Food Research International 35 (2002) 23–30

www.elsevier.com/locate/foodres

Removal of phenolic compounds in the production of high-quality

canola protein isolates
L. Xu, L.L. Diosady *
Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street,
Toronto, ON, Canada M5S 3E5

Received 9 June 2000; accepted 17 August 2000

Abstract
Phenolic compounds are a major cause of the dark colour and undesirable taste of canola protein isolates. Based on our previous
study of phenolic–protein interactions, a variety of treatments were tested for the removal of phenolic compounds from canola
proteins. The combined use of these treatments was able to reduce the phenolic content of the products by 80–90%. Thus modified,
the process produced two canola protein isolates, both of high protein content (>85%), low in phenolic acids (200 mg/100 g), and
essentially free of condensed tannins. Colour measurement and sensory evaluation confirmed that the removal of phenolic com-
pounds improved the colour and taste of canola protein isolates. # 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Canola; Protein isolates; Phenolic compounds; Protein extraction; Isoelectric precipitation; Membrane processing; Colour; Sensory eva-
luation

1. Introduction products, it seemed prudent to investigate the quantity

Based on yield and purity, the membrane-based pro-
tein isolation process developed in our laboratories
(Tzeng, Diosady & Rubin, 1990) is commercially pro-
mising in rapeseed/canola protein production. The
process consists of five key steps: alkaline extraction,
isoelectric precipitation, ultrafiltration followed by
diafiltration, and drying. Two protein isolates were
produced with a combined protein recovery of over
70% of the protein present in the seed. Both products
were of high protein content (>90%), low in phytates
(<1%), essentially free of glucosinolates (<2 mol/g),
and with desirable functional properties for a variety of
food applications (Igor, Diosady & Rubin, 1993; Xu &
Diosady, 1994). Unfortunately, the use of these isolates
in food has been prevented by their undesirable orga-
noleptic properties. Both the precipitated protein isolate
and the aqueous solutions of soluble protein isolate
were dark in colour, and had an unpleasant bitter taste.
Since phenolic compounds are known to cause
discolouration and off-flavours in vegetable protein
* Corresponding author. Tel.: +1-416-978-4137; fax: +1-416-978-
8605.
E-mail address: diosady@chem-eng.toronto.edu (L.L. Diosady).
0963-9969/02/$ - see front matter # 2002 Elsevier Science Ltd. All rights reserved.
PII: S0963-9969(00)00159-9
and effects of phenolic compounds in these canola pro-
tein isolates.
Rapeseed contains about 10 times the quantity of
phenolic compounds found in soybean (Zadernowski &
Kozlowska, 1983). Most phenolic compounds com-
monly identified in canola are phenolic acids (Durkee &
Thivierge, 1975; Zadernowski et al., 1983) and con-
densed tannins, which are polymeric phenolics based on
flavonoids (Leung, Fenton, Mueller & Clandinin, 1979).
As common canola and rapeseed varieties have similar
phenolic components within a reasonably narrow con-
centration range, no distinction will be made between
canola and rapeseed in this paper, unless needed for
clarity. The major phenolic component in rapeseed and
canola was reported to be sinapine the choline ester of
sinapic acid (Sosulski, 1979). It constitutes about 1% of
the meal mass, well above the taste thresholds of the
phenolic acids in oilseed meals (40–500 ppm). Con-
densed tannins as polymeric phenolics may cause
astringency due to their ability to precipitate the
proteins in the mouth (Shahidi, 1992). On oxidation
phenolic compounds can cause the development of dark
colours in oilseed protein products. Under alkaline
conditions they readily undergo enzymatic and non-
enzymatic oxidation to form quinones which can then
react with protein, resulting in dark green or brown
24 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30

