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Effect of freeze-dried soybean curd (tofu) on various bodily functions
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The present study was designed to examine the effect of freeze-dried soybean
curd (tofu) on various bodily functions. The dietary experiment consisted of 4 days of a
non-prescribed ordinary diet, 10 days of an experimental diet that contained 190 g of meat
contributing about 38 g of protein (hereinafter referred to as the “meat period”), and 39
days of an experimental diet that contributed vegetable protein, i.e., freeze-dried tofu,
corresponding to the protein that 190 g of meat would provide (hereinafter referred to
as the “freeze-dried tofu period”). Three eggs (about 150 g) and 180 ml of cow’s milk were
prescribed for the daily diet, and staple and other foods were dispensed freely during both
the meat and the freeze-dried tofu periods. The results showed that serum cholesterol
levels and diastolic blood pressure significantly decreased in the freeze-dried tofu period
as compared with the meat period. Freeze-dried tofu was found to be a valuable food for
preventing lifestyle-related chronic diseases.
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Effect of freeze-dried soybean curd (tofu) on various bodily
functions
Masanobu Fujii?, Tadataka Fukui†, Tamotsu Miyoshi‡
?Division of Medical Technology, The School of Medical Sciences, The University of Tokushima,
Tokushima, Japan ; †The Preventive Arteriosclerosis Research Association, Tokyo, Japan ; and
‡Department of Public Health, The University of Tokushima School of Medicine, Tokushima, Japan
Abstract : The present study was designed to examine the effect of freeze-dried soybean
curd (tofu) on various bodily functions. The dietary experiment consisted of 4 days of a
non-prescribed ordinary diet, 10 days of an experimental diet that contained 190 g of meat
contributing about 38 g of protein (hereinafter referred to as the “meat period”), and 39
days of an experimental diet that contributed vegetable protein, i.e., freeze-dried tofu,
corresponding to the protein that 190 g of meat would provide (hereinafter referred to
as the “freeze-dried tofu period”). Three eggs (about 150 g) and 180 ml of cow’s milk were
prescribed for the daily diet, and staple and other foods were dispensed freely during both
the meat and the freeze-dried tofu periods. The results showed that serum cholesterol
levels and diastolic blood pressure significantly decreased in the freeze-dried tofu period
as compared with the meat period. Freeze-dried tofu was found to be a valuable food for
preventing lifestyle-related chronic diseases. J. Med. Invest. 46 : 67-74, 1999
Key words : freeze-dried soybean curd, vegetable oil, serum cholesterol, blood pressure
INTRODUCTION
EXPERIMENTAL METHOD
With the recent developments in dietetics con-
A. Experimental Period and Subjects
cerning proteins, amino acids, and fatty acids, the
The periods of the experiment were enacted over
value of soybean protein, as well as animal protein
a total of 71 days in the following order : ordinary diet
that contains a healthy balance of essential amino
period I (4 days), meat period (10 days), freeze-dried
acids, is receiving much attention. On the other hand,
tofu period I (19 days), freeze-dried tofu period II
it has been shown that a large intake of animal fat
(20 days), and ordinary diet period II (18 days). The
contained in animal food can contribute to arterio-
subjects were 4 healthy male and 2 female adults
sclerosis (1-3). Thus, much attention is being paid
(Table 1).
to the nutritional value of soybean products that con-
tain vegetable protein (4-5). Fueling this interest,
Table 1
Michi
. Age, sex and physical characteristics of subjects
et al. (6) report that freeze-dried soybean curd
(tofu), a soybean product, decreases serum choles-
Subject
Age
Sex
Height
Body weight
terol levels.
(No.)
(years)
(cm)
(kg)
In the present study, a dietary experiment was
1
62
male
155.4
55.0
performed to determine the effect of freeze-dried
tofu on various bodily functions.
2
45
male
162.4
66.7
3
40
male
170.2
68.5
Received for publication November 26, 1998 ; accepted January
6, 1999.
4
32
male
174.6
58.0
Address correspondence and reprint requests to Masanobu
5
40
female
157.8
55.0
Fujii, Ph.D., Division of Medical Technology, The School of
Medical Sciences, The University of Tokushima, Kuramoto-cho,
6
27
female
157.6
46.0
Tokushima 770-8509, Japan and Fax : +81-88-633-9070.
The Journal of Medical Investigation
Vol.46
1999
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
B. Measurement Items and Nutrient intakes
ing drinks and staple food, without any restrictions.
1. Body weight, condition of fatigue, and blood
During the freeze-dried tofu period, the previously
pressure
administered meat protein was replaced by vegetable
Body weight was measured before breakfast once
protein, i.e., freeze-dried tofu (about 20 g/tofu x 4
a day. The condition of fatigue was assessed before
units). Similarly, three eggs (about 150 g) and 180
lunch once a day by determining flicker values
ml of cow’
s milk were prescribed during the tofu
(obtained using Ohshima’
s measurement apparatus
period, but other animal protein intake was for-
(7), blinker values obtained using Fukui’
s measure-
bidden. Staple food, other foods, and drinks were
ment apparatus) (7), and grasp values (obtained using
allowed without any restrictions.
dynamometer of Smedlay’
s method) (7). Blood pres-
The aim of this study was to examine the effect
sure was measured with a sphygmometroscope.
of freeze-dried tofu on various functions of the body.
