International Journal of Gastronomy and Food Science 32 (2023) 100701

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International Journal of Gastronomy and Food Science

journal homepage: www.elsevier.com/locate/ijgfs

Influence of temperature and time in sous-vide cooking on physicochemical

and sensory parameters of beef shank cuts
Adriana Gámbaro a, Luis Alberto Panizzolo a, *, Natalia Hodos a, Gonzalo da Rosa a,
Sofía Barrios b, Gabriela Garmendia c, Cintia Gago c, Javier Martínez-Monzó d
a
Food Science and Technology Department, School of Chemistry, Universidad de la República (UdelaR), Uruguay
b
Chemical Engineering Institute, School of Engineering, Universidad de la República (UdelaR), Uruguay
c
Microbiology Laboratory, Biosciences Department, School of Chemistry, Universidad de la República (UdelaR), Uruguay
d
Food Technology Department, UniversitatPolitècnica de València (Spain), Spain

A R T I C L E I N F O A B S T R A C T

Keywords: Sous-vide is an appropriate technique to be used in cuts of meat such as beef shank, which are inexpensive and
Sous-vide unappetizing due to their low tenderness. In the present work, time (2, 5, 8, 12, and 24 h) and temperature (55,
Meat 65, and 75 ◦ C) combinations were applied to the beef shank cut for sous-vide cooking. Its influence on physi­
Texture
cochemical and sensory parameters was studied and compared with a reference meat cut (loin) cooked for 5 h at
Sensory
65 ◦ C. The results show that at higher temperatures and time-cooking, sous-vide reaches a greater tenderness on
beef shank cuts, but with juiciness loss. Therefore, a compromise between these two parameters must be reached.
A sous-vide treatment at 55 ◦ C for 24 h resulted in meat with microbioal safety and an instrumental texture,
tenderness, stringiness, chewiness, juiciness, and sensory color like loin which is the reference cut.

1. Introduction it would be pertinent to commercially take advantage of all the cuts,

World meat consumption from 2018 to 2020 was 42.7 kg/capita, of
which 9.5 kg corresponded to bovine meat. The projections for the
decade 2021–2030 indicate that the beef consumption per capita will
increase by 2.3 kg in developed countries and 1.7 kg in developing
countries (OECD/FAO, 2021).
Uruguayan inhabitants have a great meat consumption, reaching
85.6 kg/inh/year in 2020 of which 45.7 kg was bovine meat. However,
bovine meat consumption patterns have been changing over the years
and price is not the only determinant of its preference (INAC, 2020). In
this context, several cuts currently have low or no direct consumption.
This would seem to be due to the lack of supply by butcheries and the
lack of culinary solutions for their use. Among them, is the beef shank
cut which has been mainly used for animal feed.
Shank cut is found on the region of the bovine leg. Its bone base is the
distal end of the femur and tibia, calcaneal tuberosity, and tarsus bones.
The main muscular planes are the flexor and extensor muscles of the leg
and foot. Of all commercialized cuts, beef shank cut is one of those with
less fat and hardness (Campbell and Hunt, 1996; Jeremiah et al., 2003).
As the demand for meat worldwide is increasing (OECD/FAO, 2021),
even those which are unappetizing. For this reason, it is necessary to
apply a process that allows reducing the sensory characteristics which
make them unappetizing. In the case of beef shank, it is the lack of
tenderness.
Sous-vide is an appropriate technique to take advantage of low-priced
and unappetizing meat cuts due to their lack of tenderness. The sous-
videcooking process is defined as “raw foods which are cooked under
controlled conditions of temperature and time into heat-stable vac­
uumed pouches” (Schellenkens, 1996). This cooking method consists of
packing the product in plastic pouches with low oxygen permeability
and high-temperature resistance. The product is usually cooked for a
long time in a thermostatic water bath or a convection oven to be
immediately refrigerated after being cooked (Ruiz et al., 2013). One of
the aspects that is characteristic of this technique is the use of sensibly
lower temperatures than those of traditional use. The controlled appli­
cation of low temperatures and longer time allows meat cooking under
conditions that maximize hardness reduction. This also allows collagen
denaturation without reaching the temperatures at which protein
retraction starts to occur (Ruiz et al., 2013). At the same time, an
improvement in sensory quality is achieved due to the retention of the

* Corresponding author.
E-mail address: apanizzo@fq.edu.uy (L.A. Panizzolo).

