LWT - Food Science and Technology 44 (2011) 299e306

Contents lists available at ScienceDirect

LWT - Food Science and Technology

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

Influence of blanching and low temperature preservation strategies

on antioxidant activity and phytochemical content of carrots, green beans
and broccoli
A. Patras a, *, B.K. Tiwari b, N.P. Brunton a
a
Teagasc, Ashtown Food Research Centre, Ashtown, Dublin 15, Ireland
b
Biosystems Engineering, UCD School of Agriculture, Food Science and Veterinary Medicine, University College, Belfield, Dublin 4, Ireland

a r t i c l e i n f o a b s t r a c t

Article history: The objective of this study was to investigate the effect of blast freezing and blanching in combination
Received 9 February 2009 followed by chilling, on the antioxidant activity (ARP), phenols, ascorbic acid and colour of broccoli,
Received in revised form carrots and green beans. No significant changes (p > 0.05) in ARP of blanched frozen (BLFR) broccoli,
19 April 2010
carrot and green beans were observed. In contrast, UBFR (unblanched frozen) treatments caused
Accepted 15 June 2010
a significant decrease (p < 0.05) in ARP and ascorbic acid content of vegetable samples. BLFR treated
Work dedicated to my grand parents samples had better retention of antioxidant activity and ascorbic acid as compared to UBLR counterparts
at chill storage (4
C) for 7 days. However, no significant changes were observed in phenol content for all
Keywords: vegetables. Ascorbic acid decreased exponentially for both blanched and unblanched samples. The
Ascorbic acid reaction rate constant (k) increased from 1.06  101 day1 to 1.17  101 day1 for blanched and
Phenols unblanched broccoli florets and from 4.6  103 day1 to 1.98  101 day1 for blanched and unblanched
Kinetics carrots during 7 days storage. Result presented here indicates greater stability of nutritional parameters
Blanching for BLFR samples compared to UBFR samples during 7 days storage at 4
C for all vegetables.
Freezing Ó 2010 Elsevier Ltd. All rights reserved.
Chill storage

1. Introduction enzymatic reactions, senescence and microbial growth; it does not

fully stop these processes (Bahceci, Serpen, Gokmen, & Acar, 2005).
Fruits and vegetables contain range of phytochemicals, in The result can be the development of off-odours, off-colours, off-
addition to well-known antioxidants, such as vitamins C and E, or flavours, changes in texture and nutrient loss. Fruits and vegetables
polyphenols, which significantly contribute to their total antioxi- are blanched prior to freezing mainly to inactivate enzymes, reduce
dant activity (Cao, Sofic, & Prior, 1996; Prior et al., 1998). Epidemi- microbial load, remove gases from the plant tissue, cause shrinkage
ological and clinical investigations have associated diets rich in of the product to facilitate packaging, fix texture, colour, and clean
fruits and vegetables with reduced risk of heart, cardiovascular, the surface of the vegetable (Bahceci et al. 2005; Barrett &
neurological and chronic diseases, and various forms of cancer Theerakulkait, 1995).
(Ames, Shigenaga, & Hagen, 1993). Due to the seasonal and Typically, blanching is carried out by treating the vegetables
perishable nature, raw vegetables are subjected to some form of with steam or hot water for 1e10 min at 75e95
C, the time/
preservation in order to make them available for later consumption temperature combination depending on the type of vegetable
(Lin & Brewer, 2005). Freezing of fruits and vegetables is generally (Cano, 1996). Blanching of foods involves mild heating in water and
regarded as superior to other food preservation techniques such as serves. Blanching can have negative effect on nutrients such, as
canning and dehydration, with respect to retention in sensory vitamins and phenolic compounds which are relatively unstable
attributes and nutritive properties. Freezing is often employed to when subjected to heat treatments (Prochaska et al., 2000). Apart
maintain fresh-like characteristics with minimal loss of nutrients from processing, storage conditions and domestic cooking and
such as vitamins, and antioxidant content over long periods preparation have significant effect on phytochemicals such as
(Prochaska, Nguyen, Donat, & Piekutowski, 2000). However, while ascorbic acid and phenols (Patras, Brunton , Tiwari, & Butler.,2009a;
freezing on its own helps to preserve food through the slowing of Rawson, Koidis, Patras, Tuohy,& Brunton, 2010; Vallejo, Tomás-
Barberán, García-Viguera, 2002; Verkerk & Dekker, 2004).
* Corresponding author. Tel.: þ353 879600673; fax: þ353 18059550. Consumers are now becoming aware of the need to consume
E-mail addresses: [email protected], [email protected] (A. Patras). a variety of fresh vegetables to maximise their intake of beneficial

0023-6438/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2010.06.019
300 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306

antioxidants (Prochaska et al., 2000). Several studies show the Fresh Vegetables
effects of freezing on green peas and spinach, carrots, broccoli,
green beans (Howard, Wong, Perry, & Klein, 1999; Korus, Lisiewska, Washed, Peeled, Sliced
& Kmiecik, 2002; Murcia, López-Ayerra, Martinez-Tomé, Vera, &
García-Carmona, 2000; Saccani et al., 2001). In addition, freezing
has, in some studies, been shown to influence the quality and Blanching treatment (Batch A) Without-Blanching treatment (Batch B)
nutritive value of foods (Srinivasan, Xiong, & Blanchard, 1997).
There is very little information on how these processes in combi-
nation influence the quality and nutritive value of foods despite the
Freezing treatment
fact that they are commonly used together to prolong shelf life.
Although freezing is an effective method of preserving foods
some deterioration in frozen food quality occurs during storage. Chill storage upto 7 days
Therefore the objective of the current study was to examine the
effect of blast freezing and blanching in combination followed by Freeze dried
chilling, on the ARP, phenols, ascorbic acid and colour of broccoli,
carrots and green beans with a view to their use as components of
ready-meals. Extracted and chemically
analysed

2. Materials and methods Fig. 1. Overview of the steps involved in blanching, freezing and chill storage
treatments.
2.1. Vegetable material and sample preparation

