LWT - Food Science and Technology 86 (2017) 385e392

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LWT - Food Science and Technology

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Instrumental quality attributes of single washed surimi gels of tilapia:

Effect of different washing media
Bhargavi Priyadarshini a, K.A. Martin Xavier a, Binaya Bhusan Nayak a, Kandan Dhanapal b,
Amjad Khansaheb Balange a, *
a
Department of Post-Harvest Technology, ICAR-Central Institute of Fisheries Education, Versova, Mumbai 400061, Maharashtra, India
b
Department of Fish Processing Technology, College of Fishery Science, Muthukur, India

a r t i c l e i n f o a b s t r a c t

Article history: The effects of single washing cycle with different washing media on the quality of tilapia mince in
Received 3 June 2017 comparison with the quality of conventionally washed surimi from tilapia were investigated. From the
Received in revised form results, it was observed that compared to conventional washed surimi, alkaline saline washed surimi
1 August 2017
with single washing cycle exhibited significantly (p < 0.05) highest gel strength of 60.72 N.mm. Thermal
Accepted 7 August 2017
Available online 8 August 2017
transition of alkaline saline washed surimi exhibited no endothermic transition, while all other treat-
ments showed a shift in the transition peak from surimi to paste. The single washing cycle with different
washing media promoted the heat induced conformational transition from a-helix to b -sheet, b -turn
Keywords:
Tilapia surimi
and random coil structures which are positively correlated with gel strength except for surimi washed
Texture with cold water. Therefore, it can be concluded from the present investigation that single washing with
Gel strength alkaline saline treatment yielded good quality tilapia surimi.
FTIR © 2017 Elsevier Ltd. All rights reserved.
DSC

Chemical compounds studied in this article:

Sodium chloride (PubChem CID: 5234)
Calcium chloride (PubChem CID: 24486)
Sodium bicarbonate (PubChem CID:
16212373)

1. Introduction sustainable manufacturing of surimi. Tilapia is considered as the

Surimi is stabilized myofibrillar protein obtained by mechanical
deboning, mincing and repeatedly washing with cold water
(5e10
C) to remove lipids and water soluble proteins. In general
white lean fish such as Alaska pollock, Pacific whiting and threadfin
bream which are low in lipid content are widely utilized in the
preparation of surimi. Because of the over-exploitation of lean fish
and decline in the surimi resources, there have been attempts to
exploit new fish resources for surimi industry such as dark flesh
and low cost fishes. However, it is difficult to acquire high quality
surimi from dark flesh fish species because of the high content of
lipids and myoglobin present in dark muscle (Arfat & Benjakul,
2013). As an alternative to regular raw material, low cost and
underutilized freshwater fish can be utilized effectively for
* Corresponding author.
E-mail address: amjadbalange@cife.edu.in (A.K. Balange).
food fish of the 21st century and one of the second most farmed fish
in the world (FAO, 2014, pp. 10e11) and India is also an emerging
producer of tilapia. Tilapia is a prolific breeder, omnivorous, very
hardy species and grows faster which makes this species as one of
the candidate species for aquaculture. The limiting factor of tilapia
for surimi production is the red colour of the mince and being
freshwater fish possess poor textural properties.
Gel-forming ability of myofibrillar proteins is the prerequisite to
provide the excellent quality of surimi-based products. In order to
augment the gel forming ability of myofibrillar proteins, elimina-
tion of endogenous proteolytic enzymes is essential. Washing is the
basic stride in the creation of surimi, which enhances the quality by
removing fat and undesirable substances. The number of required
wash cycles depends on species, condition, type of wash, and the
desired quality of the surimi end product (Carvajal, Lanier, & Mac
Donald, 2005). In generation and production of 1 kg of surimi
nearly around 15 kg of water is utilized (Granata, Flick Jr., & Martin,
2012). The extensive utilization of freshwater in surimi production

