Effects of different thawing methods on

physicochemical properties and structure of

largemouth bass (Micropterus salmoides)
Luyun Cai , Jiangli Wan, Xiuxia Li, and Jianrong Li

Abstract: To evaluate the physicochemical properties and structure of thawed fillets, following six treatments were
used: conventional thawing, microwave thawing, microwave or ultrasound combined with vacuum thawing, magnetic
nanoparticles combined with microwave or far-infrared thawing. The thawing loss, cooking loss, pH, color, texture
change, water-holding capacity, and water migration of fish fillets were determined. The total volatile base nitrogen

Integrated Food Science

and thiobarbituric acid were used to determine the degree of protein degradation and lipid oxidation. Microscope
observations and scanning electron microscope (SEM) were used to observe fiber microstructure. Results indicated that
both microwave combined with vacuum thawing and far-infrared combined with magnetic nanoparticles thawing had
desirable physicochemical properties as compared to other thawing methods. The lower total volatile base nitrogen and
thiobarbituric acid values had less effect on protein degradation and lipid oxidation of thawing process. Besides, both
microwaves combined with vacuum thawing and far-infrared combined with magnetic nanoparticles thawing samples had
no significant difference in immobilized and bond water compared with fresh sample. SEM and microscopic observation
showed that the myofibril bundles were arranged regularly and smoothly in microwave combined with vacuum thawing
and far-infrared combined with magnetic nanoparticles thawing samples compared with other thawed methods. Thus, the
microwave combined with vacuum thawing and far-infrared combined with magnetic nanoparticles thawing as potential
thawing methods could be used to maintain the quality of thawed fish fillets.

Keywords: far-infrared thawing, largemouth bass, magnetic nanoparticles thawing, microwave thawing, vacuum thawing

Practical Application: Largemouth bass (Micropterus salmoides) is a good source of animal protein. The fish needs to be
frozen for circulation because of geographical and seasonal factors, so thawing methods can directly affect the quality of
thawed fish. The results showed that the microwave combined with vacuum and the magnetic nanoparticles combined
with far-infrared thawing could better maintain the quality of thawed fish.

1. INTRODUCTION
Aquatic products are becoming vital dietary component in
humans’ daily life (Zhou, Ding, & Wang, 2012) due to its
high protein and low fat, especially rich in polyunsaturated fatty
acids (omega-3 and omega-6) that are good for cardiovascu-
lar health such as eicosapentaenoic (EPA, 20:5n-3) and docosa-
hexaenoic (DHA, 22:6n-3) groups (Nieva-Echevarrı́a, Manzanos,
Goicoechea, & Guillén, 2017). The circulation and distribution
of aquatic products depend on freezing and thawing techniques.
Fresh fish can be stored for its nutritional value through freez-
ing, which can extend the shelf life to avoid shortage of supply
caused by seasonal changes. Consequently, the frozen fish should
be thawed before further processing. There are different thawing
methods in foods reported, such as air thawing, water thawing,
JFDS-2019-1448 Submitted 9/5/2019, Accepted 12/3/2019. Author Cai is with
Ningbo Research Inst., Zhejiang Univ., Ningbo 315100, China. and College of
Biosystems Engineering and Food Science, Natl. & Local Joint Engineering Labo-
ratory of Intelligent Food Technology and Equipment, Zhejiang Univ., Hangzhou
310058, China Authors Wan, X. Li, and J. Li are with Natl. & Local Joint En-
gineering Research Center of Storage, Processing, and Safety Control Technology for
Fresh Agricultural and Aquatic Products, College of Food Science and Engineering,
Bohai Univ., Jinzhou 121013, China Direct inquiries to author Cai and Li (E-mail:
lycai515@163.com; li34008@126.com).
refrigerator thawing (Zhang, Li, Diao, Kong, & Xia, 2017), mi-
crowave thawing (Phinney, Frelka, Wickramasinghe, & Heldman,
2017), ultrasonic thawing (Li, Sun, Ma, Cai, & Li, 2019), high-
voltage electric field thawing (Mousakhani-Ganjeh, Hamdami,
& Soltanizadeh, 2015), high-pressure thawing (Backi, 2018), and
so on. However, each thawing method has its own disadvantages,
which can lead to drip loss, nutritional value loss, lipid and protein
oxidation, and microbial reproduction (Duygu & Ümit, 2015).
Therefore, it is necessary to explore a better thawing method that
is beneficial for maintaining quality of frozen fish.
Compared with other aquacultured fishes, largemouth bass (Mi-
cropterus salmoides) has higher economic value and delicious taste.
Moreover, largemouth bass meat has about 5% lipid content and
more than 50% of polyunsaturated fatty acid (Neff et al., 2014),
so the fish is oxidation-sensitive during storage and processing.
Therefore, the frozen largemouth bass fillets need better thawing
methods to maintain their original quality.
Microwave thawing (MT) has many advantages, such as short
thawing time, high efficiency, and simple controls (Virtanen,
Goedeken, & Tong, 1997). It is widely used in industry and
household processes. However, it has also unexpected appearance
in heating includes overheating and low thermal conversion due
to nonuniform temperature distribution during thawed processing
(Vadivambal & Jayas, 2010). MT combined with refrigerated air
could prevent overheating (Campañone & Zaritzky, 2010). In