colour in the protein solutions, and when the proteins
are precipitated at their isoelectric points, the colour
cannot be washed from the protein isolates (Sosulski,
1979). Accordingly, in order to produce protein isolates
that can be incorporated into food formulation, most of
these compounds must be removed.
Phenolic compounds in canola bind to the proteins
through a variety of mechanisms in aqueous media,
including hydrogen bonding (Loomis & Battaile, 1966),
covalent bonding (Mason, 1955), hydrophobic interac-
tions (Hagerman & Butler, 1978), and ionic bonding
(Rubino, Arntfield, Nadon & Bernatsky, 1996). The
separation of proteins from phenolic compounds is
complicated by these complex-forming mechanisms.
Numerous processes have been proposed and tested for
the removal of phenolic compounds from oilseed pro-
teins, but all were reported to have problems with
incomplete removal, loss of protein, or excessive cost.
An effective search for efficient means for the removal of
phenolic compounds required a detailed study of the
interactions between canola proteins and phenolic
compounds.
We developed a technique for the quantitative char-
acterization of canola protein–phenolic bonding in
aqueous media, which combined a series of chemical
treatments which broke specific types of protein–
phenolic bonds, followed by membrane separations to
remove the released phenolics for analysis (Xu & Dio-
sady, 2000). We found that some 50% of the extracted
phenolic compounds formed complexes with canola
proteins by several distinct mechanisms, among which
ionic bonding was predominant, accounting for about
30%. The phenolic fractions bound by hydrophobic
interactions, hydrogen bonding, and covalent bonding
were relatively small: each constituting less than 10% of
the total extractable phenolics.
Based on these findings, in the current study we
modified the process developed by Tzeng et al. (1990) by
including specific treatments to remove phenolic com-
pounds attached to the protein by different mechanisms.
Needham Heights, MA) and the supernatant polished
by filtration. The residual solids were washed twice with
6 volume of distilled water. The washing liquids were
combined with the original extract.
The resulting alkaline extract was diafiltered at a dia-
volume of 5 directly (Run 2), or after pretreatment with
0.05 M NaCl (Run 3), or with 0.05 M NaCl and 0.1%
sodium lauryl sulphate (SDS) (Run 4). The diafiltra-
tions in all runs were done at pH 12, and in Runs 3 and
4 0.05 M NaCl was also added to the diafiltration
solution.
For comparison, a control run (Run 1) was performed
with no treatment prior to isoelectric precipitation.
The pH of the treated extract was reduced to 3.5 with
6 M HCl. The precipitated proteins were recovered by
centrifugation (4000g, 15 min). The precipitate was
washed with 5 times its weight of distilled water (on
wet basis) and centrifuged again for separation. The
acidic protein solution from the isoelectric precipitation
was combined with the washing liquid and polish-fil-
tered. It was ultrafiltered at a concentration factor of 10
and then diafiltered again at a diavolume of 5 to
concentrate and purify the proteins remaining in the
solution.
Both the washed precipitate and the membrane-pro-
cessed solution were freeze-dried for 48 h using a
Labconco freeze Dryer-18 (Labconco Corp., Kansas