Fresh carrots (Daucus carota L. cv. Nairobi), green beans (Pha- Saufen, Germany). DT20 tube with a rotor stator element with
seolus vulgaris L cv. Emerit) and broccoli (Brassica oleracea L cv. a dispersing element was utilized. The stator diameter was 19 mm
Monaco) were purchased from a local supplier (Donnelly’s Ltd., and rotor diameter of 12.7 mm with gap between rotor and stator
Dublin, Ireland). The carrots were washed, peeled (using a hand of 0.3 mm with shaft length of 19.2 cm was used. The samples were
held domestic peeler) and sliced into discs of 5 mm thickness using then vortexed with a V400 Multitude Vortexer (Alpha laboratories,
a mechanical slicer (Berkel Ltd, Ontario, Canada). The green beans North York, Canada) for 20 min at 800 g and centrifuged for 15 min
were washed, topped and tailed. Broccoli was separated into florets at 2000 g (MSE Mistral 3000i, Sanyo Gallenkamp, Leicestershire,
and leaves and inedible stems were removed. UK). 10 mL of the sample was filtered through PVDF Acrodisc
syringe filters (pore size 0.45 mm, Sigma, Ireland,) and stored
2.2. Blanching and blast freezing experiment at 20
C for subsequent analysis. Total antioxidant activity was
measured using the DPPH assay as described by Goupy, Hugues,
Two different pre-treatments were used in this trial. Carrot, Boivin, and Amiol (1999). Briefly 500 mL of diluted sample and
green beans and broccoli were equally divided (200 g each) into 500 mL of the DPPH (0.238 mg/mL1) working solution were mixed
two batches [Batch A and Batch B]. Batch A was subjected to in a microcentrifuge tube. After vortexing, the tubes were left in
blanching (BL), whereas no blanching treatments (UB) were given the dark for 30 min at room temperature after which the absor-
to the batch B. Blanching was carried out by placing samples in bance was measured against methanol at 515 nm using a spectro-
water at 95
C for 3 min. The proportion of water to the raw photometer (UV-1700 Pharma Spec, Shimadzu, Milton Keynes).
material used was 5:1 by weight. Following this, samples were The decrease in absorbance of a sample was calculated in
cooled in distilled in water for 2e3 min. Prior to blast freezing, comparison to a blank sample (500 mL methanol and 500 mL
samples (200  1 g) were vacuum-sealed in 20 cm  30 cm in Poly- DPPH). The relative decrease in absorbance (PI) was then calcu-
propylene pouches (thickness- 75 mm, gas permeability- lated as follows: PI (%) ¼ 1  (Ae/Ab), with Ae ¼ absorbance of
2.7 g m2 d, sealing temperature- 100e180
C, Packex Industries sample extract and Ab ¼ absorbance of blank. The PIs used to
Ltd., Wicklow, Ireland using Vac-star S220 vacuum sealer (Vicquip calculate the related antioxidant activity were superior (PI1) and
Ltd., Dublin, Ireland)). The blanched and unblanched vegetables inferior (PI2) to the value estimated at 50%. Antioxidant activities
were blast frozen at 30
C for 2.5 h (Nu-Avon, Wiltshire, England; were expressed as the IC50 i.e. the concentration of antioxidant
air speed 3.8 m s1) (Fig. 1). required to cause a 50% reduction in the original concentration of
DPPH. For ease of interpretation, antiradical powers were also
2.3. Chill storage calculated and defined as the inverse of the IC50 value Eq. (1)
and (2). Finally, the antioxidant activity of the extracts was
Blanched frozen [BLFR] and unblanched frozen [UBFR] samples compared to that of a synthetic antioxidant (Trolox) and expressed
were placed in different propylene pouches and stored in air at 4
C as Trolox equivalent antioxidant activity (TEAC) values. The anal-
for 0, 3, 5, 7 days. Each treatment was replicated three times. At yses were done in triplicate and all quantitative results are
each sampling point, samples were removed; freeze dried (Frozen reported on a dry weight basis.
in Time Ltd., York, UK) at a temperature and pressure of 50
C and
0.03 mbar respectively for more than two days and tested for
ðC1  C2 Þ  ðPI1  50Þ
antioxidant indices and instrumental colour. DC ¼ (1)
PI1  PI2
2.4. Measurement of total antioxidant activity
IC50 ¼ C1  DC (2)
Methanolic extracts were prepared by adding 25 mL of HPLC
grade methanol to 1.25 g of freeze dried powder. Samples were
then homogenised for 1 min at 24,000 rpm using an Ultra-Turrax 1
ARP ¼ (3)
T-25 Tissue homogenizer (Janke & Kunkel, IKAÒ-Labortechnik, IC50
 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306 301

2.5. Measurement of phenolic content 3. Statistical analyses

Total phenols (TP) in vegetable samples were determined using A two-way analysis of variance (ANOVA) was performed using
the FolineCiocalteu reagent according to the method of Singelton, the GenStat Release 10.1 software. Values were considered signifi-
Orthofer, and Lamuela-Raventos (1999). Briefly 100 mL of meth- cant at p < 0.05. Three replications were carried and mean values
anolic extract, 100 mL of MeOH, 100 mL FolineCiocalteau reagent were reported. The degradation of ascorbic acid loss in blanched
(FC) and 700 mL of Na2CO3 was added to 1.5 mL microcentrifuge and unblanched samples was calculated by using the standard
tubes and the samples were vortexed briefly. The tubes were then equation for first-order reactions and degradation rate constants
left in the dark for 20 min at room temperature. Following this, the were determined by fitting (Eq. (6)) to experimental data.
samples were centrifuged (Eppendorf, Centrifuge 5417R., Germany)
at 10,000 g for 3 min. The absorbance of the sample was read at C ¼ C0 ekt (6)
735 nm using aqueous Gallic acid (10e400 mg L1) as a standard.
where, C is the studied parameter (AA) at any given reaction time,
Results were expressed as mg of Gallic acid equivalent per 100 g of
C0 are initial values of treated samples (day 0) and k are rate
dry weight of sample.
constants. Data fitting was considered significant at a probability
level of 95%.
2.6. L-Ascorbic acid analysis (AA)

