Meat Science 96 (2014) 1–4

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Meat Science
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TBARS predictive models of pork sausages stored at

different temperatures
Fan Wenjiao a,b, Zhang Yongkui a,⁎, Chen Yunchuan b, Sun Junxiu b, Yi Yuwen b
a
School of Chemical Engineering, Sichuan University, Chengdu 610065, China
b
Department of Food Science, Sichuan Tourism University, Chengdu 610100, China

a r t i c l e i n f o a b s t r a c t

Article history: 2-Thiobarbituric acid reactive substances (TBARS) is an important quality index for pork sausages. To study
Received 8 April 2013 this in pork sausages during storage, kinetic models were developed to predict TBARS content changes of
Received in revised form 14 June 2013 pork sausages at different temperatures. The predictive models of TBARS content with respect to storage
Accepted 18 June 2013
time and temperature were developed based on primary and Arrhenius equations. The regression coefficients
Available online 22 June 2013
(R2 N 0.95) indicated the acceptability of the primary reaction and Arrhenius model for predicting TBARS
Keywords:
content changes of pork sausages. The activation energy (EA) of TBARS is 14.12 kJ mol−1, and the corre-
TBA content sponding rate constant (k0) is 9.262 × 1010. Relative errors between predicted and measured values of
Predictive models TBARS content are all within ± 8%. Thus, the established model could effectively predict the TBARS content
Pork sausage of pork sausages between 5 and 35 °C during storage.
Temperature © 2013 Elsevier Ltd. All rights reserved.

1. Introduction Silva, Nascimento, & Fukuma, 2003), is widely used as an indicator

Pork sausages are one of the oldest processed meat products
enjoyed by millions of consumers all over the world. Traditionally,
pork sausages are made of pork meat and fat, which are chopped
and mixed thoroughly with seasonings, such as pepper powder,
spice paprika, salt, rice wine, sugar, monosodium glutamate and gin-
ger. After stuffing the meat mixture in natural casings from the
cleaned small intestine of pigs, the sausages are dried at room tem-
perature (Sebranek, Sewalt, Robbins, & Houser, 2005; Qiu, Zhao,
Sun, Zhou, & Cui, 2013).
Fat is an important constituent of processed pork sausage products
because it affects tenderness and juiciness. A high fat content is tradi-
tionally associated with succulence and flavor (Vural, 2003). However,
fat content is associated with a higher degree of lipid oxidation during
storage, and consumers' acceptability of pork sausages declines due to
the development of rancidity caused by lipid oxidation and/or microbial
growth (Bradley et al., 2011). Therefore, oxidative rancidity is consid-
ered the major cause of reduced quality of pork sausages. Generally
speaking, rancidity occurs through the reaction of unsaturated fats
with oxygen, which is influenced by heat, light, and pro-oxidant com-
pounds found in the ingredients, such as salt and sodium lactate
(Cheng, Wang, & Ockerman, 2007; Sallam & Samejima, 2004).
TBARS content, which reflects the content of malonaldehyde, one
of the degradation products of lipid hydro peroxides and peroxides
formed during the oxidation of polyunsaturated fatty acids (Gomes,
⁎ Corresponding author. Tel./fax: +86 28 85408255.
E-mail addresses: wenjiaolc@163.com (F. Wenjiao), zhangyongkui@scu.edu.cn
(Z. Yongkui).
of the degree of lipid oxidation and is considered an important quality
index for pork sausages (Tang, Sheehan, Buckley, Morrissey, & Kerry,
2001). Though the TBA assay is sensitive and widely used, it is not
specific and in high oxidative conditions, TBA reacts with a number
of components such as the decomposition products of aldehydes in
volatile compounds, which are present in the pork sausage at the
end of storage. There are a number of studies that investigate the var-
ious chemical changes that take place, including changes of TBARS
content in pork sausages. However, studies on the predictive models
for quality parameters of pork sausages are limited. In order to obtain
useful information to optimize pork sausage storage management
and enhance the safety of pork sausages, this study investigated the
TBARS content changes of pork sausages at different temperatures.
Primary and Arrhenius equations were combined to develop models
to predict the TBARS content of pork sausages during storage.
2. Materials and methods
2.1. Sausage sample preparation
Fresh boneless pork hams and backfat were purchased from a
local meat market. Lean tissue and pork backfat were ground through
an 8–12 mm plate. Approximately 65% lean tissue and 35% pork
backfat were mixed thoroughly with non-meat ingredients, 3% salt,
2% rice wine, 0.5% monosodium glutamate, 0.5% sugar, and 0.05% pep-
per powder. Then, the meat mixtures were stuffed into natural pork
casings, previously soaked in water. Raw sausages were manually
tied at approximately 15 cm-intervals, dried at 40 °C for 6 h, then