https://doi.org/10.1016/j.ijgfs.2023.100701
Received 13 December 2022; Received in revised form 22 February 2023; Accepted 1 March 2023
Available online 4 March 2023
1878-450X/© 2023 Elsevier B.V. All rights reserved.
A. Gámbaro et al.
aromatic compounds responsible for the aroma and flavor (Kathuria
et al., 2022).
The development of a ready-to-eat (RTE) product using a meat cut of
no direct consumption and low price, which is inserted among the so-
called convenient products and designed to save time, energy and
transfer culinary skills to consumers (Costa et al., 2007), will allow of­
fering a tender, nutritious, practical and low-cost product.This kind of
product can be targeted to either, the general population that seeks
practical solutions for everyday life but also those with chewing prob­
lems, such as hospital patients, the elderly and fourth age, individuals
with teething problems, etc.
To determine if sous-vide processing achieves a substantial
improvement in the sensory quality of the beef shank cut, it would be
reasonable to compare it with loin cuts whose degrees of tenderness are
among those most appetizing. Loin cut is located on the sub-lumbar
region; its bone base is the six lumbar hemivertebra, the last two dor­
sal vertebrae, and ilium bone; its main muscular planes are the psoas
major, psoas minor, illiacuslateralis, and quadratuslumborum.
There are multiple works on different cuts of beef cooked with sous-
videtreatment (Ayub and Ahmad, 2019; García-Segovia et al., 2007;
Ismail et al., 2019; Ruiz-Carrascal et al., 2019; Uttaro et al., 2019; Yang
et al., 2020), but the authors have not found references to the hind
shank.
To achieve an RTE product, the objective of the present work was to
assess the effects of different time-temperature combinations on beef
shank cut quality by sous-vide cooking and to study its influence on
physicochemical and sensory parameters compared with reference meat
cuts such as loin.
2. Materials and methods
2.1. Samples and processing
For this study, two cow meat cuts were used: shank (S) and loin (L)
provided by Instituto Nacional de Carne de Uruguay (INAC). All samples
came from grass-fed adult cows (number of permanent incisors present
= 8), Hereford breed. Meat from six animals was used in this work. Cuts
from different cows were randomly distributed for each treatment. The
meat cuts were cut into 3 cm thick portions to be vacuumed packed in
individual pouches with an approximate weight of 200 g (10–12 cm long
and 3 cm thick). The objective was that the consumer manages the
concept of “a portion”. Cuts were performed maintaining a similar
orientation in muscle fibers.
2.1.1. Previous preparation of samples
To improve sample appearance (Dominguez-Hernandez et al., 2018),
samples were marked/sealed with an electric grill plate (Ufesa GR 7451,
Power 2000W, hidden resistance, Dimensions: 59.0 × 31.7 cm,
Non-stick plates with tray collect fat)for 1 min each side at 250 ◦ C before
vacuum packing.
2.1.2. Sample cooking
After sealing, each meat portion was packed in polyethylene bags
− 40 ◦ C–120 ◦ C heat resistant (Cuder S.A., Montevideo, Uruguay) and
thermally sealed in a packing machine (Evox 30, Orved, Venecia, Italy).
The level of vacuum was adjusted at 90% as the highest level allowed
per equipment.Pouches were introduced in a thermoset bath (SV
THERMO, Orved, Venecia, Italy) for their cooking. Sample internal
temperature was controlled and registered using a PT 100 thermocouple
with an HH501DK (Omega Engineering, Inc., Stamford, CTUSA) regis­
ter. Thermocouples were introduced in three of the pouches located in
different places of the bath through pipe valves. After finishing the
cooking process, packs were removed from the water bath and sub­
merged in cold water (2 ◦ C) for 30 min.
Shank samples (S) were cooked at 55, 65, and 75 ◦ C. The cooking
times assayed were 2, 5, 8, 12, and 24 h, except for the samples treated at
2
International Journal of Gastronomy and Food Science 32 (2023) 100701
55 ◦ C, where the 2-h time was not considered as it did not meet the
minimum pasteurization requirements. Minimal sous-vide cooking time
was determined according to Baldwin’s (2012) tables which indicate
enough time to pasteurize meat in a water bath according to the product
thickness. These tables are based on the internationally accepted time
for one million to one reduction in Listeria monocytogenesand are applied
to all foods (FDA, 2011).
Each treatment was done in a single cooking batch. A batch was
made up of all the samples cooked at the same temperature. Samples
were removed at different cooking times. The order of treatments was
also randomized. Three repetitions were done for each condition.
Samples were maintained under refrigeration (5 ◦ C) until the analyses.
The reference cut (loin) was cooked at 65 ◦ C for 5 h (L5/65) (con­
ditions were determined according to previously unpublished studies).
2.2. Physicochemical analyses
Protein concentration was calculated by analyzing nitrogen con­
centration, using the Kjeldahl method (Vapodest 50s®, Gerhardt Labo­
ratory Systems GmbH, Koenigswinter, Germany) with factor 6.25 (ISO
937, 1978).
Fat was determined after acidic hydrolysis and the extraction by a
Soxhlet (LAT GmbH, Garbsen, Germany) (ISO 1443, 1973) equipment.
Ash concentration was analyzed from the weight difference before
and after combustion (600 ◦ C, 6 h) in a muffle (Carbolite®, LAT GmbH,
Garbsen, Germany) (ISO 936: 1998).
Moisture content was calculated as the weight difference before and
after drying in a heater (Binder GmbH, Tuttlingen, Germany) at 105 ◦ C
for 4 h (ISO 1442: 1997).
pH values were measured following the procedures described in ISO
2917:1999.
Hydroxyproline analysis was performed according to Kolar (1990),
using the collagen extraction suggested by Hill (1966). Hydroxyproline
is quantitatively determined as measure of collagenous material in meat
and meat products. Collagenous connective tissue contains 12.5% hy­
droxyproline (when nitrogen-to-protein factor of 6.25 is used) or 14%
(when factor is 5.55). Samples were hydrolyzed in H2SO4 at 105 ◦ C,
filtered, and diluted. Hydroxyproline was oxidized with chloramine-T to
pyrrole. Red-purple color that develops after addition of 4-dimethylami­
nobenzaldehyde was measured photometrically at 560 nm.
All the chemical analyses were done in triplicate.
2.3. Instrumental color measurement
Color readings were taken at five randomly selected non-overlapping
points on the surface of each cooked sample. The color was measured
using a Minolta CR-300 colorimeter (Konica Minolta Business Tech­
nologies Inc., Tokyo, Japan). Sample color was measured in the CIELAB
space, with standard illuminant D65, observer angle 10◦ , and zero and
white calibration. L* (lightness), a* (redness), and b* (yellowness) were
measured and the Chroma (C*) and Hue angle (h*) index values were
calculated as C* = (a*2 + b*2)0.5 and H* = arctan (b*/a*). The indi­
vidual differences in L*, a*, and b* values of each cooking treatment
concerning the color of the reference sample (r) were evaluated using ΔE
according to Eq. (1). For each experimental condition, twelve mea­
surements were taken.
√̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅
ΔE ∗ ab = ΔL∗2 + Δa∗2 + Δb∗2 (Eq. 1)
2.4. Instrumental texture analysis
For texture analysis, rectangular-shaped samples (2 × 2x5 cm) were
cut from beefs cooked at temperatures and times defined (cf. section
2.1.2). Samples were maintained under refrigeration conditions (4 ◦ C)
until texture assay. Texture assay was conducted at room temperature
(20 ◦ C) using a Texture Analyzer TA-XT2 (Stable Microsystems Ltd.,
A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