2. Materials and methods

2.1. Removal of phenolic compounds prior to isoelectric

precipitation

The experimental scheme for the removal of free phe-

nolic compounds by diafiltering the alkaline extract
prior to isoelectric precipitation is presented in Fig. 1.
The protein in 50 g defatted prepressed canola meal
(CanAmera Foods, Hamilton, ON) was extracted by
aqueous NaOH at pH 12.0 and a water–meal ratio of 18
for 30 min. After the extraction, the meal residue was
separated by centrifugation (4000g, 15 min) with a B- Fig. 1. Processing scheme with treatments to remove phenolic com-
22 centrifuge (International Equipment Company, pounds prior to isoelectric precipitation.
 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30
City, MO) to obtain two products: precipitated and
soluble protein isolates (PPI and SPI).
2.2. Combined process with further treatments
The process was modified to the scheme presented in
Fig. 2 with further treatments. To reduce the effects of
oxidation on the product flavour, Na2SO3 at a concentra-
tion of 0.1% was added to the extraction solution as a
reducing agent. The volume of the extract was reduced by
a factor of 3 by ultrafiltration. This was followed by dia-
filtration, and then isoelectric precipitation at pH 3.5, as
described earlier. The protein solution was treated with
different levels of insoluble polyvinylpyrrolidone (PVP), a
specific adsorbent for polyphenols (Runs 5 and 6), and the
isolates were then recovered as described in Section 2.1.
2.3. Colour measurement
The colour of all PPIs was measured using a D25A-9
Hunter Colorimeter. The instrument consisted of two
sections — the optical sensor and the signal processor.
The optical sensor used light from a quartz halogen
cycle lamp, which was filtered to closely approximate
CIE illuminant D65. The light was directed upward to
the sample port at an angle of 45 from the perpendicular.
Fig. 2. The combined process with treatments to remove phenolic
compounds for the production of high-quality canola protein isolates.
25
The light receptor was placed directly below the sample
port. The signal processor converted the photometric sig-
nals to the standard Hunter L, a, b scale. For each mea-
surement approximately 4 g of sample were used.
The colour of SPI samples was evaluated in aqueous
solution using a Beckman DU-7 UV-visible spectro-
photometer (Beckman Instruments Inc., Irvine, CA).
The sample was first dissolved in distilled water at a
concentration of 1% (w/v), and centrifuged (4000g, 15
min) with a Centra 4 centrifuge (International Equip-
ment Company, Needham Heights, MA). The super-
natant was decanted into a quartz cuvette and scanned in
the range of 385 to 700 nm against distilled water as blank.
2.4. Sensory evaluation
A simple preliminary comparison of the samples’ taste
was performed using a descriptive sensory test method,
unstructured scaling, also known as line or visual ana-
logue scaling (Poste, Mackie, Butler & Larmond, 1991).
A panel was set up consisting of 13–14 people, including
the staff, graduate, and undergraduate students in food
engineering. The samples were presented in jars wrap-
ped with aluminum foil to mask colour differences, thus
avoiding stimulus error. The order of presentation of
the samples was randomized to minimize central ten-
dency error. Drinking water was offered for mouth
rinsing between samples to control contrast effect. To
minimize expectation error, all panelists were given only
enough information to conduct the test, and the person
directly involved in making the products was not inclu-
ded on the panel.
The most commonly used unstructured scale consists
of a horizontal line 15 cm long with two anchor points
on both ends and a mid point. Each anchor point is
labeled with a word or expression. A separate line is
used for each sensory attribute to be evaluated. In this
study a technical and a hedonic attribute were investi-
gated: taste intensity and pleasantness of the products.
Panelists recorded their evaluation by making a vertical
line across the horizontal line at the point that best
reflects their perception of the magnitude of that prop-
erty. Numerical scores were then given to the ratings by
measuring the distance of the marks from the left end of
the line in units of 0.1 cm. One score was an equivalent
of 1 cm on graphical scale.
2.5. Chemical analyses
Crude protein (N6.25) was determined by the Kjel-
dahl method, American Association of Cereal Chemists
(AACC, 1976, Method 46-12), using a Büchi 425 digester
and a Büchi 315 distillation unit (Brinkmann Instru-
ments Inc., Mississauga, ON). The analytical method of
Xu and Diosady (1997) was used for determination of
total phenolic acid content with results expressed as
26 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30

