PROCESSING OF INSTANT SOFT TOFU WITH HIGH CONTENT OF ISOFLAVONE AND APPROPRIATLY HIGH CONTENT OF Dinh Huu Donga, Dong Thi Anh Daoa aDepartment of Food Technology, Ho Chi Minh City University of Technology, Viet Nam E-mail: dinhhuudong06@gmail.com, dtanhdao@hcmut.edu.vn
The traditional process of instant tofu production was adjusted to produce a high
content of isoflavone and calcium instant tofu. The soaking time, the ratio of soy bean to
water in soaking step and grinding step were studied. The isoflavone (Daidzein and
Genistein) in tofu was increased when increased the soaking time and reached the highest
content at 3.5 hours (1960,38μg Daidzein/g and ,2490,39 μg Genistein/g) . The ratio of
soy bean to water in soaking step affected the isoflavone in the instant tofu. At the soy
bean to water ratio of 1:4.5, the tofu had the highest isoflavone and calcium content. In
addition, the additives had strong influence on the calcium content in tofu. The gluconate
calcium at the concentration of 0.3% produced a tofu with high content.
Keywords: tofu, coagulation, isoflavone, calcium gluconate
in soybeans improve the nutritional value of
soybeans. The main isoflavone in soybeans
naturally exist in the form of aglycones
Isoflavones of soybeans exist in forms of
aglycone (daidzein, genistein and glycitein),
their glycoside forms (daidzin, genistein and
isoflavone has received strongly glycitin) and their malonyl glycoside and concentration due to their bioactivity. The
structural of isoflavone is similarity with
17-β-estradiol , therefore isoflavone is
called phytoestrogen and act like a weak
amount over the world, especially in Asia.
2011). Isoflavone has protective effect on
Soybeans have processed to many kinds of
products such as tofu (soybean curd), miso
soybean. The process conditions strongly
2010). In addition, isoflavone has shown
influenced the isoflavones of soybeans (Lee
and Lee, 2009; Kao et al., 2004; Molamma
Fiechter et al., 2011; Kazuhiro Nara et al.,
conditions also affected the yield of tofu
and the isoflavone content in tofu (Kao et
influence on the yield and quality of tofu
ground nut. The high content of isoflavone
(Kao et al., 2004). The aim of this study
and to suggest an adjustment for process
condition to produce soft tofu with high
2.2.2.1. Yield, solid recovery and proximate
content isoflavone and appropriately high
2.MATERIALS AND METHODS 2.1 Materials
soymilk. Moisture content was determined
by drying 5 g of fresh tofu at 105oC in an air
oven to constant weight (Tsai, Lan, Kao &
calcium chloride, acid lactic) used in this
determined by drying 10 ml ofthe whey at
Kjeldahl method (AOAC,1975) on air-dried
standards used in the HPLC analysis were
tofu samples and using the factor N_ 6.25 to
in forms of aglycone: Genistein (EC. 207-
tofusamples was measured with a digital pH
standards were supplied by Sigma-Aldrich
meter with a glass electrode (Metrohm AG,
2.2 Methods 2.2.1 Soy milk and soy curd (tofu)
The total isoflavone content in tofu was
analyzed by HPLC. The isomeric glucoside,
acetyl glucoside, Maloney glycoside were
hydrolyzed in to free isoflavones: genistein
and daidzein. The isoflavone acetonitril
58% was used as the extraction solvent and
filtered to collect soymilk. In the soaking
step, the influence of the ratio of soybeans
tofu was studied at four levels :1:3, 1:3.5,
Kjeldahl method using a protein conversion
method, respectively. The ash contents were
step, the influence of the ratio of soybeans
– to – water (w/v) on REDM was studied
digestion of the sample on a mixture of 4
curd were quantified in each experiment.
included: calcium citrate, calcium 560 instrument (Perkin-Elmer Corp.). fumarate, calcium gluconate. The most
Results were expressed on dry-matter basis.
appropriate coagulant was further studied
for the optimum concentration used. Each
difference) intervals (p < 0.05) were calculated to evaluate significantly different 3. RESULTS AND DISCUSSION
shape were cut ver-468 Eur Food Res Technol (2008) 226:467–472123 tically
3.1.Effect of soaking time on the recovery
from a block of curd using a cylindrical
efficiency of dry matter (REDM) and isoflavone concentrations. The soaking time was strongly affect the
height with a metal disc (60 mmdiameter).
