Con Ins!. Food Sci. Techno!. J. Vol. 19. No.2. pp. 53-56.
1986
RESEARCH
Effect of Boiling Treatment of Soymilk on the
Composition, Yield, Texture and Sensory Properties of Tofu
Elias E. Escueta 1, Malcolm e. Bourne and Lamartine F. Hood
Institute of Food Science
Cornell University
Geneva, New York 14456
Abstract
Soymilk was boiled for 0, 12, 30 and 60 min before coagulation
with CaS04 to make tofu, and a texture profile analysis was per-
formed on the product obtained. Hardness and gumminess increased
slightly up to 12 min of boiling but decreased significantly after 30
min and 60 min of boiling the soymilk. Chewiness decreased sig-
nificantly after 30 min and 60 min of boiling. Springiness was not
affected by boiling. Cohesiveness increased slightly after 12 min boil-
ing and then decreased slightly but not significantly after 30 min
and 60 min of boiling. Composition and sensory qualities of tofu
were not significantly affected by the length of time soymilk was
boiled prior to coagulation.
Resume
L'influence du temps d'ebullition du lait de soja avant sa coagu-
lation au C0 4Ca dans la fabrication du tofu a ete etudiee il 0, 12,
30 et 60 min en rapport avec les qualites texturales des produits obte-
nus. La fermete et la tendance il coller augmenterent legerement
jusqu'il 12 min d'ebullition, mais diminurent significativement apres
30 min et 60 min d'ebullition du lait de soja. La spongiosite ne fut
pas afffectee par ('ebullition. La cohesion augmenta legerement apres
12 min d'ebullition pour diminuer ensuite un peu, non significati-
vement apres 30 min et 60 min d'ebullition. La composition et les
qualites sensorielles du tofu ne furent pas significativement affec-
tees par Ie temps d'ebullition du lait de soja avant la coagulation.
Introduction
Heat unfolds the polypeptide chains of soybean pro-
teins dispersed in water (Wolf, 1972; Huang and Rha,
1974). This unfolding of the globular proteins before
aggregation is a requirement and an initiating step for
the formation of the three dimensional network in the
protein curd (Saio and Watanabe, 1968; Aoki and
Sakurai, 1969; Matsumoto, 1975; Furukawa et al.,
1979). Furthermore, heating increases the sulfhydryl
groups in soybean protein (II S) and results in harder
tofu (Saio et al., 1971). It is suggested that most part
of the sulfhydryl groups exists as disulfide bonds and
I PresentAddress: Institute of Food Science and Technology, Univer-
sity of Philippines at Los Banos, College, Laguna, 3720, to whom
all inquiries should be addressed.
Copyright © 1986 Canadian Institute of Food Science and Technology
53
a small part exists as free sulfhydryl groups. The opti-
mum heating time for the preparation of tofu was
reported by Saio (1979) to correspond to the maximum
range of sulfhydryl groups. On the other hand, a soy-
bean protein coagulated without heating (no unfold-
ing of the polypeptide chains) exhibits a globular struc-
ture rather than a three dimensional network and
produces a softer texture (Saio and Watanabe, 1968).
Aoki and Sakurai (1969) stated that boiling the milk
for 15 min gave a harder curd than the curd from the
milk just brought to boil prior to coagulation.
eatsimpoolas and Meyer (1970) reported that com-
plete dissociation of the protein into subunits, by
cleavage of disulfide bonds, is one of the conditions
favoring aggregation. This is brought about mainly by
the cleavage of intramolecular disulfide bridges which
facilitates disruption of quaternary structure and
exposure of reactive groups due to dissociation and
unfolding of subunits resulting in enhancement of gel
strength. Gel formation during heating of a protein
paste is prevented by addition of reducing agents hav-
ing S-S bond cleaving ability (Aoki and Sakurai, 1968).
Hashizume et al. (1975) heated the water extract
from defatted soybean flakes at 60, 70, 80, 90 and
100°C and then subjected the samples to disc elec-
trophoresis. The 7S protein decreased at 60°C and dis-
appeared at 70°C. The lIS protein did not show
changes up to 80°C, decreased at 90°C and disap-
peared at 100°C. The decrease in the lIS protein con-
tent was accompanied by an increase of new, fast mov-
ing components thought to be protein subunits,
although they were not yet found when the 7S protein
had disappeared. In the earlier studies, Watanabe and
Nakayama (1962) and Saio et al. (1968), reported that
heating a solution of water extracted proteins of soy-
bean flakes at 80°C for 10 min caused the main com-
ponents to disappear and new ones to form with higher
sedimentation velocities. The 7S protein fraction was
found to be more sensitive to heating than the lIS pro-
tein fraction. King (1977) exposed the 7S fraction to
temperatures in the range of 70 to 100°C and reported
that sedimentation coefficients of the 7S aggregate
increased with incubation temperature. Shemer and
Perkins (1975), however, reported partial degradation
of soy protein polypeptide chains in boiling water.
