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Article
Surimi Production from Tropical Mackerel: A Simple Washing
Strategy for Better Utilization of Dark-Fleshed Fish Resources
Worawan Panpipat 1 , Porntip Thongkam 1 , Suppanyoo Boonmalee 1 , Hasene Keskin Çavdar 2 and
Manat Chaijan 2, *

1 Food Technology and Innovation Research Center of Excellence, School of Agricultural Technology and Food
Industry, Walailak University, Thasala 80160, Thailand; [email protected] (W.P.);
[email protected] (P.T.); [email protected] (S.B.)
2 Department of Food Engineering, Faculty of Engineering, Gaziantep University, TR-27310 Gaziantep, Turkey;
[email protected]
* Correspondence: [email protected]; Tel.: +66-7567-2316; Fax: +66-7567-2302

Abstract: Mackerel (Auxis thazard), a tropical dark-fleshed fish, is currently a viable resource for the
manufacture of surimi, but the optimal washing procedure for more efficient use of this particular
species is required right away. Washing is the most critical stage in surimi production to ensure
optimal gelation with odorless and colorless surimi. The goal of this study was to set a simple
washing medium to the test for making mackerel surimi. Washing was performed three times with
different media. T1 was washed with three cycles of cold carbonated water (CW). T2, T3, and T4
were washed once with cold CW containing 0.3%, 0.6%, or 0.9% NaCl, followed by two cycles of
cold water. T5, T6, and T7 were produced for three cycles with CW containing 0.3%, 0.6%, or 0.9%
NaCl. For comparison, unwashed mince (U) and conventional surimi washed three times in cold tap
water (C) were employed. The maximum yield (62.27%) was obtained by washing with T1. When
varying quantities of NaCl were mixed into the first washing medium (T2–T4), the yield decreased
with increasing NaCl content (27.24–54.77%). When washing with NaCl for three cycles (T5–T7), the
yield was greatly decreased (16.69–35.23%). Conventional surimi washing (C) produced a yield of
roughly 40%, which was comparable to T3. Based on the results, treatments that produced lower
Citation: Panpipat, W.; Thongkam, P.; yields than C were eliminated in order to maximize the use of fish resources and for commercial
Boonmalee, S.; Çavdar, H.K.; Chaijan, reasons. The maximum NaCl content in CW can be set at 0.6% only during the first washing cycle
M. Surimi Production from Tropical (T3). Because of the onset of optimal unfolding as reported by specific biochemical characteristics
Mackerel: A Simple Washing Strategy such as Ca2+ -ATPase activity (0.2 µmol inorganic phosphate/mg protein/min), reactive sulfhydryl
for Better Utilization of Dark-Fleshed
group (3.61 mol/108 g protein), and hydrophobicity (64.02 µg of bromophenol blue bound), T3
Fish Resources. Resources 2023, 12,
washing resulted in surimi with the greatest gel strength (965 g.mm) and water holding capacity
126. https://doi.org/10.3390/
(~65%), with fine network structure visualized by scanning electron microscope. It also efficiently
resources12100126
removed lipid (~80% reduction), myoglobin (~65% reduction), non-heme iron (~94% reduction), and
Academic Editors: Zoltán Lakner and trichloroacetic acid-soluble peptide (~52% reduction) contents, which improves whiteness (~45%
Anita Boros improvement), reduces lipid oxidation (TBARS value < 0.5 mg malondialdehyde equivalent/kg),
Received: 17 September 2023 and decreases the intensity of the gel’s fishy odor (~30% reduction). As a result, washing mackerel
Revised: 11 October 2023 surimi (A. thazard) with CW containing 0.6% (w/v) NaCl in the first cycle, followed by two cycles of
Accepted: 20 October 2023 cold water washing (T3), can be a simple method for increasing gel-forming capability and oxidative
Published: 23 October 2023 stability. The mackerel surimi produced using this washing approach has a higher quality than that
produced with regular washing. This straightforward method will enable the sustainable use of
dark-fleshed fish for the production of surimi.

Copyright: © 2023 by the authors.

Keywords: washing; mackerel; fish; surimi; gel; protein content
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
1. Introduction
creativecommons.org/licenses/by/ Fishery and aquaculture production varies according to species, manufacturing, and
4.0/). product type and is mostly used for food. In 2018, around 88% of the total global fish

Resources 2023, 12, 126. https://doi.org/10.3390/resources12100126 https://www.mdpi.com/journal/resources

Resources 2023, 12, 126 2 of 16

production of 179 million mt was used as human food, while the remaining 12% was
employed for non-food reasons [1]. The use and processing of fish varies greatly across
regions. A significant majority of fish output in Asia is sold live or fresh to consumers,
whereas fish production in Europe and North America is predominantly offered frozen or
preserved [1].
Surimi is one of the fishery products that is intensively manufactured in Asian coun-
tries for further processing [2]. Surimi is a refined sort of fish flesh with special techno-
functionalities, such as the capacity to generate gels and bind water and oils, making
it an important ingredient in a wide range of processed foods [3,4]. Surimi processing
technology includes washing minced fish to clean and concentrate the myofibrillar proteins,
which are then transformed into additional commodities or stabilized by the incorpora-
tion of cryoprotectants, frozen, and kept for use afterward [4]. Optimizing the techno-
functionality and quality of surimi generated from raw materials, which are typically under-
used, low-value fish species, is a general objective of surimi research studies and production
operations [5–7]. Surimi may possibly be made from any fish, but the gelling features of
the surimi are determined by the nature of myofibrillar proteins, which are impacted by
the species and freshness of the fish, in addition to the processing conditions, primarily in
terms of protein concentration, ionic strength, pH, and temperature [5]. Threadfin bream
(Nemipterus spp.), bigeye snapper (Priacanthus spp.), croaker (Pennahia and Johnius spp.),
and lizardfish (Saurida spp.) are the principal white fish employed in Southeast Asia for the
manufacturing of surimi [2]. The supply of raw materials, however, is the fundamental
issue facing the Asian surimi industry [8]. Dark muscle fish have garnered more interest as
a potential replacement raw material for the manufacture of surimi owing to the restricted
white muscle fish supplies and the excessive harvesting of white fish in Thailand [8]. Ac-
cording to reports, the Gulf of Thailand has seen a rise in the catch of pelagic fish [5,9].
As a result, one of the most difficult ways to convert fish resources into human consum-
ables, especially protein-gated products, is the usage of this small pelagic fish for the
manufacturing of surimi. Chaijan et al. [5] examined the gel-forming capacities of surimi
from the dark-fleshed mackerel species Restrelligar kanagurta, Restrelligar brachysoma, and
Auxis thazard.
According to Singh et al. [10], dark-fleshed fish species naturally have a high con-
centration of dark muscle that contains significant amounts of lipids and sarcoplasmic
proteins. When compared to light muscle, dark muscle has poorer gelation qualities be-
cause it contains more sarcoplasmic proteins and lipids [11]. Fish myofibril protein gels’
strength, deformability, and color were negatively impacted by sarcoplasmic proteins [5,11].
According to Chaijan et al. [5], myoglobin predominates among the pigment proteins in
the sarcoplasmic fraction and is a factor in the surimi gel’s decreased whiteness. According
to Ochiai et al. [12], removing as much dark muscle as possible will result in high-quality
surimi that has better whiteness and a stronger gel. It is challenging to completely remove
the dark meat from fish with dark flesh, though. Therefore, the washing procedure is still
required for surimi made from the whole muscle of dark-fleshed fish in order to improve
color and gel strength.
Washing is the most critical stage in surimi manufacturing to achieve optimal gelling
along with colorless and odorless surimi [13]. When minced meat is washed, most of the
issues related to color, flavor, and odor are reduced or removed. Approximately two-thirds
of minced fish meat is myofibrillar proteins, which are the primary protein in the develop-
ment of a gel structure. The other one-third is made up of blood, myoglobin, lipids, and
sarcoplasmic proteins, which impair the surimi gels’ final quality. Consequently, washing,
which removes the undesirable one-third, improves the surimi quality by concentrating the
myofibrillar protein and extends the frozen storage life [11,13]. Washing involves combin-
ing minced fish with cold water and eliminating water using screening, dehydrators, or
centrifugation to around 5–10% solids. This procedure is carried out two or three times [14].
Surimi’s color can be brightened by extending the washing cycle, washing time, and
volume of washing medium [15]. Chen et al. [16] proposed that leaching mince with
Resources 2023, 12, 126 3 of 16