C 2020 Institute of Food Technologists

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doi: 10.1111/1750-3841.15029 Vol. 0, Iss. 0, 2020 r Journal of Food Science 1

Further reproduction without permission is prohibited
 Effects of different thawing methods . . .
comparison with MT, the ultrasound thawing is more uniform
because the ultrasound absorption rate is much higher in the
frozen area than in the thawed area (Cai, Zhang, Cao, Cao, &
Li, 2019). There was higher water-holding capacity in tuna fish
fillets thawed in ultrasonic at 280 W than control (0 W), leading
to the lowest surface hydrophobicity (S0 -ANS) and improved the
microstructure of thawed fish fillets (Li et al., 2019). Far-infrared
is universally used in heating food because of its lower cost and
higher quality of product compared with traditional thawing (Sakai
& Hanzawa, 1994). Hong, Shim, Choi, and Min (2008) thawed
pork by combining air blast thawing with infrared radiation and
found that there was minor effect on physical properties of pork.
Magnetic nanoparticles (MNP) had super paramagnetism, which
Integrated Food Science
was used to generate magnetothermal effect. Under the action of
external magnetic field, it can be magnetized quickly, and there
was no residual magnetism after removing the external magnetic
field (Ramı́rez, 2015). There is also a change in paramagnetic
state after being heated, which accelerates the conduction of
heat caused by the vigorous movement of polar molecules in the
thawed fish fillets. Because of the heat generated by frozen fish, the
temperature could increase rapidly. Some researchers reported that
there is a potential positive effect on the thawed processing, such as
thawing rate and temperature distribution by the MNP combined
with microwave or radio frequency (Etheridge et al., 2014).
To improve microwave or ultrasound thawing, there are many
methods, for example increasing the thawing time in microwave
or the frequency of ultrasound. However, these methods would
lead to higher cost and inconvenience. Vacuum thawing requires
lower temperature compared with other thawing methods, which
can reduce energy consumption and has less effect on oxidation
reaction.
The purpose of this study was to evaluate the effects of dif-
ferent thawing methods (vacuum combined with microwave or
ultrasound thawing, the MNP combined with microwave or far-
infrared thawing) on the physicochemical properties, and structure
of largemouth bass fillets after thawing.
2. MATERIALS AND METHODS
2.1 Materials
MNPs were purchased from Shanghai Aladdin Biochemical
Technology Co., Ltd. (Shanghai, China). All other chemicals
were obtained from Sinopsin Group Chemical Reagent Co., Ltd.
(Shanghai, China) and were at least of analytical grade. Deionized
water was used in all experimental work.
2.2 Sample preparation
Live largemouth basses weighing 700 ± 30 g with 30 ± 5 cm
length were directly purchased from local aquaculture base in
Jinzhou, Liaoning Province, China. The fishes were transported to
the laboratory of School of Food Science and Engineering within
0.5 hr under room temperature. The percussive stunning method
(according to the method of Sun et al., 2019) was used to kill fishes,
then their skin was removed. Two back fillets were obtained and
cut into approximately 20 g weight (3 × 3 × 2 cm3 ). Each fillet was 2.4 Determination of physicochemical properties
individually packed in a polythene bag to prevent moisture loss and
frozen in storage at −20 °C for 48 hr assure completed frozen. The the weight differential of the frozen fillet before and after thawing
fillets were randomly divided into seven groups and thawed by dif- and expressed as:
ferent treatments as follows: fresh sample (FS), conventional thaw-
ing (CT), microwave thawing, vacuum combined with microwave
thawing (MVT) or ultrasonic thawing (UVT), the MNP com-
2 Journal of Food Science r Vol. 0, Iss. 0, 2020
bined with microwave thawing (MMT) or far-infrared thawing
(FMT).
2.3 Thawing methods
2.3.1 Conventional thawing. One frozen fillet was put
into a refrigerator at 4 °C, the core temperature was recorded
by a thermocouple probe (Testo 905-T1, Germany detu company
Co., Germany). The thawing process was terminated when the
temperature reached to 0 °C.
2.3.2 Microwave thawing. The microwave oven (NN-
DF392B, Panasonic, Osaka, Japan) was used to thaw the
frozen fillets that were put in a beaker (250 mL). The thaw-
ing process included the following conditions: a frequency of
2,450 MHz, and a power of 300 W. The temperature detec-
tion and process termination were carried out in accordance with
Section 2.3.1.
2.3.3 Vacuum combined with microwave thawing.
One frozen fillet was placed on a microwave tray, then the tray was
placed in a microwave vacuum drying oven (ORW08S-3Z, Nan-
jing Aorun Microwave Technology Co., Ltd., Nanjing, Jiangsu,
China). The process was performed with the following condi-
tions: power, 300 W; frequency, 2,450 MHz; temperature, 10 °C
(in order to prevent moisture loss according to the manufacturer).
The core temperature was recorded and thawing process was ter-
minated as explained in Section 2.3.1.
2.3.4 Vacuum combined with ultrasonic. A vacuum
flask was placed in the ultrasonic cleaning machine (KQ-400KDE,
Kunshan Ultrasonic Instrument Co., Ltd, Jiangsu, China) and
about two-thirds of the vacuum flask was immersed in the exter-
nal water. The thawing process was implemented as the following
conditions: power, 200 W; frequency, 40 kHz. The water tem-
perature was controlled at 10 °C by adding ice. The core temper-
ature of fillet and water temperature were recorded by a tempera-
ture sensor. The thawing process was terminated as explained in
Section 2.3.1.
2.3.5 The MNP combined with microwave thawing.
One frozen fillet was sealed in a polyethylene bag and put into
a 250 mL beaker containing 0.1 mg/mL MNP solution (Fe3 04 ,
10 to 30 nm). The fillet was fully immersed in the solution,
and the beaker was placed in the microwave oven. The thawing
conditions were as follows: frequency, 2,450 MHz; power, 300 W.
The temperature of the fillet was recorded and the thawing process
was terminated as explained in Section 2.3.1.
2.3.6 The MNP combined with far-infrared thawing.
Two infrared tubes (4 to 14 µm, 12 × 300 mm, 300 W, Yuanxiang
Electric Appliance Co., Ltd, Lianyungang, China) were placed
on the two sides of the tomographic ark (CX-1, 250 L, Jiapeng
Technology Co., Ltd, Shanghai, China). The frozen fillet was
sealed in a polyethylene bag and fully immersed in a 250 mL
beaker that included MNP solution (0.1 mg/mL, Fe3 O4 , 10 to
30 nm), and the beaker was placed on the tomographic ark. The
temperature was set at 10 °C to prevent moisture loss. The core
temperature was recorded and the thawing process was terminated
as explained in Section 2.3.1.
2.4.1 Thawing loss. The thawing loss was determined by
m1 − m2
Thawing loss (%) = × 100% (1)
m1
Effects of different thawing methods . . .