2. Blood studies
To measure this effect, during the experimental
Blood samples were collected before breakfast.
portion of testing, while a sufficient total protein
Whole blood gravity (Gb), hemoglobin (Hb) and
intake was prescribed throughout testing, animal
hematocrit (Ht) levels were measured using the
protein was replaced by vegetable protein, i.e.,
copper sulfate method, the cyanomethemoglobin
freeze-dried tofu, and animal protein intake, other
method, and the capillary method, respectively.
than the prescribed three eggs and 180 ml of cow’
s
3. Liver function tests and measurement of serum
milk, was forbidden.
protein levels
Serum was separated from the blood samples
collected before breakfast to measure serum levels
RESULTS
of total protein (TP) and liver function under the
parameters such as GOT, GPT, ALP, albumin, and
1. Nutrient intakes
ZTT. A kit for liver function tests was supplied by
Table 2 shows the mean nutrient intakes of the
Chugai Pharmaceutical Co., Ltd., and a specific
six subjects in each experimental period. The meat
apparatus was used for the tests.
period was associated with the highest protein
4. Measurement of serum lipids
intake (108.4 g), which was about 38% higher than
Serum was separated from the blood samples
that in the ordinary diet period I. Protein intake was
collected early in the morning before the subjects
about 90 g in freeze-dried tofu periods I and II, which
had ingested any food, to measure serum total cho-
was slightly higher than that in ordinary diet period
lesterol and serum free cholesterol levels. A kit for
I. All subjects had consumed an amount of protein
measuring serum lipids was supplied by Chugai
that satisfied the protein requirement in all experi-
Pharmaceutical Co., Ltd., and a specific apparatus
mental periods. As for the nature of protein consumed,
was used for the test. Blood studies were performed
animal protein alone increased by about 70% in the
on Day 3 of ordinary diet period I, on Day 8 of the
meat period as compared with the ordinary diet
meat period, on Days 9 and 18 of freeze-dried tofu
period. In the freeze-dried tofu period, however, it
period I, on Day 18 of freeze-dried tofu period II,
decreased to about half that in the ordinary diet
and on Day 18 of ordinary diet period II.
period, and there was a two-fold increase in vegeta-
5. Nutrient intakes
ble protein. Consequently, the animal/vegetable
Based on individual food intake data that was
protein ratio was about 3 : 7 in the freeze-dried tofu
recorded for a selected two-day span during each
periods I and II, as opposed to 7 : 3 in the meat
experimental period, nutrient intakes were calcu-
period. Animal fat intake was 2.5 times higher in the
lated according to the Standard Tables of Food
meat period than in the ordinary diet period I, reflect-
Composition in Japan (8).
ing changes in the nature of protein. On the other
hand, whereas animal fat intake was only slightly
C. Conditions for Dietary Intake in Experimental
lower in freeze-dried tofu periods I and II than in the
Periods
ordinary diet period I, there was a two-fold increase
During the ordinary diet period, the subjects
in vegetable fat intake during freeze-dried tofu peri-
were given an unregulated diet. During the meat
ods. In the freeze-dried tofu periods, calcium intake
period, they were required to ingest meat (190 g),
was also particularly high, reaching 1.6 times (980
three eggs, and 180 ml of cow’
s milk per day, but
mg) the calcium requirement. Cholesterol intake
the subjects were also allowed other foods, includ-
was the highest (1100 mg) in the meat period in
The Journal
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Vol.46
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1999
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which animal protein was predominant, followed by
body weight gain was seen in the shift from the
the freeze-dried tofu period in which eggs were
ordinary diet period I to the meat and freeze-dried
given to supply animal protein (over 800 mg). In the
tofu periods. This may be attributable to an increase
ordinary diet periods I and II, cholesterol intake was
in caloric intake. There were almost no differences
about half that in the meat period.
in systolic blood pressure between ordinary diet
2. Changes in body weight and blood pressure
period I and the meat period, but it was lower in the
The mean body weight and blood pressure of the
freeze-dried tofu periods than in the meat period.
six subjects are shown according to the experi-
On the other hand, there was a statistically signifi-
mental period of measurement in Table 3. Slight
cant decrease in diastolic blood pressure from the
Table 2. Nutrient intakes in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Energy (kcal)
2139.7±388.0
2685.2±349.3
2268.7±381.5
2068.8±267.8
1994.8±469.6
Protein total (g)
78.8±12.7
108.4±10.4
90.0±9.6
88.2±5.3
84.6±16.2
Animal (g)
46.3±10.9
78.3±5.4
26.0±0.1
23.6±2.4
54.9±12.3
Animal protein
58.4±8.2
72.4±3.1
29.1±2.8
26.8±3.4
64.7±5.8
ratio (%)
Lipid total (g)
51.5±19.3
101.1±12.8
71.8±8.5
63.3±4.5
48.9±21.2
Animal (g)
31.9±18.1
78.6±5.3
30.5±3.1
26.1±0.5
34.6±18.7
Animal lipid
60.0±14.6
78.2±5.5
42.6±2.3
41.3±4.2
69.6±12.4
ratio (%)
Carbohydrate (g)
314.9±67.3
302.4±54.0
298.0±50.4
280.9±53.2
272.0±73.1
Minerals (mg)
Ca
559.2±121.4
64.0±89.4
991.2±124.0
968.5±71.4
480.0±191.1
Fe
12.2±2.3
18.7±1.7
18.3±2.1
18.2±1.8
12.7±2.5
Vitamins A (IU)
1425.5±521.4
3258.3±526.0
2519.5±597.8
3035.2±668.5
1809.8±486.8
B1 (mg)
1.47±1.05
2.33±0.35
1.66±0.28
1.21±0.09
1.30±1.14
B2 (mg)
1.19±0.24
1.72±0.14
1.21±0.11
1.21±0.09
1.12±0.27
C (mg)
59.2±25.9
87.0±20.5
86.7±28.9
103.7±29.1
128.2±57.1
Cholesterol (mg)
479.11±?12.4
1111.1±40.7
848.0±13.0
796.8±113.3
510.6±221.8
Table 3. Changes in body weight and blood pressure in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Body weight (kg)
55.5±14.7
56.8±7.7
58.6±7.6
58.8±7.5
58.1±8.8
1)
SBP (mmHg)
114.3±15.4
114.4±11.0
111.7±10.6
111.2±11.0
115.8±12.0
2)
DBP (mmHg)
71.4±6.0
72.9±7.6
69.1±7.1**
69.9±6.2*
72.2±6.3
1) Systolic blood pressure
2) Diastolic blood pressure
Values are expressed as mean±standard deviation of six subjects.
