Univeristy of Cartagena (UPCT), Paseo Alfonso XIII, 48. 30203, Cartagena, Murcia, Spain
Institute of Plant Biotechnology, I+D+I Building, UPCT, Campus Muralla del Mar. 30202, Cartagena, Murcia, Spain
c
Instituto de Investigacin y Formacin Agraria y Pesquera. Avda. Menndez Pidal, s/n. 14071, Crdoba, Spain
b
a r t i c l e
i n f o
Article history:
Received 11 February 2011
Accepted 16 June 2011
Keywords:
Brassica oleracea
Italica Alboglabra group
Tenderstem
Minimal processing
Phenolics
Antioxidant capacity
Bioactive compounds
a b s t r a c t
The effects of several UV-C pre-treatments (1.5, 4.5, 9.0 and 15 kJ m2 ) on changes in physiological, sensory and microbial quality and health promoting bioactive compounds over 19 days at 5 and 10 C of
fresh-cut Bimi broccoli were studied. Non-irradiated samples were used as controls. Bimi broccoli
(Brassica oleracea Italica Group Alboglabra Group) is characterised by a long stem with a small oret
with a mild and sweeter avor than conventional varieties well adapted for fresh-cut purposes. Low and
moderate UV-C doses (1.5 and 4.5 kJ m2 ) had inhibitory effects on natural microora growth. In relation
to sensory quality, all treatments resulted in a shelf-life of 19 and 13 days at 5 and 10 C respectively
with the exception of 15 kJ UV-C m2 treated samples which resulted in a shorter shelf-life. These doses
immediately increased total polyphenols contents up to 25% after 19 days at 5 C compared to the initial
value. All the hydroxycinnamoyl acid derivates were immediately increased after UV-C treatments, with
values 4.8- and 4.5-fold higher for 4.5 and 9.0 kJ UV-C m2 treated samples respectively over the control.
Changes in phenolic compounds were highly inuenced by the storage temperature throughout shelflife. Total antioxidant activity generally followed the same pattern: the higher the UV-C doses, the higher
total antioxidant capacity values. Generally, UV-C slightly reduced initial total chlorophyll content but
delayed its degradation throughout shelf-life. It is concluded that a pre-treatment of 4.5 kJ UV-C m2 is
useful as a technique to improve epiphytic microbial quality and health promoting bioactive compounds
of fresh-cut Bimi broccoli.
2011 Elsevier B.V. All rights reserved.
1. Introduction
Broccoli is an extremely valuable horticultural product, not only
in economic terms but also for its excellent health claims. This
brassica species has been described as a vegetable with high nutritional value owing to its exceptionally high levels of Zn, folic acid,
antioxidants, glucosinolates, ber, vitamin C and high antioxidant
activity (Jeffery et al., 2003; Freshfel, 2006; Moreno et al., 2006).
After harvest, overall quality of broccoli is greatly reduced due to
several detrimental changes such as loss of green colour and sepal
yellowing as a consequence of chlorophyll catabolism (Funamoto
et al., 2002), tissue disruption, lipid peroxidation, protein degradation and the loss of antioxidant and health promoting bioactive
compounds, which decreases nutritional value (Page et al., 2001).
328
was washed for 1 min with tap water at 5 C to remove traces of soil
and organic matter. The following UV-C radiation treatments were
applied: 0 (Control), 1.5, 4.5, 9.0 and 15 kJ UV-C m2 . UV-C exposure times ranged between 37 and 375 s. Such doses were selected
based on previous reports and our preliminary experiments. The
UV-C equipment has been earlier described (Arts-Hernndez et al.,
2010).
For passive modied atmosphere packaging (MAP), samples of
about 200 g of broccoli per treatment were randomly placed in
1.2 L polypropylene (PP) baskets and thermally sealed on the top
with a 50 m microperforated bi-oriented PP lm (BOPP) (Plsticos del Segura, Murcia, Spain). The O2 and CO2 transmission rates
at 23 C and 0% RH were similar with 11,000 cm3 m2 d1 atm1
(data provided by the supplier). Three replicates of one basket per
treatment and MAP storage duration (processing day and after 7,
13, 16 and 19 days) were prepared and stored in dark cold rooms
at 5 or 10 C. The 5 C temperature was selected as the maximum
recommended during shelf-life for fresh-cut broccoli and 10 C as
an abuse temperature during storage, distribution and retail sale.
2.3. Analysis and determinations
329
330
Fig. 2. Respiration rate of fresh-cut Bimi broccoli pre-treated with several UV-C
radiation doses and stored up to 19 days at 5 C (A) and 10 C (B). Data represent
means of three replicates (n = 3 SD).