where m1 is the weight of the fillet before thawing and m2 is the
weight of the fillet after thawing.
2.4.2 Cooking loss. The cooking loss was related to the
weight of the thawed fillet before and after cooking. The frozen
fillet was put into retort pouch and heated in an 85 °C water bath
for 10 min until the core temperature of fillet was at 75 °C (Jia,
Sha, Meng, & Liu, 2019). The cooking loss was calculated with
the following equation:
m3 − m4
Cooking loss (%) = × 100%
m3
where m3 is the weight of the fillet before cooking and m4 is the
weight of the fillet after cooking.
were weighed, and were then put into 750 mL digestive tube. To
determine TVB-N, the procedure was chosen and the unit was
mg N/100 g. Each experiment was performed three times.
2.7 Thiobarbituric acid
The thiobarbituric acid (TBA) measurement procedure was per-
formed as reported by Latip, Zzaman, and Yang (2018) with some
modifications. Ten grams of minced fish meat was taken into the
beaker, 25 mL of deionized water and 10% of trichloroacetic acid
solution (TCA) were added, and homogenized for 1 min. After
(2)
letting it stand at room temperature for 30 min, 5 mL of the
supernatant was added into a colorimetric tube (25 mL, 20 mm
diameter) that included 5 mL of 0.02 mol/L TBA solution. The
miscible liquids were heated at a constant temperature (80 °C) in

