https://doi.org/10.52973/rcfcv-e33242
Received: 06/03/2023 Accepted: 15/04/2023 Published: 14/05/2023
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Revista Científica, FCV-LUZ / Vol. XXXIII, rcfcv-e33242, 1 – 7
ABSTRACT
Four meat thawing techniques that are most commonly used in
daily life were used: refrigerator thawing, microwave thawing,
ambient temperature thawing, and water thawing, to evaluate the
physico-chemical and histological alterations in thawed beef. After
thawing, the structural, chemical, and physical characteristics of
beef meat were evaluated. The results showed that meat thawed
in the refrigerator at 4°C was characterized by the highest pH value
(5.65 ± 0.02) and a signicant difference (P<0.05) compared to meat
thawed by other thawing methods. Also for the electrical conductivity,
it reached the highest value (1.442 ± 1,012) in the microwave oven
(P<0.05); meanwhile, water activity decreased signicantly after
thawing regardless of the thawing method (P<0.05). On the other
hand, refrigerator thawing resulted in the least amount of water loss
(1.23%) with P<0.05, while high levels of microwave energy caused
signicant water loss, represented by thawing loss and cooking
loss (4.37% and 44.47%), respectively, with P<0.05. Among different
thawing methods, microwave thawing had the highest level of TBARS,
with a mean of 0.25 ± 0.034 mg·kg
-1
(P<0.05). Regarding the color, the
lightness (L*) value in the microwave-thawed samples decreased
signicantly (P<0.05) compared to the fresh control. Histologically,
samples that were thawed in a refrigerator preserved the integrity of
the bers' structure after thawing better than other methods; samples
thawed in a microwave, however, caused more structural damage.
To ensure that it thaws uniformly and to retain the meat's quality as
close to its fresh quality as possible, it is typically advised to thaw
meat in a slower, more gradual manner, such as in the refrigerator.
Key words: Freezing; thawing methods; quality; beef; microstructure
RESUMEN
Se emplearon cuatro técnicas de descongelación de carne que son
las más utilizadas en la vida diaria: descongelación en el refrigerador,
descongelación en el microondas, descongelación a temperatura
ambiente y descongelación en agua, para evaluar las alteraciones
físico-químicas e histológicas en la carne de res descongelada. Después
de descongelar, se evaluaron las características estructurales, químicas
y físicas de la carne de res. Los resultados mostraron que la carne
descongelada en el refrigerador a 4°C se caracterizó por el valor
de pH más alto (5,65 ± 0,02) y una diferencia signicativa (P<0,05)
en comparación con la carne descongelada por otros métodos de
descongelación. Además, para la conductividad eléctrica, alcanzó
el valor más alto (1,442 ± 1,012) en el horno de microondas (P<0,05);
mientras tanto, la actividad del agua disminuyó signicativamente
después de la descongelación, independientemente del método
de descongelación (P<0,05). Por otro lado, la descongelación en el
refrigerador resultó en la menor cantidad de pérdida de agua (1,23%)
(P<0,05), mientras que los altos niveles de energía de microondas
causaron una pérdida signicativa de agua, representada por la
pérdida de descongelación y la pérdida de cocción (4,37 y 44,47%),
respectivamente, con P<0,05. Entre los diferentes métodos de
descongelación, la descongelación en el microondas tuvo el nivel
más alto de TBARS, con una media de 0,25 ± 0,034 mg·kg
-1
(P<0,05).
Con respecto al color, el valor de luminosidad (L*) en las muestras
descongeladas en el microondas disminuyó significativamente
(P<0,05) en comparación con el control fresco. Histológicamente,
las muestras que se descongelaron en el refrigerador conservaron
mejor la integridad de la estructura de las fibras después de la
descongelación que en los otros métodos; las muestras descongeladas
en el microondas, sin embargo, causaron más daño estructural.
Para asegurar que se descongelaron uniformemente y se conserva
la calidad de la carne lo más cercana posible a su calidad fresca,
generalmente se recomienda descongelar la carne de manera más
lenta y gradual, como en el refrigerador.
Palabras clave: Congelación; métodos de descongelación; calidad;
carne de vacuno; microestructura
Physicochemical and structural properties of beef meat thawed using
various methods
Propiedades sicoquímicas y estructurales de la carne de vacuno descongelada mediante distintos
métodos
Saliha Lakehal
1
* , Aicha Lakehal
2
, Salima Lakehal
3
, Omar Bennoune
1
and Ammar Ayachi
1
1
University of Batna 1, Department of Veterinary Medicine. Batna, Algeria.
2
University of Batna 2, Faculty of Technical Sciences, Batna, Algeria.
