Journal of the Science of Food and Agriculture J Sci Food Agric 83:13891402 (online: 2003)

DOI: 10.1002/jsfa.1589
Blanching and long-term freezing affect
various bioactive compounds of vegetables
in different ways
Riitta Puupponen-Pimi a,
1
Suvi T H akkinen,
1
Marjukka Aarni,
1
Tapani Suortti,
1
Anna-Maija Lampi,
2
Merja Eurola,
3
Vieno Piironen,
2
Anna Maria Nuutila
1
and
Kirsi-Marja Oksman-Caldentey
1
1
VTT Biotechnology, PO Box 1500 (Tietotie), FIN-02044 VTT, Finland
2
Department of Applied Chemistry and Microbiology, PO Box 27 (Latokartanonkaari 11), FIN-00014 University of Helsinki, Finland
3
MTT, Agrifood Research Finland, Chemistry Laboratory, FIN-31600 Jokioinen, Finland
Abstract: An extensive study on the effects of blanching/freezing and long-term freezer storage on various
bioactive compounds of more than 20 commonly used vegetables was performed. Effects were strongly
plant species-dependent. Contents of dietary bre components either were not affected or increased
slightly. Minerals in general were also stable, but some losses of soluble minerals by leaching were
observed. Phenolic antioxidants and vitamins were clearly more sensitive. Signicant losses (2030%) of
antioxidant activity and total phenolics were detected in many vegetables. A qualitative HPLC proling
method for phenolic antioxidants was developed which proved to be very useful when evaluating the
complex behaviour of phenolics during food processing. Up to one-third of vitamin C contents were lost
during blanching, and further slight losses were detected during storage. Folic acid turned out to be very
sensitive to blanching, with more than half of the vitamin being lost, but was stable during freezer storage.
Carotenoids and sterols were not affected by blanching or freezer storage. The usefulness of the applied
screening methods for evaluation of the effects of processing on vegetables is shown.
2003 Society of Chemical Industry
Keywords: processing; blanching; frozen vegetables; phenolics; antioxidants; dietary bre; vitamins; sterols;
carotenoids
INTRODUCTION
Increasing knowledge of the relationship between diet
and health leads to new insights into the effects of
bioactive food components on physiological conditions
and human health. Diet is known to play an important
role in many major diseases of our society, such as
cardiovascular diseases, cancer, hypertension, diabetes
and obesity. The degree to which diet is important in
the prevention of these diseases is not known, but
a commonly accepted estimate is that at least one-
third of cancer cases and perhaps half of the cases
of heart and artery diseases and hypertension are
related to diet. One of the major factors associated
with cardiovascular diseases and cancer is the under-
consumption of bre, vegetables and fruits.
1
Certain bioactive compounds in food plants have
been known for a long time for their benecial effects,
whereas others have only recently been recognised.
Dietary bre, vitamins and minerals, for example,
belong to the rst group. Prebiotic oligosaccharides
are a relatively new concept for modulating colon
microora and thereby also physiological functions
in a positive way.
2
Several different types of
phenolic compounds are synthesised in plants, but
until now they have not been considered necessary
from the nutritional point of view. Plant phenols
possess many benecial biological properties, such
as their antioxidant and antimicrobial properties, and
extensive studies are being carried out on their effects
on human health.
35
Plant sterols are another group
of compounds which are currently being extensively
studied. Recently, plant sterols have been shown to
decrease serum cholesterol in several studies,
6,7
and
they have been suggested also to have a benecial role
in the prevention of colon cancer.
8,9
Their potentially
protective role in the prevention of cardiovascular
diseases had led to great interest in developing
functional foods enriched with plant sterols.
All these various components of plant raw material
offer a good basis for the development of functional

Correspondence to: Riitta Puupponen-Pimi a, VTT Biotechnology, PO Box 1500 (Tietotie), FIN-02044 VTT, Finland
E-mail: riitta.puupponen-pimia@vtt.
Contract/grant sponsor: Tekes
(Received 6 September 2002; revised version received 20 February 2003; accepted 4 August 2003)
2003 Society of Chemical Industry. J Sci Food Agric 00225142/2003/$30.00 1389
R Puupponen-Pimi a et al
foods and ingredients. However, food processing has a
conclusive role in the preservation and bioavailability
of these compounds. Relatively little has been reported
about the concurrent changes in different kinds of
bioactive compounds during processing. Most of the
work so far has been carried out on single compounds
such as vitamins. Nevertheless, it is most probable
that the benecial health effects attributed to fruits
and vegetables are a result of combined effects of
several compounds.
The aims of the present research were to exten-
sively elucidate the occurrence of various bioactive
compounds in vegetables used by the food industry
and to study their preservation during processing and
storage. The production of frozen vegetables and their
subsequent freezer storage were chosen as an example.
The intention was to screen various types of phyto-
chemicals, namely dietary bre components, minerals,
vitamins, phenolic antioxidants and sterols, in order to
nd out which compounds are sensitive to processing,
and to develop screening methods which could be used
as indicators of sensitivity for food processing when
developing plant-based functional foods. Our research
material consisted of more than 20 commonly used
vegetables.
MATERIALS AND METHODS
Vegetable material and sample preparation
This study was carried out in close co-operation
with the Finnish food industry. L annen Tehtaat Plc
supplied all vegetable and process samples. Depending
on the vegetable, different pretreatments were carried
out before blanching. Briey, peas were mechanically
podded already in the eld. Carrots were peeled and
cut into cubes (10 10 10mm
3
) or slices (4.5 mm).
Cauliowers were cut and inorescences (2540mm)
were used in the study. Broccoli (inorescences,
2040mm) was an imported product (from Ecuador)
and was already blanched and frozen when it arrived
at the factory. Cabbages were cut into slices (8 mm).
Spinach leaves were used as such. Potatoes were either
used shortly after harvesting (potato 1) or stored in the
farmers cold store (about 5

C) for 3 months before

processing (potato 2). Potatoes were peeled and cut
into slices (4.5mm). Swedes were peeled and cut into
cubes (10 10 10mm
3
). After these pretreatments,
defective vegetable pieces were removed by colour
sorter. Samples were taken before blanching (fresh
samples). The industrial blanching/freezing procedure
differed slightly depending on the vegetable in
question. In general, vegetables were blanched for
a few minutes in water and steam and cooled using
either water or air. The following blanching conditions
were used: pea, 95

C, 2min; cauliower, cabbage,

spinach and potato, 96

C, 3min; carrot, 97

C,
3 min; swede, 99

C, 4min. All vegetables except

potatoes and swedes were water cooled. Both cooling
methods were used for carrots. The cooling time for
all vegetables was less than 5min. Spinach leaves were
nely ground (4mm) after blanching and cooling.
Blanching conditions for broccoli were not available.
After blanching and cooling, vegetables were quickly
frozen to 40