Integrated Food Science

2.4.3 pH. After thawing, 5 g of minced fish meat was taken water bath for 40 min. After cooling to room temperature, the UV
into the beaker, 45 mL of deionized water was added, ho- spectrophotometer was used to get the absorbance of the sample
mogenated for 1 min, and let it stand for 30 min and filtered. at 532 nm. The TBA value was expressed as milligram of malon-
The pH value was determined by the filtrate. aldehyde (MDA) equivalents per kilogram of samples (mg MDA
2.4.4 Color measurement. The chroma meter (CR-400, equivalents/kg).
Konica Minolta, Tokyo, Japan) was used to determine the color
of thawed fillet. The color meter system presents the values of 2.8 Microstructure
three different color components (L∗ , a∗ , and b∗ ), lightness (L∗ ),
2.8.1 Microscopic observation. The microscopic obser-
redness/greenness (a∗ ), blueness/yellowness (b∗ ). Color measure-
vation was measured using a microscope (Nikon80i, Nikon Ltd,
ments were stochastically carried out at three different locations
Japan) according to the method of Li et al. (2019) with minor
of each thawed fillet and were averaged. The equipment was cal-
modifications. Fish fillets were cut into 10 × 10 × 10 mm3 along
ibrated using a white standard plate before measurements (Feng
the fibers and stored in a refrigerator at −20 °C for 15 min. Then,
et al., 2018).
the samples were cut into 10-µm thick slices using a microtome
2.4.5 Texture. A TA-XT plus texture analyzer (Stable Micro
(CM-1850, Leica, Germany) and stained with hematoxylin–eosin
Systems Ltd, Godalming, UK) was used to measure the texture of
(HE).
fillet according to the method of Wachirasiri, Wanlapa, Uttapap,
2.8.2 Scanning electron microscope. A scanning electron
Puttanlek, and Rungsardthong (2019) with slight modifications.
microscope (SEM) (S-4800, Hitachi, Tokyo, Japan) was used to
Largemouth bass thawed fillets were cut into 2 × 2 × 2 cm3
observe the fillets structure. SEM measurements were done ac-
cubes and weighed about 7.5 ± 0.05 g. Texture profile analysis
cording to the method of Oujifard, Benjakul, Ahmad, and Sey-
(TPA) mode of a texture analyzer and (P/50) stainless steel probe
fabadi (2012) with slight modifications. Largemouth bass fillets
were used to analyze the texture properties of the samples. The
were cut into small rectangular sections (5 × 5 × 5 mm3 ) fol-
fillet samples were tested in two cycles, the test conditions were
lowing the fibers, immersed in 2.5% glutaraldehyde solution at
as follows: the pre-test, test, and post-test speeds were 1.0 mm/s,
4 °C to fix for 24 hr, and then rinsed with 0.2 mol/L phosphate
respectively. The shape variable was 30%, the induction force was
buffer (pH 7.0) for 10 min. To prevent fixation residual, the sample
5.0 g, and the time between the two compressions was 5 s. All
was washed with deionized water, and then dehydrated in ethanol
samples were done in triplicates and averaged before statistical
gradient with the concentrations of 50%, 60%, 70%, 80%, 90%,
analysis.
and 100% for 10 min per solution. After that, the samples were
freeze-dried using a freeze dryer (FreeZone 2.5 L, Labconco, MI,
2.5 Low-field nuclear magnetic resonance (LF-NMR) USA) for 24 hr. Finally, dried slices were coated with 10 nm of
A LF-NMR analyzer (NMI20-060H, Suzhou Niumag Ana- gold for observation.
lytical Instrument Co., Suzhou, China), which have 60 mm coil
diameter and 0.5 T permanent magnet corresponding to a proton 2.9 Data analysis
resonance frequency of 21.16 MHz at 32 °C, was used to measure All experiments were performed in triplicates. All data were
the water of thawed fillets. The samples were cut into 1 × 1 × expressed as mean values and standard deviations (SDs) (SPSS 22.0,
1.5 cm (2.0 ± 0.05 g), and put in nuclear magnetic resonance tubes Chicago, IL, USA), which were compared using one-way analysis
(15 × 20 mm). The LF-NMR measurement processing was per- of variance (ANOVA) following a Duncan’s multiple range test.
formed as the method reported by Zhang et al. (2017) with some A P value of less than 0.05 was considered significant. All figures
modifications. Transverse (T2 ) relaxation time was measured by were obtained by OriginPro 9.0 (OriginLab Co., Northampton,
the Carr–Purcell–Meiboom–Gill pulse sequence (CPMG). The MA, USA).
T2 was obtained using the following conditions: a delay time of
150 µs between the 90° and 180° pulse (τ value), the repetition
3. RESULTS AND DISCUSSION
time was 3.5 s, and 8,000 echoes were acquired from eight scan
repetitions. Five replicates were performed for each sample. 3.1 Thawing loss, cooking loss, and pH
Table 1 shows the effects of different thawing methods on thaw-
2.6 Total volatile base nitrogen ing loss, cooking loss, and pH of fish. The thawing loss of all groups
Automatic kjeldahl apparatus (Kjeltec 8400, Denmark) was used from low to high were MT, CT, FMT, MVT, UVT, and MMT,
to determine the total volatile base nitrogen (TVB-N) of fillets. respectively. MT had significantly (P < 0.05) lower thawing loss
Ten grams of chopped fish meat and 1 g of magnesium oxide (1.59 ± 0.14%) than other thawing methods except CT, which