http://dx.doi.org/10.1016/j.lwt.2017.08.022
0023-6438/© 2017 Elsevier Ltd. All rights reserved.
386 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392
threatens the availability of this resource for other users and the
negative impact on the environment as a result of discharging
untreated processing water. So there is a need for eco efficient
production of surimi with reduced number of washing cycles not
only to prevent environment pollution but also to maximize the
yield and decrease the wash water volume. The various research
contemplated and found that all the distinctive washing media
with increasing number of washing cycles is effective against
increasing the quality parameters of surimi. Conventional washing
in common carp, grass carp, silver carp, tilapia was studied (Luo,
Kuwahara, Kaneniwa, Murata, & Yokoyama, 2001; Rawdkuen, Sai-
Ut, Khamsorn, Chaijan, & Benjakul, 2009). Alkaline saline washing
in sutchi catfish (Priyadarshini et al., 2016) and silver carp (Zhou,
Chong, Ding, Gu, & Liu, 2016) were studied with three washing
cycles. Generally it is found that freshwater fish have substandard
gel quality contrasted with that of marine fish. But these results
differ accordingly by fish species, age, size, season, habitat, etc.
There are no reports on the impact of single washing cycle on the
quality of freshwater fish surimi. Therefore, the objective of the
present study is to investigate how different washing media with
single washing cycle affects the quality of the surimi and their
corresponding gels.
2. Materials and methods
2.1. Fish samples
Tilapia (Oreochromis mossambicus), with a standard length and
weight of 25.66 ± 3.27 cm and 718.78 ± 225.44 g, were obtained
from a fish farm (Talegaon, Pune, Maharashtra, India). The fish were
packed in an insulated container filled with ice (fish/ice ratio of 1:2
(w/w)) and transported to the Department of Post Harvest Tech-
nology, Central Institute of Fisheries Education, Mumbai within 4 h,
where they were stored and covered with ice until processed.
2.2. Preparation of mince and surimi
2.2.1. Preparation of fish mince
Upon the arrival, fish were immediately washed, gutted, cleaned
and subjected to deboning using a mechanical deboning machine
(Baader 694, Lubeck, Germany) with a counter rotating belt and
drum mechanism having a hole diameter of 5 mm. The deboned
mince obtained was placed in low density polyethylene (LDPE)
pouches and imbedded in ice until further use.
2.2.2. Preparation of conventional washed surimi
The conventional washed surimi (CW) was prepared according
to the method of Rawdkuen et al. (2009). Fish mince was washed
with cold water (4
C) using a water/mince ratio of 3:1 (v/w) and
the mixture was stirred gently for 3 min in a Hobart mixer (Hobart
AE 200, London, England) and allowed to settle for 2 min. The slurry
was strained with a double-layer muslin cloth, and the excess water
was manually squeezed out. The washing and straining processes
were repeated three times, with the last washing containing 0.5%
NaCl in the deionised water to facilitate the dewatering step.
Finally, the washed mince was centrifuged at 2200 rpm for 15 min
in a basket centrifuge (Model 60-5, AIM Industries, Mumbai, India).
The mince obtained was referred to as ‘CW surimi.’ The surimi was
packed in LDPE pouches and stored in ice.
2.2.3. Preparation of single washed surimi with different treatments
Fish mince was washed with 3 different washing media with
different pH measured i.e with cold water only (pH 7.47), alkaline
saline (0.15% NaCl and 0.2% NaHCO3; pH 8.69) solution was pre-
pared according to the method described by Balange and Benjakul
(2009) and with calcium chloride and salt (0.1% NaCl and 0.2%
CaCl2; pH 5.86) maintained at 4
C using a water/mince ratio of 3:1
(v/w). The mixture was stirred gently for 3 min in a Hobart mixer
and allowed to settle for 2 min. The slurry was strained with
double-layer muslin cloth, and the excess water was manually
squeezed out. The washing and straining process was done for one
time. Finally, the washed mince was centrifuged in a basket
centrifuge. The resulted surimi was referred to as ‘single washed
surimi (T-1), washed with cold water’, ‘alkaline saline washed su-
rimi (T-2)’ and ‘calcium saline washed surimi (T-3)’. The obtained
surimi was packed in LDPE and stored in ice until further use.
2.3. Gel preparation
The surimi was chopped in a Philips Food Processor (India) at
low speed for 2 min. A homogenous surimi paste was obtained by
extracting surimi myofibrillar protein with 2.5 g/100 g of NaCl and
chopping at low speed for 1 min at 4
C. The paste was stuffed into