Fig. 3. Gas changes within packages of fresh-cut Bimi broccoli treated with several
UV-C doses and stored up to 19 days at 5 C (A) and 10 C (B). Data represent means
of three replicates (n = 3 SD).
which, after the rst week of storage, a strong initial decrease was
found. After 19 days, UV-C treated samples showed higher RR than
control ones, ranging from 29 to 41 and 32 to 51 mg CO2 kg1 h1
at 5 and 10 C respectively.
The gas atmosphere changes throughout shelf-life within MAP
packages showed similar patterns at both temperatures. The
equilibrium gas partial pressures from the 4th until the 19th
day of storage were 78 kPa O2 + 1011 kPa CO2 and 34 kPa
O2 + 1516 kPa CO2 at 5 and 10 C respectively. This gas composition was similar to that recommended for broccoli orets by Gorny
(2001), being favourable, when combined with low temperature
and high RH, for slowing the rate of senescence and extending
storage life. Due to the physiological stress induced by the UV-C
treatment, slight changes in the equilibrium gas partial pressures
were found (Fig. 3).
3.2. Microbial analysis
Immediately after UV-C treatment, initial microbial counts were
lowered. This effect was more marked for mesophilic and yeast
and moulds counts (Fig. 4A and D respectively), with a reduction
of around 1 log CFU g1 (for 4.5 and 9.0 kJ UV-C m2 ). Moreover,
enterobacteria and psychrophilic counts had slightly lower reductions (Fig. 4B and C respectively). These data agree with those from
331
Fig. 4. Mesophilic (A), enterobacteria (B), psycrophilic (C) and yeast and moulds (D) counts (log CFU g1 ) of fresh-cut Bimi broccoli treated with several UV-C doses and
stored up to 19 days at 5 C and 10 C. Data represent means of three replicates (n = 3 SD).
high doses of UV-C would help the growth of some bacteria, probably owing to an increase in damage of supercial tissues that makes
nutrients available for microbial growth, as reported in freshcut spinach (Arts-Hernndez et al., 2009), and observed in the
332
Fig. 5. Sensory scores for overall quality, off-odours and stem softening of fresh-cut Bimi broccoli treated with several UV-C doses and stored up to 19 days at 5 and 10 C.
Data represent means of seven replicates (n = 7 SD).
bending was almost not detected in any treatment at any sampling time (data not shown). During the rst week of storage at
10 C, slight to moderate unpleasant odours for 4.5, 9.0 and 15 kJ
UV-C m2 treated samples were detected (Fig. 5). After 13 days
at 10 C all treatments reached the limit of usability mainly due
to the moderate unpleasant odours and stem softening, except for
15 kJ UV-C m2 treatment which showed a maximum shelf-life of
12 days. However, at 5 C all samples reached a shelf-life of 19 days
with the exception again of the 15 kJ UV-C m2 treatment, which
registered a shorter shelf-life. This fact could be due to high UV-C
provoking cell damage that could have helped microbial growth
inducing softening.
Fig. 6. Total polyphenols content changes of fresh-cut Bimi broccoli treated with
several UV-C doses and stored up to 19 days at 5 C (A) and 10 C (B). Data represent
means of three replicates (n = 3 SD).
333
40, 33, 25, 22 and 10% increases for 15, 9.0, 4.5, 1.5 kJ UV-C m2
and control treatments respectively, with respect to the initial
content. These different UV-induced responses could be explained
by the specic enzyme activities involved in phenylpropanoid
metabolism, including phenylalanine ammonialyase (PAL) which
catalyzes the rst committed step in the phenolic biosynthesis
pathway, after which individual branch pathways make possible
a range of phenylpropanoid secondary compounds (Schutte, 1992;
Cisneros-Zevallos, 2003) as phenolics.
The high total phenolics content increase registered after 13
days at 5 C occurred earlier in samples stored at 10 C (Fig. 6B)
probably due to the acceleration of the metabolism of broccoli at
such higher temperature. Although the product exceeded the limit
of usability according to sensory attributes, the highest increase
(54%) in total phenolics content after 19 days at 10 C was reached
for 4.5 kJ UV-C m2 treated samples while increases in the remaining treatments ranged from 12 to 43% initial values (Fig. 6B).
Lemoine et al. (2007) also reported higher total phenolics contents in broccoli orets after illumination with 8 kJ UV-C m2 . Other
research has shown enriched total phenolics contents after UV-C
radiation in fresh and fresh-cut mangoes (Gonzlez-Aguilar et al.,
2007) and strawberries (Erkan et al., 2008). All these ndings agree
with the hypothesis of Cisneros-Zevallos (2003) that UV-C radiation can be considered an abiotic stress favourable for enhancing
phytochemicals compounds.