Surrey, United Kingdom) equipped with a Warner–Bratzler shear
attachment (1 mm thick). Each sample was cut perpendicular to the
longitudinal orientation of the muscle fibers. Test condition settings
were: pre-test speed 1 mm s-1; test speed 1 mm s-1; post-test speed 5 mm
s-1; distance 55 mm. The peak of positive force (PPF), representing
hardness, and positive area under the curve (PA), representing rigidity,
were evaluated using Texture Expert Exceed software version 3.1 (Stable
Microsystems Ltd., Surrey, United Kingdom). For each experimental
condition, six measurements were taken.
2.5. Sensory analysis
The panel of sensory judges was integrated by 9 individuals between
25 and 58 years old. Judges were selected by using ISO 8586 (2012)
standard. Judges had previous experience (minimum 250 h) in
discriminative tests and the evaluation of various foods using descriptive
analysis. To generate the appropriate descriptors to carry out the
descriptive analysis, each judge was individually presented with a piece
of approximately 30 g from 9 samples of the shank with different
cooking times and temperatures (5, 12, and 24 h at 55 ◦ C; 2, 12, and 24 h
at 65 ◦ C and 2, 12 and 24 h at 75 ◦ C). Together with these samples, a loin
sample was presented for the judges to select the most appropriate at­
tributes to describe them. Then in a group session with the panel leader,
there was consensus that the most appropriate descriptors were: color
intensity, tenderness, stringiness, chewiness, juiciness, boiled flavor,
and greasy flavor.
The judges were trained to be able to assess each attribute. 15
samples were assessed using 10 cm of length unstructured scales with
nil/high ends, except for color attribute whose ends were pink/brown
and chewiness whose ends were low/high. "Nil" means that the sensory
attribute is not perceived by the judges. Samples were presented in
balanced order and assessments were carried out in duplicate in 6
different sessions (5 samples per session). Still water and cookies
without salt were used as drafts. All assessments were carried out in a
sensory assessment laboratory designed according to ISO 8589 (2007)
standard.
Samples were assessed at 5 days of preparation, once safety was
guaranteed through the results of microbiological analyses.
determined immediately after cooking and at 7, 14, 21, and 28 days of
incubation.
2.7. Statistics analysis
Physicochemical and instrumental data were analyzed by the anal­
ysis of variance (ANOVA), using, time, temperature, meat cut, and their
interaction as fixed sources of variation. ANOVA on sensory data (color,
tenderness, stringiness, chewiness, juiciness, boiled flavor, and greasy
flavor) was carried out considering time, temperature, meat cut, and
their interaction as fixed sources and assessor and repetition as random
sources of variation. The means were compared by the Tukey test to
determine significant differences (P <0.05). Then, principal component
analysis (PCA) was carried out with the variables measured. Statistical
analyses were performed using Statgraphics Centurions (Statgraphics
Technologies, Inc., The Plains, Virginia).
3. Results and discussion
3.1. Physicochemical analyses
Table 1 shows mean values for moisture content, protein, fat, ash,
and pH values for shank samples subjected to different temperature/
time conditions as well as those of the loin sample taken as reference.
According to the variance analysis, the loin sample presented a
significantly lower content (P < 0.001) of ashes, fats, and significantly
higher (P < 0.001) of proteins than all processed shank samples, as
Table 1 shows.
Cooking temperature significantly influenced (P < 0.01) on moisture
Table 1
Physicochemical parameters (mean values ± standard errors) and ANOVA re­
sults of the hind shank (S) cooked by sous vide technique at different temper­
atures (55, 65 and 75 ◦ C) and times (2, 5, 8, 12, and 24 h), and reference sample
beef (L) cooked at 65 ◦ C for 5 h). Means with a common letter in the same
column are not significantly different (P > 0.05)according to Tukey test.
Samples* MOISTURE ASHES FAT (%) PROTEINS pH
(%) (%) (%)

2.6. Microbiological analyses S2/65 69.0 ± 0.8 3.0 ± 11.6 ± 15.9 ± 6.0 ±
ab
0.1 b 0.3 bc 0.1 b 0.2 abc

S2/75 67.2 ± 0.5 3.0 ± 11.9 ± 15.8 ± 0.1 5.5 ±

To guarantee the process viability after the cooking process and ab
0.1 b 0.1 de ab
0.3 a

sample blast-cooling, it was determined, for each assessed temperature, S5/55 69.7 ± 0.5 b 3.0 ± 11.8 ± 15.8 ± 0.1 5.6 ±
the count of total aerobic microorganisms, sporulated aerobic microor­ 0.1 b 0.1 cde ab
0.3 a

S5/65 68.3 ± 0.9 3.0 ± 11.8 ± 15.8 ± 0.1 5.6

ganisms, sporulated anaerobic microorganisms, and the presence of ab
±
0.1 b 0.1 cde ab
0.3 a
Escherichia coli, Staphylococcus aureus, and Salmonella spp in 10 g of the S5/75 67.8 ± 0.5 3.0 ± 11.9 ± 15.7 ± 0.1 5.5 ±
sample according to FDA (2020). Each assay was carried out in ab
0.1 b 0.1 de a
0.3 a

duplicate. S8/55 69.8 ± 0.7 b 3.0 ± 11.4 ± 15.8 ± 0.1 5.7 ±

0.1 b 0.1 b ab
0.2 a

S8/65 68.1 ± 0.5 3.0 ± 11.8 ± 15.7 ± 0.1 5.6

2.6.1. Microbiological challenge
±
ab
0.1 b 0.2 cde a
0.3 a

According to the results, for the time/temperature combination S8/75 65.9 ± 0.5 a
3.0 ± 11.8 ± 15.7 ± 0.1 5.7 ±
chosen, a microbiological challenge was performed against Clostridium 0.1 b 0.1 cde a
0.4 a

sporogenes ATCC 19404 (Health Canada, 2010) and Listeria innocua S12/55 68.0 ± 0.5 3.0 ± 11.6 ± 15.8 ± 0.1 5.7 ±
ab
0.1 b 0.1 bcd ab
0.3 a
ATCC (Health Canada, 2012). 10 samples were prepared for each
S12/65 68.3 ± 0.2 3.0 ± 11.7 ± 15.8 ± 0.1 6.6 ±
challenge. The time-temperature combination selected to carry out the ab
0.1 b 0.2 bcde ab
0.3 c

microbiological challenge was shank cooked at 55 ◦ C for 24 h (according S12/75 68.1 ± 1.0 3.0 ± 11.9 ± 15.7 ± 0.1 5.6 ±
to previous results). The samples were inoculated after sealing in an ab
0.1 b 0.1 de a
0.4 a

electric grill plate and before vacuum packaging with 1 × 103 spores/g S24/55 70.0 ± 0.7 b 3.0 ± 11.7 ± 15.8 ± 0.1 5.6 ±
0.1 b 0.1 cde ab
0.3 a
of sample in the case of the challenge against C. sporogenes. For the case
S24/65 68.6 ± 0.7 3.0 ± 11.8 ± 15.8 ± 0.1 6.3 ±
of the challenge against L. innocua, with 1 × 103 cells/g of the sample. ab
0.1 b 0.1 cde ab
0.1 bc