ozonized water during a short washing period can improve the color of dark-fleshed
fish surimi, such as horse mackerel. However, lipid and protein oxidation can limit its
applicability. Somjid et al. [17] developed a new method for obtaining gel-forming surimi
from dark muscle fish by employing cold carbonated water (CW) as a washing medium
for tropical mackerel (A. thazard) surimi manufacturing. The most efficient method for
increasing the total quality of mackerel surimi gel was a one-cycle washing with cold CW
and then a two-cycle rinsing with cold water [17]. However, myoglobin removal was not
enhanced and remains a difficulty. Thus, the objective of this study was to increase the
capacity for myoglobin removal from mackerel mince and to enhance the quality of the
resulting surimi by incorporating NaCl into the CW washing medium. This assumption
was made on the premise that increasing the ionic strength of the washing medium by
incorporating NaCl will aid in improving the leaching capacity of myoglobin during
washing of mackerel mince, as recently reported by Wang et al. [18]. To reduce myofibrillar
protein loss and yield reduction, the concentration of NaCl to be included with CW, as well
as the cycle of washing with NaCl, should be tuned. The biochemical features, myoglobin
and associated species contents, lipid content, and oxidative stability, as well as gelling
characteristics of the resulting surimi, were extensively evaluated in order to finalize a
suitable washing solution for straightforward mackerel surimi production.

2. Materials and Methods

2.1. Fish Sample
Mackerel (Auxis thazard) weighing 100–120 g was purchased from a local market in
Thasala, Nakhon Si Thammarat, Thailand. The fish were taken approximately a twelve-
hour period after capture and wrapped in ice with a fish/ice weight ratio of 1:2 (w/w)
before being transported within 20 min to Walailak University. Following that, the fish
were headed, eviscerated, cleaned, filleted, and skinned. The fish flesh was then thoroughly
minced with a meat processor (a Panasonic MK-G20MR, Tokyo, Japan, with a 4 mm
aperture diameter).

2.2. Surimi Production and Analysis

To study the effect of NaCl concentrations in cold CW washing on properties of
mackerel surimi, fresh fish mince was washed with cold CW (Chang® , Cosmos Brewery
Co., Ltd., Wangnoi, Ayutthaya, Thailand) in the presence of NaCl (0%, 0.3%, 0.6%, and
0.9%, w/v) in the 1st washing cycle with a medium/mince ratio of 3:1 (v/w). The 2nd
and 3rd cycles were performed using cold tap water without or with NaCl at different
concentrations (0.3%, 0.6%, and 0.9%, w/v). The maximum NaCl concentration was fixed
at 0.9% to prevent the loss of myofibrillar proteins. The mixture was gently stirred for
10 min in a cold environment (4 ◦ C), and the rinsed mince then went through a thin layer
of nylon screen. With an ultimate moisture level of 80%, hydraulic press dewatering was
applied. As controls, conventional washing with cold tap water for 3 cycles and unwashed
mince were employed [17]. All samples were mixed thoroughly prior to being placed in an
air-blast freezer with 4% (w/w) sucrose and 4% (w/w) sorbitol. The frozen samples were
maintained at −18 ◦ C until they were analyzed. Figure 1 depicts a schematic illustration of
surimi production and analysis. Table 1 illustrates the washing media employed in each
treatment as well as their yields.
 T4 CW + 0.9% NaCl CTW CTW 27.24 ± 1.4 f
T5 CW + 0.3% NaCl CTW + 0.3% NaCl CTW + 0.3% NaCl 35.23 ± 2.1 d
T6 CW + 0.6% NaCl CTW + 0.6% NaCl CTW + 0.6% NaCl 30.38 ± 1.0 e
T7 CW + 0.9% NaCl CTW + 0.9% NaCl CTW + 0.9% NaCl 16.69 ± 2.3 g
Conventional washing (C) CTW CTW CTW 39.04 ± 1.9 c
Resources 2023, 12, 126 4 of 16
* Values are given as mean ± standard deviation from 3 determinations. Different letters in the same
column represent significant differences (p < 0.05).

Figure 1.
Figure A schematic
1. A schematic illustration
illustration for
for the
the surimi
surimi production
production and
and analysis.
analysis.

2.2.1. 1.
Table Measurement of used
Washing media Yield,
in Moisture, andand
each treatment pH their yields.
The yield of surimi
Treatment
from all washing
1st Cycle
procedures was3rd
2nd Cycle
calculated
Cycle
usingYield
the weight
(%) *
of
the raw fish material in relation to the weight of minced fish. The moisture content and
Unwashed
pH of unwashed mince and - surimi were determined
- by the AOAC
- [19] and Benjakul
- et
mince (U)
al. [20] techniques, respectively.
Cold carbonated Cold tap water
T1 CTW 62.27 ± 1.1 a
water (CW) (CTW)
2.2.2. Measurement
T2 of Reactive
CW SulfhydrylCTW
+ 0.3% NaCl (SH), Ca -ATPaseCTW
2+ Activity, and54.77
Trichloroace-
± 2.3 b
tic AcidT3 CW Peptide
(TCA) Soluble + 0.6% NaCl CTW CTW 39.64 ± 2.0 c
T4 CW + 0.9% NaCl CTW CTW 27.24 ± 1.4 f
The reactive SH content and Ca2+-ATPase activity of natural actomyosin (NAM) iso-
CTW + 0.3% CTW + 0.3%
T5 unwashed
lated from CWmince
+ 0.3% and
NaClsurimi were measured by the methods 35.23 ± 2.1 d[21]
of Ellman
NaCl NaCl
and Benjakul et al. [20]. TCA-soluble peptide
CTW + was
0.6%also measured using the Benjakul et al.
CTW + 0.6%
T6 CW + 0.6% NaCl 30.38 ± 1.0 e
[20] technique. NaCl NaCl
CTW + 0.9% CTW + 0.9%
T7 CW + 0.9% NaCl 16.69 ± 2.3 g
2.2.3. Measurement of Protein HydrophobicityNaCl NaCl
Conventional
The hydrophobicity CTW
of nonsolubilizedCTW CTW
myofibrils was measured using39.04 ± 1.9 c
bromophenol
washing (C)
*blue (BPB)
Values for as
are given electrophoresis
mean ± standardand reported
deviation from 3 in terms of BPB
determinations. bound,
Different asinhighlighted
letters by
the same column
Chelh
representetsignificant
al. [22]. differences (p < 0.05).

