Vol. 13(12) pp.

287-296, December 2019

DOI: 10.5897/AJFS2019.1826
Article Number: F9C284462446
ISSN: 1996-0794
Copyright ©2019
Author(s) retain the copyright of this article African Journal of Food Science
http://www.academicjournals.org/AJFS

Full Length Research Paper

Effect of coagulants on the physicochemical properties

of fresh tofu
Ezeama, C. F. and Dobson, G. N.*
Department of Food Science and Technology, College of Applied Sciences and Tourism,
Micheal Opara University of Agriculture, Umudike, Abia State, Nigeria.
Received 31 May, 2019; Accepted 16 October, 2019

Soy flour from soybean was processed to produce fresh tofu using three coagulants namely lime,
Epson salt and tamarind. The results obtained showed that the pH of the samples were 7.20 for Epson
salt, 5.00 for lime and 5.50 for tamarind. There were significant differences (p< 0.05) in the weight
increase of all the samples with Epson salt (46.62 kg) as the highest. Epson salt had the least
coagulation time of 6 min. The result of the proximate analysis shows that there were significant
differences (p< 0.05) in protein, carbohydrate and moisture content in the three samples with the value
of protein in Epson salt (12.40) as the highest. Further analysis revealed that there were significant
differences (p<0.05) in emulsifying capacity, oil absorption capacity and swelling index in all the
samples. The value of the water absorption capacity of Epson salt (4.70 g/ml) was significantly different
(p< 0.05) from lime (3.80 g/ml) and tamarind (3.70 g/ml). In the mineral composition there were
significant differences (p< 0.05) in Ca, Mg, K, P, Na and Fe in all the samples though K had the lowest
value. There were significant differences (p<0.05) in the total fungal and viable counts. The total fungal
3
count was least in Epson salt (2.00 x 10 CFU/g) while the total viable count was also least in Epson salt
3
(2.6 x 10 CFU/g) after 6 days of ambient storage; however, it was highest in tamarind coagulated tofu.
Epson salt coagulated fresh tofu was highly appreciated for ranking highest in protein, Ca, Mg and
lowest in total fungal and viable count.

Key words: Tofu, coagulants, physical characteristics, functional properties, proximate and mineral
composition, microbial.

INTRODUCTION

Tofu is a meat analogue derived from soybean. The term possesses a three dimensional structure which makes it
meat analogue means substitute for animal proteins. possible to be used as a textured protein product. One of
Soybean (Glycine max) is an important oil seed those textured protein product is “Tofu”, a meat analogue.
belonging to the family of leguminosae .It is usually Tofu is a fermented product which serves several
grown as a food crop in Nigeria. There are three species functions such as enrichment of food substrate with
of soybean. They include G ussuriensis, G max and G proteins, essential amino acids, essential fatty acids and
gracilis. G max is commonly grown throughout the world vitamins; this improves the digestibility and acceptability
(Salunkhe et al., 1992). Soybean protein molecule of foods, detoxification of toxic substances in foods,

*Corresponding author. E-mail: giftdobson@gmail.com.

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution
License 4.0 International License
288 Afr. J. Food Sci.