Extraction of L-ascorbic acid was carried out using metaphos- 4. Results and discussion
phoric acid (6 g/100 mL) and 1.25 g of freeze-dried powder as
described for antioxidant extractions above. Ascorbic acid deter- 4.1. Effect of pre-treatment (BL/UB) and subsequent blast freezing
mination was carried out by reverse phase high performance liquid on antioxidant activity, phytochemical content of broccoli, carrots
chromatography (HPLC) according to the method of Lee and Coates and green beans
(1999) with slight modifications (Patras, Brunton, Da Pieve, Butler,
& Downey 2009b; Patras, Brunton, Da Pieve, & Butler, 2009c). A ARP of fresh, BLFR and UBFR broccoli, carrots and green beans
20 ml of aliquot was injected into the chromatographic column. The are presented in Table 1. ARP values of all BLFR vegetables were not
chromatographic system (Shimadzu Model no SPD- M10A VP, significantly lower than fresh samples indicating that blanching
Mason Technology, Dublin 8, Ireland) consisted of a pump, with subsequent blast freezing did not affect the antioxidant
a vacuum degasser, a DiodeeArray Detector and it was controlled activity of any of the vegetables. These results agree with the
through EZ Start 7.3 software (Shimadzu) at 40
C. A hypersil ODS findings of Hunter and Fletcher (2002), they demonstrated that
column (150 mm  4.6 mm, 5 mm, Supelco., US) fitted with frozen vegetables have similar antioxidant activities to their fresh
hypersil ODS guard column(Gemini C18 [4 mm L  3.0 mm ID], counterparts. Myojin et al. (2008) suggested that blanching of bitter
Phenomenex., UK) was utilised with a mobile phase (isocratic) of gourd improves the retention of antioxidant activity and total
25 mMol L1 monobasic potassium phosphate adjusted to pH 3 at phenolics during subsequent frozen storage. Puupponen-Pimia
a flow rate of 1 mL min1 at 40
C. The detector was set at 245 nm. et al. (2003) found that total phenolics and DPPH index of cauli-
For quantification external calibration curves for ascorbic acid in flower decreased slightly during blanching, but no further changes
metaphosphoric acid (6 g/100 mL) were prepared at concentra- were observed during storage. Similarly, Cooper, Chen, and King
tions from 25 mg mL1 to 500 mg mL1. The total run time was (1983) studied the effects of laboratory-scale blanching and
4.0 min. freezing on folate levels of spinach. They found that blanching in
water at 100
C for 3 min caused a 33% loss compared with fresh
spinach. Ge˛ bczyn ski and Lisiewska (2006) studied the process of
2.7. Colour measurements freezing (20, 30
C) in broccoli florets and found significant
decrease in the antioxidant activity. In the present study, non-
The colour of the samples was measured using a Hunter Lab inclusion of blanching step with blast freezing did significantly
colour meter (Hunter Lab DP-9000 colour difference meter, Hunter affect ARP values for all vegetables investigated. It is important to
Associates Laboratory, Virginia, USA) fitted with a 2.5 cm diameter note that the retention of antioxidant activity (ARP) is significantly
aperture. The instrument was calibrated using the black and white improved by blanching, indicating possible involvement of enzy-
tiles provided. Colour was expressed in Hunter Lab units L*, a* and matic reactions (PPO) persisting significantly in unblanched
b*. Three replicate measurements were performed and results samples and in a less extent still in blanched samples (Table 1).
were averaged. In addition, Chroma and total colour difference Conversely levels of phenols in all vegetables were unaffected by
(TCD) were calculated using the following equations, where L0, a0, BLFR and UBFR treatments. Ge˛ bczyn  ski and Lisiewska (2006)
b0 are the control values (untreated) for vegetables. TCD indicates studied the process of freezing (20, 30
C) in broccoli florets
the magnitude of colour difference between stored and control and found significant decrease in the level of polyphenol content.
samples. Differences in perceivable colour can be analytically Myojin et al. (2008) found that at subsequent frozen storage at
classified as very distinct (TCD > 3), distinct (1.5 < TCD < 3) and 18
C, radical scavenging activity and total phenolic content of
small difference (TCD < 1.5) (DrLange, 1999; Tiwari, unblanched and blanched bitter gourd underwent little change for
Muthukumarappan, O’Donnell, & Cullen, 2008). It should be 90 days and then gradually declined. The authors also reported that
noted that combinations of colour parameters may be more radical scavenging activity and total phenolic levels, practically
effective to evaluate the overall colour changes of processed remained unchanged throughout the entire storage period
vegetables than the individual L*, a*, b* parameters. at 40
C.
Levels of ascorbic acid were unaffected by BLFR for broccoli and
pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Chroma ¼ a2 þ b2 (4) carrot samples but decreased significantly (p < 0.05) by UBFR
treatments compared to fresh samples. This was particularly
evident for broccoli florets with an 83% reduction in AA levels being
h i1=2 detected. Davey et al. (2000) compared processing techniques most
DE ¼ ðL*  L0 Þ2 þða*  a0 Þ2 þðb*  b0 Þ2 (5)
relevant to vegetables, i.e. canning, freezing and dehydration, they
302 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306

Table 1
Influence of pre-treatment (BLFR/UBFR) and subsequent blast freezing [day 0] on antioxidant activity, ascorbic acid, phenols and colour of broccoli, carrots and green beans.