BPB bound
2.2.1. Measurement of Yield, (µg)and
Moisture, pHµg ×
= 200 (1)
The yield of surimi from all washing procedures was calculated using the weight of
where A = absorbance at 595 nm.
the raw fish material in relation to the weight of minced fish. The moisture content and pH
of unwashed mince and surimi were determined by the AOAC [19] and Benjakul et al. [20]
techniques, respectively.

2.2.2. Measurement of Reactive Sulfhydryl (SH), Ca2+ -ATPase Activity, and Trichloroacetic
Acid (TCA) Soluble Peptide
The reactive SH content and Ca2+ -ATPase activity of natural actomyosin (NAM)
isolated from unwashed mince and surimi were measured by the methods of Ellman [21]
and Benjakul et al. [20]. TCA-soluble peptide was also measured using the Benjakul
et al. [20] technique.
Resources 2023, 12, 126 5 of 16

2.2.3. Measurement of Protein Hydrophobicity

The hydrophobicity of nonsolubilized myofibrils was measured using bromophenol
blue (BPB) for electrophoresis and reported in terms of BPB bound, as highlighted by Chelh
et al. [22].
Acontrol − Asample
 
Resources 2023, 12, 126 BPB bound (µg) = 200 µg × (1) 5 of 1
Acontrol
where A = absorbance at 595 nm.
2.2.4. Measurement
2.2.4. Measurement of Myoglobin,
of Myoglobin, Non-Heme Non-Heme
Iron, and Iron,
Lipidand Lipid Contents
Contents
The concentration
The concentration of myoglobin
of myoglobin was measured
was measured spectrophotometrically
spectrophotometrically at 525 nmatby
525 nm by
Benjakul and Bauer’s approach [23]. Schricker et al. [24] devised a method
Benjakul and Bauer’s approach [23]. Schricker et al. [24] devised a method for measuring for measuring
non-hemenon-heme iron concentration.
iron concentration. The BlighThe
andBligh
Dyerand Dyer procedure
procedure [25] was employed
[25] was employed for the for the
extraction of
extraction of the lipid. the lipid.

2.3. Gel Preparation

2.3. Gel Preparation
To defrostTo defrost
frozen frozen running
samples, samples,taprunning
watertapwaswater
usedwasuntilused until thetemperature
the interior interior temperature
approximated ◦
approximated 0 °C. Following
0 C. Following that, the were
that, the samples samples
cut were cut into
into small small fragments,
fragments, and the and th
moisture moisture
level waslevel
fixedwas
to fixed
80%. toDry80%. Dry(2.5%
NaCl NaClw/w)
(2.5%was
w/w)introduced
was introduced to thetosamples
the samples and
chopped
and chopped for 5for
min5 min to generate
to generate a homogeneous
a homogeneous sol.Next,
sol. Next,the thesol
sol was
was inserted in in a polyvi
nylidene casing
a polyvinylidene casing ((⌀ ==2.52.5cm),
cm),and
andboth
bothsides were
sides weretightly
tightlysealed.
sealed.Following
Followinga 30amin incu
bation period
30 min incubation period ◦ C,the
atat4040°C, thesample
sampleunderwent
underwent aa 20 min heating
heating period
periodatat9090◦ C.
°C. Conse
Consequently, the the
quently, gelsgels
werewerechilled in ice
chilled in water before
ice water being
before stored
being ◦
at 4atC4 for
stored 24 h24[7].
°C for h [7].

2.4. Gel Analyses

2.4. Gel Analyses
The gel strength
The gel (breaking force × deformation)
strength (breaking of unwashed
force × deformation) mince and
of unwashed surimi
mince andgels
surimi gel
was reported after the texture was evaluated using a texture analyzer (TA-XT2, Stable
was reported after the texture was evaluated using a texture analyzer (TA-XT2, Stable Mi
Micro Systems, Godalming,
cro Systems, Surrey, Surrey,
Godalming, UK) mounted with a spherical
UK) mounted plunger (diameter
with a spherical 5 mm; 5 mm
plunger (diameter
depression speed 60 mm/min/trigger force 0.05 N) [7]. The expressible moisture
depression speed 60 mm/min/trigger force 0.05 N) [7]. The expressible moisture of gels of gel
was calculated as a percentage
was calculated of sampleofmass
as a percentage using
sample theusing
mass procedure of Phetsang
the procedure et al. [7]. et al. [7]
of Phetsang
The values of L*, a*, and b* were recorded with a colorimeter (Hunter Assoc. Laboratory;
The values of L*, a*, and b* were recorded with a colorimeter (Hunter Assoc. Laboratory
Reston, VA, USA),VA,
Reston, andUSA),
the whiteness was computed
and the whiteness as followsas[7]:
was computed follows [7]:
L*)−2 [(100
100 − [(100=−100
Whiteness = Whiteness + a*2 −+L*)
b*22]+1/2a*2 + b*2]1/2 (2) (2
The thiobarbituric
The thiobarbituric acidsubstance
acid reactive reactive substance
(TBARS) (TBARS) assay developed
assay developed by Buegeby andBuege and
Aust [26] Aust [26] was performed
was performed using theusing
ground thesample.
ground sample.
A scanningA scanning
electronelectron microscope
microscope (SEM)(SEM) (GeminiSEM,
(GeminiSEM, CarlCarl Zeiss
Zeiss Microscopy, Ober
Microscopy,
kochen, Germany) operating at a 10 kV accelerating voltage was
Oberkochen, Germany) operating at a 10 kV accelerating voltage was used to examine used to examine
the the mi
crostructures of
microstructures of gels [27].gels [27].
Tenpanelists
Ten trained trained panelists with extensive
with extensive experienceexperience
evaluatingevaluating off-flavors
off-flavors of seafoodof seafood
evaluatedevaluated theseverity
the gel. The gel. Theof severity of fishy
fishy odor wasodor was on
assessed assessed on ascale
a 5-point 5-point scalefrom
ranging ranging from
none to
none to intense intense
(score 0–4)(score 0–4)
[8]. The [8]. TheUniversity
Walailak Walailak University HumanEthics
Human Research Research Ethics Commit
Committee
authorizedteethe
authorized the study
study procedure procedure (WUEC-21-125-02).
(WUEC-21-125-02).