C and samples were taken (process

samples). Frozen vegetables were stored in the factory
freezing room at 20

C and samples were taken after

6, 12 and 18months (storage samples). All samples
were taken as 2 1kg portions.
Analysis of bioactive compounds
Before analyses, all samples were freeze-dried and
ground to a ne powder using a Rotor-Speed Mill
(Fritsch GmbH, Idar-Oberstein, Germany), with a
0.5mm sieve ring. Only vitamin C and carotenoids
were analysed directly from frozen samples. Bioactive
compounds were analysed at least in duplicate.
Moisture content was determined by gravimetry as
the mass loss of a 5g sample at 105

C. Residual
moisture was determined by the Karl Fischer titrimet-
ric method using a Mettler DL 18 Titrator (Mettler
Instruments, Greifensee, Switzerland) according to
the manufacturers instructions.
Dietary bre components
Contents of water-insoluble and water-soluble dietary
bre were determined by the enzymatic/gravimetric
method of Asp et al.
10
The digestive enzymes used
were -amylase (pH6.0, 95

C, 15min), pepsin
(pH1.5, 40

C, 60min) and pancreatin (pH6.8,

40

C, 60min). For the analysis, 0.30.6 g of sample

was taken.
For the determination of pentosan content, samples
were hydrolysed with sulphuric acid as described by
Karppinen et al,
11
and arabinose and xylose contents
were quantitated by high-performance anion exchange
chromatography (Dionex) with pulsed amperometric
detection. Pentosan content was calculated from
arabinose and xylose contents as follows: pentosan =
0.88 (arabinose +xylose).
Pectin content was determined, after hot water
extraction, by the spectrophotometric m-phenylphenol
method of List et al.
12
The method was modied
as described by Carbonell et al.
13
Briey, 1015mg
of sample was suspended in water and digested
in a boiling water bath for 10min. After cooling
and ltering, sodium tetraborate (0.0125M) in
concentrated sulphuric acid was added, and the
mixture was kept for 10min in a boiling water bath.
After cooling, m-phenylphenol was added and the
colour was allowed to develop for 5min, after which
the absorbance was measured in a spectrophotometer
at 520nm.
Contents of fructo-oligosaccharides were deter-
mined by a specic enzymatic kit (Megazyme, Bray
Business Park, Bray, Co Wicklow, Ireland). Briey,
sucrose was rst hydrolysed by sucrase, and starch
and maltosaccharides were hydrolysed by -amylase,
pullulanase and maltase. The resultant reducing sug-
ars were then reduced to sugar alcohols by treat-
ment with alkaline borohydride. Fructan was hydrol-
ysed to fructose and glucose by fructanase, and
1390 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
reducing sugars were determined by the PAHBAH
(p-hydroxybenzoic acid hydrazide) reducing sugar
method. For pea and potato samples, inclusion of
Aspergillus niger -galactosidase was used in order to
remove rafnose series oligosaccharides.
Vitamins and carotenoids
Folates were extracted fromfreeze-dried plant material
using a modied method of Tamura.
14
The folates
were extracted from the food matrix by autoclaving
(121

C, 5min). Ascorbic acid and 2-mercaptoethanol

were used as antioxidants. To degrade conjugated
folates, conjugase and amylase enzymes were added.
Before incubation in the dark (30

C, 16h) the samples

were vortexed and ushed under nitrogen. After
incubation the enzymes were inactivated by boiling
and the samples were ltered. Folic acid contents
were determined microbiologically by a modied
method of Molloy and Scott.
15
Briey, the extracts
as well as the medium inoculated with bacteria
were pipetted into 96-well plates, and the growth
of Lactobacillus rhamnosus NCIMB 10463 in sample
wells was compared with that in folic acid standard
wells. The sample extracts were diluted with 0.5%
sodium ascorbate, and 100l of a suitable dilution
of sample or standard (050pg) was added to each
well. Liquid medium inoculated with cryopreserved
bacteria was pipetped into each well, resulting in 250l
total volume per well. The growth was determined
spectrophotometrically at 595nm after anaerobic
incubation (37

C, 42h, dark). Each sample was

analysed in three replicates.
Vitamin C was determined as dehydroascorbic acid
according to the method of Speek et al.
16
Vitamin C
was analysed using an HP1090 Series HPLC(Hewlett
Packard, Waldbronn, Germany) equipped with a
uorescent detector. The analytical column was
a Waters Spherisorb column (125mm4.0mmid,
5 m; Waters, Taunton, MA, USA) operating at 35

C.
The isocratic mobile phase consisted of methanol and
0.08 M phosphate buffer (pH7.8).
Contents of - and -carotene were determined
according to H agg et al.
17
Briey, the method con-
sisted of acetone extraction, ltration and concen-
tration followed by liquid/liquid partitioning with
hexane/diethyl ether (7:3). After evaporation of the
organic phase, samples were dissolved in acetoni-
trile/dichloromethane/methanol (7:2:1). Determina-
tion of the analytes was accomplished using an
HP 1090 Series HPLC equipped with a diode
array detector set at 410nm. A 201TP54 col-
umn (250mm4.4mmid, 5m, Vydac, Hesperia,
CA, USA) was used with acetonitrile/methanol (1:9)
as the mobile phase.
Radical-scavenging activity
The antioxidant activity of methanolic extracts
(50mg ml
1
, freeze-dried material) was measured
using a modied DPPH (1,1-diphenyl-2-picryl-
hydrazyl) radical-scavenging method
18
in microw-
ell plates. To each well, 200l of DPPH solution
(0.23mM) and 30l of extract were added. After
5min of incubation the absorbance was measured at
515nm. Results were expressed as DPPHindex values
obtained after comparison with pyrogallol. Duplicate
extracts were prepared and four replicate measure-
ments were performed for every sample.
Electrochemically active compounds (qualitative screening
method)
Relative amounts of electrochemically active com-
pounds such as phenolics were determined using a
rapid HPLC screening method. Freeze-dried samples
were extracted with either water or methanol contain-
ing 0.1% (v/v) of 85% phosphoric acid. Samples were
analysed using a reverse phase, 150mm3.9mmid,
5m C
18
Symmetry column (Waters). The linear gra-
dient programme (15min, from 100% A to 100% B)
consisted of water with 1% acetic acid (solvent A)
and methanol with 1% acetic acid (solvent B). An
electrochemical detector (Waters M464) with a glassy
carbon electrode was used. The oxidation potential
of the detector was 0.8V. An additional diode array
detector (Waters M996) was used in the measure-
ment range 220350nm. An electrochemically active
gradient prole was obtained.
Phenolic compounds
Total phenolics in methanolic extracts (50mg ml
1
,
freeze-dried material) were analysed spectropho-
tometrically by the FolinCiocalteu procedure.
19
Briey, 100l of suitably diluted sample, 100l of
FolinCiocalteu reagent and 700l of 20% (w/v)
Na
2
CO
3
were mixed and centrifuged (14 000rpm,
3min; Microlitre centrifuge, Hermle Z231M,
Gosheim, Germany). After 20min of incubation in
the dark the absorbance was measured at 735nm. The
total phenolic content was expressed as mg gallic acid
equivalents (GAE) g
1
dry material. From each sam-
ple, two methanolic extracts were prepared, and for
each extract, three replicate analyses were performed.
Quercetin and kaempferol contents of vegetables
were determined according to Nuutila et al.
20
Briey,
freeze-dried and ground material (50mg) was hydrol-
ysed in 5ml of 1.2 M HCl in 50% aqueous methanol.
To the hydrolysis mixture, 2 mg of ascorbic acid was
added. After reuxing (80