sinapic acid equivalents. Condensed tannin content was Table 1

determined by the method of Shahidi and Naczk (1989) Mass and protein recoveries of the products from different runs
as catechin equivalents. The residual SDS was deter- Treatments Products Mass Protein
mined using a method based on dissociation precipita- (%)a (%)b
tion and gravimetric determination of the sulphate,
Run 1. Control PPIc 15.2 33.6
(Igor et al., 1993). The precipitated sulphate was then SPId 10.0 23.9
quantitated using AACC Method 40-66 (1976). Total 25.2 57.5
Run 2. Diafiltration PPI 14.9 34.3
SPI 9.4 22.6
3. Results and discussion Total 24.3 56.9
Run 3. 0.05 M NaCl PPI 15.1 35.0
3.1. Removal of phenolic compounds prior to isoelectric SPI 8.1 20.0
precipitation Total 23.2 55.0
Run 4. 0.1% SDSe PPI 15.5 35.4
The quantitative distribution of phenolic-protein with 0.05 M NaCl
complexes (Xu et al., 2000) suggested that about half of SPI 8.0 19.4
the extractable phenolic acids and condensed tannins Total 23.5 54.8
were actually unbound at extraction pH, and could be a
Percentages of mass recoveries were calculated based on 50 g
removed by diafiltration. The literature also indicated starting meal.
that treatment with 0.05 M NaCl could break phenolic– b
Percentages of protein recoveries were calculated based on the
protein complexes bonded ionically, which represented total amount of protein in 50 g starting meal, determined to be 19.05
g.
some 30% of the total phenolic acids present in the c
Precipitated protein isolate.
extract. The phenolic acids thus released would then be d
Soluble protein isolate.
removed by diafiltration. Further treatment with 0.1% e
Sodium lauryl sulphate.
SDS would release about 25% of the condensed
tannins, bound to proteins by hydrophobic interactions. All products contained 85–95% protein (Table 2).
An experimental scheme incorporating these treatments The remaining 5–15% were likely polysaccharides. The
was developed as shown in Fig. 1. existence of glycoproteins in rapeseed/canola has been
All the processes in the scheme were run to produce previously reported (Jones, 1979). The treatments
four pairs of samples, each consisting of a PPI and a removed some low-molecular-weight impurities, includ-
SPI. The mass yields and protein recoveries of all the ing phenolics, before isoelectric precipitation, thus
products are shown in Table 1, and their protein, phe- increasing the protein content of the products over the
nolic acid, and tannin contents are presented in Table 2. control run. In all cases, the SPIs were higher in protein
Mass recoveries for all runs were about 15% for PPI than the PPIs, which was consistent with previously
and 9% for SPI. The total mass yields of the protein reported results (Tzeng et al., 1990).
isolates were lower than what was previously reported The phenolic acid and condensed tannin contents
by our laboratories for canola protein preparation. showed a distinct descending trend with increased
Tzeng et al. (1990) obtained a total isolate yield of number of treatments, confirming that each treatment
32.5% with hexane-defatted air-dried canola meal. The indeed removed some phenolic compounds (Table 2).
inferior mass recoveries in this case were the direct From the control run (Run 1) where no treatment was
result of the low protein extractability of hexane-defat- employed, both protein isolates were high in phenolic
ted prepressed canola meal, due perhaps to partial pro- compounds. Their phenolic acid contents were over
tein denaturation by heat during prepressing. The total 1000 mg per 100 g sample (Table 2), approximately
protein recoveries as protein isolates were approxi- 65% of the value of the starting meal (1596 mg/100 g),
mately 55%, and the recovery ratio of PPI to SPI was and the condensed tannin contents even exceeded that
about 1.5 (Table 1), indicating that most of the extrac- of the starting meal (676 mg/100 g). Since no treatment
ted canola proteins could be precipitated at pH 3.5. was used in the control run, only those phenolic com-
Although neither mass nor protein recoveries varied pounds not bound to canola protein in the pH 3.5
much, all the runs with the treatments for the removal solution were eventually removed by the membrane-
of phenolics (Runs 2, 3 and 4) gave slightly lower total processing (ultrafiltration followed by diafiltration).
mass recoveries than the control run (Run 1). This was Although the free phenolic compounds made up more
likely a result of nitrogen losses in the membrane pro- than half of the total amount in the solution, as shown
cessing of the alkaline extract in the form of non-protein earlier by the fractionation, the remaining phenolic
nitrogenous compounds including short peptides and compounds that were bound to the proteins could still
free amino acids. give rise to high phenolic contents in the final products.