All of these studies cited showed that heat produces
changes in soybean proteins that affect the quality of
the resulting tofu. There is a need for further studies
on the effects of heat on soybean proteins and soy-
bean protein products. This study was conducted to
determine the effect of boiling soymilk prior to tofu
coagulation on the yield, composition, textural proper-
ties and sensory qualities of the resulting tofu.
Materials and Methods
Materials
Philippine grown soybeans of the L-I 14 variety were
used in this. study.
Preparation of Soymilk
Two kg of soybeans were soaked in tap water (three
times the amount of soybeans) at room temperature
for 8 h, then drained and washed with fresh tap water.
The soaked beans were then ground in batches using
a one gallon capacity stainless steel Waring Blender
for 5 min at high speed. The ratio of the beans to water
was maintained at I: 10 (w/ w). The slurry was filtered
through cheesecloth to obtain the soymilk. All extrac-
tions were done at room temperature.
The term "soymilk" as used in this paper, means
the water extract from whole soybeans with most or
all of the insolubles removed. This definition is impor-
tant because most studies on water extractable soybean
proteins have used hexane-defatted flakes, grits or
flour as the starting material. These materials had been
exposed to varying degrees of heat treatment before
the extraction began.
Preparation of Tofu
The soymilk was first pre-heated to about 85°C in
a stainless steel bucket immersed in vigorously boil-
ing water in a steam jacketed kettle to minimize scorch-
ing of the soymilk during boiling. The boiling water
was discarded, the pre-heated soymilk was poured into
the kettle and covered with aluminum foil except when
the soymilk was being stirred. At time intervals of 0,
12,30 and 60 min, 1.5 kg of soymilk was withdrawn
and placed in a stainless steel beaker immersed in cold
tap water to bring the temperature to about 80°C. A
water suspension of calcium sulfate was added at a
level of 0.25% (w/w) of CaS0 4 • The mixture was
allowed to coagulate for I h after a short but vigorous
stirring to mix the calcium sulfate with the soymilk.
The curd was poured into a wooden box (approxi-
mately 13 x 15 x 25 cm.) lined with cheesecloth and
the supernatant was allowed to drain through the 1.3
cm dia. holes in the sides and bottom of the box for
I h. The curd was allowed to set overnight at IO-15°C
under a weight of approximately 6.65 g/cm 2 , before
analyses and sensory evaluations were performed.
Textural Analysis
Texture Profile Analysis using an Instron Univer-
sal Testing Machine and following the method of
54 / Escueta et al.
Bourne (1968) was used to determine the textural qual-
ities of tofu. The Instron crosshead was set to cycle
with a vertical reciprocating movement at a constant
speed of 50 mm per min and with a stroke length of
7.5 mm. The maximum clearance between the mov-
ing horizontal plate and the stationary horizontal bed-
plate of the machine was 10 mm and the minimum
clearance was 2.5 mm giving a 751Jfo compression. Each
piece of tofu was compressed two times to give a first
and second bite. The chart speed was set at 50 cm/min
and the full scale load was adjusted to hardness of the
sample. Cylinders of tofu (2 cm dia. by 1 cm height)
used in the analyses were prepared using a cork borer.
Each test was replicated eight to ten times at room tem-
perature after equilibrating the samples for two h.
Hardness was measured as the peak of the first bite
and springiness was the distance of recovery between
the two bites. Areas under the curves were measured
by a planimeter. Based on the method of Bourne
(1968), cohesiveness, gumminess and chewiness were
calculated as representative textural parameters.
Chemical Analyses.
Analyses of samples from each treatment were made
in duplicate. Total nitrogen was determined by the
micro Kjeldahl method (A.O.A.C., 1975, method no.
47.021) on solid samples and the protein content was
calculated using a factor of 6.25 x N. Crude protein
content of liquid samples was determined by the
method of Lowry et al. (1951). Fat was determined
by petroleum ether extraction in the Goldfisch fat
extractor (A.O.A.C. 1975, method no. 14.080) and
moisture and total solids data were obtained in a
forced draft hot air oven at 100-I02°C for 24 h.
Recovery Calculations
Percent recovery of tofu was calculated by dividing
the weight of tofu by the original weight of soymilk
and multiplying the value by 100. The recoveries of
the total solids, protein and fat components of tofu
were calculated by dividing the weight of each of the
above components by its respective weight in the origi-
nal soymilk.