3
University of Batna 2, Institute of Earth Sciences and Universe. Batna, Algeria.
*Corresponding Author: saliha.lakehal@univ-batna.dz
Properties of beef meat thawed using various methods / Lakehal et al. _____________________________________________________________
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INTRODUCTION
Beef is one of the most important sources of protein known and
widely available in the World [26]. As, it is known a highly perishable
product, it is necessary apply a rigorous method to ensure its
nutritional value [32, 36]. Freezing and frozen storage are one of
the most important methods used in the export and import of meat,
and it is widely spread to extend the shelf life and maintain the quality
of meat for as long as possible [13, 18, 20]. It has been used for many
years to prevent the growth and reproduction of microorganisms and
to minimize metabolic activity during long-term storage or distribution
and sale [7, 15]. Thawing is a necessary process prior to consumption
and further processing [2]. The term "thawing" refers to the process of
bringing frozen meat from freezing to a temperature between -5 and
0 degrees Celsius so that it can be cut or sliced for further use [43].
Thawing is a more dicult process to perform safely than freezing;
it is generally slower than freezing [10]. However, as the thaw begins,
a layer of water forms, which slows down the process. However, the
thawing process has received less attention compared to refrigeration
or freezing [21], although this process is a very important and integral
step in frozen foods before further processing or consumption [37].
Many factors, including the thawing process, can affect meat quality
[7]. Several studies have examined the effects of different thawing
methods on changes in meat quality, such as water retention, tenderness,
color, and avor [15, 16, 42]; lipid oxidation [22], and microbial growth [8],
but there is little information on the microstructural effects of different
thawing techniques on histomorphological techniques. Ocial standards
do not provide information on the assessment of the thawing process
and its impact on food.
The ocial journal of the democratic and popular republic of
Algeria, April 16, 2017, Art. 47, stipulates that the temperature of
thawed meat should be within 4°C as a maximum to reduce the risk
of multiplication of microorganisms that cause many diseases and
thus become unt for consumption [28]. Moreover, it is prohibited
to re-freeze thawed foods intended for the consumer. On the other
hand, home users may use various thawing methods that will affect
the quality of beef, including refrigerator thawing, room temperature
thawing, water thawing, and microwave thawing. The aim of this
study was to determine how different thawing techniques affect
the structure of meat and its physical and chemical properties by
Comparing thawed meat to fresh meat allows us to determine how the
freezing and thawing processes impact the meat's quality. Therefore,
by understanding how different thawing techniques affect meat
quality, consumers can choose the best thawing method to minimize
damage and preserve the quality of the meat.
MATERIAL AND METHODS
A fresh portion of biceps femoris was obtained from four beef
carcasses (24 hours –h– post mortem) from a local slaughterhouse
(Batna, Algeria). Fresh beef muscle samples were cut into blocks,
and each sample was snap frozen separately by polyethylene bags
and frozen at -23°C for 2 months in a home freezer (CRF-NT64GF40,
Condor, Algeria). Samples were thawed until the meat medium
temperature reached 2°C. Four methods used for thawing are the
most common in the experiment as follows:
1.
Thawing in the refrigerator (R) (CRF-NT64GF40, Condor,
Algeria) at +4°C;
2. Thawing at room temperature (A) (23°C);
3. Thawing in water immersion (W) (15°C);
4. Microwave (M) (MWM100, Kenwood, 800W, UK).
A minimum of ve blocks were for each processing thawing methods.
Determination of physico-chemical changes pH
According to the procedure of Zhu et al.[42], a sample of 5 grames
(g) minced beef is mixed with 45 mililiters (mL) of distilled water in
order to measure the pH of the mixture. The pH was measured using
an digital pH meter (INOLAB WTW 720, Germany) and the results were
recorded. It is important to ensure that the pH meter is calibrated
correctly and that the sample is thoroughly mixed with the distilled
water before the pH measurement is taken.
Electrical conductivity (Ec)
The measurement procedure described by Jia et al. [12] with
some modications. Beef samples were homogenized and stirred
for 10 minutes –min– after being homogenized with 30 mL distilled
water. After that, a digital EC meter (FE20/EL20; Mettler Toledo, 139
Shanghai, China) was used to measure the combination.
Water activity (Aw)
To measure the Aw of a meat sample using the Hygroscope (BT-RS1
Rotronic, Germany) according to the procedure described by Lakehal
et al. [17], the sample should be chopped into small pieces and placed
in a sample cup with a volume of three quarts. The probe of the
device should be inserted into the sample cup and the humidity and
temperature data should be allowed to stabilize. Once the data has
stabilized, the result can be read from the device.