Samples ARPd (g/L)-1 Phenols AAg (mg/100g) L* TCDf Chroma

(mgGAEe/100g)
Broccoli
Fresh 0.53  0.01 446.0  12.21 374.1  2.12 29.5  3.78 0.00 12.8  1.21
Blanched Frozen [BLFR]a 0.52  0.02 448.0  0.98 373.2  5.67 28.1  5.89 0.2  1.21 11.2  0.04
Unblanched Frozen [UBFR]b 0.45  0.04 521.0  2.32 62.7  2.34 18.3  0.01 10.9  0.76 8.8  0.56
LSDc 0.06 87 12.57 2.68 0.04 2.24

Carrots
Fresh 0.12  0.45 88.4  1.78 43.9  0.05 56.3  6.89 0.00 36.2  1.78
Blanched Frozen 0.12  0.01 89.0  1.56 40.6  4.89 56.2  1.78 0.9  2.78 36.2  1.56
Unblanched Frozen 0.09  0.03 92.5  3.78 20.7  3.90 54.2  2.13 1.8  6.78 36.1  4.21
LSD 0.02 15.79 12.37 3.81 3.32 3.78

Green beans
Fresh 0.24  0.01 225.6  1.89 ND 43.6  2.67 0.00 18.7  0.01
Blanched Frozen 0.22  0.56 200.5  3.21 ND 40.2  2.78 5.1  2.78 15.1  0.98
Unblanched Frozen 0.11  0.34 186.6  4.32 ND 23.4  3.01 19.3  5.01 13.1  0.21
LSD 0.06 40.27 4.45 0.19 2.16

All values are means of three replicates of three batches and expressed in dry weight basis.
a
Blanched Blast frozen.
b
Unblanched Blast frozen.
c
least significant difference (p ¼ 5%).
d
Total antioxidant activity.
e
Gallic acid equivalent.
f
Total colour difference.
g
L- Ascorbic acid.

found that L-ascorbic acid losses were greatest during dehydration values for unblanched samples were significantly lower than for
and lowest during freezing. Similarly, Favell (1998) reported blanched samples. However, it is well known that processing of any
negligible losses in ascorbic acid of carrots during freezing. It is kind induces physical, structural, chemical and nutritional modifi-
envisaged that ascorbic acid losses can occur, particularly during cations in different vegetables (Di Scala & Crapiste, 2008). For
blanching (Lathrop & Leung, 1980) but slight change may be example Ismail and Lee (2004) blanched five species of cruciferous
expected thereafter during deep frozen storage. Several studies vegetables and observed a 9e40% decrease in the total antioxidant
demonstrated that effects of freezing on the product quality are activity. In contrast, Nicoli, Anese, Parpinel, Franceschi, and Lerici
minimal (Howard et al., 1999; Murcia et al., 2000). (1997) reported that inspite of a reduction in antioxidant phyto-
chemicals brought about by thermal process treatments, the overall
4.2. Effect of pre-treatment (BL/UB) and subsequent blast freezing antioxidative activity of food products can be preserved or even
on colour parameters of carrots and green beans and broccoli increased owing to the possibility of other antioxidants arising,
particularly as a result of the Maillard reaction. Similar results were
Table 1 shows the colour attributes of fresh, BLFR, UBFR carrots, reported by (Patras et al., 2009a). This was not the case for water
green beans and broccoli. In general, TCD values for this study were blanching in the present study and this may be related to the fact
found to be very distinct after UBFR treatment but not following that blanching is a mild process and preserves antioxidant
BLFR. For both broccoli florets and green beans TCD values were components whilst also protecting them from enzyme mediated
significantly different for UBFR samples compared to fresh degradation by inactivating enzymes such as polyphenol oxidase.
samples. The lightness values (L*) for fresh broccoli, carrots and ARP values for UBFR samples were significantly lower than for
green beans were 29.5, 56.3, and 43.6 respectively. UBFR treatment than for BLFR for all vegetables types after 7 days of storage. This
had more pronounced affect on lightness of vegetable samples indicates that initial effect of blanching in retaining more antioxi-
whereas no significant reduction in L* was observed for BLFR dants at the beginning of the storage period was maintained even
samples. Chroma, one of the important colour attribute decreased after 7 days chill storage. In the present study, heat treatment in
significantly for broccoli and green beans with the exception of blanching may induce some losses, but inactivation of oxidative
carrots. BLFR treatment maintained colour intensity of the samples enzymes prevents further losses during chill storage. Subsequent
better than UBFR treated samples. A study conducted by Lin and chill storage for up to 7 days had significant effect on ARP values of
Brewer (2005) demonstrated that blanched peas were visually broccoli, carrot disks and green beans with significantly low values
lighter green than unblanched peas immediately after blanching being present in samples stored for 7 days as compared to fresh
and after 12 weeks of frozen storage whereas they did not observe samples for all vegetable types and treatments. Puupponen-Pimia
any significant difference for a* and b* value of blanched frozen et al. (2003) also observed a pronounced decrease in the DPPH
green peas. index during the refrigerated storage of broccoli. It is also very
important to note that decrease in ARP is due to the loss of anti-
4.3. Effect of treatment (BLFR/UBFR) and short term chill storage on oxidant components such as ascorbic acid, polyphenols and glu-
antioxidant activity, phytochemical content of broccoli, carrots and cosinolates etc in green vegetables. In the present study, it is
green beans difficult to make direct statements on the relationship between
ascorbic acid and antioxidant activity in methanolic extract as
ARP values for broccoli, carrots and green beans with or without ascorbic acid is not sparingly soluble and therefore would not be
blanching and followed by chill storage are presented in Table 2. present in the extract. Linear correlations demonstrate that the
Significant effect of treatment and storage days on ARP levels for all antioxidant activity determined by DPPH assay, was well correlated
vegetable samples was observed (Tables 2 and 4). In all cases ARP to total phenolic content (r ¼ 0.76 p < 0.001).
 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306 303

Table 2
Influence of treatment (BLFR/UBFR) and subsequent chill storage on antioxidant activity, ascorbic acid, phenols of broccoli, carrots and green beans.

