Journal of Food Engineering 266 (2020) 109693

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Journal of Food Engineering

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

Non-destructive prediction of texture of frozen/thaw raw beef by Raman T

spectroscopy
Qingmin Chena,b, Yichi Zhangc, Yahui Guoa,b, Yuliang Chenga,b, He Qiana,b, Weirong Yaoa,b,
Yunfei Xiea,b,∗, Yukihiro Ozakid
a
State Key Laboratory of Food Science and Technology, Jiangnan University, China
b
School of Food Science and Technology, Jiangnan University, No.1800 Lihu Avenue, Wuxi, 214122, Jiangsu Province, China
c
Economics and Management School, Wuhan University, Wuhan, 430072, Hubei Province, China
d
Department of Chemistry, School of Science and Technology, Kwansei Gakuin University, Gakuen 2-1, Sanda, Hyogo, 669-1337, Japan

A R T I C LE I N FO A B S T R A C T

Keywords: To date, the main methods of texture measurement are sensory testing and instrumental testing. These two
Raw beef testing methods are time-consuming and often destructive respectively. In this study, Raman spectroscopy was
Frozen/thaw used to predict the texture of different frozen/thaw raw beef from continuous freezing and repeated freeze-thaw
Texture treatments. The effect of repeated freeze-thaw treatment on beef texture was significantly different (p < 0.05)
Raman spectroscopy
when the number of freeze-thaw cycles exceeded three times. Quantitative models were developed with opti-
Non-destructive
mized spectra and texture parameters of the samples based on partial least squares analysis. The result showed
Prediction
that Raman spectroscopy exhibited good performance in predicting tenderness, chewiness, firmness, and
hardness with R2p of 0.81, 0.80, 0.81, 0.82 respectively, and weaker performance for springiness with R2p of
0.53. Therefore, Raman spectroscopy has potential for the quantification of texture parameters of frozen/thaw
beef. Besides, the PCA loadings plots for PC1 and PC2 revealed that the main variables of prediction equations
were located at approximately 960–1060 cm−1, 1370–1490 cm−1, and 1550–1680 cm−1. These regions are
significantly influence by changes in hydrophobic properties and secondary structure composition of meat
protein.

1. Introduction main parameter related to the freshness of chicken meat, which is

The texture of meat is undoubtedly the most important quality that
consumers appreciate. It influence consumers' decision to make a re-
purchase and affect consumer acceptability (Bekhit et al., 2014; Chen
and Opara, 2013; Shackelford et al., 2001). For example, tenderness
was the most important palatability attribute for beef consumers and
most consumers are willing to pay a higher price for tender beef
(Huffman et al., 1996). Besides, tenderness is also regarded as an im-
portant quality of beef, along with juiciness and flavour, for assessing
beef eating satisfaction (Wu et al., 2012). Hence, meat texture studies
prefer to study the tenderness of meat. Additionally, other texture
characteristics such as hardness, springiness and firmness also play
important roles in the overall meat quality (Wu et al., 2014). Hardness
is one of mechanical characteristics used to describe food texture
(Szczesniak, 1963). Firmness is one of the main quality attributes of
freshness and mouthfeel in fish, which is a critical texture parameter
related to consumer acceptability (Ando et al., 1992). Springiness is a
mainly influenced by muscle structure such as characteristic of con-
nective tissues and physicochemical characteristics such as contents of
moisture, liquid neutral lipids, and myofibrillar proteins (Xiong et al.,
2015).
So far, freezing is the main storage method for meat preservation
due to it contributes to inhibit microbial spoilage. However, the quality
of frozen/repeated freeze-thaw meat is generally considered inferior to
fresh meat (Leygonie et al., 2012). Especially repeated freeze-thaw
cycles exert a strong effect on the meat texture due to ice crystal for-
mation, which damages the structural integrity of cell membranes
(Leygonie et al., 2012). Extracellular ice crystals destroy the physical
structure of meat, breaking myofibrils and reducing meat texture (Liu
et al., 2010). Hence, meat texture prediction is vital to meat industry, as
their main goal is to produce products with high commercial value that
are appealing and enjoyable to the consumer.
There are various methods available to measure meat texture, in-
cluding sensory testing and instrumental methods. Sensory testing of