C, 2h), samples were son-

icated and ltered through a 0.45m lter for organic
solvent. Samples were analysed using a Waters HPLC
system. Reverse phase separations were carried out
at room temperature using a 150mm3.9mmid,
5m C
18
Symmetry column (Waters) tted with a
20mm3.9mmid, 5 m C
18
Symmetry guard col-
umn (Waters). The mobile phase was a 25min,
2060% gradient of methanol in water with 300l
l
1
triuoroacetic acid, eluted at a ow rate of 0.8ml
min
1
. The eluted components were monitored at
280 and 340nm using a dual-wavelength absorbance
detector (Waters 2487).
J Sci Food Agric 83:13891402 (online: 2003) 1391
R Puupponen-Pimi a et al
Sterols
Plant sterols were analysed by gas chromatography
using a method developed by Toivo et al
21
and
modied by Piironen et al.
22
The sample preparation
method used included both acid and alkaline
hydrolysis to liberate sterols from their derivatives and
from the freeze-dried food matrix, purication of the
unsaponiable matter by silica solid phase extraction,
and derivatisation to trimethylsilyl ethers. All sample
materials were analysed in triplicate. When analysing
spinach samples, acid hydrolysis was omitted, because
spinach contains mainly
7
-unsaturated sterols which
readily decompose under acidic conditions.
Minerals
Mineral and trace elements were determined by induc-
tively coupled plasma (ICP) emission spectrometry.
Freeze-dried samples (0.52g) were digested in con-
centrated nitric acid (pa Baker) on an aluminium
heating block, diluted to 50ml with Milli-Q puri-
ed water and ltered. The entire digestion proce-
dure is described by Tahvonen and Kumpulainen.
23
Elemental concentrations were measured in a high-
resolution ICP emission spectrometer (Thermo Jarrel
Ash, IRIS Advantage Franklin, MA, USA). Measure-
ment conditions were: RF power 1150W, auxiliary
ow 0.5 l min
1
, nebuliser ow 0.57l min
1
, pump
rate 200rpm, ush time 40s, integration time 10s,
replicates 2. The accuracy of the analytical method
was tested by determining certied reference materials
in every batch of samples. The method is accredited
for all the elements studied except sodium.
RESULTS
In this study, comprehensive data on the behaviour
of various bioactive compounds of vegetables during
food processing and storage were accumulated over
a period in excess of 4years. In the following the
most interesting results concerning the production
of frozen vegetables and long-term freezer storage
are presented. Since the contents and behaviour of
bioactive compounds varied greately in different types
of vegetables, the results are grouped according to
different vegetable species.
Dietary bre components and minerals
Contents of dietary bre components varied greatly
within the studied vegetables (Tables 14). Variety
and harvest year seemed to have a marked effect on
soluble bre, as seen for peas in Table 1. During
the harvest years 1999 and 2000 the variation in
soluble bre was 3.75.2g per 100g dry weight
(dw) (data not shown). The soluble bre content
of eg the variety Snake was 5.2g per 100g dw in the
year 2000 compared with 1.4 g per 100g dw in the
year 1998. The mineral prole was also highly plant
species-dependent. Spinach was the richest source of
all minerals studied (Tables 14).
Effects of processing and storage were strongly
plant species-dependent. In general, dietary bre
components seemed to be rather stable during
blanching and freezer storage. Either they were not
affected by this processing or their contents increased
slightly. Pentosans and pectins behaved in much the
same way as dietary bre. Fructo-oligosaccharides
and minerals were not affected by blanching and
freezer storage. However, potassium contents were
often decreased during blanching, especially in leafy
vegetables. Because of the stable nature of dietary bre
components as well as most minerals, and in order to
save space, effects of freezer storage were studied only
for a few vegetables.
Table 1. Contents of dietary bre components and minerals in peas (dry weight basis)
Sample
Soluble
bre
Insoluble
bre
Total
dietary
bre Pentosans Pectins
Fructo-oligo-
saccharides Ca Mg K P Na Cu Mn Fe Zn
Bikini, 1997
Fresh 5.2 20.7 25.9 3.7 1.3 1.7 1.0 1.4 11.3 4.6 0.04 7.4 15.9 71.2 41.5
Processed 4.8 21.5 26.3 3.8 0.7 1.4 1.1 1.3 8.8 4.3 0.03 6.1 15.8 71.1 37.3
Stored 6months 3.8 1.2 1.7 1.1 1.4 9.0 4.5 0.06 6.4 15.1 71.0 39.5
Bikini, 1998
Fresh 1.2 21.3 22.5 3.9 1.9 0.9 1.2 1.6 11.3 5.4 0.05 8.9 10.4 62.4 31.6
Processed 1.2 21.7 22.9 3.8 1.6 0.8 1.2 1.4 8.6 5.0 0.05 6.7 10.3 46.9 28.8
Stored 6months 3.7 1.6 1.5 1.2 1.5 9.1 5.1 0.02 7.3 10.2 78.0 30.6
Avola, 1998
Fresh 0.8 23.1 23.9 4.1 1.1 1.8 1.2 1.3 11.5 5.5 0.03 7.3 12.7 60.0 31.4
Processed 1.7 22.9 24.6 4.0 1.2 1.8 1.4 1.4 9.6 5.4 0.05 6.7 13.6 66.3 31.0
Stored 6months 3.7 0.9 1.8 1.3 1.4 9.7 5.5 0.02 6.5 12.8 32.6
Snake, 1998
Fresh 1.4 18.6 20.0 3.7 1.8 1.0 0.7 1.1 11.0 4.4 0.05 5.3 9.7 68.6 29.9
Processed 2.7 22.1 24.8 5.0 1.6 1.1 0.9 1.1 9.1 4.2 0.09 10.1 84.6
Stored 6months 4.7 1.9 1.8 0.8 1.1 9.2 4.2 0.01 4.4 9.8 93.1 28.1
Dietary bre components in g per 100 g; Ca, Mg, K, P, Na in g kg
1
; Cu, Mn, Fe, Zn in mgkg
1
; , not determined.
1392 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
Table 2. Contents of dietary bre components and minerals in carrots (dry weight basis)
Sample
Soluble
bre
Insoluble
bre
Total die-
tary bre
Pento-
sans Pectins
Fructo-oligo-
saccharides Ca Mg K P Na Cu Mn Fe Zn
Cubes, water cooling
Fresh, 1997 11.4 12.3 23.7 2.0 2.1 1.0 1.9 1.0 16.7 2.2 1.7 3.6 16.9 38.4 24.0
Processed 14.1 17.6 31.7 3.1 2.8 1.0 2.8 1.2 15.8 2.5 2.1 3.0 22.9 42.0 22.2
Stored 6months 2.7 1.2 15.9 2.5 1.9 2.8 25.9 40.0 23.4
Stored 12months 2.7 1.3 16.3 2.5 2.3 3.4 23.0 35.0 24.6
Cubes, air cooling