Vol. 0, Iss. 0, 2020 r Journal of Food Science 3

 Effects of different thawing methods . . .
Table 1–The thawing loss, cooking loss, and pH of thawed fillets significantly decreased (P < 0.05) compared to FS except the
by different thawing methods. MMT, but the E∗ of them had contrary trend, which was due
Thawing
treatments Thawing loss (%) Cooking loss (%)
FS – 8.19 ± 0.46b
CT 2.19 ± 0.52cd 12.54 ± 0.40a
MT 1.59 ± 0.14d 12.47 ± 0.71a
MVT 2.92 ± 0.03b 13.59 ± 0.53a
UVT 3.89 ± 0.70a 13.29 ± 2.02a
MMT 4.05 ± 0.16a 13.61 ± 0.14a
FMT 2.49 ± 0.13bc 11.97 ± 1.99a
Values represent means ± standard deviation. Different letters in the same column
indicate significant difference (P < 0.05).
Integrated Food Science
may be attributed to the shorter thawing time. There was no sig-
nificant (P > 0.05) difference between UVT and MMT but higher
(P < 0.05) than MVT and FMT, indicating that there was more
water loss during thawing process in UVT and MMT. From the
results of texture (below), those samples that had a higher thawing
loss also had greater hardness and lower elasticity. For MT sam-
ples, which had less thawing loss but had highest hardness and
lower elasticity, the reason was that the protein was denaturated
due to local overheating during MT. As the volume of ice crys-
tals increased during freezing, some mechanical damage would be
caused to the tissues of fish, leading to juice loss after thawing.
Additionally, ice crystals melted into water, which cannot recom-
bine with denatured protein molecules, leading to the decreasing
water-holding capacity of fillet (Xu, Guo, Ding, An, & Wang,
2014).
There were no significant differences (P > 0.05) in cooking
loss among all thawing groups (Table 1), indicating that cook-
ing loss was not affected by the different thawing methods. But
the cooking loss was significantly (P < 0.05) increased in thawed
fillets compared to FS. Xu et al. (2014) and Leygonie, Britz, and
Hoffman (2012) researched that the cooking loss was related to the
proteins aggregation and the amount of water lost during cooking
process. In the freezing and thawing process, the formation and
melting of ice crystals would cause certain damage to the cell struc-
ture and reduce the binding force between cells. Under the action
of external force (heating treatment), the water in the muscle was
easy to flow out, leading to the increase of cooking loss. Besides,
this decrease of binding ability would also have a negative effect
on the texture of thawed fillets (hardness, chewiness, elasticity, and
so on). Actually, in addition to water loss during the cooking pro-
cess, some water-soluble proteins, free amino acids, nucleotides,
and other nutrients flew out with water (Oz & Zikirov, 2015).
These results were similar to Li et al.’s (2019) research.
Table 1 shows that there was an increasing trend from 6.71
to 7.04 in pH. The UVT and MMT samples had no significant
differences with the FS. But the CT, MT, MVT, FMT samples
had significantly higher pH compared to control, indicating the
thawing methods can affect the pH. The increased pH of the
thawed fillet may be due to the breakdown of chemical bonds
involving imidazole, sulfhydryl, and hydroxyl groups (Sánchez,
Gázquez, & Ruiz-Carrascal, 2012).
3.2 Color measurement
The color of the fillet is the intuitive indicator when the
consumer selects, and the color is closely related to the water
content and distribution (Hughes, Oiseth, Purslow, & Warner,
2014). Table 2 shows the effects of different thawing methods
on color of the fillets. The L∗ values of all thawed fillets were
4 Journal of Food Science r Vol. 0, Iss. 0, 2020
to protein aggregation during the thawing process (Hughes et al.,
pH 2014). The a∗ values of thawed fillets were significantly (P < 0.05)
6.71 ± 0.04e
lower than that of FS, and the b∗ values of thawed fillets were
6.92 ± 0.03c significantly higher than that of FS. The changes in fillets color
7.00 ± 0.01b was related to the oxidation of myohemoglobin. The activity of
7.04 ± 0.01a metmyoglobin reductase was inhibited in frozen and then the
6.74 ± 0.00e metmyoglobin, which does not transform into oxymyoglobin,
6.71 ± 0.01e causes an accumulation of metmyoglobin, and affects the color
6.84 ± 0.01d
of fish (Thanonkaew, Benjakul, Visessanguan, & Decker, 2006).
In general, the FMT had little effect on color change of the fillet
and maintained the similar color characteristic to fresh fillet.
3.3 Texture profile analysis
Following are the four representatives of TPA index: hardness,
elasticity, chewiness, and resilience were selected to reflect the
change of fillets quality. The hardness, elasticity, chewiness,
and resilience of thawed fillets all showed a decreasing trend in
Table 3. The thawing process had a great effect on the chewiness
and resilience of fish fillets, but had less effect on the hardness
and elasticity between some thawed fillets and fresh fillets. The
hardness of the MT and MMT samples were similar to the FS, the
reason was that the fillets were overheated in microwave leading
to the oxidation and aggregation of fish protein (Cai et al., 2018).
The texture of fish fillets is mainly related to the fiber state (such as
orderly/disorderly, compact/loose, smooth/uneven, and so on) of
muscle tissue. Protein denaturation and microbial decomposition
in fish lead to the degradation of texture (Hernández et al., 2009).
For elasticity, there were no significant differences (P > 0.05) in
FS, MVT, and FMT samples, but much higher than other samples,
indicating that both MVT and FMT treatments maintained the
original elasticity properties of fillets. MVT treatment which
was in oxygen-free thawing environment can reduce the protein
oxidation of fillets. For FMT treatment, the far-infrared generated
heat that was firstly absorbed by MNP solution, then the whole
frozen fillets were thawed by the MNP that was heated by conduc-
tion effect (Cao et al., 2018). The change trend of chewiness in
all thawed fillets was similar to the hardness. All thawed fillets had
significantly lower (P < 0.05) resilience compared to FS, but there
were no significant differences (P > 0.05) in all thawed treatments.
3.4 Low-field nuclear magnetic resonance
The water mobility and distribution of all samples can be de-
termined by the LF-NMR measurement. Four types of water
distribution are shown in largemouth bass fillets (Figure 2A), 0 ms
< T2b < 1 ms, 1 ms < T21 < 10 ms, 10 ms < T22 < 100 ms,
100 ms < T23 < 1000 ms. T2b and T21 represented the bound
water that was closely bonded with the macromolecules. T22 was
the immobilized water that was located within the myofibrillar
protein network. T23 represented the free water, which outside
in cell space, could be easily lost (Zou, Zhang, Kang, & Zhou,
2018). The total area percentage of the integral area of different
T2 intervals can be expressed by P2 , as shown in Figure 2E.
From Figure 2A, the peak value of immobilized water in CT
sample was the lowest, indicating that the immobilized water con-
tent in CT sample was lowest compared with other treatments.
This result may be due to the more opportunities contributed to
immobilized water transformed into free water during CT thawed
processing, resulting in the decrease of immobilized water peak.
The MVT and FMT treatments all showed more immobilized
water than others, which remained closer to FS. This suggested
Effects of different thawing methods . . .