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2 F. Wenjiao et al. / Meat Science 96 (2014) 1–4

stored at 5, 10, 15, 20, 25, 30 and 35 °C, respectively. Three sausage
samples were taken randomly every 6 days for analysis of TBARS.
2.2. TBA analysis
TBARS content was determined colorimetrically by the method of
Pokorny and Dieffenbacher (Kirk & Sawyer, 1991), which is based on
the spectrophotometric determination of the pink complex formed
after the reaction of one molecule of malondialdehyde (MDA) with
two molecules of 2-thiobarbituric acid. TBARS was expressed as mg
MDA/kg pork sausages. Samples (10 g) were homogenized in 20 ml
of TBARS solution (prepared by dissolving 200 mg of 2-TBA in
100 ml 1-butanol, filtered, stored at 4 °C for not more than five
days), and the mixture was heated for 10 min in boiling water
(95 °C). After being cooled with running water, the absorbance (As)
was measured at 530 nm against a water blank. A blank reagent
was run and the absorbance (Ab) was recorded. The TBA value (mg
of malonaldehyde per 100 g of tissue) was obtained by: TBA =
50 ∗ (As − Ab) / 200.
2.3. Statistical analysis
sausages. Differences among the initial TBA content in pork sausages
may be due to the type of seasoning, fat composition, as well as the
type of pork muscles used in each type of sausage. TBARS for all sau-
sages increased throughout storage, and the increase in TBARS may
be attributed to the partial dehydration of the sausages and increased
oxidation of unsaturated fatty acids (Mendes, Pestana, & Gonçalves,
2008). In addition, the increasing rate of TBARS during storage was
higher as storage temperature increased. Low temperatures are
known to inhibit the oxidation of lipids.
3.2. Determination of TBARS models
To validate the applicability of the models, TBARS changes of the
sausage samples stored at 5, 15, 25, 30, and 35 °C were used to estab-
lish the predictive model, and the samples stored at 10 and 20 °C
were treated as controls, verifying the application techniques as
well as demonstrating the validity of the approach (Zhang, Li, Lu,
Shen, & Luo, 2011). The TBARS data at constant temperature was
first fitted by a conventional first-order kinetic model (Maskan,
2001).
First-order kinetic model:

All analyses were run in triplicate. Data were analyzed using the
general linear model (GLM) of Statistical Analysis System's Proce-
dures (SAS Institute Inc., 1999). Duncan's multiple range tests were
used to determine differences between means. Significance level
was set at 5%.
3. Results and discussion
3.1. Changes in TBA content of pork sausages during different storage
temperatures
TBARS has been used to determine the degree of lipid oxidation,
and the presence of TBA reactive substances is due to the second
stage of auto-oxidation, during which peroxides are oxidized to alde-
hydes and ketones (Li et al., 2013). According to Connell (1990), a
TBA value of 2 mg MDA/kg is the limit. The changes in TBARS of
pork sausages at different temperatures are shown in Fig. 1. The ini-
tial TBARS of the sausages was 0.22 mg MDA/kg. This value is close
to the 0.24 mg MDA/kg reported by Martínez, Djenane, Cilla,
Beltrán, and Roncalés (2006) for Spanish fresh pork sausages but
higher than the 0.17 mg MDA/kg reported by Kuo and Chu (2003)
for Chinese pork sausages and lower than the 0.29 mg MDA/kg
reported by Tan, Liao, Jhan, and Liu (2007) for Taiwanese pork
3.0
kB t
B ¼ B0 e ð1Þ
where B is the value of TBARS indicators, B0 is the initial TBARS value,
and k is the rate constant (day−1) at a given temperature.
The TBARS values of sausages stored at different temperatures
were analyzed to determine the reaction order. The values of k
were obtained from the slope of regression of ln [B / B0] versus
time. First-order equations for TBARS values of pork sausages at dif-
ferent temperatures are given in Table 1. Regression coefficients of
the first-order kinetic equations for TBARS values of pork sausages
at different temperatures are all higher than 0.95, the highest value
being 0.9946. These high regression coefficient values indicate the ac-
ceptability of the first-order reaction.
The data was then applied to the Arrhenius model and integrated
as described by Giannakourou and Taokis (2003).
Arrhenius equation:
 
E
KB ¼ K0 exp − A ð2Þ
RT
where K0 is the frequency factor, EA is the activation energy (J mol−1), T is
the absolute temperature (K), R is the gas constant (8.3144 J (mol K)−1),
and K0 and EA are constants related to the nature systems.
The modified Logistic Arrhenius equation is given by:
 