preservation of food through production of anti-bacterial were purchased from Aba Market Ahia Ohuru in Abia State in
compounds such as lactic and acetic acid, and Nigeria.
enrichment of the diet through the development of
flavours aroma, taste, palatability and texture of the foods Sample preparation
(Steinkraus,1995).
Other benefits include decrease in risk of cancer, used The soybean sample was dehulled and milled with an attrition mill
as antioxidant, and increase in availability of nutrients to obtain flour. The milled soybean was then allowed to cool before
such as folate, vitamin K, calcium, magnesium, iron and storing in an air tight container. The lime was carefully washed and
the liquid squeezed out into an airtight container. The tamarind
fibre. A lot of phytonutrients such as flavonoids and seed was made into 20% concentrated solution and also stored in
isoflavonoids, phenolic acids, protein and peptides are airtight container.
also obtained from Tofu. Tofu is also used in the
prevention and treatment of obesity, type 2 diabetes, and
other chronic obstructive pulmonary diseases. Therefore, Tofu production
tofu as a meat analogue possesses all the essential
The processes involved in tofu production are: grinding to flour,
characteristics of a complete protein which contains all soaking, boiling, coagulating, pressing and cutting into shapes.
the indispensible amino acids in the approximate amount Four hundred and fifty grams (450 g) of soybean flour was weighed
needed by humans (Iwe, 2003). and soaked in 4500 ml of sterilized water for 5 h in a sterile
A report comparing the cost of soybean with other container, according to the method described by Sack et al. (2006).
protein sources shows that 1.25 kg of soybean and 0.25 Thereafter, the soaked soybean was sieved with muslin cloth and
soy milk collected. The total soy milk collected was 9000 ml. The
kg of beef could be purchased at the same price 9000 ml soymilk realized was divided into three to get a 3000 ml
(Dashiell, 1993). Moreover, the report from FAO shows soy milk sample. Each 3000 ml soymilk was given different
that animal protein contributed 6.5 g of an estimated 53.4 treatment from different coagulants.
g daily per capita protein intake. This figure is far below The first 3000 ml soymilk was allowed to boil for 15 min. On
the 34 g / person/day or 40% of an average 85.9 /day of reaching the boiling point, lime solution of 0.314 mol/dm-3 was
total protein in developed countries of the world. In added and allowed to coagulate the soymilk. After coagulation, the
coagulated milk was poured in cloth bag to drain water. In the same
Nigeria, the animal-protein intake is grossly inadequate. manner, the other 3000 ml soymilk was allowed to boil for 15 min. A
In Nigeria, tofu consumption is increasing due to an weighed quantity (10 g) of Epson salt was added until coagulation
increase in Asia immigrants, greater acceptance of soy took place. The coagulated milk was collected in a cloth bag to
foods by the general population and increased drain water. Lastly, the other 3000 ml sample was treated with a
recognition of health benefits of soy foods, especially by 0.42 mol/dm-3 tamarind solution.
The three samples were tied in the cloth bag and the whey was
those who wish to reduce their consumption of animal
pressed out using a presser and allowed for 12 h to drain.
products. Secondly, many who are now vegetarians and The following tofu was obtained: tofu coagulated with lime, tofu
transitioning from fresh meat to meat of plant protein coagulated with Epson salt and Tofu coagulated with tamarind
(tofu) need this analogue to feel as if animal meat is been (Plates1 to 3).
consumed. Coagulation of protein and oil (emulsion)
suspended in the boiled soy milk is the most important
Physical/ chemical properties
step in the production of tofu. This process is
accomplished with the aid of coagulants. Two types of The coagulated samples from the 3000 ml of soy milk obtained from
coagulants (salts and acids) are used commercially different coagulants were weighed using electronic weighing
(Berk, 1992). The use of different coagulants in the balance. Stop clock was used to determine the coagulation time.
preparation of tofu determines the nutritional composition pH was determined with the use of pH meter placed in the
of tofu (Oboh and Omotosho, 2005). Tofu is coagulated supernatant of the different coagulated sample (Njoku et al., 1990).
with different coagulants contain different minerals,
textures as well as yield (Prabhakaran et al., 2006). Proximate analysis
Hence, there is need for vegetarians to be
knowledgeable with the best preparation method of tofu, Proximate analyses of the fresh tofu made from the three
a plant protein which will give rise to better storage time coagulants were determined. The moisture content, crude protein,
and an improved taste. The objective of the study is to fat, crude fibre and carbohydrate were determined.
determine the nutritional and functional properties of soy
flour, to compare the physicochemical properties and the Moisture content determination
microbial population of fresh tofu coagulated with Epson
salt, lime and tamarind . The moisture content of the fresh tofu was determined by the
conventional method (AACC, 2000). Moisture cans were dried in
the oven and put in the desiccators to cool. The cans were weighed
MATERIALS AND METHODS after cooling and 2 g of the samples was put in each moisture can
and placed in the oven and heated at 105°C for 3 h. The samples
Sources of raw materials were weighed after each hour until a constant weight was obtained
and thereafter the cans were cooled in the desiccators before
The soybean and the coagulants (Epson salt, lime and tamarind) weighing. The difference was determined and expressed as a
 Ezeama and Dobson 289

desiccators before weighing quickly to prevent moisture absorption.

The crucibles containing the sample were covered and placed into
the muffle furnace whose temperature was set at 550°C. This
temperature was maintained for about two hours until a whitish ash
was obtained. After two hours, the muffle furnace was switched off
and the crucibles containing the sample weighed and percentage
ash content determined

Crude fiber determination

The crude fiber determination was determined by the method

described in Onwuka (2005). Two grams (2 g) of sample was
treated with 150 ml of 1.25% H2SO4 solution for 30 min. It was
washed with several portion of hot distilled water using a two-fold
muslin cloth to retain the sample particles. When drained dried, the
washed sample was carefully transferred back to the flask and 150
Plate 1. Fresh tofu coagulated with lime. ml of 1.25% NaOH solution was added to it. It was again boiled for
30 min, washed as before with hot water and allowed to drain dry. It
was then very carefully transferred quantitatively to be weighed in
porcelain crucible. The sample in the crucible was dried for one
hour, at 105°C in the oven, cooled in a desiccators and re-weighed.
It was then finally burnt to ashes in a muffle furnace, cooled and
weighed crude fiber content was determined.

Fat determination

Fat content was determined by the continuous solvent extraction in

Soxhlet reflux apparatus (AOAC). Two grams (2 g) of each sample
was wrapped in a porous filter paper and placed in a Soxhlet reflux
filter. An oil extraction flask was weighed and filled with 200 ml
petroleum ether. The reflux flask was mounted on the extraction
flask and connected to a condenser. The solvent was brought to
boil on an electro thermal heater. The vaporized solvent (petroleum
ether) was condensed into the reflux flask to completely submerge
the wrapped sample and extract the fat therein. When the reflux
Plate 2. Fresh tofu coagulated with tamarind. flask was filled, it refluxed carrying the extracted oil back into the
extraction flask while the boiling continued. The cycle of boiling,
vaporization, condensation extraction and reflux was allowed to go
on for four hours before the solvent was recovered leaving the
extracted oil in the flask. The flask (with oil) was further dried in the
oven at 60°C for 30 min to remove any remaining solvent. It was
cooled in the desiccators and weighed.