*p<0.05, **p<0.01 Significantly different from the Meat diet.
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
meat period to the freeze-dried tofu periods I (p<
cholesterol and serum free cholesterol in the six
0.01) and II (p<0.05), as opposed to the increase in
subjects according to the experimental period of
diastolic blood pressure that occurred between the
measurement. There were significant increases in
ordinary diet period to the meat period. Both systolic
both serum total cholesterol (p<0.01) and serum
and diastolic blood pressures were lower in the
free cholesterol levels (p<0.05) in the meat period
freeze-dried tofu periods than in the ordinary diet
as compared with ordinary diet period I. However,
period I.
both levels decreased between the meat period and
3. Measurement of fatigue
freeze-dried tofu periods. In particular, there was a
Mean flicker values, blinker values, and grasp
statistically significant difference in total cholesterol
values in six subjects are shown according to the
levels (p<0.05) and free cholesterol levels (p<0.01)
experimental period of measurement in Fig.1. There
between the freeze-dried tofu period II and the
were minimal differences in all measured values
meat period. Free cholesterol levels were lower in
among the experimental periods.
freeze-dried tofu periods I and II than in the other
4. Changes in blood findings and liver function
experimental periods, and the ester rate tended to
findings
be high in the freeze-dried tofu periods.
As shown in Table 4, there were almost no dif-
ferences in Gb, Hb, or Ht among the experimental
periods. On the other hand, there were slight differ-
DISCUSSION
ences in serum proteins and liver function findings
among the periods, which may have been attrib-
The present study of nutrient intakes revealed
utable to the different quantity of protein intake.
that approximately 80 g of protein was consumed
However, these findings were all within physiologi-
and the animal/vegetable protein ratio was 6 : 4
cally normal limits.
during the ordinary diet periods. In the meat period,
5. Changes in serum total cholesterol and serum
during which the volunteers had to take 190 g of
free cholesterol
meat, the prescribed three eggs (about 150 g), 180
Table 5 shows the mean levels of serum total
ml of cow’
s milk, and freely distributed staple and
Fig.1. Flicker, Blinker, and Grasp values in each experimental diet
The Journal
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1999
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Table 4. Changes in blood parameters in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Blood examinations
GB
1.0567±0.0046
1.0567±0.0035
1.0558±0.0032
1.0557±0.0031
1.0555±0.0024
Hb (g/dl)
14.6±1.3
14.3±1.5
13.9±1.3
14.5±1.3
14.3±1.4
Ht(%)
43.8±3.9
43.8±4.0
43.4±3.2
43.1±3.6
43.2±3.4
TP (g/dl)
7.5±0.6
7.9±0.5
7.5±0.5
7.8±0.2
7.7±0.5
Liver function tests
GOT (Karmen Unit)
14.8±12.4
16.2±8.5
17.5±7.2
10.7±8.0
14.3±11.9
GPT (Karmen Unit)
11.8±8.2
14.3±6.4
15.2±5.0
10.2±4.3
9.5±6.4
ALP (King-
8.7±1.2
9.0±1.5
9.3±1.8
10.5±2.1
11.8±2.7
Armstrong Unit)
ZTT (Kunkel Unit)
6.9±7.0
5.8±1.9
7.6±2.8
6.5±2.6
5.0±2.2
Albumin (g/dl)
4.2±0.4
4.4±0.3
4.1±0.3
4.4±0.3
4.4±0.2
Values are expressed as Mean±Standard deviation of 6 subjects.
Table 5. Changes in serum cholesterol in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Total cholesterol
184.8±15.4
220.0±19.5??
199.0±20.6
188.3±19.0*
201.4±14.0
(mg/dl)
Free cholesterol
57.7±5.8
70.5±9.1?
56.8±7.9*
54.3±3.5**
62.2±4.7??
(mg/dl)
Cholesterol ester
68.9±3.6
68.1±1.7
71.5±1.1**
71.1±1.4**
69.3±2.8
(%)
?p<0.05, ??p<0.01 Significantly different from the Ordinary diet I.
*p<0.05, **p<0.01 Significantly different from the Meat diet.
??p<0.01 Significantly different from the Freeze-dried tofu diet II.
other foods, total protein was about 110 g. Animal
intake was dependent on the nature of protein. In
protein intake alone increased by about 70% as
the ordinary diet periods, total fat intake was about
compared with the ordinary diet period I. Conse-
50 g, and the animal/vegetable lipid ratio was 6 : 4.
quently, the animal/vegetable protein ratio was 7 : 3.
The meat period brought about a 2.5-fold increase
In the freeze-dried tofu periods, the 38 g of protein
in animal fat intake, and the animal/vegetable lipid
that originated in the previously administered 190 g
ratio became 8 : 2. In the freeze-dried tofu period,
of meat was replaced by about 72 g of (vegetable
animal fat intake was almost similar to that in the
protein) freeze-dried tofu, and three eggs and 180
ordinary diet periods, but vegetable protein intake
ml of cow’
s milk were administered to contribute
was two times higher (about 40 g) than in the ordi-
animal protein. The results showed that total pro-
nary diet periods. As a result, the animal/vegetable
tein intake was slightly higher (about 90 g) in the
lipid ratio was 4 : 6, opposite to that in the ordinary
freeze-dried tofu periods than in the ordinary diet
diet periods. The discrepancy in such nutrient in-
periods, but that the animal/vegetable protein ratio
takes led to changes in blood-test results. There was
was 3 : 7, opposite to that in the meat period. Fat
a statistically significant increase in serum total
?