E
ln KB ¼ ln K0 − A : ð3Þ
RT

2.5
In the calculated rate constants at different temperatures, the slope
of the regression line (−EA / R) was obtained through the use of ln KB
TBA(mg MDA/kg)

2.0 on the reciprocal (T−1) plot. Activation energy (EA) was obtained
through the slope of the regression line, and K0 was obtained through
1.5 the intercept of the regression line. The temperature dependence of
the rates on TBARS changes is described adequately by Arrhenius

1.0
Table 1
First-order kinetic equation for TBARS values of pork sausages at different temperatures.
0.5
Temperature (K) First-order kinetic equation R2 (Regression coefficient)
0.0288t
0.0 278.15 y = 0.2072e 0.9946
0 14 28 42 56 70 84 288.15 y = 0.2516e0.033t 0.9874
298.15 y = 0.2745e0.0428t 0.9778
Time (days)
303.15 y = 0.2999e0.0439t 0.9587
308.15 y = 0.303e0.053t 0.9516
Fig. 1. Changes in TBARS content of pork sausages at different storage temperatures.
 F. Wenjiao et al. / Meat Science 96 (2014) 1–4 3

kinetics in the studied temperature range (Fig. 2). The results from Fig. 2 Table 2
show that the slope of the regression line was −1.6977, so the values of Predicted and measured TBARS contents of pork sausages stored at 10 and 20 °C.

EA and rate constant (K0) for the pork sausages in TBARS were Temperature Storage time Measured value Predictive value Relative error
14.12 kJ mol−1 and 9.262 × 1010 respectively, and the corresponding (°C) (d) (mg MDA/kg) (mg MDA/kg) (%)
R2 in TBA was 0.9960. 10 7 0.28 0.26 −7.14
The equation of the TBARS predictive model: 10 21 0.49 0.52 6.12
10 35 0.83 0.77 −7.23
20 7 0.36 0.38 5.56
 
10 20 21 0.62 0.58 −6.45
B ¼ B0 exp 9:262  10 t expð−14120=RTÞ ð4Þ 20 35 1.15 1.24 7.83

where B are the predictive values in the TBARS value of the pork sau-
sages stored for a particular time, and B0 are the initial values in
TBARS content of the sausages.
3.3. Validation of the TBARS models and discussion
In order to validate the TBARS predictive models, the comparison of
the predicted and measured values of the pork sausages stored at 10
and 20 °C on the 7th, 21st, and 35th day was evaluated (Table 2). The
results show that the established TBARS predictive models give a realis-
tic prediction of the TBARS content, relative errors are all within ±8%.
Kaymak-Ertekin and Gedik (2005) claimed that a predictive model is
acceptable if relative errors are below 10%. Therefore, according to the
predictive models, TBARS values can be quickly and accurately predict-
ed in pork sausages stored at 5–35 °C. It is well known that traditional
quality evaluations involve a large number of experimental determina-
tions that not only are time-consuming, but also require trained labor
and high cost. Predictive modeling has the potential to reduce the num-
ber of experiments and provide an important tool for optimizing the
quality of the processed food product during storage (Boekel, 1996).
Several studies have developed kinetic models to predict the quality
changes in aquatic products. A predictive model made by Yao, Luo,
Sun, and Shen (2011) was based on electrical conductivity and fresh-
ness indicators to predict the freshness of crucian carp (Carassius
carassius), and the relative errors of this model between predicted and
actual values of electrical conductivities were within ±5%. Hong, Luo,
Zhu, and Shen (2012) applied the general stability index method to pre-
dict quality deterioration in bighead carp (Aristichthys nobilis) heads,
and the relative errors of that method were all below 10% before the
15th day at −3 and 15 °C. Liu and Tu (2012) also developed a support
vector regression (SVR) model based on the electronic nose to predict
the TVB-N content in eggs, the average prediction accuracy of which
was 94.62%. Hence, the experimental data has an acceptable fit with
the established TBARS content predictive model and the application of
this model can lead to effective management systems for lipid oxida-
tion, which will optimize the quality of pork sausage products.
Fig. 2. Arrhenius curve of changes in the rate of TBARS contents.
4. Conclusions
TBARS content of pork sausages increased during storage, and the
amount increased as storage temperature increased. TBARS value is
consistently correlated to storage time and temperature by a
first-order kinetic model with high regression correlations
(R2 N 0.95). The description of TBARS content also came with high
regression correlation (R2 = 0.9660) values using the Arrhenius
equation. The established TBARS content predictive models give a re-
alistic prediction of the TBARS content, all relative errors are within
± 8%. Therefore, this model is effective for TBARS predictions in
pork sausages stored at 5–35 °C.
Acknowledgments
This work was supported by the Scientific Research Fund of
Sichuan Provincial Education Department of China.
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