Protein content determination

The protein content was carried out by the Kjeldahl methods

(AOAC, 1975) modified by Onwuka (2005). Half a gram (0.5 g) of
the sample was digested by boiling in 10 ml of conc. H2SO4 in the
presence of one selenium catalyst tablet. Digestion was done under
a fume cupboard until a clear solution (digest) was obtained. The
digest was diluted with distilled water and transferred to a 100 ml
volume flask where it was made up to mark with the distilled water.
An aliquot (10 ml) of the digest was mixed with equal volume of
Plate 3. Fresh Tofu coagulated with Epson salt. 45% NaOH solution in a Kjeldahl apparatus and distilled. The
distillate was collected in 10m/s of 4% boric acid solution into which
three drop of mixed indicator (methyl red/bromocresol green) was
added. A total of 50 ml distillate was collected and titrated against
percentage of the original sample weight. 0.02N H2SO4 from green to a deep red endpoint. A reagent blank
without sample was also digested, distilled and titrated as described
above. The total N2 was calculated and the protein content was
obtained.
Ash content determination

In determining the ash content of the samples, the method Determination of carbohydrate
recommended by AACC (2000) was used. Porcelain crucibles were
thoroughly dried by heating at 600°C and then cooled in the This was calculated by difference as the Nitrogen free extraction
290 Afr. J. Food Sci.

(NFE) (Onwuka, 2005). The formula below was used. gelatinization as the gelatinization temperature.

Percentage CHO (NFE) = 100% - (Protein + Fat + Fiber + Ash +

Mc) Determination of minerals

The mineral composition of the samples was determined by the

Functional properties of fresh Tofu methods described by Onwuka (2005). The minerals determined
were Ca, K, Na, P, Mg and Fe using Atomic Absorption
Bulk density Spectophotometer. One gram of the dried sample weighed out for
each of the minerals to be determined. Then 20 ml of the acid
The bulk density was determined using the methods described in mixture (650 ml Conc HNO3, 80 ml PCA, 20 ml Conc H2SO4) was
Onwuka (2005). A 10 ml capacity graduated measuring cylinder added in the digestion flask with 1 g weighed dried sample. The
was weighed. The sample (10 g) was weighed into 10 ml flask was heated until a clear digest was obtained. The dilute was
measuring cylinder and the volume was recorded as the bulk digested with distilled water made up to the 500 ml mark.
volume (v). The bottom of the cylinder was raised about 10 cm Appropriate dilutions were made for each element.
above the slab and made to fall in the platform continuously until a
constant volume was observed. The packed volume was recorded.
The loose density (LBD) and packed bulk densities (PBD) were Microbiological analysis
calculated as the ratio of the sample weight to the volume occupied
by the sample before and after tapping. Tryptone Soy Agar (TSA) and Sabourand Dextrose Agar (SDA)
were used for determination of bacterial and fungal population of
the samples. The two media were weighed according to the
Swelling index manufacturers’ specifications. They were sterilized at 121°C for 15
min at 15 psi using the autoclave. After sterilization, they were
The swelling index was determined using the methods described in allowed to cool and poured into sterile petri dishes for solidification.
Onwuka (2005). The sample (3 g) was weighed into a clean dry and
graduated cylinder (50 ml). The flour samples were gently leveled
and the volume was noted before addition of 30 ml distilled water Microbial population determination
(27°C). The cylinder was swirled manually and allowed to stand for
60 min and 24 h while change in volume (swelling) was recorded. A dilution was obtained from the three samples of tofu by dissolving
The swelling index was calculated as the ratio of the initial volume 25 g each in 225 ml of 0.1% (w/v) peptone water (pH 7.2±0.2), to
of sample in cylinder to the final sample. get a 1:10 dilution (Ezeama, 2007) under strict aseptic conditions.
From these, subsequent dilutions were made by introducing 1.0 ml
each of the samples into liquefied Tryptone Soy Agar (TSA) and
Water / oil absorption capacity Sabourand Dextrose Agar (SDA) and mixed by rotating the plates.
These were allowed to solidify on a level surface. The TSA plates
The water/oil absorption capacity was determined by the methods were incubated at 35- 37°C for 18-24 h for bacteria and aerobically
of Onwuka (2005). One gram of sample was weighed into a at 28 ±2°C for 4-5 d for fungi. This process was done in duplicate
chemical graduated centrifuge tube. Using a warring whole mixer, for each sample. The samples include: two samples from tofu
10 ml distilled water/oil was added and mixed thoroughly. The coagulated with lime, two samples coagulated with Epsom salt and
sample was allowed to stand for 30 min at room temperature (28°C) two samples coagulated with tamarind. These processes were
and this centrifuged at 5,000 x g for 30 min. The volume of free repeated on SDA media to determine fungal count. On
water/ oil was read from the graduated centrifuge tube. The water/ establishment of growth, discrete colonies were counted (Ezeama,
oil absorption capacity was calculated by multiplying the amount of 2007), and recorded as colony forming unit per gram (CFU/g).
water /oil absorbed (total minus free) by the density of water / oil (1
g/ml).
Statistical analysis
Emulsification capacity (EC) Treatment mean were compared using Duncan’s Multiple Range
Tests (DMRT) at 5% significant level.
The emulsification capacity of the flour was determined by the
method of Onwuka (2005) by blending 2 g flour sample with 25 ml
distilled water at room temperature for 30 s in a warring blender at
1600 rpm. After complete dispersion, 25 ml of vegetable oil (soy oil) RESULTS AND DISCUSSION
was added and blended together for 30 s. The mixture was
transferred into a centrifuge tube and centrifuged at 1,600 rpm for 5 The result of Table 1 shows the physical characteristics
min. The volume of oil separated from the sample after centrifuge of fresh tofu treated with different coagulants, The pH
was read directly from the tube. Emulsion capacity is expressed as
the amount of oil emulsified and held per gram of sample value of Epson salt used as coagulant was significantly
(Padmashree et al 1987) different (P<0.05) from that of lime and tamarind. There
was no significant difference (P>0.05) between the pH
value of tamarind and lime. The pH value ranged from
Gelatinization temperature 2.30 to 7.40. Epson salt had the highest value followed
by lime and tamarind. The components of these
The gelatinization temperature was determined by the methods of
Onwuka (2005), by preparing a 10% suspension of the flour sample coagulants affected their pH values. The addition of lime
in a test tube. The suspension was heated in a boiling water bath and tamarind used as coagulants in soymilk samples
with continuous stirring. The temperature was recorded 30 s after reduced the acidity of lime treated tofu and tamarind
 Ezeama and Dobson 291