2.5. Statistical Analysis Analysis

2.5. Statistical
Throughout the study,the
Throughout a completely randomized
study, a completely design was
randomized employed.
design ANOVA was
was employed. ANOVA wa
utilized toutilized
manage the data, and mean comparisons were made using Duncan’s
to manage the data, and mean comparisons were made using Duncan’s Multipl Multiple
Range Test. For the
Range statistical
Test. For the analysis,
statisticalthe statistical
analysis, theprogram (SPSS
statistical 23.0, SPSS
program Inc.,
(SPSS Chicago,
23.0, SPSS Inc., Chi
IL, USA) cago,
was employed.
IL, USA) was employed.
3. Results3.and Discussion
Results and Discussion
3.1. Yield
3.1. Yield
Table 1 shows the yield of surimi made with various washing mediums. It has
Table 1 shows the yield of surimi made with various washing mediums. It has been
been reported that around 50% of total protein disappears during washing, but this
reported that around 50% of total protein disappears during washing, but this can vary
depending on the washing conditions [28]. The maximum yield was obtained by washing
with CW in the first cycle and water in the second and third cycles (T1) (p < 0.05). When
varying quantities of NaCl were mixed into the first washing medium (T2–T4), the yield
decreased with rising NaCl content (p < 0.05). When washing with NaCl for three cycle
Resources 2023, 12, 126 6 of 16

can vary depending on the washing conditions [28]. The maximum yield was obtained
by washing with CW in the first cycle and water in the second and third cycles (T1)
(p < 0.05). When varying quantities of NaCl were mixed into the first washing medium
(T2–T4), the yield decreased with rising NaCl content (p < 0.05). When washing with NaCl
for three cycles (T5–T7), the yield was greatly decreased (p < 0.05). In this case, the drop in
yield was caused by the leaching of myofibrillar proteins with increasing ionic strength,
particularly with extensive washing by elevating the round of washings in the presence
of NaCl. Conventional surimi washing (C) produced a yield of roughly 40%, which was
comparable to T3. Too much salt in the wash water has been observed to promote the
solubilization of myofibrillar proteins, leading to a higher loss of myofibrillar proteins and
lower yield [29,30]. As NaCl rose by more than 1.0% in the washing medium, it reduced
the protein recovery of grass carp surimi [18]. It has been discovered that washing grass
carp with 0.25–0.5% NaCl at pH 5–7 can significantly improve the yield of surimi [18]. The
application of NaCl in the washing solution was chosen because NaCl, one of the least
expensive substances, is a critical component and a processing aid for a variety of food
products [31]. Other NaCl alternatives can be researched in the future to increase the yield
and quality of mackerel surimi. However, the proposed compounds’ cost and efficiency
should be carefully assessed.
Myofibrillar proteins are the majority of muscle proteins that possess desirable gelling,
emulsifying, and film-forming properties [32]. Myofibrillar proteins are thought to be
salt-soluble proteins [33]. Muscle tissue’s physiological ionic strength is predicted to be
0.15–0.30 M [34]. Additional salts, particularly NaCl, are usually employed during the
processing to enhance the flavor, color, texture, and shelf-life of muscle-based foods. To
solubilize the myofibrillar proteins and attain the appropriate functional characteristics,
2–3% (0.47–0.68 M) NaCl is usually required [35]. Although the salt concentration in the
washing media was maintained at no more than 0.9% to prevent myofibrillar protein loss,
some myofibrillar proteins were still eliminated during washing, especially with higher
salt content and the presence of salt in the first and second washing cycles. The DLVO
theory, which claims that colloidal stability is determined by a compromise of attractive
van der Waals forces and repulsive double-layer electrostatic bonds, can be used to explain
the impact of ions on the solubility of the protein in colloidal research [36]. Protein can
be thought of as a macroion. Under a salt solution environment, it is enclosed by more
counterions than co-ions, which shields the protein surface charge and allows it to solubilize.
As the salt content rises, the protective role of protein charge causes the electric double
layers to compress and the repulsive term of the DLVO theory to diminish [36]. Protein
solubility is projected to reduce at high salt concentrations [37,38].
Based on the results, treatments that produced lower yields than conventional washed
surimi (C) were eliminated in order to maximize the use of fish resources and for commercial
reasons. The maximum NaCl content in CW can be set at 0.6% only during the first washing
cycle (T3).

3.2. Biochemical Features

3.2.1. pH
The pH of mackerel mince was 5.54, and the pH of surimi increased significantly,
depending on the medium employed (Table 2). The highest pH value of 6.60 was found in
T1 surimi (p < 0.05). Surimi conventionally washed with water (C), T2, and T3 showed a
comparable pH value (pH ~ 6.1) (p > 0.05), which was in agreement with Chaijan et al. [39],
who reported a pH value of 6.17 in mackerel (R. kanagurta) surimi produced with 0.5%
NaCl leaching. In the case of CW washing (T1–T3), the pH tended to be lower, with NaCl
content in the washing medium, regardless of concentration. It has been reported that the
ultimate pH value of surimi may be related to the pH of the medium applied and can be
impacted by the effectiveness of leaching along with the adsorption of washing media into
surimi. Upon the washing step, both acidic and alkaline elements can be leached out of
fish mince, leading to a new muscle pH equilibrium [17].
Resources 2023, 12, 126 7 of 16

Table 2. Effect of cold carbonated water and NaCl washing on biochemical properties of mackerel
surimi in comparison with unwashed mince and conventional surimi.

Ca2+ -ATPase TCA-Soluble

Reactive Sulfhydryl Surface
Activity Peptide (µmol
Treatment pH Content (mol/108 g Hydrophobicity;
(µmolPi/mg Tyrosine/g
Protein) BPB Bound (µg)
Protein/min) Sample)
U 5.54 ± 0.01 c 2.70 ± 0.36 bc 0.47 ± 0.02 a 34.09 ± 0.50 e 0.23 ± 0.00 a
T1 6.60 ± 0.22 a 3.45 ± 0.24 ab 0.14 ± 0.02 e 52.66 ± 3.51 c 0.12 ± 0.01 b
T2 6.11 ± 0.14 b 2.19 ± 0.06 c 0.29 ± 0.04 b 43.17 ± 3.46 d 0.12 ± 0.00 b
T3 6.06 ± 0.04 b 3.61 ± 0.58 a 0.20 ± 0.03 cd 64.02 ± 3.78 b 0.11 ± 0.01 b
C 6.05 ± 0.06 b 3.46 ± 0.34 ab 0.24 ± 0.01 bc 76.27 ± 3.18 a 0.11 ± 0.01 b
Values are given as mean ± standard deviation from three determinations. Different letters in the same column
indicate significant differences (p < 0.05). BPB = bromophenol blue. TCA = trichloroacetic acid. U = unwashed
minced. T1 = cold carbonated water washing in the 1st washing cycle and cold tap water washing in the 2nd
and 3rd cycles. T2 = cold carbonated water washing in the presence of 0.3% NaCl (w/v) in the 1st washing cycle
and cold tap water washing in the 2nd and 3rd cycles. T3 = cold carbonated water washing in the presence of
0.6% NaCl (w/v) in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. C = conventional
washing with 3 cycles of cold tap water.