Thawing loss
The method for determining the thawing loss was based on Xia et
al.'s approach [38]. Initially, the samples were weighed with Scale (Kern
EW 620-3NM, Germany) before freezing, and subsequently, the frozen
samples were thawed with different thawing methods. After thawing, the
samples were dried with paper towels and weighed again immediately.
To calculate the thawing loss, the following equation was used:
Thawingloss(%)
initialweightofraw material
initialweightofraw material weightafter thawing
100
=
-
#
^h
<
F
Cookingloss(%)
thawedmeatweight
thawed meat weight meat sample weight aftercooking
100
=
-
#
^h
<
F
Cook loss
The methods given by Choi et al. [8] was used to calculate cooking
loss. The thawed sample (50 g) was placed in a polyethylene bag and
cooked for 25 min at 80°C in a water bath (GFL 1052, Germany) until it
reached 75°C. Cooking loss was determined using the following equation:
Color
In the method developed by Minz and Saini [24], color was quantied
using a computer vision system (CVS) that measures three color
parameters: lightness (L*), redness (a*), and yellowness (b*). The
experimental setup involved using a Canon DS126621 digital camera
placed vertically at a distance of 30 centimeters (cm) from the sample.
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To illuminate the CVS, two lamps with standard illumination (6500K)
were employed. These lamps, measuring 60 cm in length, were
positioned at a 45° angle above the samples. A cubical wooden box
was used to house both the lamps and the camera, with the internal
walls of the box coated in black opaque photographic cloth to reduce
background light. Adobe Photoshop CS3 software was used for
measuring and analyzing color values in images.
Lipid oxidation
Thiobarbituric acid-reactive compounds (TBARS) were used to
assess lipid oxidation, according to a protocol described by Buege
and Aust [6]. For 1 min, 5 g of minced beef was combined with 50 mL
of distilled water. Two mL of a 20 millimolar (mM) 2-thiobarbituric
acid/15 percent trichloroacetic acid/chlorhydric acid (TBA/TCA/HCl)
solution were mixed with one mL of the sample solution. Afterwards,
sample solutions were transferred to a water bath at 90°C for 20
min. The resultant solutions were chilled for 10 min under running
water. The absorbance of the resultant top layer was measured at 532
nanometers (nm) by a UV-vis spectrophotometer (Shimadzu, Japan).
The concentrations of TBARS were represented as nanomolars (nmol)
of malondialdehyde per g of beef using a molar extinction value of
1.56×10
5
·M
-1
·cm
-1
.
Microstructure observation under light microscope
Histological analysis was performed to investigate possible changes
in beef microstructure during different thawing methods compared
to fresh samples. For histological examination, specimens were
prepared according to Lakehal et al. [17], with some modications.
The samples were xed in 10% formalin for 48 h and then dehydrated
with graduated ethanol for 10 h. After dehydration, the samples were
claried by soaking them in xylene for 45 min, twice. Then, after
embedding the samples in a paran bath at 58°C for 8 h, each sample
was embedded in a block of paran and sectioned transversely to the
muscle ber over a thickness of 6 micromolars –μm– on a microtome
(Leica Jung-histocut 820, Germany) into thin slices. Glass slides were
used to mount the selected sections and for staining, it was used
hematoxylin and eosin by soaking for 2 min in each staining solution.
After marking out the intracellular ice crystals (white voids), it was
calculated the location of each ice crystal within the cell and the
number and average area of each intracellular ice crystal.
Statistical analysis
The current study's results were statistically evaluated using the
SPSS software version 20 (IBM SPSS Statistics v22). Analysis of
variance (one-way ANOVA) techniques, as well as Tukey multiple
comparison tests were employed to examine differences for the data
acquired in the experimental study. The information was presented
as a mean value with a standard deviation.
RESULTS AND DISCUSSION
pH
The pH is a determining factor for the organoleptic characteristics
of the meat [25]. Normal pH levels in living muscle are around 7.4.
After slaughter, the pH of the meat decreased from 5.6 to 5.7 within
6 to 8 h [30]. In this study (FIG. 1A), the pH of beef meat ranged from
5.53 to 6.65, indicating that freezing and thawing had an effect on
pH. By comparison between different thawing methods, meat thawed
in the refrigerator at 4°C was characterized by the highest pH value
(5.65 ± 0.02) with a signicant difference (P<0.05), while meat thawed
by other thawing methods showed no signicant difference. Meat
protein denaturation can be attributed to higher pH values of frozen or
thawed meat than fresh control meat [23]. According to Ho et al. [11],
accumulation of free amino acids, ammonia and organic sulphides
derived from the hydrolysis of proteolytic amines might be considered
to be the primary cause of the elevated pH. However, other studies
have not reported any change in pH after thawing [19, 41, 43].