Table 2–The color of thawed fillets by different thawing methods.

Thawing treatments L∗ a∗ b∗ E∗

FS 44.25 ± 0.10b 0.53 ± 0.11a −3.33 ± 0.53c 50.92 ± 0.09d
CT 40.42 ± 0.18d −0.19 ± 0.01c −1.51 ± 0.10b 54.50 ± 0.17bc
MT 40.00 ± 0.53d −0.22 ± 0.02c −1.94 ± 0.29b 55.11 ± 0.63b
MVT 41.90 ± 0.41c 0.07 ± 0.09b −2.31 ± 0.05b 53.11 ± 0.42c
UVT 37.68 ± 0.93e −0.15 ± 0.10c −2.01 ± 0.36b 57.27 ± 0.91a
MMT 46.18 ± 1.04a 0.11 ± 0.08b 0.19 ± 0.21a 48.61 ± 1.04e
FMT 43.27 ± 1.48bc 0.20 ± 0.03b −0.01 ± 0.92a 51.53 ± 1.52d
Values represent means of three samples ± standard errors. Different letters in the same column indicate significant difference (P < 0.05).

Table 3–The TPA of thawed fillets by different thawing methods.

Integrated Food Science

Thawing treatments Hardness Elasticity Chewiness Resilience
FS 1,966.80 ± 605.28a 0.79 ± 0.01a 1,082.90 ± 328.70a 0.52 ± 0.01a
CT 934.05 ± 305.73b 0.66 ± 0.04c 360.75 ± 107.71c 0.34 ± 0.02bc
MT 2,030.79 ± 503.22a 0.64 ± 0.02c 731.88 ± 194.28b 0.34 ± 0.01bc
MVT 1,042.53 ± 206.54b 0.75 ± 0.08ab 463.64 ± 100.05bc 0.33 ± 0.02c
UVT 1,612.40 ± 370.89ab 0.68 ± 0.02bc 616.83 ± 105.95bc 0.33 ± 0.03bc
MMT 1,889.83 ± 222.95a 0.56 ± 0.03d 584.30 ± 39.42bc 0.37 ± 0.02b
FMT 1,200.10 ± 119.14b 0.74 ± 0.02ab 495.52 ± 68.58bc 0.35 ± 0.02bc
Values represent means of three samples ± standard errors. Different letters in the same column indicate significant difference (P < 0.05).

Figure 1–The experimental sequence diagram

of this experiment.

that there was a minimal damage to water mobility during MVT
and FMT thawing processes.
T2 results are shown in Figure 2B–D. The T21 and T22 of thawed
samples had an increasing trend compared to FS, indicating that
the water mobility was increased. The decreased bound water may
be affected by protein aggregation and conformation changes.
Additionally, changes and losses of specific hydrophilic groups
could also cause fluctuation in bound water during thawing. For
Figure 2D, the T23 of thawed fillets was significantly (P < 0.05)
lower than FS, but no significant difference (P > 0.05) was found
among thawed samples, this was correlated with less water mobility
in thawed samples than FS.
As shown in Figure 2E, there was no significant difference
(P > 0.05) in bound water (P21 ) of all samples, while the im-
mobilized water (P22 ) and free water (P23 ) had more significant
changes in thawed fillets. The P22 in MVT and FMT samples had
no difference (P > 0.05) and slightly lower compared to FS sam-
ple, however higher than other four thawed methods. There was
higher value of P23 in MVT and FMT samples than FS, however
obviously lower compared with other thawed samples. These re-
sults were consistent with SEM, which indicated that there was
little disruption of the fiber structure with MVT and FMT treat-
ment. But the lowest P22 and highest P23 were found in the MT
sample, indicating more immobilized water changed to free water.
This result was due to microwave vibrations that loosen hydro-
gen bonds between water and other macromolecules (Ersoy &
Özeren, 2009). In addition, the ruptured microstructure in my-
ofibrils occurred if using unbefitting thawing methods (Xia, Kong,
Liu, Diao, & Liu, 2012). Generally, the MVT and FMT thawing
treatments can hold the water capacity similar to FS.
3.5 Total volatile base nitrogen
The changes in TVB-N values of largemouth bass fillets
by different thawing methods are shown in Figure 3A. The
TVB-N values significantly (P < 0.05) increased in all thawed
samples compared to FS, and there was no significant difference

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Integrated Food Science Effects of different thawing methods . . .