C. sporogenes spore concentration was estimated by direct microscopic S24/75 68.1 ± 1.4 3.0 ± 11.9 ± 15.7 ± 0.1 5.6 ±
ab
count and was confirmed through plate count, through soil cultivation. 0.1 b 0.1 de a
0.4 a

L5/65 68.3 ± 1.5 1.1 ± 5.7 ± 0.2 20.5 ± 0.1 5.9

L. innocua cell concentration was estimated by comparing with Mac ab
±
0.1 a a c
0.1 ab
Farland scale and confirmed through plate count through soil cultiva­
tion. Samples were subjected to the thermal treatment chosen and later Significance 0.0193 <0.0001 <0.0001 <0.0001 <0.0001
Level
stored at 4 ◦ C in a cold chamber. C. sporogenes or L. innocua count were

3
A. Gámbaro et al.
and fat contents of processed shank samples. At higher cooking tem­
peratures, moisture decreased (from 69.6 % on cooked samples at 55 ◦ C
to 67.4 % on those cooked at 75 ◦ C) and fat content increased (from 11.7
to 11.9 %). The cooking temperature did not significantly influence (P >
0.05) on the protein content of processed shank samples. As expected,
the sous-vide treatments carried out did not involve major changes in
physicochemical properties.
Fig. 1 shows the hydroxyproline proportion that was present in all
processed shank samples at different study conditions (55, 65, 75 ◦ C and
2, 5, 8, 12, and 24 h). It can be observed that for the treatments at 65 and
75 ◦ C there was a significant increase of hydroxyproline proportion with
cooking time. The collagen content was estimated from the amount of
hydroxyproline present, since it was an amino acid that forms, almost
exclusively, part of collagen (Bonnet and Kopp, 1984). In this case, it can
be observed that the hydroxyproline amount increased with cooking
time and temperature. These data indicate that soluble collagen
increased with time and temperature of treatment.
3.2. Color changes
Table 2 shows color coordinates obtained from sous-vide cooked
samples. Regarding lightness, SV cooked hind shank samples were not
significantly affected by cooking temperature, time, or their interaction
(P > 0.05). Some authors related changes in luminosity with the quan­
tity of water on the sample surface (Becker et al., 2016) with the water
loss, or with a higher level of myofibrillar alterations and sarcoplasmic
protein aggregation with increasing light scattering using higher tem­
perature (Christensen et al., 2012). In our study, as mentioned before,
only a few differences in water content of samples between the different
cooking treatments were observed. Similar results were obtained by
other authors (Botinestean et al., 2016) working with sous-vide cooked
meats treated with papain.
Concerning redness (a*), hind shank samples were significantly
affected by cooking temperature and cooking time, and by the interac­
tion between cooking temperature and cooking time (P < 0.05). Redness
intensity in cooked meat is inversely related to the level of denatured
myoglobin, a denaturing process which takes place between 55 ◦ C and
65 ◦ C although it continues until 75 ◦ C or 80 ◦ C. Consequently, beef
samples cooked at lower temperatures revealed a more intense red color
(higher a* values) than those cooked at higher temperatures, which
indicated higher myoglobin degradation as cooking temperature
increased (Hunt et al., 1999).
This loss of redness with increasing cooking temperature was
following the results obtained by García-Segovia et al. (2007), who
Fig. 1. Hydroxyproline proportion in hind shank fillets sous-vide cooked at 55, 65, and 75 ◦ C and different times (2, 5, 8, 12, and 24 h). Dash lines represent reference
values (sample L5/65). Means with a common letter in the same column are not significantly different (P > 0.05) according to Tukey test.
4
International Journal of Gastronomy and Food Science 32 (2023) 100701
cooked beef samples at 60 ◦ C–80 ◦ C for 15–60 min, Sánchez del Pulgar
et al. (2011), who cooked pork samples at 60 or 80 ◦ C for times as long as
12 h and Roldán et al. (2015), who cooked lamb samples at 60 ◦ C–70 ◦ C
– 80 ◦ C for 6–12 – 24 h.
Yellowness (b*) of the hind shank samples decreased with both
cooking time and temperature (P < 0.05), but their interaction was not
significant (P > 0.05). Other authors (Christensen et al., 2012; Gar­
cía-Segovia et al., 2007; Roldán et al., 2015) found an increase of yel­
lowness in sous-vide cooked samples with cooking temperature or
cooking time. These authors correlated yellowness increase with met­
myoglobin development, which resulted in a brownish color in the
cooked product. Our study presents the opposite tendency. This could be
explained by the protein composition of the hind shank, with more
quantity of other proteins (collagen) and less quantity of metmyoglobin
(Honig et al., 2020).
Sample Chroma (C*) and Hue angle (H*) values, as a dependent of a*
and b* parameters, also were significantly affected by cooking temper­
ature and cooking time, and by the interaction between cooking tem­
perature and cooking time (P < 0.05). C* decreased with the increase of
cooking temperature and cooking time while Hue angle (H*) increased
with cooking temperature and decreased with cooking time.
To evaluate the color differences of the hind shank with beef loin, the
total color difference (ΔE) was calculated between each cooked hind
shank and the beef loin cooked at 65 ◦ C for 5 h (Table 2). Total color
differences (ΔE) ranged between 5.7 and 13.4, values higher than 3
units and, therefore humanly perceptible (Bodart et al., 2008). Cooking
temperature and the interaction between cooking temperature and
cooking time showed significant effects (P < 0.05).
3.3. Instrumental texture analysis
Fig. 2 shows (a) the peak shear force values (N) and (b) positive area
(N⋅s) of hind shank fillets sous-vide cooked at 55, 65, and 75 ◦ C and
different times (2, 5, 8, 12, and 24 h). When hind shank samples were
compared, peak shear force was only significantly affected by cooking
temperature (P < 0.05), and the difference was between treatments at
55 ◦ C and treatments at 65/75 ◦ C. On the other hand, positive area
values were significantly affected by cooking temperature and time (P <
0.05) but their interaction was not significant P > 0.05). Cooking times
longer than 8 h and temperatures higher than 55 ◦ C produced similar
values of positive area. This parameter is related to the work that is
necessary to break the sample during mastication. Cooking plays a sig­
nificant role in the heat-induced denaturation of meat proteins, myofi­
brillar and connective tissue. The shrinkage of collagen fibers between
A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