polyvinylidene casings (diameter: 2.5 cm, length: 17.5 cm) by using
stainless steel sausage stuffer (Kitchener, 5 lb, China) and casings
were tightened from both sides. Surimi pastes were cooked in a
temperature controlled water bath (Steroglass strike 300, Perugia,
Italy) at 40
C for 30 min and followed by heating at 90
C for
20 min. The gels were then cooled in iced water and stored for
overnight at 4
C prior to analysis.
2.4. Textural parameters
2.4.1. Determination of gel strength
Gels held at 4
C were equilibrated at room temperature (25
C)
prior to test and cut into cylinders (2.5 cm in height). The breaking
force (maximum penetration force, g) and deformation (penetra-
tion depth, cm) were measured using a Rheo Tex (Type SD-700, Sun
Scientific Co Ltd., 4-Chome, Kamiyoga, Setagaya-KU, Tokyo, Japan)
equipped with a spherical probe (5 mm diameter, 60 mm/min)
with load cell of 2 kg. All determinations were carried out in
triplicates.
2.4.2. Determination of textural properties
Textural profile analysis (TPA) of surimi gel was performed using
a TVT 6700 texture analyser (Perten Instruments, Sweden with
software TexCalc version 4.0.2.50) equipped with a stainless steel
cylindrical probe of 20 mm diameter. The gels were cut into a
cylinders (diameter of 20 mm, height of 25 mm), then compressed
at compression degree of 40% with the pre-test, test and post-test
speed of 2 mm/s, 1 mm/s and 5 mm/s. Trigger type was set at
auto with 50 mN trigger force. The data acquisition rate was 200
pps.
2.5. Determination of whiteness
All gels were subjected to whiteness measurement using a
Hunterlab (ColorFlex, Hunter Associates Laboratory, Reston, VA).
Illuminant C was used as the light source of measurement. L*
(lightness), a* (redness/greenness) and b* (yellowness/blueness)
were measured and whiteness was calculated as described by Park
(1994) as follows:
Whiteness ¼ 100  [(100  L*)2 þ a*2 þ b*2]1/2
2.6. SDSepolyacrylamide gel electrophoresis (SDSePAGE)
Protein patterns of gels were analyzed by SDSePAGE according
 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392
to the method of Laemmli (1970). Samples (5 mg protein) were
loaded onto polyacrylamide gels comprising a 10% running gel and
a 4% stacking gel and subjected to electrophoresis at a constant
current of 15 mA/gel using a Hoefer unit (Hoefer, Inc., San Francisco,
CA, USA).
2.7. Differential scanning calorimetry
The thermal transition of surimi and surimi pastes was deter-
mined using differential scanning calorimetry (DSC) (Model DSC
822, Mettler, Toledo). About 15e20 mg of surimi and surimi paste
were placed in DSC hermetic aluminium pans (40 mL). An empty
hermetic pan was used as a reference. The samples were scanned at
10
C/min over a range of 20e100
C. The onset (Tonset), maximum
transition temperature was measured and the enthalpy (D H) was
estimated by measuring the area under the DSC transition curve
using STAR e Software. The nitrogen flow was maintained at 80 mL/
min and system was calibrated using idium.
2.8. FT-IR spectroscopy
Protein's secondary structures in surimi and their gels were
studied using a FTIR spectrometer (Model: 3000 Hyperion mi-
croscope with vertex 80 FTIR system, Bruker, Ettlingen, Germany)
equipped with a micro attenuated total reflectance (ATR) accessory.
1 mg of surimi mince and gel was mixed with 100 mg of potassium
bromide (KBr) and ground gently with an agate pestle and mortar
under a lamp to form a very fine powder and compressed to disc
using a hydraulic pellet press (Type KP, Kimaya Engineers, Mumbai,
India) into a thin disc. The spectrometer was controlled by Opus
Software-version 6.5 to collect spectra over the wavenumber range
of 4000 to 400 cm1, by accumulating 32 scans with a resolution of
4 cm1. Data collection for each sample took less than 2 min.
2.9. Scanning electron microscopy (SEM)
The microstructure of the surimi gels was analyzed by SEM (Liu,
Zhao, Xiong, Xie, & Qin, 2008). Surimi gels with a thickness of
2e3 mm were fixed with a 3% glutaraldehyde solution. The samples
were then rinsed for 1 h in distilled water before being dehydrated
in a gradient ethanol series of 50, 70, 80, 90, and 100% (v/v). The
dried samples were mounted on a bronze stub and sputter-coated
with gold. The specimens were observed using SEM (JEOL JSM-
6390 LV, Tokyo, Japan) at an acceleration voltage of 15 kV.
2.10. Statistical analysis
The data obtained from this study were subjected to one-way
analysis of variance (ANOVA) to establish significant differences be-
tween the measured parameters of treatments and determined at the
0.05 probability level and differences between means were resolved