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
cholesterol and free cholesterol levels in the meat
and a higher proportion of vegetable oil contained
period as compared with the ordinary diet periods.
in freeze-dried tofu (11-12). Besides the vegetable
On the other hand, both parameters significantly
oil in freeze-dried tofu, we consider that soybean
decreased in the freeze-dried tofu period II as
protein also had an effect on lowering the serum
compared with the meat period. The proportion of
cholesterol (13). As shown in Fig. 4, there was an
animal fat intake, as well as cholesterol intake, are
inverse correlation between the proportion of animal
dietary factors influencing serum cholesterol levels,
protein and the proportion of animal fat/vegetable
and the proportion of animal fat intake is known to
oil. While it is important to maintain a certain level
influence the rate of absorption of cholesterol intake
of animal protein in terms of the quality of protein
(9-10). Accordingly, the correlation between the
intake, an extremely high animal protein intake
cholesterol intake and the proportion of animal fat
leads to an excessive intake of saturated fatty acid,
intake in the experiment was expressed as the
resulting in an increase in serum cholesterol. Con-
following formula : cholesterol intake x animal fat
sidering these factors, the proportion of animal
intake/vegetable oil intake. As shown in Figs. 2
protein intake is recommended to be within 40% -
and 3, there was a statistically significant correlation
50% (14). In the present study, the proportion of
between the proportion of animal fat intake and both
animal protein intake to the entire nutrient intake
serum total cholesterol and serum free cholesterol.
was 60% -70%, which exceeded the recommended
The fact that the meat period was associated with
range, even in the ordinary diet period as well as
not only higher cholesterol intake but also a higher
in the meat period. This result should be carefully
proportion of animal fat intake appears to reflect the
considered, because it may lead to an excessive
appearance of higher serum cholesterol levels. On
intake of saturated fatty acid and an increase in
the other hand, although the cholesterol intake was
serum cholesterol. On the other hand, because the
1.5 times higher in the freeze-dried tofu periods
proportion of animal protein intake was found to be
than in the ordinary diet periods, and although there
low (about 30%) during the freeze-dried tofu periods,
was little difference in cholesterol intake between
freeze-dried tofu appears to prevent an excessive
the freeze-dried tofu period and the meat period, the
intake of saturated fatty acid and to restrain an
fact that serum cholesterol decreased may be ex-
increase in serum cholesterol. It is also thought
plained by a lower proportion of animal fat intake
that limiting amino acids do not appear if there is
Fig. 2. Correlation between serum total cholesterol and values of multipling cholesterol intake by ratio of animal
lipid intake to vegetable oil intake
The Journal
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1999
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sufficient protein intake, even of the proportion of
and N/E ratio in the present study, there were no
animal protein intake is only 30% (15). When the
differences in these ratios between the freeze-dried
amount of amino acids contained in the food con-
tofu period and the ordinary diet period. This indi-
sumed was calculated to determine the E/T ratio
cates that the quality of protein is not reduced by a
Fig. 3. Correlation between serum free cholesterol and values of multipling cholesterol intake by ratio of animal lipid
intake to vegetable oil intake
Fig. 4. Relationship between percent of animal protein intake and ratio of vegetable oil intake to animal lipid intake
?
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
freeze-dried tofu diet. This can also be explained
6.
Michi K, Matsushita K, Ezawa I : Effect of
by the fact that there were no differences in blood
dehydrofrozen soy bean curd, Koritofu, and
findings, such as Gb, Hb, Ht, and TP, or liver func-
dried soy bean product, Yuba, on the plasma
tion findings among the experimental periods. From
cholesterol. Nippon Eiyo Shokuryo Gakkaishi
the standpoint of preventing lifestyle-related chronic
(in Japanese) 19 (6) : 449-452, 1967
diseases, it is important to maintain the normal
7.
Kirihara H : Functional tests of fatigue evalu-
limits of blood pressure. In the present study, dias-
ation (in Japanese). Nippon Sangyo Eisei Kyokai,
tolic blood pressure, which is frequently associated
Tokyo, 1967
with high fat intakes and high cholesterol intakes,
8.
Resources Council, Science and Technology
tended to increase in the meat period (16). On the
Agency, Japan. Standard Tables of Food Com-
contrary, freeze-dried tofu decreased both systolic
position in Japan, 4 th ed (in Japanese). Daiich
blood pressure and significantly decreased diastolic
Syuppan, Tokyo, 1983
blood pressure. It has already been shown that
9.
Keys A, Anderson JT, Grande F : Prediction of
there is a correlation between blood pressure and
serum cholesterol response of man to changes
dietary calcium and magnesium intakes (17), and
in fat in the diet. Lancet 2 : 959-966, 1957
as freeze-dried tofu is rich in minerals it deserves
10. Hegsted DM, Mcgardy RB, Myers ML, Stare
further attention. In conclusion, when animal pro-
FJ : Quantitative effects of dietary fat on serum
tein intake was restricted to 30% and freeze-dried
cholesterol in man. Amer J Clin Nutr 17 : 281-
tofu administered to maintain a total protein intake
295, 1965
similar to that of a meat-rich diet, excessive eating
11. Michi K, Ezawa I : The effect of vegetable oil
can be prevented, and blood findings, blood pressure,
on the plasma, liver and adrenal cholesterol.
and serum cholesterol levels can be maintained within
Nippon Eiyo Shokuryo Gakkaishi (in Japanese)
physiologically normal limits. Therefore, freeze-dried
18 : 220-203, 1965
tofu appears to be a valuable food for preventing
12. Kaneda N : Cholesterol and food. Shoku no
lifestyle-related chronic diseases.
Kagaku (in Jpanese) 17 : 115-121, 1974
13. Yamamoto T, Inoue G : Effect of soybean pro-
tein on plasma cholesterol concentration in
REFERENCES
humans. Nippon Eiyo Shokuryo Gakkaishi (in
Japanese) 44 : 155-162, 1991
1.
Kannel WB, Catelli MD, Gordon T, Mcnamara
14. Yamaguchi M : Recommended dietary allow-
PM : Serum cholesterol, lipoproteins, and the
ances for the Japanese protein. Rinsho Eiyo
risk of coronary heart disease. Ann Intern Med
(in Jpanese) 85 : 164-168, 1994
74 : 1-12, 1971
15. Matsuno N : On the essential amino acid intake
2.