Table 1. Physical characteristics of fresh Tofu treated with different coagulants during processing.

Treated tofu pH Coagulation Weight

samples Coagulant Sample time (min) Initial (kg) Final (kg) % Increase
a a c a a a
Epson salt 7.40 7.20 6.00 1.50 2.81 46.62
b b b a c c
Lime 2.30 5.00 10.00 1.50 2.51 40.23
b b a a b b
Tamarind 2.80 5.50 15.00 1.50 2.67 43.82
SD 2.16 1.18 1.170 0.00 0.12 0.12

Table 2. Proximate composition (%) of fresh tofu treated with different coagulant.

Treated Tofu samples Protein Fat Fiber Ash Moisture Carbohydrate

a b a b a c
Epson salt 12.40 2.10 0.12 1.80 65.80 17.00
b b a a c a
Lime 12.20 2.20 0.11 1.00 58.00 26.00
c a a a b b
Tamarind 11.80 1.80 0.11 1.00 62.00 23.30
SD 0.20 0.12 0.004 0.3 3.18 3.74

treated tofu respectively. The isoelectric pH of soybean which first weakens the gel strength. The high acidity
ranges from 4.2 to 5.6, but the lowest solubility occurs at precipitated the protein without forming gel. The
the protein isoelectric point of 4.6 where molecule net weakening of the gel led to a longer time in precipitating
charge is zero. The addition of acid coagulant (lime and protein by insolubility. At extreme pH, soy protein has
tamarind) which contains citric and tartaric acid been observed to become insoluble according to Iwe
respectively lowered the pH of the mixture to isoelectric (2003).
point where the solubility is at minimum. This led to the The final weight ranged from 2.51 to 2.81 kg (Table 1).
precipitation of protein. This agrees with that of Shurtleff There was significant difference (p<0.05) between the
and Aoyagi (2000), that the addition of lime lowered the fresh tofu sample treated with Epson salt and lime. There
pH of the samples. The pH of lime lowered the sample to was also a significant difference (p<0.05) between the
5.00 while the pH of tamarind lowered the pH of the sample treated with lime and tamarind in weight gain.
sample to 5.50 (Table 1). This falls within the isoelectric The weight gain was higher in Epson salt treated tofu
range of soybean but not at the isoelectric point of 4.6 followed by tamarind treated tofu than lime treated tofu.
where the pH net charge is zero. This weight gain of Epson salt treated tofu could be
In the case of the Epson salt treated tofu the addition of attributed to the water binding capacity which varies with
the coagulant slightly increased the acidity. The Epson protein source, composition and presence of
salt coagulated the soymilk by metallic cation (Mg++) carbohydrate, lipid, pH and salt. According to Iwe (2003),
which reacted with the various proteins in the soymilk and the water binding is dependent on pH. At the pH of 7, the
precipitated it to form curds. According to Shurtleff and water binding capacity of soy protein was much higher
Aoyagi (2000), the protein precipitated by magnesium than in pH of 5 and 6. Therefore at high pH protein tend
salt is 80% of the soymilk protein. to bind more water. This explains why the weight of the
The coagulation time (min) of the sample was greatest Epson salt treated tofu with pH of 7 was higher than
in tamarind, followed by lime then Epson salt. The tamarind treated tofu and lime treated tofu.
coagulation time ranged from 6.00 to 15.00 min (Table 1). The higher weight in Epson salt treated tofu could also
There was a significant different (p<0.05) in the be as a result of the salt being capable of coagulating
coagulation time of tofu coagulated with Epson salt and protein concentration of below 3%, (Iwe, 2003). Therefore
tofu coagulated with lime. There was also a significant enough protein was precipitated soymilk due to the ionic
difference (p<0.05) in tofu coagulated with tamarind and attraction. In the case of lime and tamarind coagulation,
lime. The difference in the coagulation time could be as a some proteins such as globulins were still soluble at that
result of the difference in the ionic strength. The ionic pH during the time of coagulation. This gave rise to
strength is very high in Epson salt; therefore the addition lowered weight (Liu, 2000).
of salt increased the ionic strength of the protein which The proximate composition of fresh tofu as presented
promoted quick aggregation by the divalent cations. The in Table 2 shows that the protein content in the three
ionic strength also enhanced gel formation and gel samples ranged from 11.80 – 12.40% with the protein of
strength lowering coagulation time (Iwe, 2003). But the Epson salt treated tofu ranking highest followed by lime
use of lime and tamarind lowered the pH of the protein treated tofu. There were significant differences (p<0.05)
292 Afr. J. Food Sci.