Days of storage ARPb (g/L)1 Phenols mgGAEc/100g AAd mg/100g

BLe UBf BL UB BL UB
Broccoli
0 0.51  0.02 0.45  0.04 422.0  3.35 521.0  5.06 211.8  9.81 62.7  1.38
3 0.53  0.02 0.40  0.02 404.0  11.25 671.0  0.89 153.6  15.21 41.1  0.51
5 0.52  0.03 0.46  0.05 403.1  4.56 679.1  2.26 110.3  0.37 31.7  0.51
7 0.29  0.02 0.17  0.02 387.0  9.56 636.0  12.61 96.7  0.64 30.5  0.40
LSDa 0.25 29.4 16.14

Carrots
0 0.11  0.01 0.09  0.80 92.5  5.90 92.5  2.45 39.0  2.67 20.6  1.89
3 0.09  0.01 0.07  1.23 110.1  6.90 110.1  7.89 38.5  3.67 7.5  1.78
5 0.08  0.02 0.04  0.90 104.1  1.32 104.1  0.03 38.6  7.21 7.2  0.04
7 0.06  0.01 0.03  0.05 90.0  8.90 90.0  0.06 38.6  0.02 6.4  4.89
LSD 0.01 19.16 0.70

Green beans
0 0.24  0.89 0.11  0.21 224.2  6.01 186.6  2.89 nd nd
3 0.25  0.78 0.07  2.67 226.5  8.21 187.9  6.69 nd nd
5 0.11  0.12 0.07  0.08 220.5  0.34 219.4  2.78 nd nd
7 0.09  4.78 0.06  0.01 198.3  0.81 188.4  1.32 nd nd
LSD 0.07 40.94

All values are means of three replicates of three batches and expressed in dry weight basis.
nd ¼ non-detectable.
a
least significant difference (p ¼ 5%).
b
Total antioxidant activity.
c
Gallic acid equivalent.
d
L- Ascorbic acid.
e
Blanched.
f
Unblanched.

Ascorbic acid levels ranged from 30.5 to 211.8 mg/100 g DW and was found decrease exponentially with R2 > 0.74 (Fig. 2) with
6.40e39.0 mg/100 g DW for UBFR and BLFR broccoli and carrots degradation rate constant of 1.1  101 day1 and 1.2  101 day1
samples respectively as illustrated in Table 2. Observed values were for blanched and unblanched broccoli samples respectively. Simi-
within the range of those reported elsewhere for these vegetables larly, higher degradation rate constant was observed for
(Howard et al., 1999). Ascorbic acid levels were below the limit of unblanched carrot samples (2.0  101 day1) as compared to
detection in green beans. Following chill storage, ascorbic acid blanched samples (0.05  101day1).
decreased significantly for unblanched samples, whereas better Phenolic content (TP) of broccoli, carrots, green beans ranged
retention of ascorbic acid was observed for blanched broccoli from 387 to 679 mg GAE/100 g; 90.0e110.1 mg GAE/100 g;
samples. In contrast no changes in ascorbic acid content was 186.6e226.5 mg GAE/100 g, respectively among all the samples
observed for BLFR carrots whereas 30% ascorbic acid was lost for analysed (Table 2). The observed values were within the range for
UBFR samples (p < 0.05). The retention of ascorbic acid is often used these vegetables by other authors (Leja, Mareczek, Starzyn  ska, &
as an estimate for the overall nutrient retention of food products Rozék, 2001; Puupponen-Pimia et al., 2003; Zhang & Hamauzu,
(Murcia et al., 2000). Ascorbic acid is by far the least stable nutrient 2004). Phenolic content of blanched broccoli samples were fairly
during processing; it is highly sensitive to oxidation and leaching consistent throughout the chill storage. Results also demonstrated
into water-soluble media during processing, storage and cooking of that phenolic content of unblanched broccoli samples increased
fresh, frozen and canned fruits and vegetables (Franke, Custer, significantly from day 0 to day 5 and than decreased thereafter.
Arakaki, & Murphy, 2004). Higher losses of vitamin C in vegeta-
bles due to blanching in water are reported in the literature (Murcia
et al., 2000; Sikora, Cieslik, Leszczyn  ska, Filipiak-Florkiewicz, & 1.20

Pawe1, 2008). Some authors also suggested that steam blanching

(96
C for 3 min) prior to freezing storage decreased vitamin C 1.00

concentration by about 32% and these results are similar to those of

Howard et al. (1999). The authors reported a vitamin C loss of about 0.80

30% in steam-blanched broccoli (92
C for 1.8 min).

Ct /Co

In a study carried out by Albrecht, Schafer, and Zottola (1991) 0.60

reported vitamin C losses ranging from 2% to 48% for 6 different

broccoli cultivars stored at 2
C for 21 days, similarly, Howard et al. 0.40

(1999), observed vitamin C losses of 13% and 48%, respectively, after

3 wk of storage of broccoli at 4
C. The author also suggested that 0.20

AA retention during prolonged storage was better in vegetables

stored at 20
C than during refrigerated storage at 4
C. The 0.00
0 1 2 3 4 5 6 7 8
obtained results underline the important role of several pre-pro-
cessing factors on the qualitative and nutritional characteristics of Storage time (days)
carrot and broccoli samples. Fig. 2. First- order plot of ascorbic acid degradation in blanched and unblanched
In our results, non-inclusion of a blanching step, decreased vegetable samples. [ Blanched broccoli (:),Unblanched broccoli (A); Blanched carrot
ascorbic acid significantly over 7 days of storage period. AA content (-), Unblanched carrot (C)].(values are average of n ¼ 3).
304 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306

Table 3
Influence of treatment (BLFR/UBFR) and subsequent chill storage on colour parameters of blanched and unblanched (95
C, 3 min) broccoli florets, carrots and green beans
during storage for 7 days at 4
C.