Martin MJ, Hulley SB, Browner WS, kuller
of the Japanese people (report 10). The essen-
LH : Serum cholesterol, blood pressur, and
tial amino acid intake of the Japanese people
mortality implications from a cohort of 361662
calculated from the National Nutrition Survey
men. Lancet 2 : 933-939, 1986
in Japan from 1946-1971. Jap J Nutr (in Japanese)
3.
Keys A : Coronary heart disease in seven
33 : 311-320, 1975
countries. Circulation 41 : I-1-I-211, 1970
16. Komachi Y : Circulatory organic disease and nu-
4.
Imaizumi K, Mawatari K, Murata M, Ikeda I,
trition. In : Itakura H, Yoshimura M, Yasumoto
Sugano M : The contrasting effect of dietary
K, eds. Geriatric diseases and nutrition (in
phosphatidylcholine on serum lipoproteins and
Japanese). Koseikan Inc, Tokyo, 1992 pp. 61-98
liver lipids in rats. J Nutr 113 : 2403-2411, 1983
17. Yoshimura M : Hypertension and nutrition.
5.
Tanaka k, Imaizumi K, Sugano M : Effect of
In : Itakura H, Yoshimura M, Yasumoto K, eds.
dietary proteins on the intestinal synthesis and
Geriatric diseases and nutrition (in Japanese).
transport of cholesterol and apolipoprotein A-I
Koseikan Inc, Tokyo, 1992, pp. 99-143
in rat. J Nutr 113 : 2403-2411, 1983
?
?
Effect of freeze-dried soybean curd (tofu) on various bodily
functions
Masanobu Fujii?, Tadataka Fukui†, Tamotsu Miyoshi‡
?Division of Medical Technology, The School of Medical Sciences, The University of Tokushima,
Tokushima, Japan ; †The Preventive Arteriosclerosis Research Association, Tokyo, Japan ; and
‡Department of Public Health, The University of Tokushima School of Medicine, Tokushima, Japan
Abstract : The present study was designed to examine the effect of freeze-dried soybean
curd (tofu) on various bodily functions. The dietary experiment consisted of 4 days of a
non-prescribed ordinary diet, 10 days of an experimental diet that contained 190 g of meat
contributing about 38 g of protein (hereinafter referred to as the “meat period”), and 39
days of an experimental diet that contributed vegetable protein, i.e., freeze-dried tofu,
corresponding to the protein that 190 g of meat would provide (hereinafter referred to
as the “freeze-dried tofu period”). Three eggs (about 150 g) and 180 ml of cow’s milk were
prescribed for the daily diet, and staple and other foods were dispensed freely during both
the meat and the freeze-dried tofu periods. The results showed that serum cholesterol
levels and diastolic blood pressure significantly decreased in the freeze-dried tofu period
as compared with the meat period. Freeze-dried tofu was found to be a valuable food for
preventing lifestyle-related chronic diseases. J. Med. Invest. 46 : 67-74, 1999
Key words : freeze-dried soybean curd, vegetable oil, serum cholesterol, blood pressure
INTRODUCTION
EXPERIMENTAL METHOD
With the recent developments in dietetics con-
A. Experimental Period and Subjects
cerning proteins, amino acids, and fatty acids, the
The periods of the experiment were enacted over
value of soybean protein, as well as animal protein
a total of 71 days in the following order : ordinary diet
that contains a healthy balance of essential amino
period I (4 days), meat period (10 days), freeze-dried
acids, is receiving much attention. On the other hand,
tofu period I (19 days), freeze-dried tofu period II
it has been shown that a large intake of animal fat
(20 days), and ordinary diet period II (18 days). The
contained in animal food can contribute to arterio-
subjects were 4 healthy male and 2 female adults
sclerosis (1-3). Thus, much attention is being paid
(Table 1).
to the nutritional value of soybean products that con-
tain vegetable protein (4-5). Fueling this interest,
Table 1
Michi
. Age, sex and physical characteristics of subjects
et al. (6) report that freeze-dried soybean curd
(tofu), a soybean product, decreases serum choles-
Subject
Age
Sex
Height
Body weight
terol levels.
(No.)
(years)
(cm)
(kg)
In the present study, a dietary experiment was
1
62
male
155.4
55.0
performed to determine the effect of freeze-dried
tofu on various bodily functions.
2
45
male
162.4
66.7
3
40
male
170.2
68.5
Received for publication November 26, 1998 ; accepted January
6, 1999.
4
32
male
174.6
58.0
Address correspondence and reprint requests to Masanobu
5
40
female
157.8
55.0
Fujii, Ph.D., Division of Medical Technology, The School of
Medical Sciences, The University of Tokushima, Kuramoto-cho,
6
27
female
157.6
46.0
Tokushima 770-8509, Japan and Fax : +81-88-633-9070.
The Journal of Medical Investigation
Vol.46
1999
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
B. Measurement Items and Nutrient intakes
ing drinks and staple food, without any restrictions.
1. Body weight, condition of fatigue, and blood
During the freeze-dried tofu period, the previously
pressure
administered meat protein was replaced by vegetable
Body weight was measured before breakfast once
protein, i.e., freeze-dried tofu (about 20 g/tofu x 4
a day. The condition of fatigue was assessed before
units). Similarly, three eggs (about 150 g) and 180
lunch once a day by determining flicker values
ml of cow’
s milk were prescribed during the tofu
(obtained using Ohshima’
s measurement apparatus
period, but other animal protein intake was for-
(7), blinker values obtained using Fukui’
s measure-
bidden. Staple food, other foods, and drinks were
ment apparatus) (7), and grasp values (obtained using
allowed without any restrictions.
dynamometer of Smedlay’
s method) (7). Blood pres-
The aim of this study was to examine the effect
sure was measured with a sphygmometroscope.
of freeze-dried tofu on various functions of the body.