Table 2
Color parameters (mean values ± standard errors) and ANOVA results of the hind shank (S) cooked by sous vide technique at different temperatures (55, 65 and 75 ◦ C)
and times (2, 5, 8, 12, and 24 h), and reference sample beef (L) cooked at 65 ◦ C for 5 h). Means with a common letter in the same column are not significantly different
(P > 0.05)according to Tukey test.
Sample L* a* b* C* H* ΔE
a cd cde de a
S2/65 48 ±2 13 ± 2 15 ± 1 20 ±1 0.85 ± 0.09 7 ± 2 ab
S2/75 46 ±4a 7 ± 1 ab 11 ± 3 ab 13 ± 3 ab 1.0 ± 0.1 bcde 12 ± 5 d
S5/55 45 ±3a 17 ± 4 e 21 ± 2 f 27 ±4g 0.9 ± 0.1 ab 9±5b
S5/65 49 ±3a 8±1b 16 ± 2 de 18 ± 3 cd 1.13 ± 0.08 efg 7 ± 1 ab
S5/75 47 ±5a 5±1a 12 ± 2 abc 13 ± 2 ab 1.16 ± 0.08 g 12 ± 4 cd
S8/55 48 ±2a 14 ± 1 de 20 ± 1 f 25 ± 2 fg 0.94 ± 0.06 abc 5±2a
S8/65 49 ±5a 8±1b 15 ± 3 cde 17 ± 3 cd 1.0 ± 0.1 cdefg 8 ± 5 ab
S8/75 47 ±5a 5±1a 11 ± 2 ab 12 ± 2 ab 1.14 ± 0.09 fg 13 ± 4 d
S12/55 48 ±1a 13 ± 2 cd 18 ± 1 ef 22 ± 1 ef 0.92 ± 0.06 ab 5±1a
S12/65 50 ±3a 6 ± 1 ab 13 ± 3 bcd 15 ± 2 bc 1.1 ± 0.1 defg 9 ± 3 bc
S12/75 47 ±1a 6 ± 1 ab 9±1a 11 ±1a 1.01 ± 0.06 bcdef 13 ± 2 d
S24/55 48 ±2a 12 ± 3 cd 15 ± 2 de 20 ± 2 de 0.9 ± 0.1 ab 7 ± 2 ab
S24/65 46 ±3a 7 ± 1 ab 11 ± 2 ab 13 ± 2 ab 0.9 ± 0.1 abcd 12 ± 3cd
S24/75 47 ±4a 6 ± 1 ab 10 ± 2 a 12 ± 2 ab 0.9 ± 0.1 abcd 13 ± 4 d
L5/65 52 ±1a 12 ± 1 c 20 ± 1 f 23 ± 1 ef 1.04 ± 0.01 bcdefg –

Significance Level 0.0521 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001

50 and 65 ◦ C decreased the breaking strength of the perimysial con­
nective tissue, while only smaller changes in the toughness of myofibrils
were observed. When heated to temperatures of 58–64 ◦ C the collagen
molecule undergoes a transition from the helical (crystalline) state to a
randomly coiled (amorphous) structure. Unrestrained collagen fibers
shrink when heated to temperatures of 60–70 ◦ C, denaturation then
proceeds into granulation, increased solubilization, and then gelatini­
zation, in connection with the breaking of intermolecular bonds by
increasing heat (Dominguez-Hernandez et al., 2018). Increasing
tenderness in muscles containing higher collagen contents from hind
shank cooked at the prolonged cooking time could be explained by the
fact that prolonged heat treatments at higher temperatures increased the
solubilization of collagen, thus decreasing the connective tissue strength
(Christensen et al., 2012). Myofibrillar proteins such as myosin are
rather thermolabile. The globular heads of the myosin molecule start to
denature at 40 ◦ C while heating above 53 ◦ C marks a more complete
denaturation. Actin has been reported to require somewhat high tem­
peratures, between 68 and 80 ◦ C, before it starts to denature (Berhe
et al., 2014). Results obtained in the present work show that prolonged
treatments at 55 ◦ C (24 h) or shorter treatments at 65/75 ◦ C are
adequate to increase sample tenderness. In our case hind shank fillets are
cuts with a high quantity of collagen and the tenderness of these cuts
could be associated mainly with changes in collagen structure as has
been mentioned before. In Fig. 2 it can also be observed that treatments
at 65 or 75 ◦ C, or longer treatments (24 h) at 55 ◦ C allow obtaining
values of shear force like meat cuts of higher quality, such as beef. These
results support the use of sous-vide treatments to revalue lower quality
cuts from a culinary point of view. Comparing shank samples with loin
samples, the latter had a significantly lower (P < 0.0001) hardness (PPF)
and rigidity (PA) values than boiled shank samples during 5, 8, and 12 h
at 55 ◦ C, according to Table 3.
The cooking temperature had a highly significant influence (P <
0.001) regarding the hardness and rigidity of the shank samples studied.
When cooking temperature increased, PPF values decreased (from
223.09 N on boiled samples at 55 ◦ C to 110.74 N on boiled samples at
75 ◦ C) decreasing PA values (from 2661.24 N s to 1812.47 N s).
3.4. Microbiological analyses
For all the time and temperature combinations assessed, counts of
total aerobic microorganisms sporulated aerobic microorganisms, and
anaerobic microorganisms were <10 ufc/g of the sample. The absence of
Escherichia coli, Staphylococcus aureus, and Salmonella spp. was deter­
mined in 10 g of sample.
This result shows that the meat sample used was in good conditions
and the sample treatment during the cooking process was adequate
(Moragas et al., 2019).
3.5. Microbiological challenge
The time-temperature combination selected to carry out the micro­
biological challenge (shank cooked at 55 ◦ C for 24 h) was enough for
controlling L. innocua and C. sporogenes. After the thermal treatment was
selected, the absence of both microorganisms in 10 g of the sample was
determined. Due to these species behaving similarly to L. monocytogenes
and C. botulinum, it could be inferred that the thermal treatment selected
might ensure the safety of the finished product.
3.6. Sensory analysis
According to Table 4, the loin sample had different ratings of color,
tenderness, stringiness, chewiness, and juiciness than those of the shank
samples studied. Highly significant differences (P < 0.0001) were found
among color, tenderness, stringiness, chewiness, and juiciness of the
different samples assessed.
The cooking temperature had a highly significant influence (P <
0.001) on the color, tenderness, chewiness, and juiciness of the shank
samples studied. When the cooking temperature increased, the color
intensity and tenderness of samples increased whereas chewiness and
juiciness decreased.
Cooking time had a highly significant influence (P < 0.001) on the
color, tenderness, stringiness, chewiness, and juiciness of the shank
samples studied. When cooking time increased, color intensity and
tenderness of samples also increased, whereas stringiness, chewiness,
and juiciness decreased.
3.7. Relationship among sensory, instrumental, and physicochemical data
An analysis of principal components was carried out regarding
texture and color instrumental data and sensory parameters. A physi­
cochemical parameter (moisture) linked to sensory parameters was also
included in the analysis, as reported in other studies (Naqvi et al.,
2021b). Two of the first principal components represented 57.1% and
24.6% of the variance respectively. As Fig. 3 shows, the first principal
component (F1) was positively related to the juiciness and moisture, also
to the two instrumental parameters of texture (PPF y PA) and a*, b*, and
C* color parameters, but negatively with sensory color, and H* color
parameter. Therefore, the samples to the right of the first principal
component presented a greater intensity in a highly desirable attribute
in cooked meat cuts, such as juiciness. The second principal component