Figure 2–Effect of different thawing treatments on moisture migration in largemouth bass meat. Distribution of the LF-NMR T2 (A), T21 (B), T22 (C), and
T23 (D) relaxation times of fish meat with different thawing treatments. The percentage of the total area of the integral area of different T2 intervals
is shown in Figure 2E. “a–e” letters indicate significant difference (P < 0.05). Error bars show standard deviation.

(P > 0.05) between the MVT and FMT samples which were
significantly (P < 0.05) lower than other four treatments. The
higher TVB-N value indicated more destruction to methionine
and tyrosine in fish protein, leading to more nutrition loss (Lu,
2009). These results indicated that the slighter protein degradation
and the less nutrition loss occurred in MVT and FMT treatments.
Mousakhani-Ganjeh et al. (2015) had reported that TVB-N would
be reduced significantly by using the HVEF thawing method, and
with increasing electric field strength, more air was ionized (mostly
oxygen) that resulted in the enhancement of antimicrobial prop-
erties (Karaca & Velioglu, 2014). For MVT treatment, the lower
TVB-N value might be due to vacuum environment that could
isolate from oxygen.
3.6 Thiobarbituric acid
The TBA value can reflect the content of malondialdehyde
(MDA), which is a secondary lipid oxidation product that can
be used to judge the degree of oxidative rancidity of fats in fish
fillets. The TBA values in different thawed fish fillets are shown
in Figure 3B. The TBA values of thawed fillets significantly
(P < 0.05) increased compared to FS except for the FMT sample,
indicating the less oxidative rancidity of lipids in FMT treatment.

6 Journal of Food Science r Vol. 0, Iss. 0, 2020

Effects of different thawing methods . . .

Integrated Food Science

Figure 3–(A) Effects of different thawing methods on TVB-N of largemouth
bass. (B) Effects of different thawing methods on TBA of largemouth bass.

This result was similar with SEM result, which suggested that the
microstructure of FMT sample was smoother than other thawed
samples. The highest TBA value was found in UVT sample, which
may be related to the thermal, mechanical action, and cavitation
effect of ultrasonic. The degree of oxidative rancidity of fats is
positively correlated with TBA (Li, Li, Hu, & Li, 2013). The TBA
values would be increased when using inappropriate thawing that
could destroy cell structure and cause cell to release pro-oxidants,
which promotes lipid oxidation (Benjakul & Bauer, 2001).

3.7 Microscopic observation

As shown in Figure 4, the microstructural characteristics of fillets
by different thawing methods can be observed. The well-organized
arrangement of fibers was displayed in FS, these muscle fibers were
surrounded by the sarcolemma membrane along with the connec-
tive tissue (Ozuna, Puig, Garcı́a-Pérez, Mulet, & Cárcel, 2013).
The spaces between the myofibrils in all thawed samples became
wider. The MT treatment had the most destruction on the mi-
crostructure of the fillets compared to other thawed samples. This Figure 4–Effect of different thawing methods on muscular tissue of large-
result may be due to local heating of microwave on MT sample, mouth bass (microscope magnifications × 40, × 100, the length of scale
which could cause muscle fibers to break. The arrangement of mark in each figure is 100 µm).

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Integrated Food Science Effects of different thawing methods . . .

Figure 5–Effect of different thawing methods on the microstructure of largemouth bass muscle (× 1.5 K).

Table 4–The correlation analysis of indicators values.

Thawing Cooking
loss loss pH L∗ a∗ b∗ E∗ Hardness Elasticity Chewiness Resilience TVB-N TBA
Thawing loss 1
Cooking loss 0.469∗ 1
pH −0.067 0.213 1
L∗ −1.06 −0.315 −0.343 1
a∗ −0.322 −0.628∗∗ −0.426 0.730∗∗ 1
b∗ 0.568∗∗ −0.058 −0.126 0.358 −0.35 1
E∗ 0.071 0.308 −0.348 −0.998∗∗ −0.718∗∗ −0.400 1
Hardness −0.223 −0.082 −0.356 0.174 0.159 −0.124 −0.156 1
Elasticity −0.569∗∗ −0.455∗ 0.095 −0.101 0.423 −0.557∗∗ 0.122 −0.232 1
Chewiness −0.582∗∗ −0.358 0.373 0.197 0.448∗ −0.454∗ −0.165 0.839∗∗ 0.233 1
Resilience −0.668∗∗ −0.467∗ −0.523∗ 0.489∗ 0.686∗∗ −0.455∗ −0.456∗ 0.409 0.429 0.745∗∗ 1
TVB-N 0.615∗∗ 0.665∗∗ 0.161 −0.494∗ -0.810∗∗ 0.330 0.472∗ 0.076 −0.705∗∗ −0.390 −0.707∗∗ 1
TBA 0.788∗∗ 0.824∗∗ 0.019 −0.185 −0.470∗ 0.217 0.168 0.131 −0.653∗∗ −0.274 −0.529∗ 0.757∗∗ 1
∗
Correlationis significant at 0.01 level (P < 0.05).
∗∗
Correlation is significant at 0.05 level (P < 0.01).