Table 3. Functional properties of fresh tofu treated with different coagulants.

Water Oil absorption Swelling

Treated Tofu Emulsion Bulk Tapped
absorption gelling index
samples Capacity % capacity (g/ml) density density
capacity (g/ml) temp. (°C)
a a c a a a
Epson salt 23.00 4.70 1.65 0.48 0.56 54 >100
c b a a a b
Lime 21.43 3.80 3.12 0.46 0.53 1.46 >100
b b b a a c
Tamarind 22.34 3.70 1.85 0.46 0.57 1.23 >100
SD 1.05 0.45 0.65 0.003 0.01 0,01 >100
SD = Standard deviation of the mean values in column. Values in column with different superscripts are significantly different (P<0.05).

in protein, moisture and carbohydrate in all the three lowest with tofu coagulated with lime. The variation in
samples. The high protein content in Epson salt treated moisture content of tofu prepared with different
tofu (12.40%) could be as a result of the solubility of the coagulants is probably due to the tofu particles influenced
protein brought about by the pH of Epson salt (Iwe, 2003) by different anions and its ionic strengths towards the
which at extreme pH of the soy protein became insoluble. water holding capacity of soy protein gels. It may also be
It could also be attributed to the capability of Epson salt due to the unique coagulating properties of the
to coagulate as much little as 3% protein concentrate in coagulants used. This could be as a result of the high
solution, due to the ionic strength of Epson salt (Iwe, protein content in Epson salt coagulated tofu binding
2003). Hence with the pH value of Epson salt (7.4) all the water to the protein according to Iwe (2003).
protein were soluble and precipitated out compared to pH The carbohydrate content ranged from 12.00 to 23.3%
of 2.3 and 2.8 for lime and tamarind treated tofu with the lime treated tofu ranking highest followed by
respectively (Table 1). tamarind coagulated tofu then Epson salt coagulated
The fat contents were 1.80, 2.10 and 2.20% for tofu. There were significant differences (p<0.05) in the
tamarind, Epson salt and lime treated tofu respectively. three samples. These differences may be as a result of
There were significant differences (p<0.05) between an increase in protein leading to significant decrease in
tamarind treated tofu and other samples. The decrease in the non-protein constituents such as oil and sugar
fat content of tamarind coagulated tofu could be as a (Salunkhe et al., 1992).
result of the high amount of anti-oxidant contained in Table 3 shows the function properties of fresh tofu
tamarind (Friedman et al., 2000). The high amount of treated with different coagulant. Functional properties
isoflavone contained in tamarind could also help in the determine the behavior of nutrients in food during
reduction of low density lipo-protein and triglyceride in processing, storage and preparation because they affect
tofu coagulated with tamarind(Iwe,2003).The fat content the overall quality of foods and acceptability (Amajor et
of Epson salt coagulated tofu could also be low due to al., 2014). The water absorption capacity of the Epson
the higher protein content of this Epson salt coagulated salt, lime and tamarind treated tofu were 4.70, 3.80 and
tofu (Brim, 1973). 3.70 g/ml respectively. The functional properties of fresh
Epson salt treated tofu had the highest ash content. tofu treated with Epson salt, lime and tamarind were
There was significant difference in the ash content of tofu significant difference (p<0.05) in water absorption
fermented with Epson salt and other treatments. This capacity of tofu treated with Epson salt, lime and
could be due to the addition of magnesium Sulphate tamarind. The Epson salt treated tofu has the highest
(Epson salt) used as coagulant. This agrees with the type value at 4.70 g/ml. Water absorption capacity is usually
of coagulant used, which determines the mineral defined as weight increase in relation to the original dry
composition of the tofu (Oboh and Omotosho, 2005). The weight of the sample. Water absorption is reported on the
crude fiber contents of the tofu were low with values of basis of protein weight according to Apotiola and
0.12, 0.11 and 0.11% for Epson, lime and Tamarind Fashakin ((2013).
treated tofu respectively. There was no significant From Table 2, the protein content was highest on
(p>0.05) difference between the treatments. The crude Epson salt treated tofu because the solubility of protein
fibre reduced during filtration as the filtrate (soymilk) depends on pH. At a lower pH of 2, the solubility is lost
through the filter which eliminated some of the fibre in the while at pH 6.5 to 7.5, there is higher solubility of protein
filter cake. The crude fiber is important in the diet, since it according to Iwe (2003). The increase in solubility of
increases the faecal output and prevents gastrointestinal protein in Epson salt treated tofu could be as a result of
diseases in man (Amajor et al., 2014). its higher pH at 7.2. It could also be as a result of its ionic
The moisture level (58.0 – 65.8%) attained in this study strength. The use of magnesium salt promotes aggregate
showed that of tofu coagulated with Epson salt was the of protein even at low concentration of 3% as reported
highest followed by tofu coagulated with tamarind and the by Iwe (2003). This helped to increase the protein
 Ezeama and Dobson 293

Table 4. Mineral composition (%) of tofu coagulated with lime, Epson salt and tamarind.