∗
Corresponding author. State Key Laboratory of Food Science and Technology, Jiangnan University, China.
E-mail address: xieyunfei@jiangnan.edu.cn (Y. Xie).

https://doi.org/10.1016/j.jfoodeng.2019.109693
Received 20 November 2018; Received in revised form 17 August 2019; Accepted 17 August 2019
Available online 22 August 2019
0260-8774/ © 2019 Elsevier Ltd. All rights reserved.
Q. Chen, et al.
Fig. 1. Schematic illustration of the force-time plot from (A) the texture parameter 1 test and (B) texture parameter 2 test.
meat is time consuming, and has poor repeatability, and often results in
strong subjective evaluation (Sun et al., 2012). The instrumental
methods to determine texture parameters, including shear force (SF)
measurement, that provide reliable results regarding meat tenderness
are destructive (Kamruzzaman et al., 2013; Wu et al., 2012) and un-
suitable for online detection. Therefore, developing a non-destructive,
accurate, and on-line method that can predict meat texture has become
an urgent need in the meat industry. As a rapid and non-destructive
methodology, spectroscopic techniques have attracted significant at-
tention because of the non-destructive analysis, short analysis time,
high sensitivity, and low operating cost. As Raman spectroscopy allows
for the development of fibre optic components and energy saving lasers
suitable, even hand-held Raman spectrometers have allowed for a wide
range of applications. In addition, Raman spectroscopy can provide
biochemical information from tissue analysis. Thus, Raman spectro-
scopy has the potential to predict different meat quality attributes
(Herrero, 2008; Nian et al., 2017; Scheier et al., 2014). Numerous
studies have been reported regarding the prediction of meat qualities
using Raman spectroscopy for tenderness (Bauer et al., 2016; Fowler
et al., 2014a, 2014b, 2015), texture (Chen and Han, 2011; Kang et al.,
2014), pH (Fowler et al., 2015; Scheier et al., 2014), colour (Scheier
et al., 2014, 2015), water holding capacity (Phongpa-Ngan et al., 2014;
Sanchez-Gonzalez et al., 2008), and spoilage (Zajac et al., 2017).
This study was performed to examine the potential of Raman
spectroscopy for predicting frozen/thaw raw beef texture in a non-de-
structive manner based on partial least squares (PLS) analysis.
Continuous freezing and repeated freeze-thaw treatments are carried
out for sample handling. PLS models of the texture parameters in-
cluding tenderness, chewiness, firmness, hardness and springiness were
developed based on the Raman spectral data. Loading plots were used
to identify the wavenumbers associated with the highest variation
(contribution) in the prediction equations.
2. Materials and methods
2.1. Sample preparation
A total of 16 complete M. longissimus dorsi muscles from 8 Chinese
Yellow bulls (Yanbian) were purchased from the local market. As
shown in Fig. S1, the top and tail of the dorsi muscle were cut and
discarded, leaving only the middle section muscle for segmentation.
After removing the fat and connective tissue from the meat surface, the
dorsi muscle was cut into around 23 × 5 × 6 cm3 pieces as one sample
which were packed individually in Ziploc bags, a total of 130 sub-
samples were used in this study. Two different freezing methods were
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Journal of Food Engineering 266 (2020) 109693
performed, in which the repeated freeze-thaw samples were frozen at
−20 °C for 7 days and thawed at 4 °C for 24 h for one freeze-thaw cycle.
The frozen group was frozen at −20 °C, with a period of 7 day and
thawed under the same conditions before testing. 130 sub-samples were
randomly assigned to different treatments. 76 sub-samples were as-
signed to the repeated freeze-thaw treatment and 54 sub-samples were
assigned to the continuous freezing treatment. There are 10–12 samples
in each freeze-thaw cycle group, and 6–8 samples in each continuous
freezing group (Table S1).
2.2. Texture tests
All texture tests were performed using the TA.XT Plus (Stable Micro
Systems, Surrey, England) instrument. All sub-samples were tested
parallel to their myofibrils. Every sub-sample was assessed 10 times in
each texture test.
2.2.1. Texture test 1
Texture test 1 was performed using the Meulle-net–Owens razor
shear (MORS) probe and the test parameters are as follows:
Test Mode: Compression; Pre-Test Speed: 1.00 mm/s; Test Speed:
2.00 mm/s; Post-Test Speed: 10.00 mm/s; Distance: 20.00 mm; Trigger
Force: 10.0 g.
A force-by-time curve for the calculation of corresponding texture
values was shown in Fig. 1A. The tenderness (peak force), chewiness
(the positive force area above the baseline or area 1), and firmness
(peak force of area 2) values were also given.
2.2.2. Texture test 2
A cylindrical probe (P/2) with a diameter of 2 mm was used for this
test, which consisted of two procedures. The first procedure involves
the non-destructive deformation of the meat structure, measuring the
sample hardness and springiness with a fixed pressure from a certain
distance. The second procedure involves the destruction of the sample
structure.
When the probe first contacted the beef, its thickness was auto-
matically recorded by the software. The probe continuously moved
downwards to a fixed distance inside the beef, then returned to the
initial point of contact with the beef and stopped for a set period
(0.01 s). The distance between locations at which the first and second
procedures were performed was approximately 2 mm. The distance
between the different tests was greater than 5 mm. The test parameters
are as follows:
Pre-test speed: 1.00 mm/s; test speed: 3.00 mm/s; post-test speed:
3.00 mm/s; trigger force: 5.0 g; 1st distance: 5.00 mm; hold time: 0.01 s;
Q. Chen, et al. Journal of Food Engineering 266 (2020) 109693