As a result of the high phenolic contents, the protein
 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30
Table 2
Effects of treatments to remove phenolic compounds on compositions of the products
Proteina (%)
Treatments PPIb SPIc
Run 1. Control 84.7 91.1
Run 2. Diafiltration 87.7 92.6
Run 3. 0.05 M NaCl 88.5 94.9
Run 4. 0.1% SDSd with 87.2 92.3
0.05 M NaCl
ACVe (%) 0.27 0.35
a
All results are reported as is; results of protein content are means of triplicates, and others are means of duplicates.
b
Precipitated protein isolate.
c
Soluble protein isolate.
d
Sodium lauryl sulphate.
e
Average coefficient of variation.
products still exhibited undesired organoleptic proper-
ties such as dark colour and bitter taste. The high phe-
nolic acid and tannin contents in the protein isolates
from the control run confirmed that further treatments
to remove these compounds were necessary in order to
improve the quality of the products.
Therefore, diafiltration prior to isoelectric precipita-
tion was first tested in Run 2 to remove the free
fractions of both phenolic acids and condensed tannins
from the alkaline extract. It was shown in Table 2 that,
with this treatment, the phenolic acid contents in PPI
and SPI were decreased by 17 and 22%, respectively,
while condensed tannin contents were reduced by 32
and 24%, respectively (Table 2). Apparently, this addi-
tional diafiltration step was effective in removing free
phenolic compounds.
In Run 3, the treatment with 0.05M NaCl was tried
since it was shown by the fractionation to be able to
break ionic bonding between canola protein and phe-
nolic compounds (Xu et al., 2000). The phenolics released
by this treatment were then removed by diafiltration prior
to isoelectric precipitation. As expected the removal of
the ionically bound reduced the phenolic acid content of
the protein isolates to half of the level obtained in Run 2
(Table 2). The treatment, however, resulted in a less
dramatic decrease in condensed tannins in PPI and SPI
of 24 and 13%, respectively (Table 2). As the ionically
bonded fraction of condensed tannins was insignificant
(Xu et al.), these results were also expected.
The treatment with 0.1% SDS in Run 4 greatly
reduced the amount of condensed tannins in both pro-
ducts. Again, this was expected since the condensed
tannins bound by hydrophobic interactions represented
a significant fraction of extracted phenolics (Xu et al.,
2000). With the addition of SDS, condensed tannin
contents in both protein products were slashed by more
than 90% compared with Run 3 (Table 2). In fact,
condensed tannin levels in the products were so close to
27
Phenolic acidsa Condensed tanninsa
(mg/100 g sample) (mg/100 g sample)
PPI SPI PPI SPI
1094 1053 675 852
917 823 457 648
451 470 347 562
301 345 62 34
7.1 5.3 8.0 5.8
the detection limit of the analytical method (10 mg/
100g sample) that it was likely that these products were
essentially tannin-free.
Binding of SDS to canola proteins was demonstrated
previously (Igor et al., 1993). Residual SDS would ren-
der the protein products practically useless due to the
sensory and health effects. However, in this study, SDS
levels in the products from Run 4 were well below 0.5%,
thus acceptable on the basis of health safety alone
(Health Canada, 1994). To explain this, it was postu-
lated that at a high pH such as 12, the binding of SDS
to canola proteins became much weaker than in the
acidic range since both were negatively charged at the
high pH, and the electrostatic repulsion was able to
keep them apart to a certain extent, thus allowing the
removal of SDS by diafiltration, resulting in low resi-
dual SDS content in the final products.
3.2. Combined process with PVP treatment
The process was further modified to improve the
quality of the final products (Fig. 2). Oxidation of phe-
nolic compounds is known to increase covalent-binding
to the protein, hence darkening the colour of the protein
extracts or solutions (Gheysuddin, Cater & Mattil,
1970). To prevent this, Na2SO3 was added as an anti-
oxidant at a concentration of 0.1% during alkaline
extraction of the protein and diafiltration of the extract.
An ultrafiltration step was included prior to the dia-
filtration of the alkaline extract simply to reduce the
volume processed so that the amount of water for the
diafiltration could be greatly reduced, and the proces-
sing time shortened.
After isoelectric precipitation 5 g PVP solution (10%
of the starting meal) was added to the pH 3.5 solution
to further remove phenolic acids in the acidic solution
(Run 5). The results presented in Table 3 show that PVP
treatment reduced the phenolic acid content in the
28 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30
acidic SPI by more than 50% while the protein content
in the SPI was not affected by PVP. The condensed
tannin contents of these protein isolates were not deter-