using the Duncans test. The statistical analyses of data were per-
formed using SPSS (SPSS 20.0 for Windows, IBM, SPSS Inc., Chicago,
IL, USA). The data are reported as mean values ± standard deviation
(SD). The experiments were independently triplicated (n ¼ 3).
3. Results and discussion
3.1. Effect of different washing media on textural properties
3.1.1. Gel strength of the mince and surimi
Breaking force, deformation and gel strength of surimi gels from
tilapia mince washed with different washing media (T-1, T-2 and T-
3) with single washing cycle, conventional washed surimi (CW) and
unwashed mince (UM) are depicted in Table 1. The breaking force
387
increased (p < 0.05) significantly in all treatments when compared
with UM and the greatest deformation was observed in T-2 gels.
The improved characteristics of CW gels are due to washing which
enhanced the removal of tropomyosin, troponin and myosin light
chains in the first two washes that may interfere with proteine-
protein interactions involved in gel formation (Baxter & Skonberg,
2008). The significant increase (p < 0.05) in deformation of T-2 gels
when compared with UM indicates the greatest elasticity of the
gels. Different result was observed with T-3 surimi gels when
washed with CaCl2 and salt solution, where there was a significant
increase (p < 0.05) in breaking force and a decrease in deformation,
which may be due to the fact that calcium ion might have activated
some proteinases such as calpains. Similar results were observed in
surimi gel properties of Alaska pollock (Hunt & Park, 2013) and gels
(suwari and kamaboko) of grass carp, when 80e100 mmol/kg CaCl2
incorporated into surimi (Ding et al., 2011).
The gel strength is an important indicator in evaluating the
quality of fish surimi gel. The gel strength values are summarized in
Table 1. The lowest gel strength was observed in UM which can be
explained due to the presence of thermal induced proteinases and
the lipids that can interfere with the myosin cross-linking during
gel matrix formation and hence cannot form gels. Since the
sarcoplasmic protein in unwashed mince will coagulate during the
heat setting of salt added sol and does not participate in formation
of gel network (Yathavamoorthi, Sankar, & Ravishankar, 2010).
Notably the gel strength was enhanced in T-2 gels significantly
(p < 0.05) when washing were done with alkaline saline solution in
comparison with UM gels where alkaline saline solution may
attributed to the partial denaturation of protein, exposing the
reactive group (Chaijan, Panpipat, & Benjakul, 2010), decrease in
lipid content and also the tail to tail gel network which could form
to a higher extent during thermal gelation of alkaline-treated
protein, resulting in the enhanced gel strength (Luo, Yang, Zhao,
Cheng, & Jiang, 2010).
3.1.2. Textural profile analysis
TPA has been generally used for determination of a number of
textural attributes of surimi gels. The effect of single washing with
different washing media on the textural properties of surimi gels is
shown in Table 2. The hardness exhibited a significant difference
between the UM and treatments (p < 0.05) and the highest hard-
ness was found in T-3 thermal gels followed by T-1, CW and T-2. The
hardness of T-1 thermal gels or heat induced gels can be due to
increased moisture content and removal of water-soluble proteins
to some extent (Wiles, Green, & Bryant, 2004). The highest values of
cohesiveness were observed in T-2 heat induced gels and can be
correlated to the increased concentration of myofibrillar proteins.
Regarding adhesiveness, gumminess and stringiness, there were no
significant difference between UM, CW and treated samples
(p > 0.05). Cohesiveness and springiness are indicative of the
damage on gel structure from the first compression (Handa,
Takahashi, Kuroda, & Froning, 1998). However, the mean values of
cohesiveness increased slightly in T-2 heat induced gels with
respect to the UM and CW, T-1, T-2 and T-3 gels.
3.2. Changes in whiteness of gels
The unwashed mince (UM) gels had the lowest whiteness,
compared with those from CW, T-1, T-2 and T-3 and (Table 1). All
gels exhibited a significant difference (p < 0.05) in lightness (L*),
redness (a*), yellowness (b*) and whiteness. Whiteness is one of the
important quality parameter in assessing the quality of surimi and
which can be achieved by removing the myoglobin, haeme pig-
ments and lipids as much as possible. Comparatively whiteness
values in CW surimi gels were better to T-2 surimi gels (p < 0.05).
388 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392