Fresh 11.8 13.4 25.2 2.4 2.9 1.3 2.2 1.1 14.0 2.6 1.6 2.8 12.4 24.8 17.5
Processed 12.2 15.6 27.8 2.8 3.8 0.9 2.5 1.1 15.4 2.7 1.6 3.0 15.7 27.3 11.8
Stored 6months 2.6 2.6 1.1 15.1 2.7 1.7 3.1 14.7 25.5 12.9
Stored 12months 2.5 1.2 13.6 2.5 1.6 2.7 12.7 25.7 9.2
Slices, air cooling
Fresh 11.9 12.3 24.2 2.2 2.0 1.0 1.9 1.1 20.8 2.7 2.5 3.7 22.5 35.6 27.3
Processed 14.6 15.9 30.5 3.1 3.1 1.2 2.4 1.2 19.7 2.8 2.3 3.2 26.8 41.0 27.0
Stored 6months 2.3 1.1 18.9 2.9 2.6 3.3 25.7 39.8 24.5
Stored 12months 2.3 1.1 20.3 2.8 2.4 3.5 24.1 37.1 24.2
Cubes, air cooling
Fresh, 1998 12.0 12.1 24.1 2.5 1.8 1.1 2.0 0.9 20.8 2.5 2.5 2.5 29.7 20.4 19.8
Processed 13.4 13.6 27.0 3.1 1.8 1.2 1.8 1.0 19.1 2.6 0.9 2.3 60.9 14.7 25.2
Stored 6months 3.4 2.1 0.9 1.8 0.9 19.2 2.4 0.9 1.9 55.9 24.4 23.7
Stored 12months 1.8 1.1 22.0 2.7 1.1 2.1 59.8 19.8 31.4
Yellow carrot
Slices, air cooling
Fresh 12.1 11.5 23.6 3.0 2.4 0.8 2.5 1.1 20.5 1.8 1.7 4.7 13.0 28.3 14.3
Processed 12.1 12.1 24.2 3.3 2.7 0.9 3.1 1.0 15.4 2.1 1.5 2.9 13.3 21.8 14.2
Stored 6months 3.3 3.2 0.7 2.8 1.1 18.3 1.9 1.8 2.6 12.5 26.7 15.6
Stored 12months 2.9 1.2 19.7 1.9 1.8 2.7 11.8 23.1 18.0
Dietary bre components in g per 100 g; Ca, Mg, K, P, Na in gkg
1
; Cu, Mn, Fe, Zn in mg kg
1
; , not determined.
Table 3. Contents of dietary bre components and minerals in brassicas (dry weight basis)
Sample
Soluble
bre
Insoluble
bre
Total die-
tary bre
Pento-
sans Pectins
Fructo-oligo-
saccharides Ca Mg K P Na Cu Mn Fe Zn
Cauliower
Fresh 6.7 23.5 30.2 4.5 1.7 0.5 2.8 1.9 40.8 6.4 0.8 2.7 19.6 45.7 35.8
Processed 7.6 25.7 33.3 4.4 1.7 0.6 2.8 1.7 34.3 5.6 0.7 3.1 14.8 43.2 25.2
Stored 6months 2.7 1.7 32.4 5.7 0.7 2.2 16.5 48.1 29.9
Stored 12months 2.9 1.6 32.4 4.9 0.6 2.3 12.4 38.0 21.1
Broccoli
Processed 9.6 23.4 33.0 5.0 1.1 0.4 2.8 2.2 25.9 6.3 0.7 3.4 17.1 76.8 35.1
Stored 6months 2.8 2.1 25.6 6.1 0.7 3.3 15.8 74.6 31.8
Stored 12months 2.9 2.2 24.9 6.1 0.8 3.5 17.2 76.0 30.8
Cabbage
Fresh 5.4 20.8 26.2 4.0 0.7 0.9
Processed 8.3 24.5 32.8 5.0 1.0 0.8 4.6 1.3 24.2 3.2 0.2 2.7 9.8 23.5 9.4
Stored 6months 4.4 1.2 23.9 3.1 0.2 2.0 9.9 23.4 9.4
Stored 12months 4.1 1.2 23.1 2.9 0.2 2.2 9.0 23.6 8.2
Dietary bre components in g per 100 g; Ca, Mg, K, P, Na in g kg
1
; Cu, Mn, Fe, Zn in mgkg
1
; , not determined.
Peas
Dietary bre components either were not affected by
blanching or increased slightly. Signicant increases in
soluble, insoluble and total dietary bre were noticed
only with the variety Snake. Soluble bre almost dou-
bled, while insoluble and total dietary bre increased
by about 20%. Double the amount of soluble bre was
also measured in the Avola variety after blanching.
Pentosans, pectins and fructo-oligosaccharides were
also stable during processing.
Carrots
Carrot was a good source of soluble bre (more than
11g per 100g dw), which constitutes almost half of the
total dietary bre in carrots. Dietary bre components
of carrots were clearly affected by blanching. Soluble,
J Sci Food Agric 83:13891402 (online: 2003) 1393
R Puupponen-Pimi a et al
Table 4. Contents of dietary bre components and minerals in spinach, potatoes and swede (dry weight basis)
sample
Soluble
bre
Insoluble
bre
Total
dietary
bre
Pento-
sans Pectins
Fructo-oligo-
saccharides Ca Mg K P Na Cu Mn Fe Zn
Spinach
Fresh, leaves 6.5 29.4 35.9 2.4 1.7 0.7 24.5 6.8 56.2 6.3 5.0 11 50 113.7 33.0
Processed 5.7 33.0 38.7 3.2 2.1 0.6 26.7 5.0 35.9 5.5 3.0 11 38 120.0 37.0
Stored 6months 28.1 5.4 38.2 5.8 3.3 12 47 126.9 35.0
Stored 12months 26.8 4.7 35.1 5.5 2.9 12 38 126.7 30.0
Potato 1
Fresh, slices 2.9 2.6 5.5 0.3 0.5 0.6 0.1 1.1 17.6 2.1 ND 3.5 7.1 16.2 5.4
Processed 1.5 3.0 4.5 0.3 0.9 0.8 0.1 1.0 15.6 1.9 ND 3.2 6.4 15.9 4.6
Stored 6months 0.1 1.1 15.8 1.9 ND 3.5 7.1 17.6 6.3
Stored 12months 0.1 1.0 15.1 1.8 ND 3.6 6.7 16.7 5.9
Potato 2
Fresh, slices 2.1 2.7 4.7 0.3 0.3 0.7 0.2 1.1 17.3 2.0 ND 3.0 7.7 20.0 6.9
Processed 2.4 2.9 5.3 0.4 0.3 0.7 0.1 1.0 13.9 1.8 0.1 2.8 7.8 19.5 6.3
Stored 6months 0.2 0.9 13.8 1.8 ND 3.0 7.0 17.9 6.5
Stored 12months 0.1 1.0 15.6 2.9 ND 3.0 7.3 16.8 6.8
Swede
Fresh, cubes 9.3 15.3 24.6 3.2 2.2 1.8 4.1 1.0 24.0 4.0 0.9 1.6 5.1 25.4 7.6
Processed 12.1 19.1 31.2 3.9 3.4 1.8 4.8 1.1 19.7 3.9 0.7 1.7 5.9 29.3 7.3
Dietary bre components in g per 100 g; Ca, Mg, K, P, Na in g kg
1
; Cu, Mn, Fe, Zn in mgkg
1
; , not determined; ND, below detection limit.
insoluble and total dietary bre components increased
during processing, as did pentosans. The increase
was highest in cubes with water cooling. This can be
explained by the better washing out of small molecules
and concentration of bre components in water-cooled
samples compared with air-cooled samples. In water-
cooled samples the increases were 24% in soluble
bre, 43% in insoluble bre and 34% in total dietary
bre. Some increase in pentosans and pectins was also
noticed. Fructo-oligosaccharides were not affected by
blanching.
Brassicas
Different brassicas behaved in different ways when
blanched. Contents of dietary bre components in
cauliower increased slightly. Pentosans, pectins and
fructo-oligosaccharides were not affected. Soluble,
insoluble and total dietary bre contents of cabbage
increased signicantly. The increases were 54% in
soluble bre, 18% in insoluble bre and 25% in total
dietary bre. Pentosans and pectins also increased
slightly. Fructo-oligosaccharides were not affected.
Spinach
Soluble, insoluble and total dietary bre contents
were not changed signicantly by processing. Pen-
tosans and pectins, however, increased by 33 and
24% respectively. Fructo-oligosaccharides were not
affected. Almost 40% of potassium was lost during
blanching.
Potatoes
Freshly processed potatoes (potato 1) contained
similar amounts of dietary bre components as
potatoes stored at 5