Days of storage L* TCDb Chroma

BLc UBd BL UB BL UB
Broccoli
0 28.8  1.34 18.3  0.08 1.2  1.21 10.9  0.04 12.1  4.45 8.8  2.10
3 27.5  1.64 18.1  1.01 1.8  3.01 11.8  2.67 11.9  3.90 6.9  3.90
5 27.4  1.55 18.1  0.66 2.2  0.05 11.2  3.32 12.0  3.54 8.7  0.67
7 28.0  1.04 14.0  0.06 1.2  2.45 7.4  1.45 11.7  2.78 9.1  0.89
LSDa 3.67 0.92 1.35

Carrots
0 56.1  1.32 56.3  2.21 1.4  1.56 1.8  2.78 35.3  2.78 36.1  2.78
3 56.9  1.67 42.4  0.80 1.6  1.89 16.2  4.78 34.5  0.06 42.0  3.78
5 56.1  2.56 44.5  3.67 1.9  2.67 13.3  2.01 34.6  0.04 38.7  0.01
7 55.1  4.31 43.1  0.45 4.1  3.01 14.4  0.01 33.3  2.43 35.8  0.56
LSD 0.52 0.98 1.39

Green beans
0 41.3  5.90 23.4  0.34 2.9  6.61 19.3  5.89 12.9  2.78 8.8  2.90
3 41.6  0.01 28.8  4.78 4.3  5.89 13.4  0.09 11.9  4.78 6.9  0.01
5 41.0  0.45 27.0  2.45 2.5  0.91 15.6  0.20 12.0  4.17 8.7  0.02
7 41.7  0.78 26.1  2.90 2.3  5.89 16.5  1.67 11.7  3.89 9.1  2.71
LSD 3.04 1.08 1.35

All values are means of three replicates of three batches and expressed in dry weight basis.
a
least significant difference (p ¼ 5%).
b
Total colour difference.
c
Blanch.
d
Unblanch.

A similar trend was observed for carrots and green beans. Studies in samples had lower values (Table 3). A decrease in L* value is related
vegetables confirm that processing significantly alters the physical to product lightness loss. These results demonstrate the significant
and bio-chemical composition and functionality (Patras, Brunton, effect of UBFR and BLFR treated samples on the colour degradation
Tiwari, & Butler, 2008; Zhang & Hamauzu, 2004) and may play of the vegetable samples studied. The major colour changes in
a vital role in non-uniform behaviour of phenols towards different vegetable samples were largely due to the non-inclusion of
processing treatments. The total phenolic content of unblanched blanching step.
broccoli increased at a much higher rate during storage than that of Mean colour intensity (chroma) values were higher for blanched
blanched samples (Table 2). This increase could be related to the samples than unblanched counterparts. In the case of carrots, not
developmental changes and wound-like response due to freezing. much variation was observed in colour intensity for blanched or
Dixon and Paiva (1995) reported that plants respond to wounding unblanched samples as shown in Table 3. Interestingly, Chroma
with increases of phenolic compounds involved in the repair of
wound damage and in defence against microbial invasion.
Table 4
According to Sarkar and Phan (1979), the total phenolics of carrots Effect of treatment (BL/UB), storage days and interactions on quality parameters of
stored at 3  1
C and at z90% relative humidity increased steadily broccoli, carrots and green bean.
with storage time. Chubey and Nylund (1969) suggested that
Sample Treatment Storage Interaction
carrots richer in phenolics are more susceptible to browning, but days (Treatment  Storage days)
their contributions (in fruits and vegetables) for resisting parasitic Broccoli
attack could be of benefit to minimally processed carrots stored. ARPa S S NS
Phenols S NS NS
AAb S S S
4.4. Effect of treatment (BLFR/UBFR) and chill storage on L* S S S
instrumental colour parameters of broccoli, carrots and green beans TCDc S S S
Chroma S S S

Colour is often the first parameter by which a consumer judges Carrots

a food product before purchase (Gormley, 1978). Colour is used to ARP S S NS
Phenols NS NS S
judge the quality, maturity and age after harvest of many foods.
AA S S S
During chilled storage, the colour of vegetables can become grey or L* S S S
brownish, or in some cases paler (Zacharias, 1980). In our study, TCD S S S
total colour difference and the colour intensity calculated from Chroma S S S
colour parameters L*, a*, and b* varied significantly among storage Green beans
time and treatments (Table 3). Analysis of variance indicated that ARP S S S
this variation was in most cases due to differences between vege- Phenols NS NS NS
L* S S S
table types. Mean L* values decreased from 28.85 to 28.07 for BLFR
TCD S S S
broccoli samples, whereas low values (14.04e18.3) of lightness Chroma NS NS S
were observed for UBFR samples at the end of storage. (Table 3).
S ¼ significant at 5% level; NS ¼ not-significant.
Similar results were reported by (Cruz, Vieira, & Silva, 2007) for a
ARP.
watercress (Nasturtium officinale). In the case of carrots and green b
L- Ascorbic acid.
c
beans, L* value was fairly stable in BLFR samples, whereas UBFR Total colour difference.
 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306 305