mined as they were known to be essentially tannin-free
(Run 4). Despite the noticeable effects of PVP on phe-
nolic acid removal, treatment at the 10% PVP level was
not economical as PVP is expensive. The process was
repeated with 1% PVP treatment (Run 6). The phenolic
acid content of SPI obtained was similar to that with 10%
PVP addition, suggesting that 1% addition was adequate.
For commercial production of canola protein isolates,
regeneration and reuse of PVP is required for economic
reasons. However, this was outside the scope of this study.
The effect of reducing the concentration factor from
10 to 4 was investigated in the second ultrafiltration for
the acidic solution, since the solution had already been
concentrated three-fold during the ultrafiltration of the
alkaline extract. We wished to avoid increasing the
concentration to the point where permeation would be
drastically reduced by gel formation on the membrane
(Michaels, Nelson & Porter, 1971).
The mass and protein distribution among major pro-
cess streams (Table 3) was determined. They were not
greatly affected by the process modifications. While
more than half of meal protein was recovered in the
protein isolates, about 60% meal solids (mass) ended up
in the meal residue. It was impractical to analyze the
permeate, which contained small molecular weight
components. Previous work has shown that the mass of
these components was approximately 12% of the start-
ing mass, and 6–10% of the total nitrogen (Tzeng et al.,
1990). The observed solids recovery of 82.1% indicates
that the mass balance reflects an experimental uncer-
tainty of about 6%, made up from losses in transfers
during the experiments, and analytical errors.
3.3. Colour measurement
The colours of PPIs from different runs were mea-
sured and compared using a Hunter colorimeter. For
Table 3
Compositions and yields of the products from Run 5a
Compositionsb
Protein Phenolic acids
(%) (mg/100 g sample)
Starting meal 38.1 1596
Precipitated protein isolate 87.0 274
Soluble protein isolate 91.6 114
Meal residue 22.1 360
‘‘Unrecovered’’d – –
a
Modified with 0.05 M NaCl and 0.1% sodium lauryl sulphate of treatment of alkaline extract, and ultrafiltration followed by diafiltration
before isoelectric precipitation, and treatment of acidic solution with 10% polyvinylpyrrolidone after isoelectric precipitation.
b
All results are reported as is; results of protein content are means of triplicates, and others are means of duplicates.
c
Not determined.
d
‘‘Unrecovered’’ was calculated by subtraction: starting meal (100) — all products.
each measurement a 4 g sample was needed, which was
actually more than half of the amount of each sample
produced in a single run. A Hunter colorimeter mea-
sures the colour in the three-dimensional opponent-col-
our system. All L values in Table 4 were measures of
sample lightness, with 100 being white and 0 black. All a
values indicated redness varying between +100 and
80 as sample colour changed from red to green,
whereas yellowness was read by b values from +100
(yellow) to 80 (blue). It was confirmed that the sam-
ples became lighter with more intensive treatments to
remove phenolic compounds except for the treatment
with 0.05 M NaCl, which did not influence the colour
lightness. The PPI sample from the control run (Run 1)
had a much higher phenolic content than that from Run
5, hence a more intense yellow colour.
All SPIs were fluffy, and displayed a similar desirable
off-white colour. However, upon dissolution in water,
their solutions showed brown colours of different
intensities. Therefore, colour measurement of the SPI’s
was performed by scanning their aqueous solutions in
the UV-visible range. The absorbency curves are pre-
sented in Fig. 3. The colour of these solutions may also
be due to the presence of phenolic compounds as the
products with lowered phenolic contents were lighter in
colour. However, no treatment could completely elim-
inate the colour. Even the solution of SPI from Run 5
still had a light brown colour. This was probably
because of the covalently bonded phenolic–protein
complexes, which could not be removed by any treat-
ment. According to the mechanism proposed by Torch-
inskii and Dixon (1974), covalent bonds could be
formed through the reaction of SH-containing amino
acids such as cysteine with quinones that were derived
from phenolic compounds. The products of this reac-
tion are thioethers which have absorption maxima
between 420–430 nm. All SPI solutions seemed to have
a significant absorbance in this wavelength range
although no peak could be observed because of the
proteins in the solutions (Fig. 3). Like any other protein
Yields (as % of starting meal)
Condensed tannins Mass Protein
(mg/100 g sample)
677 100 100
NDc 15.3 35.0
NDc 8.5 20.4
NDc 58.3 33.8
– 17.9 10.8
 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30 29