C for 3months (potato 2).

Dietary bre components were not appreciably
changed by blanching in either case.
Swede
Soluble bre increased by 30%, insoluble bre by
25% and total dietary ber by 27% during blanching.
Pentosans increased slightly and the amount of pectins
doubled. Fructo-oligosaccharides were not affected.
Antioxidants, vitamins and sterols
There was a great variation in concentrations
of bioactive compounds in different vegetables
(Tables 58, Fig 1). The lowest folic acid content
was 39g per 100g fresh weight (fw) (cabbage) and
the highest 176g per100g fw (spinach). Vitamin C
contents varied from 5.5 to 81mg per 100g fw (carrot
and cauliower respectively). -Carotene contents
varied from 0.3 to 11mg per 100g fw (pea and carrot
respectively). -Carotene was found only in carrots,
in concentrations between 2 and 4 mg per 100g fw.
The lowest amount of total phenolics was 0.5 mg GAE
g
1
dw (potato) and the highest 7.2 mg GAE g
1
dw
(spinach). Potatoes had the lowest DPPH index,
around 10, and cauliower the highest, around 200.
Quercetin and kaempferol were quantied in spinach
and brassicas. Kaempferol was the main avonol
found in green vegetables such as spinach. Fresh
spinach contained the highest amounts of quercetin
and kaempferol, about 800 and 3000mg kg
1
dw
respectively (Fig 1). Cauliower and broccoli were
found to be very good sources of sterols, with about
500mg per 100g dw.
In general, vitamins and antioxidants were clearly
more sensitive than dietary bre components to
processing and storage. In many cases, signicant
1394 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
Table 5. Contents of antioxidants, vitamins and sterols in peas
Sample
Folic acid
(g per 100g
fresh weight)
Vitamin C
(mg per 100 g
fresh weight)
-carotene
(mg per 100 g
fresh weight)
Total phenolics
(mg GAE g
1
dry weight)
DPPH
index
Sterols (mg
per 100 g
dry weight)
Bikini, 1997
Fresh 110 30 0.6 0.9 12 119
Processed 74 22 0.4 0.7 8 150
Stored 6months 63 23 0.2 0.7 8 146
Stored 12months 63 21 0.4 0.6 7 140
Bikini, 1998
Fresh 93 23 0.3 0.8 18 119
Processed 89 22 0.4 0.6 12 150
Stored 6months 81 19 0.4 0.6 11
Stored 12months 77 20 0.3 0.6 11
Stored 18months 82 22 0.3 0.6 8
Avola, 1998
Fresh 109 22 0.3 0.8 16 119
Processed 78 22 0.4 0.7 13 144
Stored 6months 73 20 0.4 0.7 10
Stored 12months 84 20 0.4 0.7 7
Stored 18months 81 21 0.3 0.7 8
Snake, 1998
Fresh 103 33 0.4 1.2 12 114
Processed 67 26 0.3 0.9 8 140
Stored 6months 57 22 0.4 0.9 6
Stored 12months 62 23 0.3 0.9 8
Stored 18months 66 24 0.3 0.8 7
, not determined.
Table 6. Contents of antioxidants, vitamins and sterols in carrots
Sample
Vitamin C (mg
per 100 g
fresh weight)
-Carotene
(mg per 100 g
fresh weight)
-Carotene
(mg per 100 g
fresh weight)
Total phenolics
(mg GAE g
1
dry weight) DPPH index
Sterols (mg per
100 g dry
weight)
Cubes, water cooling
Fresh, 1997 5.5 11.0 3.6 1.2 23
Processed 4.0 11.8 4.2 1.2 24
Stored 6months 4.0 10.0 3.2 1.0 22
Stored 12months 3.9 3.0 1.8 1.0 22
Cubes, air cooling
Fresh 5.8 10.7 3.5 1.3 22 168
Processed 4.7 11.3 3.9 1.3 23 192
Stored 6months 3.8 9.0 2.9 1.2 24 174
Stored 12 months 4.4 1.1 23 166
Slices, air cooling
Fresh 6.1 9.8 4.4 1.1 18
Processed 4.4 10.6 4.9 1.1 23
Stored 6months 4.3 10.7 3.9 1.1 27 172
Stored 12months 5.1 3.1 1.8 1.2 26
Cubes, air cooling
Fresh, 1998 6.1 5.6 2.0 1.2 29
Processed 4.5 10.1 3.2 0.8 20 224
Stored 6months 5.3 5.1 2.2 1.0 24
Stored 12months 4.7 7.4 2.5 1.0 25
Stored 18months 4.5 6.6 2.7 1.0 26
Yellow carrot
Slices, air cooling
Fresh 9.6 0.4 ND 0.8 23 181
Processed 8.5 0.7 ND 0.8 31 201
Stored 6months 9.2 0.4 ND 0.7 31
Stored 12 months 7.1 0.4 ND 0.8 33
Stored 18 months 8.0 0.4 ND 0.7 19
, not determined; ND, below detection limit.
J Sci Food Agric 83:13891402 (online: 2003) 1395
R Puupponen-Pimi a et al
Table 7. Contents of antioxidants, vitamins and sterols in brassicas
Sample
Folic acid (g
per 100 g
fresh weight)
Vitamin C (mg
per 100 g
fresh weight)
-Carotene
(mg per 100 g
fresh weight)
Total phenolics
(mg GAE g
1
dry weight)
DPPH
index
Quercetin/kaempferol
(mgkg
1
dry weight)
Sterols (mg
per 100 g
dry weight)
Cauliower
Fresh, inorescences 122 81 ND 5.6 202 70/25 526
Processed 73 68 ND 4.9 155 190/75 489
Stored 6months 73 66 ND 4.8 162 125/30 499
Stored 12months 66 ND 4.5 156 85/15 505
Broccoli
Processed 67 83 0.5 3.2 63 370/340
Stored 6months 68 80 0.5 3.6 60 280/330 483
Stored 12months 69 3.1 57 100/105 465
Cabbage
Fresh 39 43 ND 1.9 67 12/ND 137
Processed 15 30 ND 2.4 73 21/ND 159
Stored 6months 13 26 ND 2.0 66 17/ND 171
Stored 12 months 13 21 ND 1.9 70 19/ND 153
, not determined; ND, below detection limit.
Table 8. Contents of antioxidants, vitamins and sterols in potatoes and swede
Sample
Vitamin C
(mg per 100 g
fresh weight)
-Carotene
(mg per 100 g
fresh weight)
Total phenolics
(mg GAE g
1
dry weight)
DPPH
index
Quercetin/
kaempferol (mg
kg
1
(dry weight)
Sterols
(mg per 100 g
dry weight)
Potato 1
Fresh 23 ND 0.5 10 ND 12
Processed 17 ND 0.3 6 ND 9
Stored 6months 15 ND 0.4 12 ND
Stored 12months 13 ND 0.5 13 ND
Potato 2
Fresh 15 ND 0.6 10 ND 7.3
Processed 11 ND 0.5 9 ND 7.3
Stored 6months 9 ND 0.4 10 ND
Stored 12months 9 ND 0.4 10 ND
Swede
Fresh 44
a
ND 3.2 123 171
Processed 31
a
ND 2.5 89 183
Stored 6months 25 ND 2.2 98
Stored 12months 22 ND 2.5 75
, not determined; ND. below detection limit.
a
Reference material.
losses were detected. However, in some vegetables,
even higher concentrations of bioactive components,
such as carotenoids and sterols were measured after
processing.
Vitamin C losses were strongly plant species-
dependent. Highest losses during blanching were
around 30%. Storage further decreased vitamin C
contents slightly. Folic acid was very sensitive to
blanching. In almost every vegetable, at least half
of this vitamin was lost during blanching. However,
folic acid was stable during freezer storage. Carotene
contents increased during blanching, then decreased
gradually during storage.
Total phenolics of most vegetables correlated