Individual phenolic compounds of Bimi broccoli are shown in
Table 1. Hydroxycinnamoyl acid derivates were identied by their
chromatographic behaviour and UV spectra, HPLC/MS and chromatographic comparisons with external standards. The phenolic
prole found in Bimi broccoli was very similar to that described for
other broccoli cultivars and Chinese broccoli (Vallejo et al., 2003a,b;
Lin and Harnly, 2009). The hydroxycinnamoyl acids identied, from
higher to lower content, were 1,2-diferuloylgentibiose, neochlorogenic acid, 1,2,2 -trisinapoylgentibiose, 1,2-disinapoylgentibiose,
1,2 -disinapoyl-2-feruloylgentibiose, chlorogenic acid, 1-sinapoyl2,2 -diferuloylgentibiose and 1-sinapoyl-2-feruloylgentibiose. A
characteristic phenolic prole chromatogram is presented in Fig. 7.
UV-C radiation generally increased the hydroxycinnamoyl acid
derivates content on the processing day (20 mg kg 1 fw) just after
illumination, with the exception of 15 kJ UV-C m2 which generally resulted in the same values as the controls. The highest
increases were found for 4.5 and 9.0 kJ UV-C m2 , 4.8- and 4.5-fold
higher than the control respectively (Table 2). In our experiment,
neochlorogenic acid had the highest increases on the processing
day for the 4.5 and 9.0 kJ m2 UV-C treatments (9.0- and 11.5-fold
higher with respect to the initial value respectively) followed by
1,2-diferuloylgentibiose (5.5- and 5.3-fold higher respectively), and
1,2-disinapoylgentibiose and 1,2 -disinapoyl-2-feruloylgentibiose,
both with similar increases over the control (4.4- and 4.5-fold
higher for 4.5 and 9.0 kJ m2 doses respectively).
Table 1
HPLC/DAD/MS analysis of phenolic compounds in Bimi broccoli. Phenolic compounds numbered as in Table 1.
No.
1
2
3
4
6
7
8
5
Phenolic compound
HPLC retention
time (min)
Max UV absorption
HPLC/DAD (max )
12.2
19.8
327, 295 sh
327, 295 sh
353, 179
353, 179
34.7
35.1
36.8
37.0
38.0
331
330
328
328
320
753, 529
723, 499
959, 735
929, 705
899, 705
36.0
330.4
693, 499
334
Table 2
Total and individual caffeoyl-quinic, sinapic acid and feruloyl acid derivates changes in fresh-cut Bimi broccoli treated with several UV-C doses and stored up to 19 days at
5 C and 10 C. Data represents means (n = 3).
Phenolic acid (mg kg1 fw) and UV-C treatment
Time initial
After 7 days
After 13 days
After 16 days
After 19 days
5 C
10 C
5 C
10 C
5 C
10 C
5 C
10 C
3.8
6.5
47.2
38.1
3.0
20.9
1.5
1.4
21.8
20.1
41.8
15.7
15.7
1.1
13.9
40.1
64.3
38.5
57.3
52.2
50.6
3.3
6.7
2.1
9.1
1.5
3.3
7.2
0.9
1.2
51.7
69.9
5.5
19.5
13.2
14.6
12.1
19.1
8.0
5.0
52.3
32.2
5.2
11.5
11.9
1.5
1.5
1.4
2.5
1.1
1.7
1.6
1.0
1.7
1.1
1.3
1.4
1.2
0.8
0.9
1.4
3.2
1.6
2.6
3.1
2.2
1.1
2.5
2.5
3.0
1.7
2.7
2.8
1.3
1.6
3.1
4.7
2.1
4.4
3.6
1.5
2.6
3.1
1.4
1.7
54.6
71.9
4.1
25.3
21.4
2.7
4.2
14.5
14.6
2.5
8.0
2.2
1.9
7.4
7.9
9.8
7.9
9.6
2.0
6.7
12.7
18.0
11.7
18.7
15.4
15.2
3.3
5.4
2.9
3.7
3.0
5.4
6.8
1.8
2.5
17.6
26.1
5.8
30.0
14.0
4.0
4.4
5.1
2.5
3.1
18.4
30.2
5.0