Fig. 2. Statistics of tenderness, chewiness, firmness, hardness, and springiness of beef after continuous freezing and repeated freeze-thaw. Standard error bars are
indicated. Within a measured variable, values with different letters are different (P < 0.05).

2nd type: distance; 2nd distance: 20.000 mm; 2nd strain: 5.0%.
A force-vs-time curve for calculating the corresponding texture va-
lues is shown in Fig. 1B. The hardness (peak force of the first com-
pression), springiness (ratio of the positive area input to negative area
input during the first compression or area 4/area 3 × 100).
2.3. Raman measurement
A portable RamTracer-200-HS Raman spectrometer (Opto-Trace
Technologies Co. Ltd, Silicon Valley, CA, USA) was used with the fol-
lowing parameters: excitation light source of 785 nm, laser power of
285 mW, sweep range of 100–3300 cm−1, and resolution of 4 cm−1.

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Q. Chen, et al.
The integration time was 5 s and 6 accumulations were measured per
point. Each sub-sample has three slices about 2 cm thick for Raman
measurement, and each slice was scanned randomly at 5 separate
points. Thus, the final spectrum for each sub-sample was the average of
15 points. The Raman spectra were calibrated using acetonitrile before
collection.
2.4. Data processing and analysis
SPSS software version 19.0 was used for data analysis. One-way
ANOVA followed by Duncan’ test was used to compare reflect the ef-
fects of different freeze-thaw cycles and freezing time on the different
texture parameters. The value of p < 0.05 was considered statistically
significant.
All texture data and unit conversion were implemented by Exponent
software version 6.1.16.0 (Stable Micro Systems, Surrey, England).
Among 130 sub-samples, eight damaged sub-samples from texture test
1, five damaged sub-samples from texture test 2 and six of abnormal
and damaged sample spectra were checked and removed. Finally, there
are 111 sub-samples have all texture test 1, texture test 2 and Raman
data obtained. A calibration-set (81 sub-samples) and validation-set (30
sub-samples) were used for prediction of traits using Raman spectra.
Analysis of the Raman spectra was performed using TQ Analyst soft-
ware (V9.5.0.76, Thermo Fisher Scientific, Madison, WI, USA). The
developed predictive models were calculated using PLS. Multiplicative
scatter correction (MSC), standard normal variate (SNV), and first de-
rivation (1st D) techniques were used for spectral pre-processing. MSC
and SNV are widely used to correct light scattering variation and
multiplicative noise arising from the physical structure of the samples
(Luypaert et al., 2004; Maleki et al., 2007). The 1st D technique is
commonly applied to eliminate baseline offset variations in a series of
spectra (Savenije et al., 2006). Moreover, the Raman spectral ranges
were investigated in this study. The optimal pre-treatment was chosen
considering the highest R2 and lowest square error of calibration values.
The prediction results were evaluated using correlation coefficients of
calibration (R2c), correlation coefficients of prediction (R2p), root mean
square error of calibration (RMSEC), and root mean square error of
prediction (RMSEP), these parameters were calculated using the TQ
analyst software. The performance and reliability of the prediction
equations were also evaluated using residual prediction deviation
(RPD). The RPD values of the models were calculated as the ratio be-
tween the standard deviation of the reference values to standard error
of prediction (RPD of prediction = SDref/SEP). All samples were equally
distributed over the spectral variations to cover the entire variation
range in both data sets.
3. Results and discussion
3.1. Statistical analysis of measured texture parameters
As show in Fig. 2A1, an increase in the SF values were observed
after repeated freeze-thaw cycles and was significantly higher after the
5th cycle (P < 0.05). The increase in the SF values may be attributed
to the formation of ice crystals during the freezing process, which de-
creased water-holding capacity properties of the meat (Lee et al., 2008).
In addition, freeze–thaw for 6th and 7th cycles resulted in higher
chewiness (Fig. 2A2) compared to that from 1st to 5th cycles
(P < 0.05). As shown in Fig. 2A3, a significant increasing was ob-
served since the 4th freeze-thaw cycles cycle (P < 0.05). Similarly, the
same phenomenon is also found in hardness (Fig. 2A4) during the re-
peated freeze-thaw process. This can be attributed to the decrease of