3.2.2. Reactive SH, Ca2+ -ATPase Activity, and Surface Hydrophobicity

Biochemical changes, such as reactive SH, Ca2+ -ATPase activity, and surface hydropho-
bicity in this case, can show that the structure of fish proteins has changed during the
manufacturing of surimi [27]. Those indices were altered in various ways depending on
the washing approach (Table 2). Myosin molecules’ head and tail segments frequently
contained a number of reactive SH groups, according to Buttkus [40]. When the molecular
structure of myosin changes, the reactive SH groups may become more exposed [20]. After
washing, surimi’s reactive SH concentration changed (p < 0.05; Table 2). Therefore, washing
can accelerate the protein denaturation, exposing hidden SH and accelerating the oxidation
of SH groups. Unwashed mince and T2 surimi had the next-highest SH levels, followed by
T1, T3, and conventional surimi.
Unwashed mince exhibited the highest Ca2+ -ATPase activity (p < 0.05; Table 2). Re-
gardless of the washing media, a decline in the Ca2+ -ATPase activity was seen in all surimi
(p < 0.05). In comparison with unwashed mince, Das et al. [41] and Somjid et al. [27] ob-
served that the Ca2+ -ATPase activities in mackerel, pink perch, croaker, and sardine surimi
gradually decreased. A reduction in NAM’s Ca2+ -ATPase activity suggested that myosin
integrity was lost due to denaturation and/or aggregation [42]. The structural alterations
of the myofibrillar proteins upon washing are responsible for the loss of Ca2+ -ATPase
activity [41]. Unwashed mince and surimi have Ca2+ -ATPase activity in the following
order: U > T2 ≥ C ≥ T3 > T1.
Table 2 also shows the surface hydrophobicity of surimi and unwashed mince. Since
hydrophobic amino acid sequences are usually found inside protein structures, the surface
hydrophobicity of protein molecules is typically related to their exposed hydrophobic
moieties [43]. Exposed hydrophobic residues can create contacts and enhance the gel
structure during thermal gelation [39,44]. A good gel-forming capability of protein, on
the other hand, requires a sufficient shift in surface hydrophobicity [17]. According to the
findings, unwashed mince carried the lowest surface hydrophobicity (p < 0.05). Surimi
with the greatest surface hydrophobicity (p < 0.05) was conventional surimi, followed by
T3, T1, and T2. Among the CW washes, the one with 0.6% NaCl (T3) produced surimi with
higher surface hydrophobicity than the others. Wang et al. [18] discovered that increasing
the NaCl concentration during washing elevated the α-helix content of actomyosin, an
indication of protein unfolding.

3.2.3. TCA-Soluble Peptide

Table 2 displays the TCA-soluble peptide contents of unwashed mackerel mince,
typical surimi, and surimi washed with CW-NaCl solutions. The TCA-soluble peptide
Resources 2023, 12, 126 8 of 16

in unwashed mince was 0.23 µmol tyrosine/g sample, and the value was reduced by
approximately 50% following washing, regardless of washing medium. TCA-soluble
peptide indicated proteolytic breakdown and a higher concentration indicated greater
muscle protein hydrolysis [20]. According to Somjid et al. [27], unwashed mince provided
the most TCA-soluble peptide, suggesting the presence of some proteinases that can
produce proteolytic breakdown products. During handling and storage, endogenous
proteinases in the sarcoplasmic fraction can trigger autolysis and buildup of particular
soluble peptides in mackerel mince [27]. By repeatedly washing the mince, peptides can be
removed, and proteinases may be inactivated. Due to proteolytic enzymes’ association with
myofibrillar proteins and interference with gelation, dark-fleshed fish species have weak
gelation ability [45]. Activation of myofibril-bound proteinases during the early phases of
thermal gelation can result in gel-weakening [45].

3.3. Myoglobin, Non-Heme Iron, and Lipid Contents

Unwashed mince contained about 27 mg/100 g of myoglobin. As a result, regardless
of the washing method, the myoglobin content of surimi was lower than that of unwashed
mince (p < 0.05) (Figure 2a). The results revealed that washing with cold tap water or
CW could remove some myoglobin. However, due to the association among myoglobin
and muscle components, particularly myofibrillar proteins, it is difficult to separate all
myoglobin from mackerel mince [5]. When washed with the conventional process (C), the
myoglobin content decreased by 46.76%. However, CW washing (T1) only removed 3.8% of
the myoglobin. This was consistent with a previous study in which CW washing enhanced
the gel functionality of mackerel surimi but did not increase heme protein removal [17].
When salt was added to the CW medium, the myoglobin removal efficacy increased to
36.68% for T2 and 64.63% for T3. The presence of salt at the optimal level may have aided
in weakening the association between myoglobin and myofibrillar proteins or muscle
components, hence facilitating myoglobin leaching efficacy. As previously indicated,
raising the ionic strength of the washing medium can improve the solubility of myofibrillar
proteins. This can cause myoglobin to dissociate from the myoglobin–myofibrillar complex,
allowing myoglobin to be eliminated within the washing water. Unwashed mince had
a non-heme iron level of 50.63 mg/g, which significantly decreased following washing
with all available methods (Figure 2a). Non-heme iron was eliminated by 93.72% by T3,
compared to 79.68%, 68.56%, and 65.83% by T2, T1, and C, respectively. T3 may remove
the non-heme iron through washing media due to the enhanced solubility of myofibrillar
proteins at high ionic strength. Overall, the removal of myoglobin and non-heme iron from
mackerel by T3 appeared to be the most successful washing procedure, which may have
enhanced the gel’s functionality and oxidative stability.
Figure 2b depicts the lipid content of unwashed mince and surimi obtained by con-
ventional washing and CW-NaCl washing. Lipid is regarded as an unfavorable surimi
component because it interferes with gel formation and induces gel rancidity [39,46]. Un-
washed mince included about 2 g/100 g lipid, which was removed to variable degrees
after washing depending on the washing solution employed. According to the previous
work [17], conventional washing eliminated nearly half of the lipid content in fish mince.
CW washing (T1) removed 52% of the lipid, and the removal capacity improved to 63 and
80% when washed with T2 and T3, respectively, as the NaCl level in the washing medium
increased. This was most likely due to the solubility of myofibrillar proteins, which can aid
in the removal of lipids from muscle components. The reduced yield of surimi produced
by T3 (Table 1) may potentially be related to the residual lipid content. However, it has
been noted that oxidative stability and gel-forming ability increase with decreasing lipid
concentration [46].
Resources 2023, 12, 126 9 of 16
Resources 2023, 12, 126 9 of 16

(a)
30 70
a
b

-■- Non-heme iron (mg/g)

-□- Myoglobin (mg/100 g)
25 60
a
50
20 c
d 40
15
30
e
10 b
c 20
d
5 10
e
0 0
U T1 T2 T3 C
Treatments