Sensory Evaluation
A trained panel evaluated flavor, mouthfeel, moist-
ness and overall acceptability using 7 point scales; their
ranges are indicated in the Results section. The tofu
was fried and cooled to room temperature (28-30°C)
before presenting a 40-50 g sample to the panelists for
evaluation. All samples were coded with randomly
Table I. Texture profile analysis of tofu from pre-boiled soymilk l
Boiling time (min)
o 12 30 60
Hardness (N) 4.04 c 4.07 c 3.45 b 2.77 a
Springiness(cm) 0.26a 0.24 a O.27 a 0.27 a
Chewiness (N-cm) 0.51 b 0.51 b 0.38 a 0.28 a
Cohesiveness (A 21AI) 0.48 ab 0.52 b O.4l a 0.39 a
Gumminess (N) I. 96ab 2.IO b 1.42ab 1.09a
1Row means with the same letters are not significantly different (P
= 0.05).
LSD values: Hardness, 0.34; Springiness - F value not significant;
chewiness, 0.12; Cohesiveness, 0.09; Gumminess, 1.01.
J. InSl. Can. Sci. Techno/. Alimem. Vol. 19. No.2, 1986
Table 2. Composition (070 "as is") of soymilk, tofu and whey as the increase in hardness of tofu, can occur between
affected by boiling. I
12 min and 30 min boiling.'
Boiling Time (min) Saio et al. (1971) observed the maximum amount
Component
o 12 30 60 of sulfhydryl groups after heating for 1 min at 100°C
A. Soymilk possibly because their soymilk was prepared from a
Total Solids 5.3" 5.5" defatted soybean meal. Soybean meal, unlike fresh
Fat 1.0" 1.2 b
Protein 2.4" 2.8"
whole soybeans, receives heat treatment that may
B. Tofu affect its protein as a result of the solvent extraction
Moisture 79.5" 79.8" 79.2" 80.9" process.
Fat 5.4" 5.3" 5.4" 5.4" Springiness was not affected significantly by the
Protein 11.2" 11.3" 11.9" 11.7" boiling treatment as indicated by the fairly constant
C. Whey
Total Solids 1.4" 1.5" 1.6"b 1.8 b values throughout the 60 min boiling period (Table 1).
Fat 0.2" 0.2" 0.2" 0.2" Chewiness decreased with boiling time with the
Protein 0.6" 0.6" 0.6" 0.6" decrease becoming significant after 30 min of boiling.
IRow means with the same letter are not significantly different Between 30 min and 60 min boiling, however, the
(P = .05) chewiness values did not differ significantly. Cohesive-
LSD values: A) Soymilk, total solids, 0.68; Fat, 0.19; Protein, 0.42;
B) Tofu, Moisture, Fat and Protein F-values not significant; C) ness and gumminess behaved similarly to hardness.
Whey, total solids, 0.26; Fat and Protein F-values were not sig- Both did not change significantly after 12 min boiling
nificant. but decreased significantly after 30 min boiling.
Chemical Composition
assigned numbers and presented to the panelist in a The soybean used had the following chemical com-
randomized arrangement to equalize the sample position (average of 3 analyses, "as is" basis):
sequence effect on food preference (Eindhoven et al., moisture, 10.5070; protein, 38.0%; fat, 16.2%; crude
1964). fiber, 4.8070; ash, 5.1 %; carbohydrates (by difference),
Statistical Design 25.42%. Boiling soymilk before coagulation caused no
The experiments were performed using the ran- significant effect on the composition of tofu (Table
domized complete block design (RCB). Each experi- 2). The composition of soymilk was affected signifi-
ment was done independently with 3 replications (3 cantly by boiling. Sixty min of boiling showed a highly
blocks) prepared as separate batches of soymilk. Ana- significant increase in the total solids content of soy-
lyses and measurements were made in duplicate, giv- milk over that of 0 min, 12 min and 30 min boiling.
ing a total of six values per observation. Analysis of The protein content of soymilk also increased signifi-
variance was performed using the RCB design, fol- cantly after 30 min and 60 min of boiling. The fat con-
lowed by a multiple range test using the least signifi- tent increased significantly after 12 min of boiling with
cant difference (LSD) when the F value was signifi- further, statistically non-significant increase after 60
cant (Cochran and Cox, 1957). min of boiling. These changes in the components of
soymilk during boiling were due primarily to the loss
Results and Discussion of water through evaporation which occurred during
Texture Profile Analysis the boiling treatment despite covering the kettle. It was
The data on textural qualities of tofu as affected by also shown (Table 2) that tofus of different composi-
the length of time of boiling soymilk prior to coagu- tions could not be achieved by merely prolonging the
lation are summarized in Table 1. The hardness was boiling time of soymilk prior to coagulation. Saio et
not affected by boiling for the first 12 min, but al. (1973) reported that tofu of 50% or less moisture
decreased significantly after boiling for 30 min and fur- content could be prepared from the 11 S protein frac-
ther decreased after 60 min boiling. tion but this condition cannot be achieved by boiling
An increase in the hardness of tofu might have been procedures.
observed if boiling exposures between 12 min and 30
min had been explored as it is possible that the maxi- Recovery of Tofu and Its Components
mum increase in sulfhydryl groups, which will cause Table 3 shows the effects of the different times of
boiling on the % recovery of tofu and its components.