Table 1
Effect of single washing with different treatments on textural properties and whiteness of the gels.

Treatments Breaking force (N) Deformation (mm) Gel strength (N.mm) Whiteness

UM 2.21 ± 0.21a 8.70 ± 3.03b 18.87 ± 5.30a 65.65 ± 0.71a

CW 6.78 ± 0.38b 8.53 ± 0.94ab 57.98 ± 8.83b 72.23 ± 0.75e
T-1 6.27 ± 0.66b 7.50 ± 0.32ab 47.08 ± 6.20b 70.36 ± 0.72d
T-2 6.53 ± 0.83b 9.20 ± 1.10b 60.72 ± 14.66b 68.98 ± 0.41c
T-3 7.69 ± 0.56c 6.10 ± 0.80a 47.21 ± 9.56b 67.44 ± 0.58b

Data are given as mean ± SD (n ¼ 3). Different superscript letter a-e show the significant differences (P < 0.05) among samples (within the same column). Unwashed mince
(UM); Conventional washing (CW); Single washing (T-1); Alkaline saline washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl and 0.2% CaCl2) (T-
3).

Table 2
Texture profile analysis of mince and surimi gels.

Parameters Treatments

UM CW T-1 T-2 T-3

Hardness (N) 40.31 ± 0.41a 54.52 ± 1.15c 66.25 ± 0.58d 46.60 ± 0.88b 111.71 ± 0.08e
Adhesiveness 0.74 ± 0.01c 0.04 ± 0.01a 0.04 ± 0.01a 0.01 ± 0.00a 0.52 ± 0.11b
Chewiness (N) 14.48 ± 0.21a 22.12 ± 0.53c 25.61 ± 0.25d 18.48 ± 0.41b 42.70 ± 0.39e
Cohesiveness 0.41 ± 0.00a 0.44 ± 0.00a 0.43 ± 0.00a 0.66 ± 0.19b 0.43 ± 0.01a
Gumminess (N) 16.42 ± 0.29a 23.81 ± 0.30b 29.47 ± 0.77c 35.29 ± 0.79d 47.52 ± 0.27e
Springiness (mm) 0.89 ± 0.02a 0.9 ± 0.00a 0.90 ± 0.00a 0.91 ± 0.00a 0.91 ± 0.01a
Stringiness 2.09 ± 0.41c 1.11 ± 0.36ab 1.67 ± 0.37bc 0.54 ± 0.04a 2.19 ± 0.26c

Data are given as mean ± SD (n ¼ 3). Different superscript letter a-e show the significant differences (P < 0.05) among samples (within the same row). Unwashed mince (UM);
Conventional washing (CW); Single washing (T-1); Alkaline saline washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl and 0.2% CaCl2) (T-3).