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A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

Fig. 2. (a) Peak shear force values (N) and (b) Positive area (N⋅s) of hind shank fillets sous-vide cooked at 55, 65, and 75 ◦ C and different times (2, 5, 8, 12, and 24 h).
Dash lines represent reference values (sample L5/65). Means with a common letter in the same column are not significantly different (P > 0.05)according to
Tukey test.

(F2) was positively linked to stringiness and chewiness (no desirable
attributes in cooked meat cuts) and negatively to tenderness (another
highly desirable attribute in cooked meat cuts). This figure shows that
the samples were quite spread out on both PC1 and PC2 which indicates
that they had different sensory and instrumental characteristics.
3.8. Relationship between sensory and instrumental data linked to color
As the cooking temperature increased, samples were sensory evalu­
ated as less red and darker. This can be confirmed by the lowest values of
a* obtained, which also coincides with the reported by other authors
(Becker et al., 2016). Sensory color had a negative correlation with a*
(R2 = 0.887, P< 0.0001), b* (R2 = 0.716, P = 0.0001), and C* (R2 =
0.841, P< 0.0001), and a positive correlation with H* (R2 = 0.452, P =
0.0084).
At 55 and 75 ◦ C the samples did not show a significant sensory color
increase with cooking times, or with a sensory color intensity range of
1.4–2.5 and 7.9–8.5 respectively. At 65 ◦ C a significant increase in color
between 2 and 5 h and between 12 and 24 h was found. a* value
significantly decreased at 55 ◦ C and 65 ◦ C. The difference in a* among
samples with different times of cooking at 55 ◦ C was not reflected in
sensory color values.
As it was previously indicated, the red color of cooked meat is mainly
determined by myoglobin amount, its redox state, and the denaturation
heat dependent (King and Whyte, 2006). Heating below 60 ◦ C may not
denature significant amounts of myoglobin and thus may not develop a
well-done appearance compared with heating at higher temperatures
(Becker et al., 2016). The reddest color of samples cooked at 55 ◦ C might
take potential consumers to reject it due to an undesirable raw
appearance or worries related to microbiological safety.
A positive significant correlation was found among a* and b*(R2 =
0.745, P< 0.0001). Whereas a* values decreased with heating temper­
ature and time, b* values showed a concomitant decrease which does
not coincide with what other studies reported (Becker et al., 2015,
2016). Roldán et al. (2015) assumed that the increase in b* values might
be explained due to metmyoglobin formation and a higher denaturation

6
A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

Table 3 Table 4
Instrumental texture parameters (mean values ± standard errors) and ANOVA Sensory data (mean values ± standard errors) and ANOVA results of the hind
results of the hind shank (S) cooked by sous vide technique at different tem­ shank (S) cooked by sous vide technique at different temperatures (55, 65 and
peratures (55, 65 and 75 ◦ C) and times (2, 5, 8, 12, and 24 h), and reference 75 ◦ C) and times (2, 5, 8, 12, and 24 h), and reference sample beef (L) cooked at
sample beef (L) cooked at 65 ◦ C for 5 h). Means with a common letter in the same 65 ◦ C for 5 h). Means with a common letter in the same column are not signif­
column are not significantly different (P > 0.05) according to Tukey test. PPF = icantly different (P > 0.05) according to Tukey test.
peak positive force, represents hardness. PA = positive area, represents rigidity.
Samples COLOR TENDERNESS STRINGINESS CHEWINESS
Samples PPF (N) PA (N.s)
S2/65 2.0 ± 1.1 4.2 ± 1.4 ab 6.8 ± 1.5 c
6.3 ± 1.9 cd
a
S2/65 116 ± 56 ab 1901 ± 859 abcd
S2/75 137 ± 27 abc 2454 ± 480 abcd S2/75 8.2 ± 1.4 4.5 ± 1.7 ab 6.4 ± 1.6 c
7.1 ± 1.8 d
e
S5/55 232 ± 121 bc 3188 ± 1339 d
S5/65 102 ± 46 ab 1779 ± 801 abcd S5/55 1.4 ± 1.3 3.6 ± 1.6 ab 7.0 ± 1.8 c
6.8 ± 1.4 d
a
S5/75 119 ± 45 ab 1947 ± 439 abcd
S8/55 282 ± 132 c 2960 ± 672 cd S5/65 5.3 ± 1.8 3.6 ± 1.6 ab 6.0 ± 1.8 bc
7.2 ± 1.7 d
cd
S8/65 101 ± 28 a.b 1712 ± 410 abc
S8/75 127 ± 38 abc 2019 ± 393 abcd S5/75 8.5 ± 1.6 5.3 ± 1.5 bc 6.1 ± 1.3 bc
7.4 ± 1.8 d
e
S12/55 234 ± 110 bc 2665 ± 838 bcd
S12/65 84 ± 19 a 1298 ± 223 ab S8/55 1.2 ± 1.3 4.9 ± 1.7 ab 5.6 ± 1.8 bc
5.1 ± 1.8 bcd
a
S12/75 77 ± 49 a 1106 ± 600 a
S24/55 143 ± 127 abc 1831 ± 784 abcd S8/65 6.8 ± 1.3 5.5 ± 1.6 bc 5.7 ± 1.7 bc
5.9 ± 1.7 cd
de
S24/65 95 ± 48 ab 1609 ± 561 abc
S24/75 91 ± 54 ab 1533 ± 651 abc S8/75 8.2 ± 1.7 7.3 ± 1.5 cd 5.2 ± 1.1 bc
5.5 ± 1.8 cd
e
L5/65 86 ± 19 a 1078 ± 227 a
S12/55 2.1 ± 1.8 3.1 ± 1.1 a 7.6 ± 1.3 c
6.3 ± 1.5 cd
Significance Level <0.0001 <0.0001 a