values of green beans decreased significantly as a combined effect Cooper, R. G., Chen, T. S., & King, M. A. (1983). Thermal destruction of folacin in
microwave and conventional heating. Journal of the American Dietetic Associa-
of storage and treatment as illustrated in Table 4. Proportion of grey
tion, 73, 406e410.
component characterizing colour is given by values of Chroma. As Cruz, R. M. S., Vieira, M. C., & Silva, C. L. M. (2007). Modelling kinetics of watercress
Chroma decreases, colour becomes less intense (Lancaster, Lister, (Nasturtium officinale) colour changes due to heat and thermosonication
Reay, & Triggs, 1997) changing from a vivid to a dull green in case treatments. Innovative Food Science and Emerging Technologies, 8, 244e252.
Davey, M. W., Van Montagu, M., Inze, D., Sanmartin, M., Kanellis, A., Smirnoff, N.,
of broccoli and green beans in the present study. et al. (2000). Plant L-ascorbic acid: chemistry, function, metabolism, bioavail-
Total colour difference (TCD) is a function of the three CIE L* a* ability and effects of processing. Journal of the Science of Food and Agriculture,
b* coordinates (Eq. (2)). TCD values increased from 1.2 to 2.2, 1.4 to 80, 825e860.
Di Scala, K. C., & Crapiste, G. H. (2008). Drying kinetics and quality changes during
4.1, 2.3 to 4.3 for BLFR broccoli, carrots and green beans respec- drying of red pepper. LWT: Food Science and Technology, 41(5), 789e795.
tively, whereas greater differences were observed for UBFR Dixon, R. A., & Paiva, N. L. (1995). Stress-induced phenylpropanoid metabolism.
samples. Colour change was highest in unblanched samples as Plant Cell, 7, 1085e1097.
DrLange. (1999). Fundamentals of colorimetry-application report. DrLange, 10e,
compared to blanched samples for all three vegetables (Table 3). 362, (USA).
This may be due to lower L* values in unblanched samples as level Favell, D. J. (1998). A comparison of the vitamin C content of fresh and frozen
of enzyme activity (PPO) would be high in unblanched samples. vegetables. Food Chemistry, 62, 59e64.
Franke, A. A., Custer, L. J., Arakaki, C., & Murphy, S. P. (2004). Vitamin C and flavonoid
A study conducted by (Cruz et al., 2007) demonstrated that levels of fruits and vegetables consumed in Hawaii. Journal of Food Composition
blanching by heat and a combined treatment of heat/ultrasound and Analysis, 17, 1e35.
(thermosonication) resulted in higher TCD values of 6 and 12 Gormley, T. R. (1978). Colour and its measurement in foods. Food Progress, 13, 2.
Goupy, P., Hugues, M., Boivin, P., & Amiol, J. M. (1999). Antioxidant composition and
respectively in for watercress (N. officinale) as compared to control.
activity of barley (Hordeum vulgate) and malts extracts and of isolated phenolic
Higher TCD values were reported by (Patras et al., 2009a) in heated compounds. Journal of the Science of Food and Agriculture, 79, 1625e1934.
treated carrot samples as compared to fresh sample. Hue angle Ge˛ bczyn ski, P., & Lisiewska, Z. (2006). Comparison of the level of selected anti-
oxidative compounds in frozen broccoli produced using traditional and modi-
provides more information about the spatial distribution of colours
fied methods. Innovative Food Science and Emerging Technologies, 7, 239e245.
than direct values of tristimulus measurements (Sigge, Hansmanw, Howard, L. A., Wong, A. D., Perry, A. K., & Klein, B. P. (1999). b-Carotene and ascorbic
& Joubert, 2001) but an inconsistent affect of this colour parameter acid retention in fresh and processed vegetables. Journal of Food Science, 64,
was observed in this study (data not shown). 929e936.
Hunter, K. J., & Fletcher, J. M. (2002). The antioxidant activity and composition of
425 fresh, frozen, jarred and canned vegetables. Innovation of Food Science
Emerging Technology, 3, 399e406.
5. Conclusions Ismail, A., & Lee, W. Y. (2004). Influence of cooking practice on antioxidant prop-
erties and phenolic content of selected vegetables. Asia Pacific Journal of Clinical
The retention of total antioxidant activity, total phenols and Nutrition, 13, S162eS165, (Suppl).
Korus, A., Lisiewska, Z., & Kmiecik, W. (2002). Effect of freezing and canning on the
ascorbic acid in broccoli, carrots and green beans was strongly
content of selected vitamins and pigments in seeds of two grass pea (Lathyrus
influenced by the different pre-treatment and preservation sativus L.) cultivars at the not fully mature stage. Nahrung/Food, 46, 233e237.
methods investigated. BLFR was an effective technique for Lancaster, J. E., Lister, C. E., Reay, P. F., & Triggs, C. M. (1997). Influence of pigment
composition on skin colour in a wide range of fruits and vegetables. Journal of
preserving ascorbic acid and antioxidant activity in all vegetables.
the American Society for Horticultural Science, 122(4), 594e598.
The retention of these moieties being negatively influenced by non- Lathrop, P. J., & Leung, H. K. (1980). Thermal degradation and leaching of vitamin C
inclusion of a blanching pre-treatment step. Vegetables subjected in green peas during processing. Journal of Food Science, 45(4), 995e998.
to blanching treatments had better retention of ARP and ascorbic Lee, H. S., & Coates, G. A. (1999). Vitamin C in frozen, fresh squeezed, unpasteurized,
polyethylene-bottled orange juice: a storage study. Food Chemistry, 65(2),
acid and instrumental parameters as compared to unblanched 165e168.
counterparts. Total colour change was found to be substantially Leja, M., Mareczek, A., Starzyn  ska, A., & Rozék, S. (2001). Antioxidant ability of
higher (TCD > 3) for unblanched samples. Future directions for broccoli flower buds during short-term storage. Food Chemistry, 72, 219e222.
Lin, S., & Brewer, M. S. (2005). Effects of blanching method on the quality charac-
research might include investigation of methods to prevent losses teristics of frozen peas. Journal of Food Quality, 28, 350e360.
during blanching that would be feasible in a home or industrial Murcia, M. A., López-Ayerra, B., Martinez-Tomé, M., Vera, A. M., & García-
environment. Carmona, F. (2000). Evolution of ascorbic acid and peroxidase during industrial
processing of broccoli. Journal of the Science of Food and Agriculture, 80,
1882e1886.
Myojin, C., Enami, N., Nagata, A., Yamaguchi, T., Takamura, H., & Matoba, T. (2008).
Acknowledgement
Changes in the radical-scavenging activity of bitter gourd (Momordica charantia
L.) during freezing and frozen storage with or without blanching. Journal of Food
This project was funded under the Food Institutional Research Science, 73(7), C546eC550.
Measure (FIRM) by the Irish Agriculture and Food and Fisheries Nicoli, M. C., Anese, M., Parpinel, M. T., Franceschi, S., & Lerici, C. R. (1997). Loss and/
or formation of anthioxidants during food processing and storage. Cancer
Development Authority. Letters, 114, 71e74.
Patras, A., Brunton, N. P., Tiwari, B. K., & Butler, F. (2009a). Modelling the effect of
different sterilization treatments on antioxidant activity and color of carrot
References slices during storage. Food Chemistry, 114(2), 484e491.
Patras, A., Brunton, N. P., Da Pieve, S., Butler, F., & Downey, G. (2009b). Effect of
Albrecht, J. A., Schafer, H. W., & Zottola, E. A. (1991). Relationship of total sulfur to thermal and high pressure processing on antioxidant activity and instrumental
initial and retained ascorbic acid in selected cruciferous and noncruciferous colour of tomato and carrot purees. Innovative Food Science and Emerging
vegetables. Journal of Food Science, 55, 181e183. Technologies, 10(1), 16e22.
Ames, B. N., Shigenaga, M. K., & Hagen, T. M. (1993). Oxidants, antioxidants, and the Patras, A., Brunton, N. P., Da Pieve, S., & Butler, F. (2009c). Impact of high pressure
degenerative diseases of aging. Proceedings of the National Academy of Sciences processing on antioxidant activity, phenolic, ascorbic acid and anthocyanins of
of the United States of America, 90, 7915e7922. blackberry and strawberry puree. Innovative Food Science and Emerging Tech-
Bahceci, K. S., Serpen, A., Gokmen, V., & Acar, J. (2005). Study of lipoxygenase and nologies, 10(3), 308e313.
peroxidase as indicator enzymes in green beans: change of enzyme activity, Prior, R. L., Cao, G., Martin, A., Sofic, E., McEwen, J., O’Brien, C., et al. (1998). Anti-
ascorbic acid and chlorophylls during frozen storage. Journal of Food Engi- oxidant capacity as influenced by total phenolic and anthocyanin content,
neering, 66, 187e192. maturity, and variety of Vaccinium species. Journal of Agricultural and Food
Barrett, D. M., & Theerakulkait, C. (1995). Quality indicators in blanched, frozen, Chemistry, 46, 2686e2693.
stored vegetables. Food Technology, 49(1), 62e65. Prochaska, L. J., Nguyen, X. T., Donat, N., & Piekutowski, W. V. (2000). Effects of food
Cano, M. P., (1996). Vegetables. In L. E. Jeremiah (Ed.), Freezing effects on food processing on the thermodynamic and nutritive value of foods: literature and
quality. pp. 520. New York: Marcel Dekker. database survey. Medical Hypotheses, 54(2), 254e262.
Cao, G., Sofic, E., & Prior, R. L. (1996). Antioxidant capacity of tea and common Puupponen-Pimia, R., Hakkinen, S. T., Aarni, M., Suortti, T., Lampi, M.-A., Eurola, M.,
vegetables. Journal of Agricultural and Food Chemistry, 44, 3426e3431. et al. (2003). Blanching and long term freezing affect on various bioactive
Chubey, B. B., & Nylund, R. E. (1969). Surface browning in carrots. Canadian Journal compounds of vegetables in different ways. Journal of Science of Food and
of Plant Science, 49, 421e426. Agriculture, 83, 1389e1402.
306 A. Patras et al. / LWT - Food Science and Technology 44 (2011) 299e306