2. Blood studies
To measure this effect, during the experimental
Blood samples were collected before breakfast.
portion of testing, while a sufficient total protein
Whole blood gravity (Gb), hemoglobin (Hb) and
intake was prescribed throughout testing, animal
hematocrit (Ht) levels were measured using the
protein was replaced by vegetable protein, i.e.,
copper sulfate method, the cyanomethemoglobin
freeze-dried tofu, and animal protein intake, other
method, and the capillary method, respectively.
than the prescribed three eggs and 180 ml of cow’
s
3. Liver function tests and measurement of serum
milk, was forbidden.
protein levels
Serum was separated from the blood samples
collected before breakfast to measure serum levels
RESULTS
of total protein (TP) and liver function under the
parameters such as GOT, GPT, ALP, albumin, and
1. Nutrient intakes
ZTT. A kit for liver function tests was supplied by
Table 2 shows the mean nutrient intakes of the
Chugai Pharmaceutical Co., Ltd., and a specific
six subjects in each experimental period. The meat
apparatus was used for the tests.
period was associated with the highest protein
4. Measurement of serum lipids
intake (108.4 g), which was about 38% higher than
Serum was separated from the blood samples
that in the ordinary diet period I. Protein intake was
collected early in the morning before the subjects
about 90 g in freeze-dried tofu periods I and II, which
had ingested any food, to measure serum total cho-
was slightly higher than that in ordinary diet period
lesterol and serum free cholesterol levels. A kit for
I. All subjects had consumed an amount of protein
measuring serum lipids was supplied by Chugai
that satisfied the protein requirement in all experi-
Pharmaceutical Co., Ltd., and a specific apparatus
mental periods. As for the nature of protein consumed,
was used for the test. Blood studies were performed
animal protein alone increased by about 70% in the
on Day 3 of ordinary diet period I, on Day 8 of the
meat period as compared with the ordinary diet
meat period, on Days 9 and 18 of freeze-dried tofu
period. In the freeze-dried tofu period, however, it
period I, on Day 18 of freeze-dried tofu period II,
decreased to about half that in the ordinary diet
and on Day 18 of ordinary diet period II.
period, and there was a two-fold increase in vegeta-
5. Nutrient intakes
ble protein. Consequently, the animal/vegetable
Based on individual food intake data that was
protein ratio was about 3 : 7 in the freeze-dried tofu
recorded for a selected two-day span during each
periods I and II, as opposed to 7 : 3 in the meat
experimental period, nutrient intakes were calcu-
period. Animal fat intake was 2.5 times higher in the
lated according to the Standard Tables of Food
meat period than in the ordinary diet period I, reflect-
Composition in Japan (8).
ing changes in the nature of protein. On the other
hand, whereas animal fat intake was only slightly
C. Conditions for Dietary Intake in Experimental
lower in freeze-dried tofu periods I and II than in the
Periods
ordinary diet period I, there was a two-fold increase
During the ordinary diet period, the subjects
in vegetable fat intake during freeze-dried tofu peri-
were given an unregulated diet. During the meat
ods. In the freeze-dried tofu periods, calcium intake
period, they were required to ingest meat (190 g),
was also particularly high, reaching 1.6 times (980
three eggs, and 180 ml of cow’
s milk per day, but
mg) the calcium requirement. Cholesterol intake
the subjects were also allowed other foods, includ-
was the highest (1100 mg) in the meat period in
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which animal protein was predominant, followed by
body weight gain was seen in the shift from the
the freeze-dried tofu period in which eggs were
ordinary diet period I to the meat and freeze-dried
given to supply animal protein (over 800 mg). In the
tofu periods. This may be attributable to an increase
ordinary diet periods I and II, cholesterol intake was
in caloric intake. There were almost no differences
about half that in the meat period.
in systolic blood pressure between ordinary diet
2. Changes in body weight and blood pressure
period I and the meat period, but it was lower in the
The mean body weight and blood pressure of the
freeze-dried tofu periods than in the meat period.
six subjects are shown according to the experi-
On the other hand, there was a statistically signifi-
mental period of measurement in Table 3. Slight
cant decrease in diastolic blood pressure from the
Table 2. Nutrient intakes in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Energy (kcal)
2139.7±388.0
2685.2±349.3
2268.7±381.5
2068.8±267.8
1994.8±469.6
Protein total (g)
78.8±12.7
108.4±10.4
90.0±9.6
88.2±5.3
84.6±16.2
Animal (g)
46.3±10.9
78.3±5.4
26.0±0.1
23.6±2.4
54.9±12.3
Animal protein
58.4±8.2
72.4±3.1
29.1±2.8
26.8±3.4
64.7±5.8
ratio (%)
Lipid total (g)
51.5±19.3
101.1±12.8
71.8±8.5
63.3±4.5
48.9±21.2
Animal (g)
31.9±18.1
78.6±5.3
30.5±3.1
26.1±0.5
34.6±18.7
Animal lipid
60.0±14.6
78.2±5.5
42.6±2.3
41.3±4.2
69.6±12.4
ratio (%)
Carbohydrate (g)
314.9±67.3
302.4±54.0
298.0±50.4
280.9±53.2
272.0±73.1
Minerals (mg)
Ca
559.2±121.4
64.0±89.4
991.2±124.0
968.5±71.4
480.0±191.1
Fe
12.2±2.3
18.7±1.7
18.3±2.1
18.2±1.8
12.7±2.5
Vitamins A (IU)
1425.5±521.4
3258.3±526.0
2519.5±597.8
3035.2±668.5
1809.8±486.8
B1 (mg)
1.47±1.05
2.33±0.35
1.66±0.28
1.21±0.09
1.30±1.14
B2 (mg)
1.19±0.24
1.72±0.14
1.21±0.11
1.21±0.09
1.12±0.27
C (mg)
59.2±25.9
87.0±20.5
86.7±28.9
103.7±29.1
128.2±57.1
Cholesterol (mg)
479.11±?12.4
1111.1±40.7
848.0±13.0
796.8±113.3
510.6±221.8
Table 3. Changes in body weight and blood pressure in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Body weight (kg)
55.5±14.7
56.8±7.7
58.6±7.6
58.8±7.5
58.1±8.8
1)
SBP (mmHg)
114.3±15.4
114.4±11.0
111.7±10.6
111.2±11.0
115.8±12.0
2)
DBP (mmHg)
71.4±6.0
72.9±7.6
69.1±7.1**
69.9±6.2*
72.2±6.3
1) Systolic blood pressure
2) Diastolic blood pressure
Values are expressed as mean±standard deviation of six subjects.