myofibrils in UVT sample was disordered and the myofibrils were
overlapped, suggesting that the myofibrils of thawed fillet by UVT
were destroyed. Li et al. (2019) also found that many filaments
and irregular networks at the microstructural level were found in
tuna fish by ultrasound thawed at 160 W. This may be due to the
long thawing time and strong cavitation induced by ultrasound.
For MVT, the fibers were hollow and bent compared to FS. The
fibers of CT, MMT, and FMT were straight, but the space of fibers
in MMT was wider, both the sarcolemma membrane of CT and
FMT fibers can be observed. These results indicated the minor
damage for the microstructure of fillets by the methods of CT and
FMT in thawing process.

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Effects of different thawing methods . . .

3.8 Scanning electron microscope
Different muscle fibers of vertical section in fillets by different
thawing methods are shown in Figure 5. The distinct myofib-
rils could be observed in FS, and the muscle fibers were neatly
arranged with no rupture and fracture. For CT sample, the my-
ofibrils could be also seen clearly and had some bends, while some
myofibrils were bent and twined in MMT sample. With the dis-
ruption of fibers, the surface of the muscle was in disorder and
crude in MT and UVT samples, and the myofibrils of which were
cross-linked, and the gap between myofibrils was large. But smooth
and no uneven surface appeared in MVT and FMT samples, both
of these fibers were straight and orderly.
These results of SEM were consistent with the results of mi-
certainty and unknown factors (Amenta et al., 2015). So far, the
toxic properties of magnetic nanometer are not very clear, limiting
its practical application in industrial sectors, the safety of MNPs is
still in further research to explore (Cockburn et al., 2012). There-
fore, considering the safety, economy, efficiency, and consumer
acceptance, MVT is a promising thawing method for commercial
applications.
ACKNOWLEDGMENTS
This study was supported by National Key R&D Program of
China (2018YFD0901106), the National Natural Science Foun-
dation of China (31401478), and the Aquatic Products Process-
ing and Safety Key Laboratory of Guangdong Province (GDP-

Integrated Food Science

croscopic observations. The breakdown of epimysium and en- KLAPPS1805).
domysium or outflow of water can cause myofibrils shrinkage and
fracture. These may be due to excessive dehydration of the mus- AUTHOR CONTRIBUTIONS
cle fibers or uneven thawing, which had a relation to the huge J.L.W. and X.X.L. performed the experiment, J.L.W analyzed
temperature difference between the air and the frozen fillets or the date, J.L.W. and L.Y.C. drafted the manuscript. J.R.L. modified
overheated of microwave and far infrared. At the same time, the the manuscript and L.Y.C. provided the funding.
change of microstructure in thawed fillets could also affect the
water-holding capacity (Bertram, Schäfer, Rosenvold, & Ander- CONFLICT OF INTEREST
sen, 2004). Therefore, the SEM results could verify the results
The authors declare no conflict of interest.
of LF-NMR, those disordered arrangement of fibers had larger
water loss. This indicated that the mechanical restraint of fibers
had a negative correlation with the immobilized water loss (Xia ABBREVIATIONS
et al., 2012). The less damage on muscle fibers in MVT and FMT CT conventional thawing
samples indicated the less water loss. DHA docosahexaenoic
EPA eicosapentaenoic
3.9 Correlation analysis FMT far-infrared combined with magnetic nanoparticles
The correlation analysis between thawing loss, cooking loss, pH, thawing
L∗ , a∗ , b∗ , E∗ , hardness, elasticity, chewiness, resilience, TVB- FS fresh sample
N and TBA values of fillets under different thawing methods HE hematoxylin–eosin
are shown in Table 4. There were good relationship between all LF-NMR low-field nuclear magnetic resonance
indicators with relatively high Pearson’s correlations. The Pearson’s MMT microwave combined with magnetic nanoparticles
∗
correlations between cooking loss, a , and TVB-N, also between thawing
L∗ and E∗ , between hardness and chewiness were all above MNP magnetic nanoparticles
0.80 (P < 0.01). These results indicated that there is a certain MT microwave thawing
positive and negative correlation between these indicators, and MVT microwave combined with vacuum thawing
have a mutual influence, the results of some indicators can be SEM scanning electron microscope
used to evaluate the results of other indicators that have a good S0-ANS surface hydrophobicity
correlation with them. In general, the correlation between these TBA thiobarbituric acid
indicators was more significant at the level of 0.01 than at the level TCA trichloroacetic acid solution
of 0.05. TPA texture profile analysis
TVB-N total volatile base nitrogen
4. CONCLUSIONS UVT ultrasound combined with vacuum thawing
The thawing loss of thawed fillets ranged from small to large was
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