(b)

2.5

a
2
Lipid (g/100 g)

1.5
b
c
1
d

0.5 e

0
U T1 T2 T3 C
Treatments
Figure 2. Effect of cold carbonated water and NaCl washing on myoglobin and non-heme iron
contents
Figure (a) and
2. Effect lipid carbonated
of cold content (b) of mackerel
water surimiwashing
and NaCl in comparison with unwashed
on myoglobin mince and
and non-heme iron con-
conventional
tents surimi.
(a) and lipid Bars indicate
content standard surimi
(b) of mackerel deviationin from 3 determinations.
comparison Differentmince
with unwashed letters and
on the
conven-
tional
barssurimi. Bars
represent indicate
significant standard(pdeviation
differences < 0.05). U from 3 determinations.
= unwashed minced. T1 =Different letters water
cold carbonated on the bars
washingsignificant
represent in the 1st washing
differencescycle(pand cold tap
< 0.05). U =water washing
unwashed in the 2nd
minced. T1 =and
cold3rd cycles. T2 =
carbonated cold wash-
water
ing in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. T2 =cold
carbonated water washing in the presence of 0.3% NaCl (w/v) in the 1st washing cycle and cold car-
tap water
bonated washing
water in thein2nd
washing theand 3rd cycles.
presence T3 = NaCl
of 0.3% cold carbonated
(w/v) in thewater
1stwashing
washingincycle
the presence
and cold tap
of 0.6%
water NaCl (w/v)
washing in thein2nd
the 1st
andwashing cycle T3
3rd cycles. and=cold
coldtap water washing
carbonated waterin washing
the 2nd andin 3rd
the cycles.
presence of
C =NaCl
0.6% conventional
(w/v) inwashing with 3 cycles
the 1st washing of and
cycle cold cold
tap water.
tap water washing in the 2nd and 3rd cycles. C =
conventional washing with 3 cycles of cold tap water.
3.4. Gelling Properties
3.4.1. Gel Strength, Expressible Drip, and Whiteness
Figure 2b depicts the lipid content of unwashed mince and surimi obtained by con-
Unwashed mince consisted of more lipid, myoglobin, non-heme iron, and TCA-soluble
ventional washing and CW-NaCl washing. Lipid is regarded as an unfavorable surimi
peptide, all of which had a negative impact on the ability of myofibrillar proteins to form
component because it interferes with gel formation and induces gel rancidity [39,46]. Un-
gels. From the unwashed minced gel, the lowest gel strength (Figure 3a), most expressible
washed mince3b),
drip (Figure included about
and lowest 2 g/100(Figure
whiteness g lipid,3c)
which was
can be removed
obtained. to variable
Furthermore, degrees
one of the after
washing
elements influencing the textural characteristics of surimi gel may be its pH. Unwashed work
depending on the washing solution employed. According to the previous
[17], conventional
mince had a pH of washing
5.54, whicheliminated
was close tonearly half ofpoint
the isoelectric the lipid content
of myosin in fish which
(pI 5.0–5.5), mince. CW
washing (T1) removed 52% of the lipid, and the removal capacity improved to 63 and 80%
when washed with T2 and T3, respectively, as the NaCl level in the washing medium
increased. This was most likely due to the solubility of myofibrillar proteins, which can
aid in the removal of lipids from muscle components. The reduced yield of surimi pro-
Resources 2023, 12, 126 10 of 16

is more likely to promote agglutination with water released from the matrix than other
treatments with higher pH values (pH 6–7) (Table 2). Surimi washed with all treatments
performed better in terms of gel strength, water-holding capacity, and whiteness than
unwashed mince (Figure 3). T3 produced the highest gel strength surimi, followed by
T1/T2 and conventional surimi. T3 may increase the ionic strength in the washing medium
to achieve the best conditions for protein dissociation and denaturation in the subsequent
step, hence increasing gel strength. Among CW washing, surimi rinsed with T3 tended to
exhibit the highest reactive SH content and surface hydrophobicity (Table 2), indicating that
those groups can join to create a three-dimensional network with water-binding potential.
Surimi’s SH groups may encourage the development of disulfide bonds, which are essential
for gel strengthening after gelation [47]. The T2 with the lowest SH group among the surimi
may create a gel with a higher water release. The conventional surimi had the maximum
hydrophobicity (Table 2), which may prevent the appropriate protein–protein interaction
from forming a gel. Protein unfolding has been shown to expose hydrophobic residues,
altering the surface hydrophobicity of the protein [48]. Exposed hydrophobic residues can
create contacts and strengthen the gel network during thermal gelation [39,44]. A good
gel-forming property of protein, on the other hand, requires a sufficient shift in surface
hydrophobicity [17].
Surimi gel had a higher level of whiteness than unwashed mince, and among the
surimi, the gel from T3 had the highest whiteness. Surimi’s color was influenced by its
residual pigment content and thermal gelation stability. Furthermore, components like
lipids, which can oxidize when heated, can serve as an additional source of color via the
Maillard reaction because lipid oxidation produces aldehyde, which can then combine with
amine to create brown pigment via the amine–carbonyl reaction or Maillard reaction [27,49].
T30 s gel was the whitest because it had the least amount of myoglobin, non-heme iron, and
lipid (Figure 2), as well as the lowest degree of lipid oxidation (see below).
In the investigation of another species of mackerel, Scomber japonicus, washing was
performed using 4 vol of 0.04 M phosphate buffer (pH 8.0 and 8.5) or 0.04 M bicarbonate
buffer (pH 8.0 and 8.5) at 4 ◦ C for 5 min in the first cycle, followed by two cycles of 3 vol of
chilled tap water (4 ◦ C). During the final washing, the pH of the mince was decreased to 7.0
by using a 1.0 M citric acid solution. The moisture content of washed mince was reduced
to 78% by centrifugation for 15–20 min at 1500× g [50]. Washing with alkaline phosphate
or bicarbonate buffers did not remove the pigment from minced mackerel [50]. Because
the alkaline treatment alone was ineffective at improving the color of mackerel surimi,
ozonation was attempted. Ozonation at pH 3.0 for 30 min improved the color of mackerel
surimi. To improve the gel strength, the ozonated mackerel mince could be blended with
0.1% NaHSO3 , 0.2% ascorbic, 0.15% cysteine, or their combination after regaining the pH
to neutral on the final washing [50].
According to the findings of those studies, a complex step was used in the production
of mackerel surimi [50], alkaline washing did not improve the color of dark-fleshed fish,
and the use of ozone, an oxidizing agent, may have a negative impact on gel-forming ability
and oxidative stability of lipid in the resulting surimi. Thus, washing with cold carbonated
water containing 0.6% NaCl in the first washing cycle, followed by two cycles of cold tap
water washing (T3), can be a straightforward method for producing good surimi from
mackerel mince.
 pigment content and thermal gelation stability. Furthermore, components like lipids, which
can oxidize when heated, can serve as an additional source of color via the Maillard reaction
because lipid oxidation produces aldehyde, which can then combine with amine to create
brown pigment via the amine–carbonyl reaction or Maillard reaction [27,49]. T3′s gel was
Resources 2023, 12, 126 the whitest because it had the least amount of myoglobin, non-heme iron, and lipid (Figure11 of 16
2), as well as the lowest degree of lipid oxidation (see below).