Table 4 phenolic content. Treatments that remove phenolic

Colour measurement of precipitated protein isolates (PPIs) from dif- compounds lead to lighter coloured protein isolates.
ferent runs

PPI samples L a b 3.4. Sensory evaluation

(Lightness) (Redness) (Yellowness)

Run 1 Control 52.1 3.1 21.4

Since canola protein isolates prepared as above are
Run 2 Diafiltration 61.5 3.4 14.2 intended to be eventually used as functional ingredients
Run 3 0.05 M NaCl 60.2 3.8 14.3 in food, it is desirable that they do not contribute to
Run 4 0.1% SDSa with 66.9 2.7 12.9 flavour, or provide only minimal, complementary
0.05 M NaCl flavour to food products. Therefore, their taste was
Run 5 Combined treatment 69.3 0.8 17.5
evaluated using sensory test methods to determine both
a
Sodium lauryl sulphate. taste intensity and acceptability. As PPI and SPI have
distinctly different functional properties (Igor et al.,
1993; Xu et al., 1994), and will likely be used in different
food systems, they were evaluated and compared sepa-
rately. The unstructured scaling method was chosen in
this study because it is useful for providing information
on the degree or intensity of the sensory characteristics
of concern, thus helping to identify treatments or pro-
cess variables responsible for these characteristics.
In order to determine the difference in taste among
the PPI or SPI samples made by the above processing
runs, the sensory test data were analyzed using ANOVA
(analysis of variance) method. Based on the results of
ANOVA it could be concluded that, while the effect of
human bias was insignificant, there were statistically
significant differences in taste intensity and pleasantness
among these canola protein isolates from different runs
(P40.05). To further determine whether these products
were different from one another, Tukey’s multiple com-
Fig. 3. UV-visible spectra of solutions of soluble protein isolates from parison test was performed (Snedecor and Cochran,
different runs: 1, Run 1; 2, Run 2; 3, Run 3; 4, Run 4; 5, Run 5. 1989). The results are presented in Table 5, using letters
to indicate differences. For taste intensity, the higher
soluble in water, soluble canola proteins in solution had numerical values connoted stronger taste. Pleasantness
a high aborbance in the UV range, so all absorbency was a hedonic measurement, the values of which repre-
curves rose sharply as the wavelength approached the sented the degree of acceptability or preference of taste.
UV range. Any two values not sharing a common letter are sig-
The colour measurements confirm that the dark col- nificantly different at P40.05. An ideal product from this
our of canola protein isolates is dependent on their work will have minimal or zero taste intensity. The scale

Table 5
Results of sensory test for canola protein isolatesa

Process meanb

Products Taste Run 1 Run 2 Run 3 Run 4 Run 5 Run 6 Soy protein
features Control Diafiltration NaCl NaCl and SDS Combined treatments Combined treatments (Supro 500)
(10% PVP) (1% PVP)

PPIc Taste intensity 11.5e 12.1e 8.2f 7.3fg 4.9g NAe 4.4g
Pleasantness 4.9ef 4.1e 6.0efg 7.7efg 7.9fg NAe 9.1g

SPId Taste intensity 9.9ef 10.9e 7.9fg 6.8g 6.3g 6.4g NAd
Pleasantness 4.8ef 3.6e 6.5fg 7.3fg 7.9g 7.8g NAd
a
SDS, sodium lauryl sulphate; PVP, polyvinylpyrrolidone; PPI, precipitated protein isolate; SPI, soluble protein isolate.
b
Means in each row not sharing a common letter are significantly different (P<0.05).
c
Results for PPI samples are means of data of 14 panelists.
d
Results for SPI samples are means of data of 13 panelists.
e
Not applicable.
30 L. Xu, L.L. Diosady / Food Research International 35 (2002) 23–30
for pleasantness ranges from 0 to 15, and on this scale the
completely bland product would have an ideal score of 7.5.
It was shown that, while both the PPI and SPI from
the control run (Run 1) had a distinct flavour (Table 5),
the low taste intensity of the PPI from the modified
process was comparable to that of a commercial soy
protein isolate. The difference in their pleasantness was,
however, far less significant than their taste intensity, as
the panelists did not find the blander products much
more pleasant to taste than the products from the con-
trol run. They also seemed to like the PPI from Run 5 as
much as the commercial soy protein isolate, as sug-
gested by the data. As for the SPI from Run 5, not only
did the panelists find it much blander than that from the
control run, they also had an obvious preference for it
to its counterpart from the control run (Table 5). Phe-
nolic adsorption by PVP only slightly improved the
taste of the SPI at either 1 or 10% level.
The results of sensory evaluation show that phenolic
compounds are the major contributors to the undesir-
able flavours such as bitterness or astringency of canola
protein isolates, and the taste of these products was
improved as phenolic compounds were removed by the
treatments.
4. Conclusions
The targeted dissociation of phenolic–protein com-
plexes bound by different mechanisms could be incor-
porated into the canola protein isolation process
originally developed by Tzeng et al. (1990). The result-
ing process recovered over 50% of the proteins of the
starting meal in its two products, PPI and SPI, with an
80% reduction in phenolic acids and 90% reduction in
condensed tannins. The removal of phenolic com-
pounds resulted in protein isolates that were lighter in
colour and blander in taste, thus suitable for incor-
poration into food products.
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