positively with radical-scavenging activity, indicat-
ing that our radical-scavenging assay measured
especially antioxidant activity connected with phe-
nolic compounds. The amount of total phe-
nolics and radical-scavenging activity decreased
during blanching by 2030%. Storage further
decreased phenolic contents and DPPH activity
slightly.
The HPLC prole of electrochemically active
compounds such as phenolic antioxidants often
changed during processing and storage, showing the
overall changes in these compounds. Peak heights
correlating positively with total phenolics and DPPH
activity could be detected in many chromatograms.
Thus the HPLC proling method developed in this
study proved to be a sensitive indicator for the effects
of processing on phenolic compounds. The method
could also be applied directly to industrial processes.
1396 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
0
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Rad. scav. activity
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EC-detector
Time (min)
stored 6 months in freezer
processed
fresh
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2
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Figure 1. Effects of blanching and freezer storage on various bioactive compounds of spinach: (a) prole of electrochemically active compounds;
(b) radical-scavenging activity and total phenolics; (c) vitamin C; (d) folic acid; (e) -carotene; (f) sterols; (g) quercetin and kaempferol.
Effects of processing and storage on avonols were
highly plant species-dependent. In cauliower and
cabbage, avonol contents increased during blanch-
ing, then decreased during storage (Table 7). How-
ever, in spinach, avonol contents decreased during
blanching and continued to decrease slightly during
storage.
Blanching did not decrease sterol contents of
vegetables. In many cases, higher concentrations were
measured after blanching. Sterols were also stable
during freezer storage.
Peas
Folic acid contents of peas decreased during blanch-
ing. Losses were between 12 and 35%. Freezer storage
did not have any further effects. Vitamin C contents
decreased in those pea samples in which the origi-
nal vitamin content was high, 30mg per 100g fw or
more. Losses were about 2030%. However, when
the original vitamin content was lower, 20mg per
100g fw, no losses were detected during process-
ing. -Carotene was either decreased slightly or not
affected by blanching and storage. The amount of total
J Sci Food Agric 83:13891402 (online: 2003) 1397
R Puupponen-Pimi a et al
phenolics and radical-scavenging activity decreased by
2030% during blanching. Freezer storage further
decreased phenolic contents and antioxidant activ-
ity slightly. Sterol contents were 2030% higher after
blanching. Slightly lower contents were measured after
long-term storage.
Carrots
Vitamin C contents decreased by around 30% during
blanching. No signicant further losses were detected
during storage. When comparing water- and air-
cooled samples, bigger losses where found in water-
cooled samples. Carotene contents increased during
blanching, then decreased slightly during storage. It
was interesting to observe that when carrot cubes and
slices (harvest year 1997) were stored frozen for 1 year,
carotene contents fell dramatically compared with
8 months of storage. However, such a decrease was not
found in the next harvest year (1998), when carotene
contents in the raw material were already much
lower. The amount of total phenolics and radical-
scavenging activity were not affected by blanching
and storage. Sterol contents increased slightly after
blanching. Somewhat lower contents were measured
after long-term storage.
Brassicas
Folic acid contents of cauliower and cabbage
decreased by 4060% during blanching, but no
further during storage. Vitamin C contents of
these vegetables decreased by around 2030%
during blanching. Slight losses were also detected
during storage. Total phenolics and DPPH index
of cauliower decreased slightly during blanching,
but no further during storage. Such a decrease after
blanching was not found in cabbage. Flavonol contents
of cauliower and cabbage doubled during processing,
then decreased slowly during storage. Sterol contents
of these vegetables were not signicantly affected by
blanching and storage.
In order to get an estimation of how broccoli
phenolics behave during blanching, some laboratory-
scale blanching experiments (2min in boiling water)
were carried out with another reference material (data
not shown). In these experiments, clear decreases
in total phenolics (by 37%) and DPPH index (by
37%) were detected. Also, avonol contents decreased
signicantly (quercetin by 40% and kaempferol by
70%). In industrial samples, folic acid, vitamin C, -
carotene, total phenolics, radical-scavenging activity
and sterols were not affected by freezer storage.
However, avonol contents of broccoli decreased
considerably during long-term storage.
Spinach
Spinach was very sensitive to blanching, with 70%
of folic acid being lost. Storage did not cause
any extra losses. Vitamin C contents of spinach
decreased during processing by about 30% and during
storage by a further 30%. -Carotene increased
slightly during blanching. Eight months of storage did
not further decrease concentrations. Total phenolics
and DPPH index decreased by about 30% during
blanching of spinach. Storage did not further decrease
them. Quercetin and kaempferol contents of spinach
decreased gradually during blanching and storage,
by about 30% overall. The HPLC prole of
electrochemically active components such as phenolic
antioxidants showed that some of these compounds
(methanol extract) were signicantly affected after
6months of storage. About 20%higher sterol contents
were measured after blanching. Storage did not have
any further effects. Effects of processing and storage
on spinach are shown in Fig 1.
Potatoes
Potatoes stored at 5