moisture content in beef after repeated freeze-thaw as lower-moisture
foods typically possess a higher degree of hardness (Szczesniak, 1963).
Decreasing springiness was observed after 7 freeze-thaw cycles in
Fig. 2A5. Springiness is related to quality deterioration and freshness
because it reflects the mechanical resistance of the meat to external
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Journal of Food Engineering 266 (2020) 109693
forces (Bourne, 1982). This indicates a deterioration in the quality of
the meat after the repeated freeze-thaw process.
As showed in Fig. 2B1, a little effect on tenderness of continuous
freezing storage was obtained between the first 7 days and other frozen
storage days (P < 0.05). No significant difference (P < 0.01) of ten-
derness was observed in samples from 7 to 49 days continuous freezing
storage. Lee et al. (2008) found that frozen time less than 2 months had
no effect on broiler breast billet texture. Similar results also appeared in
chewiness (Fig. 2B2). Besides, the continuous freezing of beef from 7
days to 49 days has a little effect on firmness (Fig. 2B3) and has no
effect (p > 0.05) on hardness (Fig. 2B4). As shown in Fig. 2B5, the
values of springiness in continuous freezing beef group changes greatly.
The structure and biochemistry of meat change significantly during
freezing storage, especially when meat is accidently frozen or thawed
due to improper temperature control (Coombs et al., 2016). Compared
with continuous freezing groups, repeated freeze-thaw cycles over three
times has a significant different (p > 0.05) in beef texture parameters,
the increase of the average value of tenderness, chewiness, firmness and
hardness was found. However, repeated freeze-thaw showed negative
effects on springiness parameters, resulting in a decrease in beef
springiness.
3.2. Raman spectroscopic analysis
Raw Raman spectra of the 111 frozen/thaw beef sub-samples (81
calibration sub-samples and 30 validation sub-samples) from 500 to
1800 cm−1 were shown in Fig.3A, and a summary of the Raman band
assignments was listed in Table 1. The peaks at 826 and 850 cm−1
(tyrosine doublet), 758 cm−1 (tryptophan), and 1000 cm−1 (phenyla-
lanine) can be attributed to aromatic amino acid side chains (Herrero,
2008). Tryptophan exhibits a characteristic peak at 1552 cm−1. The
intense bands at approximately 1446 cm−1 can be assigned to the CH2
bending modes of aliphatic amino acids. The amide I and III bands,
which reflect protein secondary structure, were observed at 1610–1680
and 1220–1350 cm−1, respectively. (Beattie et al., 2004).
3.3. Prediction of texture based on PLS models using Raman spectroscopy
A larger variability of these texture parameters were provided to
examine the potential of Raman for predicting tenderness, chewiness,
firmness, hardness and springiness. The optimized prediction results
were shown in Fig. 3. By setting the mean Raman spectra extracted
from all samples as the predictors and their corresponding texture
parameter values as the target values, PLS was conducted to establish
quantitative models based on full spectral range. The pre-treatment that
best contributed to the PLS quantitative calibration models for each
texture parameter was listed in Table 2.
For tenderness (Fig. 3B), good prediction result was obtained with
an R2p of 0.81 and RMSEP of 2.57 N. Similar prediction accuracy with
R2CV of 0.75 and RMSECV of 6.82 N was also found in predicting
Warner-Bratzler shear force (WBSF) of young dairy bull beef using
Raman spectroscopy (Nian et al., 2017). Both studies were processed
samples with freezing and thawing before Raman measurements.
Samples after freezing and thawing with more desirable results com-
pared to fresh meat have been reported (Schmidt et al., 2013). A pos-
sible reason was interpreted as the damage of peptide backbone and
amino acid side chains by freezing and thawing, which resulted in
measuring more sensitive of the vibrations of chemical bonds by Raman
scattering (Fowler et al., 2014b). In general, freezing and thawing
process affects water redistribution and induce extracellular ice crystal
formation, resulting in mechanical damage to muscle structures. The
micro-environments of amino acids and special protein structures are
changed by freezing and thawing cycles (Zhang et al., 2017). For-
tunately, Raman spectroscopy is a powerful technique for studying the
changes in protein structure induced by the local environments such as
those consisting of mostly aliphatic hydrophobic residues, tryptophan,
Q. Chen, et al. Journal of Food Engineering 266 (2020) 109693