Similar observations were reported in Atlantic menhaden, mack-
erel and short-bodied mackerel gels (Chaijan, Benjakul,
Visessanguan, & Faustman, 2006; Pe rez-Mateos and Lanier, 2007)
and sutchi catfish surimi gels (Priyadarshini et al., 2016). Whiteness
for CW surimi gels could be attributed because of the removal of
myoglobin and haeme pigments through repeated washings. T-2
exhibited the lowest whiteness than the CW and T-1, which may be
due to tightly bound oxidized haeme pigments which could not be
removed by washing. A comparable significant increase (p < 0.05)
in the whiteness of T-3 gels were noticed when compared to UM
gels which can be due to formation of complex by CaCl2 with some
anion in the muscle, resulting in formation of insoluble particles,
leading to the light scattering in resulting gels (Benjakul,
Visessanguan, & Kwalumtharn, 2004). The lowest whiteness was
found in UM gels which was probably due to the heat denaturation
of pigments and further lead to turbidity.
3.3. Effect of different washing media on the protein pattern of
mince and gels
Protein pattern of the unwashed mince, surimi and their gels are
shown in Fig. 1A and B respectively. Myosin, actin, and actomyosin
are the most important proteins amongst myofibrillar proteins and
myosin is primarily responsible for the functional properties of
meat products. From the figure, it can be seen that the unwashed
mince and gels showed lighter MHC bands as compared with CW,
T-1, T-2 and T-3 indicating the presence of sarcoplasmic proteins.
However, no marked differences in MHC band intensity were
noticeable in CW, T-2 and T-3 surimi. Single washing with different
solutions exhibited an increased concentration of myofibrillar
proteins and reduced the amount of soluble sarcoplasmic proteins,
indicating the successful removal of sarcoplasmic proteins from the
washed mince. Heavy chain myosin bands intensity decreased in
heat induced gels, compared to that observed in the mince and
surimi due to polymerization of proteins on setting indicating
strong gel matrix. Among all gels tested, myosin heavy chain from
CW and T-2 decreased to a greater extent, than other treatments.
The decrease in heavy chain myosin was presumed to be due to the
cross-linking of protein during setting, leading to the lower band
intensity appearing on the SDS-PAGE (Benjakul, Chantarasuwan, &
Visessanguan, 2003). The result was coincidental with an increase
in gel strength. In the present study actin was found to be the
dominant protein in the gel (45 kDa) and similar observation was
made by Balange and Benjakul (2009) and reported that actin was
more resistant to proteolysis or could not be polymerised during
gelation.
3.4. Differential scanning calorimetry
Differential scanning calorimetry (DSC) is a widely used
analytical technique for studying the thermal stability of proteins,
and was applied in this study to investigate the effects of single
washing on mince, surimi and their respective pastes (Table 3).
Typical DSC thermograms result in two distinct peaks: the first
peak represents the temperature at which myosin denatures and
the second peak represents the denaturation of actin (Taskaya,
Chen, Beamer, & Jaczynski, 2009). The thermograms in case of
unwashed mince showed a transition occurring at 53.34
C which is
due to the association of myosin tails (Wright & Wilding, 1984). The
shift of the endothermic transition of myosin peak temperature to
higher values from surimi mince to pastes and between the treat-
ments indicated the delayed denaturation (Park, 1994) and stabi-
lization of fish protein, which is due to presence of NaCl. Similar
results were reported by Ferna ndez-Martin, Fernandez, Carballo,
and Jime nez-Colmenero (2000), when salt (NaCl) is added to
pork and chicken meat batters increased the onset temperature and
extent of protein denaturation. It was observed that the first tran-
sition peak in case of CW shifted to a lower value of 59.05
C. The
peak around 59.05
C in CWP was attributed to the breaking of the
actomyosin complex (Lin, Chen, & Chen, 2005). There was no
endothermic transition observed in T-2, which might be due to the
denaturation of myosin during alkaline saline washing. A similar
finding was reported in grass carp proteins when extracted through
alkaline process (Chen et al., 2016). While their respective pastes
exhibited two major transition peaks at 61.92
C and 74.60
C,
which may due to solubilisation of proteins in the presence of salt.
 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392 389