Treated Tofu samples Calcium Potassium Magnesium Phosphorous Sodium Iron

a b a c c b
Epson salt 11.03 0.66 1.72 5.12 2.28 4.07
c c b a b c
Lime 4.00 0.55 1.22 7.63 2.56 3.22
b a c b a a
Tamarind 4.21 0.85 0.54 6.03 2.99 5.11
SD 3.25 0.85 0.48 1.04 0.63 0.77
SD = Standard deviation of the mean values in column. Values in column with different superscripts are significantly different (P<0.05).

concentration as well as the water binding capacity (Table 3). There were no significant differences (P>0.05)
according to McWaters et al. (2003). On the other hand, in the bulk density of fresh tofu treated with lime, tofu
McWater et al. (2003) reported that lower water treated with tamarind and tofu treated with Epson salt.
absorption is due to less availability of polar amino acids Bulk density gives an indication of the relative volume of
in flour. Water absorption capacity is important in bulking packaging material required and high bulk density is a
and consistency of product as well as in baking good physical attribute when determining the mixing
application (Apotiola and Fashakin, 2013). quality of a particular matter. The bulk density is a
The oil capacity ranged from 1.65 g/ml to 3.12 g/ml oil reflection of the load the flour sample can carry if allowed
absorption capacity while Epson salt treated tofu had the to rest on one matter. Therefore, tofu as a product of soy
lowest. There was significant difference (p<0.05) between flour can be packaged without breakage from the
Epson salt treated tofu and lime treated tofu. There was different coagulants. The density of processed product
also a significant difference between lime treated tofu and dictates the characteristics of the container or package
tamarind treated tofu. The oil absorption capacity is the product. Density influences the amount and strength of
measure of the amount of oil that can be absorbed by the packaging material, texture or month feel. Lower bulk
physical matrix of the food. It indicates the degree of density would be an advantage in the formulation of
hydrophobicity of the system (Hayta et al., 2002). The oil complementary foods while higher bulk density is
absorption capacity ranked highest in lime treated as a desirable for greater ease of dispensability (Apotiola and
result of low NSI (Nitrogen solubility index). According to Fashakin, 2013).
Iwe (2003) and Liu et al. (2002), soy proteins having low The swelling index ranged from 1.23 – 1.54 or the fresh
NSI value have a high fat absorption capacity. The oil tofu treated with lime, Epson salt and tamarind (Table 3).
absorption capacity is also affected by the water holding There was significant difference (p<0.05) between tofu
capacity. Because the water holding capacity was lower treated with Epson salt and tofu treated with lime and
in lime treated tofu therefore the oil capacity became tamarind. The swelling index of tofu treated with Epson
higher. salt had the highest value followed by the tofu treated
The emulsifying capacity of fresh tofu treated with with lime then tofu treated with tamarind. This implies that
Epsom salt, lime and tamarind ranged from 23.00 - the Epson salt treated tofu can increase in volume when
22.34. There were significant differences (p<0.05) in immersed in water. Swelling capacity is the measure of
Epson salt treated tofu, lime and tamarind. Epson salt the ability of starch to immobile water and swell. High
treated tofu had the highest emulsifying capacity due to swelling capacity has been reported as part of the criteria
the high protein content. Proteins are effective in for a good quality product (Apotiola and Fashakin, 2013).
promoting emulsion formation because they possess The gelation temperature was above 100°C for the
both hydrophilic and hydrophobic regions (Liu et al., fresh tofu samples treated with the three coagulants
2002). Soy protein performs two main emulsifying (lime, tamarind and Epson salt). This could be as a result
functions which include encapsulating fat droplets of the interaction between lipid and proteins which could
through the hydrophobic groups and forming a affect the gelling as well as the time, strength of pressing
continuous protein gel network through the aqueous and the particle size of the solid (Iwe, 2003).
phase (hydrophilic group) (Iwe, 2003); therefore, the Table 4 shows the mineral composition of tofu
higher the solubility of protein, the higher the emulsion coagulated with lime, Epson salt and tamarind. The
capacity. Emulsifying capacity is the formation of a stable calcium content of all the tofu ranged from 4.00 to
emulsion by the intimate mixing of two or more 11.03%. The tofu coagulated with Epson salt ranked
immiscible liquids so that one dispersed phase is formed highest (11.03%) and the tofu coagulated with lime was
into very small droplets within the second the continuous the lowest. The high calcium content of tofu coagulated
phase (Fellows, 1988). with Epson salt could be as a result of the low acidity of
The bulk density loose of the tofu ranged from 0.46 to the sample (Reddy et al., 1982).
0.47 and the bulk density tapped ranged from 0.53-0.57 There potassium found in the soybean flour used for
294 Afr. J. Food Sci.