Fig. 3. (A) Raw Raman spectra of beef samples used for PLS modeling. (B) Predictions of tenderness, chewiness, firmness, hardness and springiness based on PLS
model 3 from calibration (black circles) and validation (red circles). Samples of raw beef were for calibration (n = 81) and validation (n = 30). (For interpretation of
the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Table 1 (R2cv of 0.27 and RMSEP of 11.5 N) of lamb 1 days post-mortem

Assignment of bands in the Raman spectra of beef. (Fowler et al., 2014b), and there was no improvement in the SF pre-
Band Wavenumber(cm−1) Assignment diction after freezing and thawing (Fowler et al., 2015). Although both
Fowler et al. (2015) and Schmidt et al. (2013) focused on predicting SF
1 758 tryptophan values of lambs, the comparison of results is limited due to the differ-
2 826, 850 tyrosine doublet ences of experimental parameters and design. The background, sex and
3 890 CH3 Rocking
4 932 CC stretching (α-Helix)
age of lambs studied by Fowler et al. were unknown. In addition,
5 1000 Phenylalanine stretching Fowler et al. trimmed (removed cap muscle and m. adducto) the whole
6 1072 CC Aliphatic stretching M. semimembranosus muscles and then cut a part of it for each test.
7 1122 CC Aliphatic in-phase stretching Schmidt et al. pre-cut the right loin muscle into a 3 cm thick sample
8 1263 = C-H (cis) In-plane bending
weighing around 65 g. Therefore, so far, it is unclear whether freezing
9 1270 Amide Ⅲ (α-Helix)
10 1312 Amide Ⅲ and thawing contribute to SF prediction in fresh and freeze/thaw
11 1446 CH3, CH2, CH Bending meats. Nevertheless, these studies illustrate the complexity of the fac-
12 1552 tryptophan tors affecting the prediction of meat texture traits. In addition to
13 1650 Amide Ⅰ freezing and thawing, ageing has also been considered as one of the
important factors affecting meat texture traits prediction. The SF of
porcine M. semimembranosus was predicted using Raman spectroscopy
and tyrosine (Herrero, 2008). The intensity of Raman peaks of α-helix
with a poor correlation 24 h post-mortem (R2cv of 0.22 and RMSECV
protein, myoglobin, tryptophan, tyrosine and connective tissue was
of 7.8 N), but a better correlation (R2cv of 0.7 and RMSECV of 4 N) was
found to vary between tender and tough meat (Bauer et al., 2016).
observed with 72 h post-mortem (Scheier et al., 2014). The significant
However, inconsistent results were obtained in predicting SF values
differences in prediction performance were attributed to shortening of

Table 2
Summary of reference values of meat texture traits and PLS model performances for texture prediction by Raman spectra in beef.
Ranges Means SD Pre-processing Selected ranges(cm−1) Factors Calibration Validation