REDM and isoflavone concentration. The REDM increased when extending the
tocalculate the hardness, brittleness and
soaking time (table 1 and chart 1). The increase of soaking time from 2.5 hours to 4 hours resulted in the increase of 9.56% of REDM. However, the REDM and isoflavone
chewiness) using the instrument software
content had no significant difference when soaking soybeans in 3.5 hours and in 4 hours. The enough period of soaking time allowed the swelling of tissues, the hydration loosened and broke down the matrix inside soybean. The soluble
panellists evaluate the sensory attribute of
components increasingly released when the
the fresh tofu. Panellists were familiar
soaking time was prolonged. The table 1, chart 1 and chart 2 have shown that the daidzein content and genistein content in soft tofu also enhanced to certain level. Extending the soaking time to 4 hours did
panellists took 10–15 min to complete the
not gain the significantly high content of
rating of all the samples. Tofu was cut into
isoflavone in comparison to 3.5 hours. The soaking time of 3.5 hours was chosen for
attributes evaluated were colour, flavour , mouth feel and overall acceptability. For
Table 1. Effect of soaking time on REDM
each sample, panellists scored their liking
point hedonic scale (1 = dislike extremely,
Daidzein content Genistein content
2 = dislike very much, 3 = dislike moderately, 4 = dislike slightly, 5 = neither like nor dislike, 6 = like slightly, 7 41.625(±0.12)a 1828.22(±1.03) a 1965.76(±1.02) a = like moderately, 8 = like very much, and 44.325(±0.15)b 1855.91(±1.05) b
45.725(±0.14)c 1871.38(±1.06) c 1990.39(±1.03) c
46.025(±0.17)c 1885.82(±1.04) c 1998.65(±1.08) c
The experiments was carried out in triple. ANOVAs (STATGRAPHICS 3.0 version software) were used to evaluate significant differences between treatments (p=0.05%). The LSD (least significance Table 2. Effect of the Ratio of soybean-to-
in each column with different superscripts
water on REDM and the content of
are significantly different (p < 0:05).
isoflavones
43.8(±0.20) 1863.36(±0.36) 1985.17(±0.60)
45.3(±0.17) 1868.10(±0.18) 1989.39(±0.81)
46.3(±0.16) 1872.48(±0.14) 1991.48(±0.57)
46.8(±0.19) 1881.59(±0.15) 1995.51(±0.45)
Chart 1.Effect of soaking times and performance on REDM Chart 3: Effect of the ratio of soybeans- to- water soaked by 3.5 hours on REDM Chart 2. Effect of soaking times on concentration of isoflavones 3.2. Effect of the ratio of soybean – to – water on the recovery efficiency of dry matter (REDM) and isoflavone contents.
Soybeans were soaking in 3.5 hours with various water level and then was went into
coagulant. The table 2 demonstrated that the soybean – to – water ratio strongly
affected the REDM but had slightly effect
Chart 4.Effect of soaking times on
on and the content of isoflavones in the
concentration of isoflavones
soft tofu. The chart 2 show that the REDM
drastically increased 8.5% at the ratio of
1:4.5 in comparison to the ratio of 1.3. However the isoflavone content slightly
between two cases: the ratio 1:4 and 1:4.5.
soft tofu also enhanced about 2.5% with the increasing of water level.
Therefore the ratio of 4 was used for the
properly absorbed and health - usefull in human bodies: Calcium fumarate, calcium
citrate and calcium gluconate. The calcium gluconate showed the best recovery of
3.3. Effect of the ratio of soybean-water
calcium in soft tofu than the others when
grinding on REDM
used at the same concentration (table 3).
Table 3. The effect of coagulants to total concentration of calcium in soft tofu Chart 5: Effect of the ratio of soybeans- to- water grinding on REDM
the ratio 1:6 and 1:7. Therefore the ratio
of 1:6 was used for the next experiments.
3.4. Effect of coagulant on calcium concentration in the soft tofu
calcium were usually used to produce tofu. However, the types of coagulants
Chart 6. The effect of coagulant on total
determined the structure of tofu and the
calcium concentration in the soft tofu
concentration of calcium in the final product. The soluble content strongly loss
in the coagulation (Kao et al, 2004). Three
various concentration. The concentration of
types of coagulant with concentration of
calcium gluconate increased resulted in the
increase of calcium in the final products.
the ability of calcium retaining in soft
The calcium gluconate concentration of 0.3
and 0.4% did not gave the significant difference on the calcium content in soft
0.3% calcium gluconate was chosen for the
Conclusions Table 4. Effect of calcium gluconate
In conclusion, the isoflavone content in soft
concentration to the calcium content of
tofu depended on the process condition. The
soft tofu.
swelling and the ability of release of soluble
content in soybean. The ratio of soybeans –
to – water caused the significant difference
during the process. However, the isoflavone
content could be optimized by change the
process condition. The soaking time of 3.5
hours with the ratio of soybean – to – water
of 1:4 showed the ability to retain highest
isoflavone in soft tofu. The coagulant type
and coagulant concentration influenced the
structure and the calcium content of soft
tofu. The calcium gluconate was used at the
appropriated high calcium content in soft
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