Rawson, A., Koidis, A., Patras, A., Tuohy, M., & Brunton, N. P. (2010). Modelling the Srinivasan, S., Xiong, Y. L., & Blanchard, S. P. (1997). Effects of freezing and thawing
effect of thermal processing on polyacetylene levels in carrot disks. Food methods and storage time on thermal properties of freshwater prawns (Mac-
Chemistry, 121(1), 62e68. robrachium rosenbergii I). Journal of Science and Food Agriculture, 75, 37e44.
Saccani, G., Trifiro, A., Cortesi, A., Gherardi, S., Zanotti, A., & Montanari, A. (2001). Tiwari, B. K., Muthukumarappan, K., O’Donnell, C. P., & Cullen, P. J. (2008). Effects of
Effects of production technology and storage conditions on the content of sonication on the kinetics of orange juice quality parameters. Journal of Agri-
water-soluble vitamins in tomato purees. Industria Conserve, 76, 107e118. cultural and Food Chemistry, 56, 2423e2428.
Sarkar, S. K., & Phan, C. T. (1979). Naturally-occurring and ethylene induced phenolic Vallejo, F., Tomás-Barberán, F. A., & García-Viguera, C. (2002). Glusinolates and
compounds in the carrot root. Journal of Food Protection, 42, 526e534. vitamin C content in edible parts of broccoli florets after domestic cooking.
Sigge, G. O., Hansmanw, C. F., & Joubert, E. (2001). Effect of storage conditions, European Food Research and Technology, 215, 310e316.
packaging material and metabisulphite treatment on the colour of dehydrated Verkerk, R., & Dekker, M. (2004). Glucosinolates and myrosinase activity in red
green bell peppers (Capsicum annuum L.). Journal of Food Quality, 24(3), 205e218. cabbage (Brassica oleracea L. var. capitata f. rubra D.C.) after various microwave
Sikora, E., Cieslik, E., Leszczyn
 ska, T., Filipiak-Florkiewicz, A., & Pawe1, M. P. (2008). treatments. Journal of Agricultural and Food Chemistry, 52, 7318e7323.
The antioxidant activity of selected cruciferous vegetables subjected to aqua- Zacharias, R. (1980). Chilled meals: sensory quality. In G. Glew (Ed.), Advances in
thermal processing. Food Chemistry, 107, 55e59. Catering Technology (pp. 409e416). London, England: Applied Science.
Singelton, V. L., Orthofer, R., & Lamuela-Raventos, R. R. (1999). Analysis of total Zhang, D., & Hamauzu, Y. (2004). Phenolics, ascorbic acid, carotenoids and antiox-
phenols and other oxidation substrates and antioxidants by means of Foline idant activity of broccoli and their changes during conventional and microwave
Ciocalteu reagent. Methods in Enzymology, 299, 152e178. cooking. Food Chemistry, 88, 503e509.