14.8
12.4
1.0
1.2
1.0
1.7
1.0
1.6
1.3
1.2
1.1
1.6
2.1
2.1
2.4
1.2
1.9
1.4
2.6
2.0
2.6
1.9
2.2
1.2
1.5
1.2
1.1
1.7
2.4
2.0
1.1
1.2
3.3
3.1
2.2
6.0
4.0
1.9
2.6
2.0
1.2
1.3
3.3
3.5
2.4
6.4
4.5
4.7
8.3
30.2
30.9
3.4
16.8
3.9
2.8
15.8
17.5
23.1
16.1
20.9
2.9
13.7
26.8
42.5
26.1
41.4
35.8
32.8
6.2
9.4
4.6
6.3
5.7
11.5
11.9
2.9
4.3
36.3
60.5
11.1
81.9
33.2
13.9
23.6
15.4
5.4
7.7
41.2
55.6
12.6
97.4
40.6
2.8
2.7
4.3
4.7
1.7
3.4
2.4
1.4
1.8
2.3
2.7
3.8
4.8
1.5
2.4
2.7
3.8
2.7
3.4
3.3
2.7
1.7
2.3
2.3
1.1
4.8
6.1
3.5
1.9
2.5
4.7
6.6
2.9
10.7
7.7
6.2
7.1
3.7
2.3
3.6
5.1
7.5
3.4
12.6
8.8
2.4
4.0
13.0
13.3
1.9
7.7
1.8
2.3
7.4
8.5
11.4
8.1
8.4
1.5
6.3
12.3
22.0
17.1
20.6
17.4
14.2
2.7
3.5
1.9
2.4
2.2
4.8
5.6
1.2
1.5
17.0
31.5
5.1
27.9
14.7
11.0
13.8
11.2
6.3
2.1
19.3
34.4
8.3
30.1
17.8
1.2
1.7
3.7
4.1
1.1
2.7
1.4
1.7
2.9
3.3
3.9
3.6
6.3
2.1
3.5
3.2
5.6
3.3
5.9
5.8
4.3
1.7
4.2
3.3
2.1
1.8
3.3
3.5
1.5
1.9
5.1
10.4
3.6
13.9
5.4
2.3
4.8
3.6
2.6
2.6
5.3
12.0
5.7
12.8
6.0
20.0
30.1
115.3
109.9
15.8
62.8
16.0
13.7
59.9
62.4
96.1
58.6
69.4
13.1
494
100.6
162.2
102.9
152.4
134.9
124.2
21.2
35.4
20.9
28.8
22.5
39.4
43.3
12.5
16.7
138.8
212.8
38.3
194.3
95.9
55.4
71.0
63.1
29.8
27.1
199.5
247.4
46.5
211.0
123.4
Neochlorogenic
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
Chlorogenicb
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1,2-Disinapoylgentibiosec
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1-Sinapoyl -2 feruloylgentibiosed
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1,2-Diferuloylgentibiosee
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1,2,2 -Trisinapoylgentibiosef
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1,2 -Disinapoyl-2-feruloylgentibioseg
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
1-Sinapoyl-2,2 -diferuloylgentibioseh
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
Totali
Control
1.5 kJ m2
4.5 kJ m2
9.0 kJ m2
15 kJ m2
a
b
c
d
e
f
g
h
i
335
Fig. 7. HPLC chromatogram recorded at 320 nm of phenolic compounds present in Bimi broccoli.
Fig. 8. Total antioxidant capacity changes of fresh-cut Bimi broccoli treated with
several UV-C doses and stored up to 19 days at 5 C (A) and 10 C (B). Data represent
means of three replicates (n = 3 SD).
336
Table 3
Total chlorophylls (a + b) and carotenoids content changes in fresh-cut Bimi broccoli treated with several UV-C doses and stored up to 19 days at 5 C and 10 C. Data
represents means (n = 3).
Parameter/UV-C dose
Time initial
Day 13
Day 16
Day 19
10 C
5 C
10 C
5 C
10 C
5 C
10 C
89.5
87.7
78.5
70.9
83.2
83.8
76.2
70.1
80.1
61.6
70.0
86.7
62.6
73.2
87.9
70.7
78.5
71.1
74.3
72.2
63.8
65.2
58.9
81.7
84.7
64.2
84.2
70.2
65.2
78.4
52.2
68.3
72.2
70.2
80.9
77.3
90.5
76.2
73.6
70.2
7.5
7.9
7.3
6.7
7.9
5.1
6.5
8.4
7.0
6.0
8.2
9.4
6.8
9.2
9.5
8.0
8.0
9.9
9.8
8.2
6.5
8.1
7.4
7.9
8.2
8.5
8.1
9.6
8.4
9.4
6.2
7.5
7.9
8.2
9.0
8.4
8.6
10.9
7.5
7.8
Day 7
5 C
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