Water activity (Aw)
The Aw of a food is an important factor in determining its shelf life
and the risk of microbial contamination [40]. Foods with low Aw are less
likely to support the growth of microorganisms, while those with high
water activity are more susceptible to spoilage and foodborne illness
[4]. The Aw results of different meat samples depending on the thawing
method are shown in FIG. 1B. In fresh meat samples, the average Aw
was 0.944. After freezing/thawing, Aw decreased signicantly in all
frozen meat samples. However, Oliveira et al. [29] found no differences
in Aw in chicken (Gallus gallus domesticus) breast meat thawed using
different thawing methods. Medić et al. [22] found that variations in
Aw are related to uid migration and ice crystallization.
Electrical conductivity (Ec)
FIGURE 1C depicts the Ec of frozen beef thawed using various
methods. The value of Ec at the beginning of the experiment was
1,341 second·cm
-1
(s·cm
-1
). There was an upward trend after thawing.
The Ec of refrigerator thawing (R), immersion thawing (W), room
temperature thawing (A), and microwave thawing (M) increased
signicantly (P<0.05) to 1,436, 1,394, 1,370, and 1,442, respectively.
The reason for this may be that when the membrane permeability
of muscle bers increases, there is a greater inux of ions, such as
sodium and chloride, into the extracellular space. This leads to an
increase in the concentration of ions in the extracellular space, which
increases the Ec of the tissue. At the same time, the increase in the
uid losses after thawing of muscle bers can lead to an increase in
extracellular volume, further contributing to the increase in Ec [10].
Water losses
It is widely acknowledged that freezing and thawing have a negative
impact on the water retention capacity, often assessed as loss from
thawing and loss from cooking [40]. FIGURE 1D shows the water loss
results. High levels of microwave energy can cause signicant water
loss as represented by thawing loss and cooking loss 4.37 and 44.47%,
respectively (P<0.05), which can negatively affect humidity levels [14].
Excessive heat can cause this, altering muscle protein structure and
causing protein denaturation, resulting in a high amount of thawing
loss, while refrigerator thawing results in the least amount of water
loss (1.23%) with (P<0.05).
According to FIG. 1D, the cooking losses of the samples showed
similar trends which comparable to those in the samples' thawing
losses. Meats frozen/thawed by different methods are characterized
by higher cooking losses than fresh meat, in agreement with Xia et al.
[39]. Thawing methods had a signicant inuence on cooking losses,
which tended to be higher in microwave-thawed meat (44.47%). In
general, the highest cooking loss values of thawed meat could be
related to the aforementioned tissue damage due to the formation
of ice crystals during the freezing process.
FIGURE 1. Impact of thawing methods on pH (A), water activity (B),electrical, Electrical conductivity (C), cooking loss and thawing loss(D). F: Fresh
meat, R: refrigerator thawing (4°C), A: ambient temperature thawing (23°C), W: water immersion thawing (15°C), M: microwave thawing. (a, b, c)
differ signicantly (P<0.05)
FIGURE 2. Impact of thawing methods on color of beef meat. F: Fresh
meat, R: refrigerator thawing (4°C), A: ambient temperature thawing
(23°C), W: water immersion thawing (15°C), M: microwave thawing.
(a, b) differ signicantly (P<0.05)
Properties of beef meat thawed using various methods / Lakehal et al. _____________________________________________________________
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It should be noted that the volume lost by cooking is generally
made up of a mixture of liquid and soluble matter coming from the
muscle during cooking as reported by Zhang et al. [40]. For that
reason, differences in meat fat and protein content may be partly
responsible for the amount of cooking loss [1].
Because the consumer prefers to use color as an indicator of
freshness and health, color is a major evaluation factor affecting
the appearance, presentation, and acceptance of many foods,
particularly meat [3, 41]. Several studies have recorded a greater
proportion of metmyoglobin and less redness in thawed red meat
then fresh state [19, 27, 31, 40]. FIGURE 2 shows the inuence of
various thawing procedures on the color characteristics of beef
samples. When compared to fresh control, the lightness (L*) value in
the microwave-thawed samples reduced signicantly (P<0.05), which
was consistent with the results of Zhang et al. [41]. The (L*) values for
the thawing in a refrigerator, in Water immersion and thawed room
temperature did not differ from that of fresh meat. The thawing
at the room temperature sample had the lowest redness (a*value)
(P<0.05). Furthermore, there is signicant variation in the (b*) value
was detected in the microwave thawing and water immersion thawing
methods (P<0.05) compared to fresh control, which was different with
the results of Kim et al. [13] and Leygonie et al. [19]. Protein oxidation
and pigment degradation are major elements that contribute to the
color stability of meat during the freezing and thawing processes,
according to several researchers [19].