(a)
1200 a
1000

Gel strength (g.mm)

800 b
b
600 c
400
d
200
Resources 2023, 12, 126 11 of 16
0
U T1 T2 T3 C
Treatments
(b)
100
Expressible drip (%)

80

60
a
b c
40 c c

20

0
U T1 T2 T3 C
Treatments

(c)
100

80
a
b
Whiteness

b
60 c
d
40

20

0
U T1 T2 T3 C
Treatments
Figure 3. Effect of cold carbonated water and NaCl washing on gel strength (a), expressible drip
Figure 3. Effect of cold carbonated water and NaCl washing on gel strength (a), expressible drip (b),
(b), and whiteness
and whiteness (c) of mackerel
(c) of mackerel surimi gel surimi gel in comparison
in comparison with gelswith
fromgels from unwashed
unwashed mince and mince
con-and
conventional
ventional surimi. surimi. Bars indicate
Bars indicate standardstandard deviation
deviation from 3from 3 determinations.
determinations. Different
Different letters
letters on the
on the
bars represent significant differences (p < 0.05). U = unwashed minced. T1 = cold
bars represent significant differences (p < 0.05). U = unwashed minced. T1 = cold carbonated water carbonated water
washing in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. T2 = coldcold
washing in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. T2 =
carbonated water
carbonated washing
water in theinpresence
washing of 0.3%
the presence of NaCl (w/v) in
0.3% NaCl the 1st
(w/v) washing
in the cycle and
1st washing cold
cycle tapcold
and
water washing in the 2nd and 3rd cycles. T3 = cold carbonated water washing in the
tap water washing in the 2nd and 3rd cycles. T3 = cold carbonated water washing in the presence presence of
0.6%ofNaCl (w/v) in the 1st washing cycle and cold tap water washing in the 2nd and 3rd
0.6% NaCl (w/v) in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. cycles. C =
conventional washing with 3 cycles of cold tap water.
C = conventional washing with 3 cycles of cold tap water.

In the investigation of another species of mackerel, Scomber japonicus, washing was

performed using 4 vol of 0.04 M phosphate buffer (pH 8.0 and 8.5) or 0.04 M bicarbonate
buffer (pH 8.0 and 8.5) at 4 °C for 5 min in the first cycle, followed by two cycles of 3 vol
of chilled tap water (4 °C). During the final washing, the pH of the mince was decreased
to 7.0 by using a 1.0 M citric acid solution. The moisture content of washed mince was
reduced to 78% by centrifugation for 15–20 min at 1500× g [50]. Washing with alkaline
 ity and oxidative stability of lipid in the resulting surimi. Thus, washing with cold car-
bonated water containing 0.6% NaCl in the first washing cycle, followed by two cycles of
Resources 2023, 12, 126 cold tap water washing (T3), can be a straightforward method for producing good surimi
12 of 16
from mackerel mince.

3.4.2. Microstructure
3.4.2. Microstructure
Surimi gel’s fine network structure was visible in T1, T2, and, notably, T3 (Figure 4).
Surimi gel’s fine network structure was visible in T1, T2, and, notably, T3 (Figure 4).
Overall, T1, T2, and T3 surimi gel showed routinely continuous and smooth network archi-
Overall, T1, T2, and T3 surimi gel showed routinely continuous and smooth network
tectures due to the removal of non-gel-forming components, including lipid and myoglobin
architectures due to the removal of non-gel-forming components, including lipid and
[8] with appropriate unfolding [17,39,44], which might be linked to the proper gel network
myoglobin [8] with appropriate unfolding [17,39,44], which might be linked to the proper
formation of myofibrillar proteins. During thermal gelation, myosin and other salt-soluble
gel network formation of myofibrillar proteins. During thermal gelation, myosin and other
myofibrillar proteins undergo complicated changes in rheological features. A good surimi
salt-soluble myofibrillar proteins undergo complicated changes in rheological features. A
gel can be generated
good surimi gel can be by generated
constructing byaconstructing
regular assembled structure
a regular assembledwith astructure
well-organized
with a
three-dimensional matrix [8,17]. The network of the conventional surimi
well-organized three-dimensional matrix [8,17]. The network of the conventional surimi (C) gel had some
(C)
holes
gel hadinsome
it andholes
a discontinuous structure, which
in it and a discontinuous may have
structure, whichcontributed
may have to its reduced
contributed togel
its
strength gel
reduced compared
strengthtocompared
the surimitogelthewashed
surimi with CW-NaCl
gel washed with(T1–T3)
CW-NaCl (Figure 4). Unwashed
(T1–T3) (Figure 4).
mince had amince
Unwashed proteinhadnetwork packed
a protein with clots
network packedandwithaggregates, which
clots and was associated
aggregates, which with
was
a weaker gel and more water expelled (Figure 3). Sarcoplasmic proteins,
associated with a weaker gel and more water expelled (Figure 3). Sarcoplasmic proteins, according to Arfat
and Benjakul
according [51],and
to Arfat can Benjakul
interact with myofibrillar
[51], can interact with proteins and impede
myofibrillar theand
proteins formation
impedeof thea
robust gel of
formation structure.
a robust Residual lipids
gel structure. were reported
Residual lipids were to impair
reportedthetogel-forming capacity of
impair the gel-forming
surimi because lipids are unable to gel and tend to interfere with the protein
capacity of surimi because lipids are unable to gel and tend to interfere with the protein networks [8].
Overall, the finest structure of T3 surimi gel was associated with the highest
networks [8]. Overall, the finest structure of T3 surimi gel was associated with the highest gel strength
(Figure
gel 3a). As
strength a result,
(Figure 3a).T3
Aswas determined
a result, T3 wastodetermined
be the optimal washing
to be method
the optimal for producing
washing method
good gel-forming surimi from tropical mackerel.
for producing good gel-forming surimi from tropical mackerel.

U C

T1 T2 T3
Figure 4. Microstructure
Microstructure ofof gels
gels from
fromunwashed
unwashedmince
minceand
andsurimi
surimifrom
frommackerel
mackerelsurimi.
surimi.(Magnifica-
(Magnifi-
cation:
tion: 10,000×
10,000 EHT:5.0
× EHT: 5.0kV).
kV).UU==unwashed
unwashedminced.
minced.T1T1== cold
cold carbonated
carbonated water
water washing
washing in the 1st
washing cycle
washing cycle and
and cold
cold tap
tap water
water washing
washing in
in the
the 2nd
2nd and
and 3rd
3rd cycles.
cycles. T2
T2 == cold
cold carbonated
carbonated water
water
washing in the presence of 0.3% NaCl (w/v) in the 1st washing cycle and cold tap water
washing in the presence of 0.3% NaCl (w/v) in the 1st washing cycle and cold tap water washing washing in
the 2nd and 3rd cycles. T3 = cold carbonated water washing in the presence of 0.6% NaCl
in the 2nd and 3rd cycles. T3 = cold carbonated water washing in the presence of 0.6% NaCl (w/v) (w/v) in
the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. C = conventional wash-
in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. C = conventional
ing with 3 cycles of cold tap water.
washing with 3 cycles of cold tap water.