S12/65 5.0 ± 1.6 7.2 ± 1.7 cd 6.4 ± 1.8 c

4.6 ± 1.5 bcd
cd

of this molecule by heat, which results in a brown color. However, our S12/75 8.0 ± 1.4 7.4 ± 1.3 cd 5.8 ± 1.7 bc
6.8 ± 1.6 d

study was not able to wholly respond about what muscular proteins e

finally influenced on b* values (Roldán et al., 2015). S24/55 2.5 ± 1.0 8.5 ± 1.7 d 2.2 ± 1.6 a
2.1 ± 1.5 ab
ab
At 55 ◦ C, it took 24 h of cooking thus the color perceived by the
S24/65 7.8 ± 1.3 8.9 ± 0.9 d 1.2 ± 1.1 a
3.5 ± 1.8 abc
sensory judges did not significantly differ from the reference loin sam­ e

ple, whereas at 65 ◦ C it took between 5 and 12 h of cooking to match the S24/75 7.9 ± 1.7 8.3 ± 1.6 d 3.7 ± 1.8 ab
7.1 ± 1.5 d

color. At 75 ◦ C all cooking samples lost reddish colorations and had e

significantly higher brown color intensities than the reference loin. L5/65 4.3 ± 1.5 8.5 ± 1.0 d 1.9 ± 1.4 a
1.6 ± 1.0 a
bc

Significance <0.001 <0.000 <0.001 <0.001

3.9. Relationship between sensory and instrumental data linked to Level
tenderness Samples JUICINESS GREASY FLAVOR BOILED FLAVOR
cd a a
S2/65 5.1 ± 1.8 2.1 ± 1.3 3.9 ± 1.7
Tenderness is a multifactorial sensory attribute that is determined by S2/75 2.3 ± 1.0 ab
2.3 ± 1.4 a
5.0 ± 1.8 a

the complex interaction of different factors. Among them, are its S5/55 6.6 ± 1.5 de
3.6 ± 1.6 a
3.9 ± 1.8 a
bc a a
anatomic location and the animal muscular function (Rhee et al., 2004). S5/65 4.1 ± 1.2 4.0 ± 1.8 4.2 ± 1.7
a a a
S5/75 1.9 ± 1.3 2.7 ± 1.7 4.0 ± 1.4
In our study, cooking time and temperature influenced sensory texture cde a a
S8/55 5.7 ± 1.1 3.9 ± 1.4 4.2 ± 1.8
assessment turning meat cuts more tender when these parameters S8/65 2.2 ± 1.6 ab
2.5 ± 1.3 a
4.8 ± 1.8 a

increased. These studies coincide with what other researchers found S8/75 1.7 ± 1.3 a
3.0 ± 1.5 a
4.6 ± 1.6 a

who have indicated that sous-vide cooking improves tenderness (Boti­ S12/55 6.1 ± 1.8 cde
3.2 ± 1.8 a
3.7 ± 1.0 a
ab a a
nestean et al., 2016; Sánchez del Pulgar et al., 2011) and decreases meat S12/65 2.3 ± 0.9 3.6 ± 1.4 4.0 ± 1.7
ab a a
S12/75 2.5 ± 1.8 3.2 ± 1.3 4.5 ± 1.8
hardness which is associated with structural changes induced by the e a a
S24/55 7.3 ± 1.7 2.6 ± 1.7 4.6 ± 1.3
heat on myofibrillar proteins and connective tissue (Christensen et al., S24/65 2.0 ± 0.8 ab
2.2 ± 1.1 a
4.9 ± 1.8 a

2000; Silva et al., 2016). A minimum of 24 h of cooking at 55 ◦ C, 12 h at S24/75 2.0 ± 1.5 ab