Figure 1. Total fungal count (CFU/g) of Tofu samples treated with various coagulants during 6 days storage
at ambient temperature.

production was negligtble. This indicated that the coagulated tofu. According to Salunkhe et al. (1992), the
presence of potassium in the coagulated tofu could come amount of phosphorus is highest in tofu compared to all
from the coagulants. The potassium content ranged from other soy product. This could be as a result of heating of
0.55 to 0.85% (Table 4). The highest was from tofu the curd which liberated the phosphorus from the phytate.
coagulated with tamarind followed by tofu coagulated The sodium content ranged from 2.28 to 2.99% (Table
with Epson salt than with tofu coagulated with lime. This 4). There were significant difference (p<0.05) among the
could be as a result of high potassium content contained three samples. The sodium contents of all the samples
in tamarind. Epson salt could also contain a little were low.
potassium salt which contributed to the potassium There was significant difference (p<0.05) in the iron
content of tofu coagulated with Epson salt. content between tofu coagulated with Epson salt and tofu
Magnesium content ranged from 0.54 to 1.72% (Table coagulated with lime and as also a significant difference
4). The highest was found in Tofu coagulated with Epson (p<0.05) between the tofu coagulated with lime and tofu
salt followed by lime coagulated tofu than tamarind coagulated with tamarind. The iron ranged from 3.22 –
coagulated tofu. The magnesium content may be 5.11%. The tofu coagulated with tamarind ranked the
attributed to the addition of the Epson salt and also phytic highest due to the high content in iron. This may be as a
acid affects magnesium content according to Shimi and result of the iron contained in tamarind. The value of iron
Hasnah (2013). Since the Epson salt was used in content agrees with Salunkhe et al. (1992) which states
coagulation, there was also a reduction in acidity which that the iron content in soycurd ranges from 2 - 17%.
increased the value of magnesium in Epson salt The result in Figures 1 and 2 show the total fungal and
coagulated tofu compared to tamarind and lime total viable count of ambient stored tofu treated in Epson
coagulated tofu with high acidity value (Sareen et al., salt, lime and tamarind respectively. The total fungal
2005). count was least in Epson salt treated tofu gave(2.00x10
3
The phosphorus content ranged from 5.12 to 7.63% CFU/g) while tamarind gave the highest after 6 days of
(Table 4). The highest was found in lime (7.63%) ambient storage as shown in Figures 1 and 2. This
coagulated tofu followed by lime coagulated tofu then makes the tofu treated in Epson salt safe and acceptable
Epson salt coagulated tofu. There were significant for formulation food. The difference among the
differences (p<0.05) in all the samples. The lowest rank treatments could be as a result of lower water activities in
in Epson salt coagulated tofu could be as a result of the tofu treated with Epson salt. The lower water activity was
magnesium content of the sample. The presence of due to the presence of salt. Water activity (aw) is an
calcium and magnesium binds dietary phosphorus index of the availability of water for the growth of micro-
(Sareen et al., 2005). Phytate helps to liberate organism (Frazier and Westhoff, 2011). The salt slowed
phosphorus, therefore the presence of high acidity helped down or prevented the growth of micro-organism by
in liberating phosphorus as found in lime and tamarind causing high osmotic pressure and hence plasmolysis of
 Ezeama and Dobson 295

5
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
0 1 2 3 4 5 6
Storage Time (Days )
Epson salt treated tofu Lime treated tofu Tamarind Treated tofu

Figure 2. Total viable count (CFU/g) of the Tofu samples treated with various coagulants
during 6 days storage at ambient temperature.

cells which may harm the cell of micro-organism. It led to Magnesium and functional properties such as emulsion
dehydration of food by drawing out and tying up moisture capacity, water absorption capacity and swelling index
as it dehydrates microbial cells. Epson salt reduced the compared to Lime and Tamarind coagulated Tofu.
solubility of oxygen in the moisture, which interfered with Furthermore, Epson salt coagulated Tofu gave the least
the action of proteolytic organisms (Frazier and Westhoff, microbial attack indicating lower susceptibility to spoilage.
2011).
Acid precipitable proteins such as precipitated by lime
and tamarind are called storage protein. These storage CONFLICT OF INTERESTS
proteins do not possess biologically active proteins which
compose of trypsin inhibitors, hemaglutinine, The authors have not declared any conflict of interests.
lipoxygenases, β amylases etc. But these biologically
active proteins are contained in the supernatant or whey
ACKNOWLEDGEMENT
proteins. The presence of these biologically active protein
caused quick decomposition of tofu and soymilk since all The authors wish to acknowledge the assistance of the
the whey protein were not completely pressed out laboratory staff of the Department of Food Science And
(Salunkhe et al., 1992). Therefore the existence of all Technology MOUAU for their contribution to the study.
these enzymes reduced the storage time of the acid
coagulated tofu.
Microorganisms especially Lactobacillus spp. has the REFERENCES
inability to synthesis most of the vitamins they require,
AACC (2000). Approved Methods of the American Association of
making them unable to grow well in foods poor in
Cereal Chemist 10th Edition, AACC Press, St Paul MN.
vitamins but useful in assays for the vitamin content of AOAC (1975). Official Methods of Analysis Association of Official
food (Frazier and Weshoff, 2011). Tamarind has a lot of Analytical Chemist 12th Edition, Washington DC.
vitamins. This affected the tofu coagulated with tamarind. Amajor JU, Oti E, Ekeledo N, Omodamiro R, Amajor EE, Aniedu C
(2014) . Studies on the characteristic properties of fermented sun-
The presence of the vitamins and other rich nutrient
dried orange Fleshed Sweet potato Flour Nigeria Food Journal 32:45-
aided in the fast decomposition of tofu treated with 53.
tamarind (Table 3). Apotiola ZO, Fashakin JF (2013). Evaluation of cookies from wheat,
yam and soybean blend, Department of Hospitality Management
Lagos State Polytechnic Ikorodu Lagos, Nigeria.
Berk Z (1992). Technology of production of edible flours and protein
Conclusion products from soybeans, FAO agricultural services bulletin 97, Rome:
Food and Agriculture Organization of the United Nations.
Brim CA (1973). Quantitative genetics and breeding. In: Soybean;
Based on the result of the findings, the following
improvement, production and uses, ed. B.F. Caldwell, Madison:
conclusions were drawn. Soy flour coagulated with Epson American Society of Agronomy.
salt produced Tofu with the highest protein, Calcium, Ezeama CF (2007). Food Microbiology: Fundamentals and Applications.
296 Afr. J. Food Sci.