C for 3 months (potato 2)

contained 35% less vitamin C than fresh potatoes
(potato 1). Vitamin C contents decreased by about
25% during blanching in both potatoes. Storage
further decreased the contents slightly. There were
no differences in phenol contents of these two
potatoes. Total phenolics and DPPH index were not
signicantly affected by blanching and storage. Cold-
stored potatoes contained 40% less sterols than fresh
potatoes. Sterols were not affected by processing.
Swede
Vitamin C contents, total phenolics and DPPH
index of swede decreased by about 2025% during
blanching. Storage did not have any effects. Sterols
were not signicantly affected by blanching.
DISCUSSION
Fruits and vegetables provide an optimal mix of
phytochemicals such as natural antioxidants, bres
and other bioactive compounds. However, the health-
promoting capacity of fruits and vegetables depends
on their processing history. Processing is expected
to affect the content, activity and bioavailability of
bioactive compounds. The major variables during
processing are the endogenous enzyme levels, water
activity, oxygen pressure, and thermal and mechanical
energy. The unprocessed plant tissue is much more
stable as compared with crushed or milled material,
where enzymes become activated owing to breakage of
cell walls, bringing together substrates and enzymes.
In this study the production of frozen vegetables
and their long-term freezer storage were investigated.
Frozen vegetables are products that have been
harvested, prepared, blanched and quickly frozen
under the commercial conditions appropriate for each
vegetable. Typical commercial blanching conditions
are 9095

C for a duration of 110min, usually

achieved by exposure to hot water or steamfollowed by
a rapid cooling procedure. Typically, several months of
low-temperature storage can elapse before purchase,
possibly followed by several months of in-home
storage.
1398 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
Thermal processes such as steaming and cooking
change the properties of dietary bre in many ways.
24
Both increases and decreases in total dietary bre
content have been reported, as well as changes in
extractability, leading to redistribution between the
relative amounts of soluble and insoluble bre.
25
In
our experiments, dietary bre contents either increased
or were not affected. Increases in bre content were
most probably due to the washing out of soluble
components and small molecules, such as free sugars,
leading to the concentration of bre components in
the processed material. The mechanical distruption
of cells during processing might have resulted in
better extraction of bre components. In addition,
the enzymatic release of bre components from
the insoluble cell wall matrix might have increased
soluble bre contents. Also, hydrolysis during storage
might have released bre components from the
plant material, thus increasing their concentrations.
Svanberg et al
26
showed the effects of processing
on dietary bre in vegetables to be complex, with
heat processing generally degrading the dietary bre
polysaccharides. A reduction in molecular weight or
increased solubility of these carbohydrates inuences
their dietary properties.
The behaviour of minerals during preparation
and cooking is related closely to their solubility.
Minerals are generally stable to most of the conditions
encountered in cooking of foods, eg heat, oxidation,
acid or alkali.
27
Potassium, which is by far the most
abundant mineral in vegetables, is extremely mobile
and is easily lost by leaching during cooking because of
its high solubility in water. Calcium and magnesium
are generally present in plant tissue in bound form
and hence are not readily lost by leaching. However,
these minerals are also typically present in tap water
in areas with hard water, and they are known to be
taken up by vegetables during cooking in water.
28
Movement of calcium and magnesium into and out
of vegetables during cooking is principally observed
in leafy vegetables such as cabbage and spring greens,
reecting the large surface-to-volume ratio of these
vegetables. We also found that potassium was easily
leached during blanching, especially with spinach.
Only small uctuations in other mineral contents were
noticed in our experiments. Leaching into washing or
cooling water was the most probable reason for the
decreases. The slight increase in metal ion contents of
some vegetables can be explained by their dissolution
from processing equipment. However, these increases
and decreases might also be due to variations in raw
material.
Vitamin C has often been used as an indicator of
processing, because it is vulnerable to chemical and
enzymatic oxidation and is highly water soluble.
29,30
The losses that occur during the freezing process are
mainly due to the thermal degradation at the blanching
stage, but are also due to the leaching of nutrients into
the blanching medium. Favell
31
has recently studied
vitamin C losses in commercially frozen vegetables.
Average vitamin C losses during blanching/freezing
were approximately 30% for peas, 10% for green
beans, 30% for broccoli, and 40% for spinach, and
in three individual studies on peas, blanching losses
varied between 26 and 37%, showing the extent
of variation in a well-controlled commercial system.
Through a 12month storage period, losses were 10%
or less. In spinach a further 30% loss occurred
during deep frozen storage. Thus, for vegetables, losses
through the freezing process are typically in the range
of 1040%. The variation in the rate of losses shows
the different vulnerabilities of different vegetables, eg
surface area, mechanical damage, and sulphhydryl
contents, as well as their different enzyme activities.
Our results are in agreement with these results. We
found that the highest vitamin Closses connected with
blanching were about 30%, and storage induced some
extra losses. Favell
31
found that the nutrient status
of frozen peas and broccoli was similar to that of
typical market-purchased vegetables. The nutritional
status of frozen spinach and carrots was similar to
that of the fresh vegetables at harvest. When Davey
et al
32
compared processing techniques most relevant
to vegetables, ie canning, freezing and dehydration,
they found that losses are greatest during dehydration
and lowest during freezing.
The solubility and reactivity of folates make them
susceptible to potentially large losses during food
processing and storage.
33
The chemical stability of
folates in plant foods can be adversely inuenced by
heat, exposure to oxygen, and light intensity. Because
of their solubility, a large effect of processing may
also occur by leaching of folates into surrounding
water used for washing, blanching and cooking.
34
Cooper et al
35
studied the effects of laboratory-scale
blanching and 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. Microwave blanching without water resulted
in a 14% loss compared with fresh spinach. De
Souza and Eitenmiller
36
found, in laboratory-scale
blanching of spinach, 83 and 42% losses of folate
after water and steamblanching respectively compared
with fresh spinach. Folate losses were also studied
with broccoli. Losses were 60 and 9% after water
and steam blanching respectively compared with
fresh broccoli.
36
However, losses in these simulated
processes using spinach were higher when compared
with commercially frozen spinach. Malin
37
has studied
the stability of folates in frozen vegetables. Enzymatic
inactivation is required to prevent degradation of
folate, even when stored frozen. Also in our study,
folate was very sensitive to blanching: losses as high
as 70% were detected. Freezer storage had no further
effects on folate contents.
Food processing is reported to have both nega-
tive (loss due to oxidation) and positive (increased
bioavailability) effects on carotenoids.
38
Thermal pro-
cessing increases carotenoid concentration, perhaps
owing to greater extractability, enzymatic degradation
J Sci Food Agric 83:13891402 (online: 2003) 1399
R Puupponen-Pimi a et al
and unaccounted losses of moisture and soluble solids
that concentrate the sample per unit weight.
39
Heat
treatment also results in the inactivation of enzymes (eg
oxidases during blanching) and the breakdown of food
structures, leading to increased bioavailability.
40,41
It
may lead to signicant losses of epoxy-carotenoids,
lutein and, to a lesser extent, carotenes. Heat also
induces cis/trans isomerisation, which may lead to
the formation of different carotenoid by-products.
42,43
The degree of changes in carotenoids is dependent
upon the type of vegetable, method, and tempera-
ture and time conditions. Heat treatment in blanching
may provoke some losses, but inactivation of oxidative
enzymes prevents further losses during slow process-
ing and storage.
39
Microwaving is less destructive than
steaming and boiling.
39
However, when it is compared
with steaming and blanching, it produces signicant
losses of -carotene in leafy vegetables.
44
Freezing
generally preserves the provitamin A carotenoids, but
long thawing is detrimental.
45
These ndings are
in agreement with our results, which showed that
carotenoid contents are rst increased after process-
ing. Carotenoids of carrots were decreased during
long-term storage.
Very few data are available on the stability of
plant sterols in different food preparation procedures.
Signicant sterol losses occur only at temperatures
which are in the range of those used for deep-
frying.
46
Norm en et al
47
investigated sterol losses in
cooking of 13 vegetables and fruits. They found that
there was no signicant difference between raw and
cooked samples. Albi et al
48
have compared microwave
and conventional heating on ve oils. There was no
signicant difference in sterol contents of untreated
and treated oils. The stability of plant sterols during
food storage has been little studied. No signicant
changes in total sterol contents are likely to take
place in most common conditions. However, after
prolonged storage, some oxidation products may
be found.
46
In our study, no decrease in sterol
content was found during processing, even higher
contents were measured. Increased sterol contents
after blanching can be explained by better extraction
of these compounds or inactivation of enzymes, which
led to better stability during sample treatment. Also,
variations in raw material might have an effect.
Long-term storage seemed to decrease sterol contents
slightly.
Price et al
49
identied the main avonol glycosides
present in broccoli orets as quercetin and kaempferol
3-O-sophoroside. They studied the effect of cooking
on the composition and contents of avonol glycosides
in broccoli orets. When the orets were cooked
in boiling water until soft (15min), the amount of
avonol glycosides retained in the cooked tissue was
between 14 and 28% of that present in the raw
material. These gures are much lower than those
found for onion bulbs when subjected to similar
conditions.
50
The proportion of quercetin glycosides
retained in the cooked tissue was between 81 and
88%. These differences in different plant species
can be explained by the much larger surface area
that broccoli presents in the cooking water, thereby
allowing greater leaching of the glycosides into the
water. Our laboratory-scale experiments with broccoli
are in line with these results. However, in our
study with industrial samples, other brassicas behaved
in different ways. Flavonol contents were increased
during blanching. It can be speculated that the short
heat treatment released these compounds from the
cell matrix, resulting in better extraction. The increase
could also be explained by variations in raw material.
Gil et al
51
recently studied the effects of cooking on
avonoids of fresh-cut spinach. Boiling extracted 50%
of total avonoids in the cooking water. They also
found that avonoid glucuronides were extracted more
in the cooking water than the other glycosides. Also
in our studies, blanching clearly decreased avonol
contents of spinach. Flavonol contents of all vegetables
decreased during storage, as reported in other studies.
Free radical scavenging is the accepted mechanism
for an antioxidant to inhibit lipid oxidation. The
method of scavenging stable DPPH free radicals
can be used to evaluate the antioxidant activity of
specic compounds or extracts in a short time.
52
Chu et al
53
found that 3060s of blanching of leafy
vegetables (green leaves of sweet potatoes) at 100