TBARS, or thiobarbituric acid reactive substances, is a measure
of lipid oxidation in food. It is typically measured in milligrams per
kilogram of sample (mg·kg
-1
). The higher the TBARS value, the more
oxidized the lipids in the sample are [25].
Lipid oxidation
The results presented in the FIG. 3 suggest that thawing beef in a
refrigerator or at room temperature does not signicantly affect the
TBARS value compared to fresh beef (P>0.05), while thawing using
water immersion or microwave methods signicantly increases the
TBARS value (P<0.05). This suggests that these methods of thawing
may contribute to lipid oxidation in the beef. One possible explanation
for this is that the high temperatures generated in microwave thawing
may release more oxidative enzymes and pro-oxidants from ruptured
cellular organelles, leading to increased lipid oxidation. Similarly,
the electromagnetic heating during microwave thawing may also
contribute to lipid oxidation [9, 39].
FIGURE 3. Impact of thawing methods on TBARS of beef meat. F: Fresh
meat, R: refrigerator thawing (4°C), A: ambient temperature thawing
(23°C), W, water immersion thawing (15°C), M: microwave thawing.
(a, b) differ signicantly (P<0.05)
(A)
(B) (C)
(E)(D)
FIGURE 4. Impact of thawing methods on microstructure of beef meat:
(A) fresh meat, (B) thawing in refrigerator, (C) thawing in water, (D)
thawing in air temperature and (E) thawing in microwave
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Morphological differences under microscopic observation
The results of histological examinations on fresh and thawed beef
muscle tissues following different thawing treatments are depicted in
FIG. 4. The analysis revealed varying degrees of gaps between muscle
bers in all thawing methods, possibly due to mechanical damage
sustained during the process. Unfrozen beef exhibited uniformly
distributed and regularly-formed bers. Notably, the refrigerator
(R) thawing method had the least detrimental effect on meat
microstructure, with tight muscle bers and minimal gaps between
them closely resembling that of fresh meat, which may be due to the
small changes in ambient temperature. However, using a microwave
to thaw meat resulted in important widening of muscle ber gaps and
breaking of muscle ber bundles. These results are consistent with
previous studies about extracellular space expansion due to cellular
and myobrillar compression between ice crystals during freezing,
frozen storage, and thawing of beef muscle [33, 34, 35].
Moreover, the presence of intracellular spaces in the form of vacuoles
differs in shape and size in most muscle cells. According to Bozzetta
et al.[5], the presence of intracellular vacuoles of varying shape and
size in most muscle cells is a key indicator associated with freezing. It
was also observed in several areas of damaged and partially deformed
muscle bers in which the ber boundaries could not be determined
(FIG. 4E). This tissue damage during the processes of freezing and
thawing is an inevitable consequence of the formation of ice crystals
inside and outside the cell, resulting in structural changes.
CONCLUSIONS
With an increasing concern among people regarding their health
while consuming red meat, particularly frozen imported beef, research
into the physical, biochemical, and histological changes that occur
during beef thawing is gaining more signicance. This study aimed
to investigate how commonly used thawing procedures affect the
histological structure of beef and its physico-chemical properties.
In this study, the quality of frozen meat was signicantly affected
by thawing techniques. The pH level of thawed meat was increased
(P<0.05). In contrast, Aw decreased signicantly after being thawed
regardless of the thawing method used (P<0.05). While refrigerator
thawing resulted in the least water loss at 1.23% (P<0.05), so the
brightness was closer to fresh meat, microwave thawing caused a
loss of signicant water demonstrated by thawing loss and cooking
loss at 4.37 and 44.47%, respectively (P<0.05). Microwave thawing also
resulted in the highest level of TBARS at 0.25 ± 0.034 mg·kg
-1
(P<0.05).
The light value (L*) of the microwave-thawed samples decreased
signicantly (P<0.05) compared to the fresh control. Histologically,
upon analyzing meat samples, it was discovered that beef subjected
to refrigerator thawing seemed comparatively lesser harm to its
Properties of beef meat thawed using various methods / Lakehal et al. _____________________________________________________________
6 of 7
structural composition. Additionally, the organization of the muscle
bers was successfully preserved.
Conict of interests
The authors declared no potential conicts of interest with respect
to the research, authorship, and/or publication of this article.
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