Fig. 1. Changes in protein pattern of mince and surimi gels (SDS-PAGE). Unwashed
mince (UM); Conventional washing (CW); Single washing (T-1); Alkaline saline
washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl and
0.2% CaCl2) (T-3). A e Before heat setting; B e After heat setting.
3100e3300 cm1, indicate amides III, II, I and amide B & A vibra-
tional modes in protein structure, respectively (Ahmad, Rizawi, &
Srivastava, 2010). In general, the amide I band consists of over-
lapped band components falling in the 1650e1660, 1600e1640,
1660e1695 and 1640e1650 cm1 ranges, which are attributable to
a -helix, b -sheet, b -turn and random coil structures, respectively
(Chan, Gill, & Paulson, 1992). The present study considered a
qualitative estimation of b-sheet and a-helix secondary structure
fractions only, since they are the main structures implicated in the
gelation process (Bouraoui, Nakai, & Li-Chan, 1997). Fig. 2 shows a
typical FTIR spectrum derived from the mince, surimi and their
respective gel ranging from 4000 to 400 cm1. The distinctive
central band at 1655 and 1654 cm1, diagnosed for a-helical
structures, which indicating a more compact structure (Kong & Yu,
2007) in mince and surimi.
The pronounced differences in the secondary structure were
observed in surimi gel treatments. The disappearance of the a
-helix band after heating indicating the unfolding of protein
structures during heating and the formation of b-sheet as an or-
dered network except for T-1 gel. The highest band intensity for UM
gels was observed at 1661.42 cm1 indicating random/loop struc-
tures which correspond to partial unfolding of a secondary struc-
ture during the solubilization process and peak at 1637.79 cm1
indicates the b-sheet band (Zhou, Zhao, Su, Cui, & Sun, 2014). A
major peak in the spectra of CW gels were observed at
1644.22 cm1 resulting in a predominance of b-sheet and hence a
more stabilized structure (Mozhaev, Heremans, Frank, Masson, &
Balny, 1996). The a-helix band (1654.58 cm1) in T-1 gels, dem-
onstrates that proper gelation did not occur after setting which
coincides with the poor gel strength of the gels. In T-2 (1662.01 and
1632.60 cm1) and T-3 (1661, 1640.02 and 1606.44 cm1) gels, the
major peak bands represent the random/loop structures indicating
that the fish protein denaturation is initiated by heat (Lin, Yang, Xu,
& Wang, 2015). When surimi samples were heated to form a
definitive gel there was a reduction in an a-helix structures and a
corresponding increase in b-sheet which is occurred due to protein
solubilizing by the NaCl and is a natural response of secondary
structures to heating (Moreno et al., 2015). According to Kobayashi,
Mayer, and Park (2017), tilapia being a warm water fish it was
observed that b-sheet bands increased in surimi gels when chop-
ped for a longer duration at the higher temperature and when
incubated at 4
C for more than overnight.

3.5. Changes in the Fourier transform infrared spectra of mince and
gels
FTIR technique is widely used for estimating protein secondary
structure (Fig. 2). Infrared absorptions around 1200, 1500, 1700 and
3.6. Microstructure of surimi gels
The SEM micrographs of the surface layers of the thermal gels at
7000 x magnification are shown in Fig. 3. The gels of UM displayed
a loose and coarse surface with pores of various sizes and no clear

Table 3
The onset (Tonset) and maximum (Tmax) temperatures for the endothermic transitions and net heat energy (Enthalpy DH) for different treatments (surimi and surimi pastes).