*p<0.05, **p<0.01 Significantly different from the Meat diet.
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
meat period to the freeze-dried tofu periods I (p<
cholesterol and serum free cholesterol in the six
0.01) and II (p<0.05), as opposed to the increase in
subjects according to the experimental period of
diastolic blood pressure that occurred between the
measurement. There were significant increases in
ordinary diet period to the meat period. Both systolic
both serum total cholesterol (p<0.01) and serum
and diastolic blood pressures were lower in the
free cholesterol levels (p<0.05) in the meat period
freeze-dried tofu periods than in the ordinary diet
as compared with ordinary diet period I. However,
period I.
both levels decreased between the meat period and
3. Measurement of fatigue
freeze-dried tofu periods. In particular, there was a
Mean flicker values, blinker values, and grasp
statistically significant difference in total cholesterol
values in six subjects are shown according to the
levels (p<0.05) and free cholesterol levels (p<0.01)
experimental period of measurement in Fig.1. There
between the freeze-dried tofu period II and the
were minimal differences in all measured values
meat period. Free cholesterol levels were lower in
among the experimental periods.
freeze-dried tofu periods I and II than in the other
4. Changes in blood findings and liver function
experimental periods, and the ester rate tended to
findings
be high in the freeze-dried tofu periods.
As shown in Table 4, there were almost no dif-
ferences in Gb, Hb, or Ht among the experimental
periods. On the other hand, there were slight differ-
DISCUSSION
ences in serum proteins and liver function findings
among the periods, which may have been attrib-
The present study of nutrient intakes revealed
utable to the different quantity of protein intake.
that approximately 80 g of protein was consumed
However, these findings were all within physiologi-
and the animal/vegetable protein ratio was 6 : 4
cally normal limits.
during the ordinary diet periods. In the meat period,
5. Changes in serum total cholesterol and serum
during which the volunteers had to take 190 g of
free cholesterol
meat, the prescribed three eggs (about 150 g), 180
Table 5 shows the mean levels of serum total
ml of cow’
s milk, and freely distributed staple and
Fig.1. Flicker, Blinker, and Grasp values in each experimental diet
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Table 4. Changes in blood parameters in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Blood examinations
GB
1.0567±0.0046
1.0567±0.0035
1.0558±0.0032
1.0557±0.0031
1.0555±0.0024
Hb (g/dl)
14.6±1.3
14.3±1.5
13.9±1.3
14.5±1.3
14.3±1.4
Ht(%)
43.8±3.9
43.8±4.0
43.4±3.2
43.1±3.6
43.2±3.4
TP (g/dl)
7.5±0.6
7.9±0.5
7.5±0.5
7.8±0.2
7.7±0.5
Liver function tests
GOT (Karmen Unit)
14.8±12.4
16.2±8.5
17.5±7.2
10.7±8.0
14.3±11.9
GPT (Karmen Unit)
11.8±8.2
14.3±6.4
15.2±5.0
10.2±4.3
9.5±6.4
ALP (King-
8.7±1.2
9.0±1.5
9.3±1.8
10.5±2.1
11.8±2.7
Armstrong Unit)
ZTT (Kunkel Unit)
6.9±7.0
5.8±1.9
7.6±2.8
6.5±2.6
5.0±2.2
Albumin (g/dl)
4.2±0.4
4.4±0.3
4.1±0.3
4.4±0.3
4.4±0.2
Values are expressed as Mean±Standard deviation of 6 subjects.
Table 5. Changes in serum cholesterol in each experimental diet
Ordinary I
Meat
Freeze-dried tofu
Ordinary II
I
II
Total cholesterol
184.8±15.4
220.0±19.5??
199.0±20.6
188.3±19.0*
201.4±14.0
(mg/dl)
Free cholesterol
57.7±5.8
70.5±9.1?
56.8±7.9*
54.3±3.5**
62.2±4.7??
(mg/dl)
Cholesterol ester
68.9±3.6
68.1±1.7
71.5±1.1**
71.1±1.4**
69.3±2.8
(%)
?p<0.05, ??p<0.01 Significantly different from the Ordinary diet I.
*p<0.05, **p<0.01 Significantly different from the Meat diet.
??p<0.01 Significantly different from the Freeze-dried tofu diet II.
other foods, total protein was about 110 g. Animal
intake was dependent on the nature of protein. In
protein intake alone increased by about 70% as
the ordinary diet periods, total fat intake was about
compared with the ordinary diet period I. Conse-
50 g, and the animal/vegetable lipid ratio was 6 : 4.
quently, the animal/vegetable protein ratio was 7 : 3.
The meat period brought about a 2.5-fold increase
In the freeze-dried tofu periods, the 38 g of protein
in animal fat intake, and the animal/vegetable lipid
that originated in the previously administered 190 g
ratio became 8 : 2. In the freeze-dried tofu period,
of meat was replaced by about 72 g of (vegetable
animal fat intake was almost similar to that in the
protein) freeze-dried tofu, and three eggs and 180
ordinary diet periods, but vegetable protein intake
ml of cow’
s milk were administered to contribute
was two times higher (about 40 g) than in the ordi-
animal protein. The results showed that total pro-
nary diet periods. As a result, the animal/vegetable
tein intake was slightly higher (about 90 g) in the
lipid ratio was 4 : 6, opposite to that in the ordinary
freeze-dried tofu periods than in the ordinary diet
diet periods. The discrepancy in such nutrient in-
periods, but that the animal/vegetable protein ratio
takes led to changes in blood-test results. There was
was 3 : 7, opposite to that in the meat period. Fat
a statistically significant increase in serum total
?
?