R2c RMSEC R2p RMSEP PRD

Tenderness (N) 38.68–16.55 25.52 4.81 1st D + SNV 550–1750 7 0.83 2.78 0.81 2.57 1.87
Chewiness (g.s) 12097.86–5934.22 8706.02 1527.72 1st D + MSC 650–1650 9 0.91 625 0.80 942 1.62
Firmness (g) 109.14–28.70 60.29 20.83 1st D + SNV 1000–1700 7 0.91 8.70 0.81 11.5 1.81
Hardness (g) 140.64–46.01 80.92 20.98 Raw + SNV 900–1750 6 0.82 11.9 0.82 12.8 1.64
Springiness (%) 39.81–22.27 28.11 3.61 Raw + SNV 800–1800 3 0.71 2.75 0.53 2.26 1.60

Notes：PLS model performances for texture prediction were optimized to the best. Raw represented raw spectra; 1st D represented first derivation.

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Q. Chen, et al. Journal of Food Engineering 266 (2020) 109693

the sarcomeres during rigor mortis, resulting in distortion of the SF

values 24 h post-mortem. The result implies that ageing may be con-
tribute to improve the accuracy of prediction. However, other study
found that the WBSF predictions of aging for 2 days were more accurate
than those of aging for 8 days. Therefore, they considered that if the
sample data set could be separated according to factors such as animal
grade, sex or ultimate pH value, it would be helpful to improve the
accuracy of prediction (Leroy et al., 2004).
Furthermore, as showed in Fig. 3C, chewiness predictions were al-
most as accurate as those of the tenderness predictions with an R2p of
0.80, and RMSEP of 942 g.s. This may be related to a significant cor-
relation between chewiness and tenderness. Good predictions were
obtained using the Raman spectra for firmness (Fig. 3D) and hardness
(Fig. 3E) whose R2p values were 0.81 and 0.82, respectively, and the
corresponding RMSEP were 11.5 g and 12.8 g, respectively. A better
prediction model with a high determination coefficient (R2p of 0.941)
was obtained for predicting firmness quality of grass carp fillet after
frozen storage (Cheng et al., 2014). Regression coefficients, genetic
algorithm, and successive projection algorithm were combined to afford
an ideal prediction. In addition, the prediction of springiness (Fig. 3F)
yielded an R2p of 0.53 and RMSEP of 2.26% in this study. A high
correlation coefficient of 0.84 was obtained for predicting the springi-
ness of fresh chicken meat in a previous study (Xiong et al., 2015).
Compared to beef, the muscle fibres in chicken meat (white meat) are
thinner and denser, and the connective tissues are softer (Berzaghi
et al., 2005). This different muscle structure possesses different light
scattering, transmission, and reflection properties (Hughes et al., 2014).