Table 3. Percent yield of tofu, fat, protein and total solids from
pre-boiled soymilk I. Table 4. Sensory qualities of tofu from pre-boiled soymilk l
Boiling Time (min) Boiling Time (min)
Component Sensory parameters
o 12 30 60 o 12 30 60
Tofu 2 20.8" 2I.l" 21.2" 23.2 b Mouthfeel 5.3 5.2 5.4 5.5
Solids 3 79.6" 81.7" 88.3 c 84.9 b Flavor 5.0 4.9 5.3 5.1
Fat 3 88.5" 94.7 b 88.9" 93.6 b Moistness 5.5 5.4 5.4 5.5
Protein 3 88.5" 88.6" 95.4c n.5 b Acceptability 5.3 5.0 5.2 5.2
I Row means with the same letter are not significantly different. IMouthfeel, from I-very coarse to 7-very smooth; Flavor, from
2Percent of original weight of soymilk. I-very strong off-flavor to 7-very pleasant flavor; Moistness, from
3Percent of original amount in soymilk. I-very dry to 7-very moist; Acceptability, from I-dislike very much
LSD values: Tofu, 0.71; Solids, 2.27; Fat, 1.56; Protein, 1.71. to 7-like very much.

Can. lnst. Food Sci. Technol. J. Vol. 19. No.2, 1986 Escueta et al. / 55
Boiling soymilk for 30 min instead of 12 min did not
increase the recovery of tofu. The 070 recovery of tofu
increased significantly after boiling for 60 min. Hence,
the traditional tofu-making process which requires
10-15 min of boiling is reasonable from the viewpoint
of economy and nutrition since trypsin inhibitors and
other anti-nutritional factors in soymilk are destroyed
by 10-15 min boiling (Liener, 1972; Rackis, 1974). The
recovery of tofu can be increased by boiling for 60 min
but the product will be significantly softer as shown
in Table 1.
The fat recovery increased significantly after 12 and
60 min of boiling. The protein and solids recovery were
also significantly affected by the length of boiling. Pro-
tein and solids recoveries increased as the boiling time
was increased. The increases in yield of tofu, protein
and solids with increasing boiling time were obviously
caused by the unavoidable concentration of the soy-
milk during boiling, as shown in Table 2.
Sensory Qualities
Table 4 shows that the time of boiling soymilk prior
to coagulation did not have a significant effect on the
sensory qualities of tofu. The acceptability scores of
about 5 given to all of the tofu samples are moderate
even though tofu is a customary part of the Filipino
diet. This suggests that the way the tofu is cooked or
prepared has more influence on its acceptability than
tofu as a product itself, which is bland in flavor and
is rarely eaten alone. Future acceptability tests should
be made with tofu prepared as a traditional Filipino
dish.
Conclusion
The composition and sensory qualities of tofu were
not significantly affected by the time of boiling soy-
milk prior to coagulation. The yield increased slightly
up to 30 min and then significantly after 60 min
boiling.
Tofu of acceptable sensory quality and chemical
composition was made from pre-heated soymilk (0 min
treatment). Prolonged boiling of soymilk gave no
advantage in terms of composition and acceptability
of tofu. The increase in the yield of tofu at the longer
boiling periods has to be weighed carefully against the
higher energy requirement of the longer boiling
process. Boiling for 30 min and 60 min produced tofu
that was softer and less chewy. Cohesiveness of tofu
also decreased significantly after 60 min boiling.
Springiness was the only textural property that was not
affected by boiling treatment. Thus in terms of econ-
omy, textural quality and yield the 0 min pre-boiling
treatment of soymilk prior to coagulation is sufficient
but 12 min boiling of soymilk prior to coagulation is
still appropriate in order to destroy the trypsin inhibi-
tors and other anti-nutritional factors present in
soymilk.
Acknowledgement
This research project was supported by the National
Science and Technology Authority (NSTA) formerly
56 / Escuata et al.
National Science Development Board of the Philip-
pines.
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Submitted May 29, 1985
Revised September 8, 1985
Accepted October II, 1985
J. Ins!. Can. Sci. Technol. Aliment. Vol. 19, No.2, 1986