Peak 1 Peak 2

T onset (
C) T max (
C) Enthalpy (J/g) T onset (
C) T max (
C) Enthalpy (J/g)

UM 49.56 53.34 1.12 65.70 66.55 0.04

UMP 69.24 69.65 0.01 73.84 74.39 0.02
CW 61.04 61.95 0.04 64.86 65.25 0.01
CWP 58.53 59.05 0.03 63.42 63.84 0.01
T-1 54.00 57.18 1.39 72.20 73.01 0.00
T-1P 66.87 68.39 0.61 74.92 75.76 0.45
T-2 e e e e e e
T-2P 51.45 61.92 0.18 74.60 75.37 0.24
T-3 52.94 54.24 0.62 66.11 66.66 0.02
T-3P 68.26 68.97 0.03 75.74 76.34 0.09

Unwashed mince (UM); Conventional washing (CW); Single washing (T-1); Alkaline saline washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl
and 0.2% CaCl2) (T-3) and UMP, CWP, T-1P, T-2P and T-3P represent the solubilized surimi in NaCl.
390 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392

Fig. 2. Fourier transform infrared (FTIR) spectroscopy of samples from mince, surimi and gels. Unwashed mince (UM); Conventional washing (CW); Single washing (T-1); Alkaline
saline washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl and 0.2% CaCl2) (T-3) and UMP, CWP, T-1P, T-2P and T-3P represent the solubulised surimi
in NaCl.

network structure was observed. T-2 gels exhibited a fine structure
with an absence of voids and larger strands appeared which may be
due to the aggregated protein initiated by alkaline saline washing.
The compactness in CW can be explained that unfolding of the salt
solubilized surimi protein might have exposed reactive groups
resulting in denaturation on heating which interact to form a fine,
compact three-dimensional network structure, mainly through
hydrophobic interactions (Weng & Zheng, 2015). The strand-type
gel structure observed in T-3 can be due to reduced pH. It has
been reported that proteineprotein interactions such as associa-
tion, aggregation and polymerization in the gel network are
dependent upon temperature and pH (Totosaus, Montejano,
Salazar, & Guerrero, 2002). T-3 exhibited a compact homogenous
structure, which may be because of the presence CaCl2 that induced
the unfolding of myosin and lead to protein aggregation by pro-
moting hydrophobic interactions and forming Ca bridges (Pan, Guo,
 B. Priyadarshini et al. / LWT - Food Science and Technology 86 (2017) 385e392
Fig. 3. Scanning electron micrograph (magnification:  7000) of unwashed mince, surimi and surimi gels: Unwashed mince (UN); Conventional washing (CW); Single washing (T-
1); Alkaline saline washing (0.15% NaCl and 0.2% NaHCO3) (T-2); Calcium chloride and salt (0.1% NaCl and 0.2% CaCl2) (T-3).
Li, Song, & Ren, 2017) and this was in agreement with the higher
breaking force of the gel.
4. Conclusion
Overall, this study demonstrates that the surimi and their
respective gels can be produced from tilapia through single
washing cycle with different washing media. Improvement of the
thermal gelation properties of tilapia surimi was successfully
attained with alkaline saline washing. However, surimi prepared
from CW exhibited superior whiteness to that of other treatments.
This study found that the presence of CaCl2 decreased the gel
strength and whiteness. Manufacturing of surimi with single
washing cycle with alkaline saline washing can reduce the release
of wash water and thus prevent the detrimental effect of untreated
water into the environment. This recommendation can be useful for
the aquaculture and surimi industry.
Acknowledgment
The authors wish to thank the Director, ICAR-Central Institute of
Fisheries Education, Mumbai, Naik oceanic exports (Taloja, Mum-
bai) and CIFT (Vashi, Mumbai) for providing the facilities to conduct
the research.
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