3.4.3. Lipid Oxidation and Fishy Odor

Lipid oxidation of gels from unwashed mince and surimi washed with different media,
as monitored by the TBARS assay, is shown in Figure 5a. Generally, all the surimi gels
showed decreased TBARS levels than unwashed mince gel. This was due to the elimination
of lipid and pro-oxidants like myoglobin and non-heme iron during washing (Figure 2).
 3.4.3. Lipid Oxidation and Fishy Odor
Lipid oxidation of gels from unwashed mince and surimi washed with different me-
dia, as monitored by the TBARS assay, is shown in Figure 5a. Generally, all the surimi gels
showed decreased TBARS levels than unwashed mince gel. This was due to the elimina-
Resources 2023, 12, 126 13 of 16
tion of lipid and pro-oxidants like myoglobin and non-heme iron during washing (Figure
2). Among the surimi, conventional surimi gel had the highest TBARS content, followed
by T1/T2 and T3, respectively. The lowest TBARS value in T3 surimi gel was also linked
to Among theresidual
the lowest surimi, lipid
conventional
content assurimi
well asgel had the highest
myoglobin TBARS content,
and non-heme followed
iron contents (Fig-by
T1/T2
ure 2). and T3, respectively. The lowest TBARS value in T3 surimi gel was also linked to the
lowest residual lipid content as well as myoglobin and non-heme iron contents (Figure 2).

(a)
0.8
TBARS (mg MDA equivalent/kg)
0.7 a

0.6 b
c c
0.5 d

0.4

0.3

0.2

0.1

0
U T1 T2 T3 C
Treatments

(b)
4 a

b b b
Fishy odor score*

3 b

0
U T1 T2 T3 C
Treatments
Figure 5. Effect of cold carbonated water and NaCl washing on lipid oxidation measured as TBARS
Figure
assay5.(a)
Effect
andoffishy
coldodor
carbonated water
score (b) and NaCl
of mackerel washing
surimi gel on lipid oxidation
in comparison measured
with gels from asunwashed
TBARS
assay (a) and conventional
mince fishy odor score (b) ofBars
surimi. mackerel
indicatesurimi gel in
standard comparison
deviation fromwith gels from unwashed
3 determinations except for
mince
fishyand conventional
odor score (n = surimi. Bars indicate
10). Different letters standard deviation
on the bars from
represent 3 determinations
significant except
differences (p < for
0.05).
fishy odor score (n = 10). Different letters on the bars represent significant differences
U = unwashed minced. T1 = cold carbonated water washing in the 1st washing cycle and cold (p < 0.05). U =tap
unwashed minced.
water washing the=2nd
inT1 coldand
carbonated water
3rd cycles. T2 =washing in the 1stwater
cold carbonated washing cyclein
washing and
thecold tap water
presence of 0.3%
washing in the 2nd and 3rd cycles. T2 = cold carbonated water washing in the presence of 0.3% NaCl
NaCl (w/v) in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. T3 = cold
(w/v) in the 1st washing cycle and cold tap water washing in the 2nd and 3rd cycles. T3 = cold car-
carbonated water washing in the presence of 0.6% NaCl (w/v) in the 1st washing cycle and cold tap
bonated water washing in the presence of 0.6% NaCl (w/v) in the 1st washing cycle and cold tap
water washing in the 2nd and 3rd cycles. C = conventional washing with 3 cycles of cold tap water. A
score of 4 represented ‘very strong’ while 0 represented ‘none’.

The TBARS assay is a measure of aldehydes that can be generated and vaporized
during heating, therefore altering the TBARS value [7]. In the current investigation, the
TBARS content in all surimi gel was within the allowable threshold (TBARS < 1.5 mg
MDA/kg) [52]. Surimi gel, particularly when prepared with T3, seemed to be preserved
against lipid peroxidation, as evidenced by the lowest TBARS level (0.47 mg MDA equiv-
Resources 2023, 12, 126 14 of 16

alent/kg). Overall, T3 washing was found to successfully postpone the formation of

lipid oxidation.
Surimi received a lower fishy odor score (2.5–2.8 out of 4) from all panelists than
unwashed mince, with no significant variations between treatments (p > 0.05). Unwashed
mince received the highest fishy odor score, whereas surimi was rated the lowest due
to the removal of volatile molecules, pigments, and other odorous substances in fish
muscle, which was adjusted by washing solution [27]. Some compounds may be identified
postmortem endogenously, while others may be created during the handling and washing
stages. The washing media utilized in this investigation had no effect on the fishy odor of
the resultant surimi.

4. Conclusions
Washing with CW containing 0.6% NaCl in the first washing cycle, followed by two
cycles of cold water washing, can be a simple technique to increase the gelling capability and
oxidative retention of mackerel surimi. This medium effectively removes lipid, myoglobin,
non-heme iron, and TCA-soluble peptide contents, which reduces lipid oxidation while
also enhancing gel-forming ability in dark-fleshed fish, owing to the induction of optimal
unfolding as reported by some biochemical features such as reactive SH content, Ca2+ -
ATPase activity, and hydrophobicity. The outcomes of this study will help to establish
the practicality of tropical mackerel, a dark-fleshed fish species, as a resource for surimi
production, which will establish the sustainability of the surimi industry. In the future,
upscale production of mackerel surimi using the proposed solution can be performed to
advance its potential application in industry. It is also possible to investigate the impact of
frozen storage on the gel-forming ability and oxidative stability of the produced surimi.

Author Contributions: Conceptualization, W.P. and M.C.; methodology, W.P., H.K.Ç. and M.C.;
software, W.P. and M.C.; validation, W.P., H.K.Ç. and M.C.; investigation, P.T., S.B., W.P. and M.C.;
resources, W.P. and M.C.; data curation, P.T., S.B., W.P. and M.C.; writing—original draft preparation,
W.P. and M.C.; writing—review and editing, W.P., M.C. and H.K.Ç.; funding acquisition, W.P. and
M.C. All authors have read and agreed to the published version of the manuscript.
Funding: This project was funded by the National Research Council of Thailand (NRCT) (NRCT5-
RSA63019-01).
Institutional Review Board Statement: The study was conducted according to the guidelines of the
Declaration of Helsinki and approved by the Ethics Committee of Walailak University (WUEC-21-
125-02; date of approval 8 June 2022).
Informed Consent Statement: Informed consent was obtained from all subjects involved in the
study.
Data Availability Statement: Data are contained within the article.
Acknowledgments: The authors thank the Food Technology and Innovation Research Center of
Excellence, Walailak University, for facility support.
Conflicts of Interest: The authors declare no conflict of interest.

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