2.6 ± 1.8 a
4.9 ± 1.2 a
de a a
65 ◦ C, and 8 h at 75 ◦ C were needed for tenderness of shank samples was L5/65 6.6 ± 1.8 1.7 ± 1.3 4.4 ± 1.6
like those of loin reference. Significance Level <0.001 0.2631 0.0787
Consumer’s perception of meat quality is affected by different sen­
sory attributes, but tenderness is one of the most important attributes
which influences consumer’s satisfaction and purchase decision. Con­ cooked for 24 h at 55 and 65 ◦ C, which reflects a greater tenderness of
sumers are increasingly willing to pay a high price for tender meat these samples.
(Naqvi et al., 2021b). The inconsistent supply of tender meat not only Also, sensory tenderness had a negative correlation with PPF (R2 =
has a negative effect on consumers’ demand and acceptance but also has 0.307, P = 0.032) and PA (R2 = 0.484, P = 0.004), both instrumental
an impact on the meat industry (Naqvi et al., 2021a). The fact of parameters of texture measured by Warner-Bratzler blade, which are
obtaining time and temperature combinations for sous-vide cooking interpreted as hardness and rigidity respectively. These results show that
which allow that consumer perceives shank cut such tender as the loin of the instrumental measure similarly reflects this sensory attribute.
reference, gives a value to this cow cut which is traditionally considered Similar results were found by Holman et al. (2020), who showed that
as a “very hard” one. shear force had a negative relationship with tenderness. The meat’s
Tenderness had a negative correlation with stringiness (R2 = 0.709, sensory texture must be not only represented by tenderness but also by
P< 0.0001) and chewiness (R2 = 0.425, P = 0.008), which was to be other attributes such as juiciness which can have a positive impact on
expected since more tender meat implies a shorter chewing time and the consumer’s acceptance. This last attribute was not reflected by the
breakage of muscle fibers due to temperature and cooking times de­ texture instrumental measure.
creases stringiness perception. Comparing shank samples with those of
loin reference, loin’s stringiness and chewiness were like shank samples

7
A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

with portions of 3 cm thick with an approximate weight of 200 g. Other

portion sizes might affect the sensory and physico-chemical character­
istics of the product studied and cooked under the same time and tem­
perature conditions. Therefore, more studies are required.

4. Conclusions

This study finally confirmed that sous-vide is a cooking technique that

is particularly beneficial for adding value to low-value meat cuts by
transforming hard cuts into tender meat. In this work, it was possible to
define the time-temperature combination for sous-vide cooking, which
allows hard meat cut such as shank to improve its characteristics, mainly
in terms of instrumental and sensory texture. We concluded that a 55 ◦ C
treatment for 24 h resulted in meat with instrumental texture, tender­
ness, stringiness, chewiness, juiciness, and sensory color like the refer­
ence loin cut. Although meat cooked at a higher time and temperature
was more tender, juiciness loss implies that these meats are not neces­
sarily going to satisfy consumers’ necessities. A compromise between
these two parameters must be found to obtain a high-quality sensory
product.

Implications for gastronomy

Fig. 3. Principal component analysis carried out on sensory and instrumental
data results of the hind shank (S) cooked by sous vide technique at different
Several beef cuts currently have low or no direct consumption. This
temperatures (55, 65 and 75 ◦ C) and times (2, 5, 8, 12, and 24 h), and reference
would seem to be due to the lack of supply by butcheries and the lack of
sample beef (L) cooked at 65 ◦ C for 5 h).
culinary solutions for their use. Among them, is the beef shank cut which
has been mainly used for animal feed. The use of these cheaper cuts of
3.10. Relationship between sensory and physicochemical data linked to meat can save a lot of money to the families and restaurants.Often these
juiciness
cheaper cuts require a longer cooking time (and lend themselves to sous-
vide cooking) but are full of flavour and usually just as tasty, if not more
Juiciness is an important trait of food quality that influences con­
so than some of the more expensive cuts. Sous vide can tenderize even
sumers’ satisfaction with cooked meat after softening and is linked to the the toughest of cuts.
meat moisture content and the losses due to cooking (Naqvi et al.,
2021b). The most tender meat is not necessarily the juiciest and a bal­ Credit authorship contribution statement
ance must be found among these properties to satisfy consumers’ ex­
pectancies. In our study, the increase of cooking temperature decreased A. Gámbaro: Funding acquisition, Project administration, Investi­
the sample juiciness which might be explained by the severe longitu­
gation, Supervision, Writing – original draft, Writing – review & editing;
dinal contraction of muscular fibers (Palka and Daun, 1999). These re­ L.A. Panizzolo: Funding acquisition, Investigation, Supervision, Writing
sults coincide with the sensory evaluations reported about pigs and lamb
– original draft, Writing – review & editing; N. Hodos: Investigation,
(Becker et al., 2016; Christensen et al., 2012; Mortensen et al., 2012). Data curation; G. da Rosa: Investigation, Data curation; S. Barrios:
Cooking times did not influence sample juiciness at 55 and 75 ◦ C.
Investigation, Data curation, Writing – original draft; G. Garmendia:
Samples cooked at 55 ◦ C had high juiciness similar to those of the loin Investigation, Data curation, Writing – original draft; C. Gago: Investi­
reference. In the case of the samples cooked at 65 ◦ C, when cooking time
gation, Data curation; J. Martínez: Writing – original draft, Writing –
increased, juiciness significantly decreased which coincides with what review & editing.
Dominguez-Hernandez et al. (2018) observed.
Various authors have evaluated meat juiciness exclusively by its total Ethical guidelines
content of water during cooking (Ayub and Ahmad, 2019; Naqvi et al.,
2021a, 2021b). In our work, despite both variables having a strong This study does not require ethics approval.
correlation, moisture variation with cooking temperatures (variation of
only 2.2% average among samples cooked at 55 and 75 ◦ C) would not Declaration of competing interest
explain the great changes of sensory juiciness perceived. These results
reaffirm the importance of sensory measures of parameters that are The authors declare that they have no conflicts of interest to this
critical on the consumer’s acceptance of meat cuts and not only deter­ work.
mine them by instrumental measures.
As juiciness and tenderness are the most important sensory attributes Data availability
in cooked meat, any process must be designed to obtain optimal values
on both traits. Even some authors indicated how a juicier sensation No data was used for the research described in the article.
improved tenderness perception (Becker et al., 2016; Dinardo et al.,
1984). In our study, the only cooking time and temperature combination Acknowledgments
that resembled shank with the loin of reference was the cooking during
24 h at 55 ◦ C. This research was financially supported by the Comisión Sectorial de
Despite shank softening being achieved by the temperature and time
Investigaciones Científicas (CSIC), Universidad de la República (Ude­
combinations assayed, juiciness loss due to cooking at 65 and 75 ◦ C laR), Uruguay.
temperatures might have a negative impact on consumers’ experience.
The size of the sample is one of the factors that affect the optimiza­
tion of the conditions of the sous-vide process. In our study, we worked

8
A. Gámbaro et al. International Journal of Gastronomy and Food Science 32 (2023) 100701

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