C
still maintained high amounts of avonoids and
free radical-scavenging activities as compared with
more than 1min of blanching. In our experiments
a few minutes of blanching decreased the amount
of total phenolics and radical-scavenging activity of
peas, spinach and swede by about 2030%. On
the other hand, Talcott et al
54
showed that total
soluble phenolics increased immediately after thermal
processing of carrot puree by 70%, and antioxidant
activity by an average of 30%.
It is well known that natural antioxidants contained
in foods are signicantly lost during processing.
55
However, thermal treatments can also induce the
formation of compounds with new antioxidant
properties, eg Maillard reaction products. The
Maillard reaction occurs when sugars condense with
free amino acids, peptides or proteins, leading to the
formation of brown melanoidins. It is not clear whether
the Maillard reaction products exhibit mutagenic or
antimutagenic activity.
56,57
Antimutagenic activity has
been found to be closely related to their antioxidant
activity.
57
Nicoli et al
58
have recently reviewed the
inuence of processing on the antioxidant properties
of fruits and vegetables. They concluded that the
changes in the overall antioxidant properties of
foods can be attributed to the sum of different and
sometimes opposite effects. For short heat treatments
of vegetables a reduction in the overall antioxidant
properties due to the loss of naturally occurring
antioxidants and/or the formation of Maillard reaction
pro-oxidants can be detected. By prolonging the
heating time, this loss can be minimised by a recovery
or even an enhancement of the antioxidant activity
1400 J Sci Food Agric 83:13891402 (online: 2003)
Effect of blanching and freezing on vegetables
due to the formation of advanced Maillard reaction
products.
59
CONCLUSIONS
Effects of industrial blanching/freezing and long-term
freezer storage on various bioactive components of
a large number of different types of vegetables were
screened. Contents of dietary bre components and
minerals were not signicantly affected by processing.
Also, plant sterols proved to be stable during
blanching and freezer storage. However, phenolic
antioxidants and vitamins were more sensitive. Plant
species-dependent decreases were often detected with
phenolic compounds, vitamin C and folic acid.
The following assays proved to be very practicable
in order to get a general view of the effects of
processing on plant bioactivity: total phenolics and
DPPH radical-scavenging activity, HPLC prole of
phenolic antioxidants, vitamin C and folic acid.
Screening methods for folic acid and DPPH index
were developed using microtitre plates, making it
possible to analyse large amounts of samples. The
qualitative HPLC method for phenolic antioxidants
developed also in this study proved to be very useful
when evaluating the complex behaviour of phenolics
during food processing. Results of this investigation
and methods developed will be used by the vegetable-
processing industry in raw material choice, process
development and designing plant-based healthy foods.
ACKNOWLEDGEMENTS
The authors thank Sirpa Karppinen and Tuomo Kiu-
tamo for dietary bre component analysis. Tuulikki
Sepp anen-Laakso is also thanked for valuable dis-
cussions. The skilful technical assistance of Jaana
Rikkinen, Annika Majanen, Kari Kammiovirta and
Teija Jokila fromVTTand Tuula Kurtelius and Leena
Puura from MTT is gratefully acknowledged. Juhani
Hvitfelt, Maritta Pyysalo and Annikki Pihlava from
L annen Tehtaat Plc are warmly thanked for their help
in supplying samples and for useful discussions. This
study was nancially supported by Tekes.
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