Natural Prints Lagos. Nigeria, pp. 64-66. Reddy NR, Pierson MD, Sathe SK, Salunkhe DK (1982). Legume-
Dashiell KE (1993). Soybean Production and Utilization in Nigeria. based Fermented Foods: Their Preparation and Nutritional Quality.
Paper presented at the National Workshop on small scale and Critical Reviews in Food Science and Nutrition 17:335.
industrial level processing of Soybean held at IITA, Ibadan, 27th-29th Sacks FM, Lichtenstein AH, Linda HW, Kris-Etherton P, Winston M
July. (2006). American Heart Association Nutrition Committee Soy Protein,
Fellows PJ (1988). Food processing technology principles and practice. Isoflavones, and Cardiovascular Health. An American Heart
Woodhead publishing limited Cambridge England, P. 110. Association Science Advisory for Professionals from the Nutrition
Frazier WC, Westhoff DC (2011). Food Microbiology. The McGraw Hill Committee. Circulation 113(7):1034-1044.
Companies, pp. 341-348. Salunkhe DK, Chavan RN, Adsule KN, Kadam SS (1992). World
Friedman M, Fitch TE, Levi CE, Yokoyama WH (2000). Feeding Oilseeds, Chemistry, Technology and Utilization. An AVI Book
Tomatoes to Hamsters Reduces their Plasma LDL Cholesterol and Published by Van Nostrand Reinhold. New York.
Triglycerides. Journal of Food Science 65:897-900. Sareen SG, Jack LS, James LG (2005). Advanced nutrition and human
Hayta M, Alphaslan M, Baysar A (2002). Effect of Drying Method metabolism. Thompson Wadsworth Belmont CA., USA.
Functional Properties of Tarhana. A Wheat Flour-yougurt Mixture. Shimi G, Hasnah H (2013). Does cooking affect the phytate content in
Journal of Food Science 67(2):740-744. local soy based dishes? International Food Research Journal
Iwe MO (2003). The Science and Technology of Soybean. Published by 20(5):2873-2880.
Rojoint Communication Services Ltd, Enugu Nigeria pp. 66-100. Shurtleff W, Aoyagi A (2000). Tofu and soymilk production: a craft and
Liu K (2000). Expanding Soybean Food Utilization. Food Technology technical manual (3rd ed.), Lafayette, California: Soyfoods Center.
54(7):46-47. Onwuka GI (2005). Food Analysis and Instrumentation Theory and
McWaters HG, Hall A, Bastow RM, Davis SJ, Hanano S, Hibberd V, Practice. Naphtali Prints. Lagos, pp. 90-139.
Doyle MR, Sung S, Halliday KJ, Amasino RM, Miller AJ (2003).The Steinkraus KH (1995). Handbook of Indigenous Fermented Foods. New
Time for Coffee Gene Maintains the Amplitude and Timing of York, Marcel Dekker, Inc.
Arabidopsis Carcadias Clocks. Journal of Plant Cell 15(11):2719-
2729.
Njoku HO, Ogbulie JN, Nnubia C (1990). Biochemical changes during
natural fermentation of African oil bean for the production of ugba.
Journal of the Science of Food and Agriculture 49(4):457-465.
Oboh G, Omotosho OE (2005). Effect of types of Coagulant on the
Nutritive Value in vitro Multienzyme Protein Digestibility of Tofu.
Journal of Food Science and Technology 3(2):182-187.
Prabhakaran MP, Perera CO, Valiyaveettil S (2006). Effect of different
coagulants on the isoflavone levels and physical properties of
prepared firm tofu. Food Chemistry 99(3):492-499.
Padmashree TS, Kshmi LV, Puttaragi S (1987). Effect of Traditional
Processing on the Functional Properties of Cowpea (vigna catjang)
flour. Journal of Food Science and Technology 24:221-225.