3.4. Main variable contribution in the prediction equations

The loading plots of the best PLS models were shown in Fig. 4. The
contribution ratio of first principal component (PC1) and second prin-
cipal component (PC2) was typically greater than 65% for all models.
Loading plots were used to identify spectral regions that contributed
most significantly to the analytes (Lu et al., 2010).
As shown in Fig. 4A–C, PC1 and PC2 contributed approximately
50% and 20% to the variation between samples, respectively. The main
peaks in the loading plots are located at approximately 1000,
1420 cm−1, and from 1550 to 1680 cm−1. The contributions for PC1
and PC2 are shown in Fig. 4D and E, the variables at approximately
996, 1438, and 1642 cm−1 of PC1 exhibited contributions towards
hardness, and 1004 and 1462 cm−1 also exhibited obvious contribu-
tions towards springiness. Overall, for the prediction of beef texture,
intensity changes specifically occurred around 960–1060 cm−1,
Fig. 4. Loading plots of the first and the second principle components (PCs)
1370–1490 cm−1, and 1550–1680 cm−1. The bands at 890–1060 cm−1
obtained from PCA analysis of the Raman spectra of five texture parameters:
can be assigned to C-C stretching vibrations arising from α-helices
tenderness (A), chewiness (B), firmness (C), hardness (D), and springiness (E)
(890–945 cm−1) and β-sheets (1020–1060 cm−1). The variations in (PC1: Black solid line, PC2: Red dotted line). (For interpretation of the refer-
these structures lead to corresponding variations in the intensity of ences to colour in this figure legend, the reader is referred to the Web version of
their Raman bands (Barrett et al., 1978). In general, the freeze-thaw this article.)
process is accompanied by a decrease in α-helix content and an increase
in β-sheet content in meats (Xu et al., 2016). Phenylalanine exhibits a
sheet than tender meat (Tu, 1982). In addition, the ratio of α-helix to β-
strong band around 1000 cm−1, which is a relatively active in the
sheet has also been proved to be an important factor in shear force and
contribution to texture prediction equations. It is well known that
texture of meat (Beattie et al., 2004). Interestingly, the predictive
freezing and thawing cause protein structural deformation and increase
contribution of PC1 to beef hardness and springiness was different at
surface hydrophobicity of protein (Leygonie et al., 2012). The con-
1422–1512 cm−1 and 1594–1714 cm−1. It appears that the contribu-
tribution of the variable around 1002 cm−1 is likely related to the more
tion of changes in Raman band at 1446 cm−1 and amide I band around
exposure of phenylalanine hydrophobic residues to protein surface
1610–1680 cm−1 to the prediction of hardness and springiness was
during freeze-thaw cycles. Aspartic and glutamic acid residues exhibit a
reverse.
Raman band in the 1400–1420 cm−1 region arising from their COO−
groups. The band at 1446 cm−1 (C-H Bending) of PC1 has an obvious
contribution to hardness, and is sensitive to the hydrophobicity of the 4. Conclusions
protein environment. Increasing intensity of the band at 1446 cm−1
was observed in tough meat (Beattie et al., 2004). The band centred at In this study, continuous freezing and repeated freeze-thaw treat-
1610–1680 cm−1 arises from the amide I band of proteins, and is sen- ment were used for sample handling and had different effects on the
sitive to the Secondary structure of protein (α-helix, β-sheet, twisted texture of beef. Non-destructive texture prediction in frozen/thaw raw
and random coil, etc.) (Herrero, 2008). Tough meat contains more β- beef by Raman spectroscopy was investigated based on PLS models. The

6
Q. Chen, et al.
results demonstrated that Raman spectra exhibited good performance
in the prediction of tenderness (R2p = 0.81), chewiness (R2p = 0.80),
firmness (R2p = 0.81), and hardness (R2p = 0.82) of frozen/thaw beef.
However, the Raman spectra gave poor predictions of springiness
(R2p = 0.53). Besides, the PCA loadings plots for PC1 and PC2 revealed
that the main variables are located at approximately 960–1060 cm−1,
1370–1490 cm−1, and 1550–1680 cm−1. It suggested that the predic-
tion of meat texture is mainly affected by changes in protein physico-
chemical properties and protein composition. It is mainly reflected in
the influence of hydrophobic properties of amino acid residues, as well
as the changes in secondary structural components of proteins (α-helix,
β-sheet, etc.). The results showed that Raman spectroscopy has poten-
tial as a non-destructive and rapid method to predict frozen/thaw beef
texture. Moreover, as meat texture is a complex traits, which can be
influenced by breed, diet, physiological stress, muscle type, ageing,
storage conditions and other factors. Therefore, more factors affecting
texture should be considered in further study. Besides, it would be in-
teresting to study the mechanisms that affect predictions in fresh and
freeze/thaw meat texture.
Acknowledgements
The work described in this article was supported by the National
Key R&D Program of China (2018YFC1602300), the Natural Science
Foundation of Jiangsu Province (BK20171139), the Yangtze River Delta
Project of Shanghai (18395810200), the Forestry Science and
Technology Innovation and Extension Project of Jiangsu Province
(No.LYKJ[2017]26), the Postgraduate Research & Practice Innovation
Program of Jiangsu Province (KYCX18_1766), the National First-class
Discipline Program of Food Science and Technology
(JUFSTR20180509), the Science and Technology Project of Jiangsu
Bureau of Quality and Technical Supervision (KJ175923 and
KJ185646), the China Postdoctoral Science Foundation funded project
(2018M642165), and the Science and Technology Plan of Changzhou
City (CE20172002).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jfoodeng.2019.109693.
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