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
cholesterol and free cholesterol levels in the meat
and a higher proportion of vegetable oil contained
period as compared with the ordinary diet periods.
in freeze-dried tofu (11-12). Besides the vegetable
On the other hand, both parameters significantly
oil in freeze-dried tofu, we consider that soybean
decreased in the freeze-dried tofu period II as
protein also had an effect on lowering the serum
compared with the meat period. The proportion of
cholesterol (13). As shown in Fig. 4, there was an
animal fat intake, as well as cholesterol intake, are
inverse correlation between the proportion of animal
dietary factors influencing serum cholesterol levels,
protein and the proportion of animal fat/vegetable
and the proportion of animal fat intake is known to
oil. While it is important to maintain a certain level
influence the rate of absorption of cholesterol intake
of animal protein in terms of the quality of protein
(9-10). Accordingly, the correlation between the
intake, an extremely high animal protein intake
cholesterol intake and the proportion of animal fat
leads to an excessive intake of saturated fatty acid,
intake in the experiment was expressed as the
resulting in an increase in serum cholesterol. Con-
following formula : cholesterol intake x animal fat
sidering these factors, the proportion of animal
intake/vegetable oil intake. As shown in Figs. 2
protein intake is recommended to be within 40% -
and 3, there was a statistically significant correlation
50% (14). In the present study, the proportion of
between the proportion of animal fat intake and both
animal protein intake to the entire nutrient intake
serum total cholesterol and serum free cholesterol.
was 60% -70%, which exceeded the recommended
The fact that the meat period was associated with
range, even in the ordinary diet period as well as
not only higher cholesterol intake but also a higher
in the meat period. This result should be carefully
proportion of animal fat intake appears to reflect the
considered, because it may lead to an excessive
appearance of higher serum cholesterol levels. On
intake of saturated fatty acid and an increase in
the other hand, although the cholesterol intake was
serum cholesterol. On the other hand, because the
1.5 times higher in the freeze-dried tofu periods
proportion of animal protein intake was found to be
than in the ordinary diet periods, and although there
low (about 30%) during the freeze-dried tofu periods,
was little difference in cholesterol intake between
freeze-dried tofu appears to prevent an excessive
the freeze-dried tofu period and the meat period, the
intake of saturated fatty acid and to restrain an
fact that serum cholesterol decreased may be ex-
increase in serum cholesterol. It is also thought
plained by a lower proportion of animal fat intake
that limiting amino acids do not appear if there is
Fig. 2. Correlation between serum total cholesterol and values of multipling cholesterol intake by ratio of animal
lipid intake to vegetable oil intake
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sufficient protein intake, even of the proportion of
and N/E ratio in the present study, there were no
animal protein intake is only 30% (15). When the
differences in these ratios between the freeze-dried
amount of amino acids contained in the food con-
tofu period and the ordinary diet period. This indi-
sumed was calculated to determine the E/T ratio
cates that the quality of protein is not reduced by a
Fig. 3. Correlation between serum free cholesterol and values of multipling cholesterol intake by ratio of animal lipid
intake to vegetable oil intake
Fig. 4. Relationship between percent of animal protein intake and ratio of vegetable oil intake to animal lipid intake
?
?
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M. Fujii et al.
l
Bodily functions of
o freeze-dried
f
soybean curd
c
freeze-dried tofu diet. This can also be explained
6.
Michi K, Matsushita K, Ezawa I : Effect of
by the fact that there were no differences in blood
dehydrofrozen soy bean curd, Koritofu, and
findings, such as Gb, Hb, Ht, and TP, or liver func-
dried soy bean product, Yuba, on the plasma
tion findings among the experimental periods. From
cholesterol. Nippon Eiyo Shokuryo Gakkaishi
the standpoint of preventing lifestyle-related chronic
(in Japanese) 19 (6) : 449-452, 1967
diseases, it is important to maintain the normal
7.
Kirihara H : Functional tests of fatigue evalu-
limits of blood pressure. In the present study, dias-
ation (in Japanese). Nippon Sangyo Eisei Kyokai,
tolic blood pressure, which is frequently associated
Tokyo, 1967
with high fat intakes and high cholesterol intakes,
8.
Resources Council, Science and Technology
tended to increase in the meat period (16). On the
Agency, Japan. Standard Tables of Food Com-
contrary, freeze-dried tofu decreased both systolic
position in Japan, 4 th ed (in Japanese). Daiich
blood pressure and significantly decreased diastolic
Syuppan, Tokyo, 1983
blood pressure. It has already been shown that
9.
Keys A, Anderson JT, Grande F : Prediction of
there is a correlation between blood pressure and
serum cholesterol response of man to changes
dietary calcium and magnesium intakes (17), and
in fat in the diet. Lancet 2 : 959-966, 1957
as freeze-dried tofu is rich in minerals it deserves
10. Hegsted DM, Mcgardy RB, Myers ML, Stare
further attention. In conclusion, when animal pro-
FJ : Quantitative effects of dietary fat on serum
tein intake was restricted to 30% and freeze-dried
cholesterol in man. Amer J Clin Nutr 17 : 281-
tofu administered to maintain a total protein intake
295, 1965
similar to that of a meat-rich diet, excessive eating
11. Michi K, Ezawa I : The effect of vegetable oil
can be prevented, and blood findings, blood pressure,
on the plasma, liver and adrenal cholesterol.
and serum cholesterol levels can be maintained within
Nippon Eiyo Shokuryo Gakkaishi (in Japanese)
physiologically normal limits. Therefore, freeze-dried
18 : 220-203, 1965
tofu appears to be a valuable food for preventing
12. Kaneda N : Cholesterol and food. Shoku no
lifestyle-related chronic diseases.
Kagaku (in Jpanese) 17 : 115-121, 1974
13. Yamamoto T, Inoue G : Effect of soybean pro-
tein on plasma cholesterol concentration in
REFERENCES
humans. Nippon Eiyo Shokuryo Gakkaishi (in
Japanese) 44 : 155-162, 1991
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