Meat Science 90 (2012) 6065

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Meat Science
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m e a t s c i

Total antioxidant capacities of raw and cooked meats

Arda Serpen a,, Vural Gkmen a, b, Vincenzo Fogliano c
a
b
c

Food Research Center, Hacettepe University, 06800 Beytepe, Ankara, Turkey

Department of Food Engineering, Hacettepe University, 06800 Beytepe, Ankara, Turkey
University of Naples Federico II, Department of Food Science, I-80055 Portici, Naples, Italy

a r t i c l e

i n f o

Article history:
Received 15 February 2011
Received in revised form 28 May 2011
Accepted 31 May 2011
Keywords:
Meat
Antioxidant activity
Quencher procedure
Heating
Denaturation

a b s t r a c t
This study investigated the total antioxidant capacity (TAC) of meats (beef, chicken, pork and sh) and its
changes on thermal treatment. The QUENCHER procedure, which is performed directly on the solid material
without extraction, was selected and proved to be particularly suitable for meat samples. The ABTS+
scavenging capacity of raw meats ranged between 25.9 1.0 and 51.7 1.2 mmol Trolox Eq./kg. Raw chicken
had the highest TAC followed by pork, beef and sh. Upon heating at 180 C, TAC of meats increased to an
apparent maximum at 5 min followed by sudden decreases until 15 min, while the nal stage of heating was
characterized by slight increases. The modications of TAC during cooking can be explained considering
factors such as denaturation and exposure of reactive protein sites, degradation of endogenous antioxidants
and the formation of Maillard reaction products having antioxidant properties.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
The primary importance of meat in human nutrition is related to
its high quality proteins that provide essential amino acids upon
digestion. However, besides the high protein contents, meat also
provides valuable amounts of many essential micronutrients such as
some unsaturated fatty acids, vitamins and minerals (Tornberg,
2005).
Meat as a food has a complex physical structure and chemical
composition that is very susceptible to oxidation (Wood et al., 2008).
The oxidative stability of meats depends upon the balance and the
interaction between endogenous anti- and pro-oxidant substances
and the composition of substrates prone to oxidation including
polyunsaturated fatty acids (PUFA), cholesterol, proteins and pigments (Bertelsen et al., 2000; Decker & Xu, 1998). Endogenous
antioxidant systems are composed of non-enzymatic hydrophilic and
lipophilic compounds such as vitamin E, vitamin C, carotenoids,
ubiquinols, polyphenols, cellular thiols, and enzymes like superoxide
dismutase, catalase and glutathione peroxidase. Together, enzymatic
and non-enzymatic antioxidant systems operate to counteract the
action of pro-oxidants in muscle tissues (Chan & Decker, 1994;
Decker, Livisay, & Zhou, 2000) both in living animals and also after
slaughter.
The composition of endogenous antioxidants and pro-oxidant
compounds can differ among meat of different species, among
animals of a single species (Descalzo & Sancho, 2008; Pradhan,

Corresponding author. Tel.: + 90 312 297 7120; fax: +90 312 299 2123.
E-mail address: aserpen@hacettepe.edu.tr (A. Serpen).
0309-1740/$ see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2011.05.027

Rhee, & Hernandez, 2000) and muscle type. Also the diet of the animal
by means of pasture or grain plays a signicant role in modifying the
concentration of antioxidants, pro-oxidants and fatty acids in the
meat. (Hernandez, Park, & Rhee, 2002; Hernandez, Zomeno, Arino, &
Blasco, 2004). Depending on the consumers' preferences meat
undergoes a wide range of cooking procedures and thermal
treatments. Compositional and structural changes during cooking
could signicantly affect the TAC of meats (Palka & Daun, 1999;
Tornberg, 2005). Therefore, a reliable estimation of the TAC value of
meat can be useful to describe the capacity of muscle to resist
oxidation processes and to determine modications in TAC values
during thermal processing.
Due to its complex structure, various extraction methodologies
involving different solvents (water, aqueous buffers, alcohols,
chloroform, etc.) have been applied to assess the TAC of meats
(Descalzo et al., 2007; Sacchetti, Di Mattia, Pittia, & Martino, 2008).
Unfortunately, extraction-based methodologies can only measure the
antioxidant capacities of the soluble and extractable hydrophilic or/
and lipophilic fractions of meat. On the other hand, independent of the
cooking process, meat contains a signicant insoluble fraction.
Moreover, various factors like the type and the pH of the extraction
solvents, molecular interactions, aggregation phenomena, denaturation and oxidation can affect the extraction capacities and lead to an
underestimation in TAC values of meat.
A robust extraction-independent procedure to measure the TAC of
solid foods, based on the interaction occurring at the interface
between the solid matrix and a liquid colored radical probe has been
developed (Serpen, Capuano, Fogliano, & Gkmen, 2007). The
method, so called QUENCHER has been applied to various food
matrices including cereals, bakery products and nuts and seeds (Aar,

A. Serpen et al. / Meat Science 90 (2012) 6065

Gkmen, Pellegrini, & Fogliano, 2009; Gkmen, Serpen, & Fogliano,

2009; Serpen, Gkmen, Pellegrini, & Fogliano, 2008). The QUENCHER
procedure in principle can be used for all food matrices and adapted to
various assays commonly used to measure antioxidant activity
(Amigo-Benavent, del Castillo, & Fogliano, 2010).
In this work, the QUENCHER procedure was used to measure the
TAC of four types of meat (beef, chicken, pork and sh) and to
evaluate changes in TAC by thermal treatment.

61

assays, but not for the FRAP assay where no dilution was necessary.
After dilution with cellulose 10 mg per sample was used, an amount
ensuring good reproducibility for high antioxidant materials.
Cellulose was found to be inert towards the various colored probes
(Serpen et al., 2007): mixing cellulose powder (10.0 mg) with each
radical probe solution no discoloration (i.e. decrease in absorbance)
was observed over the 180 min of the assay.
2.4. Preparation of ABTS +, DPPH and FRAP radical solutions

2. Materials and methods

2.1. Chemicals
All chemicals and solvents used were analytical grade. Water,
potassium persulfate (di-potassium peroxdisulfate), acetic acid
(glacial), sodium acetate and ferric chloride were purchased from
Merck (Darmstadt, Germany). Cellulose (powder from spruce) and
2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were
purchased from Fluka Chemie AG (Buchs, Switzerland) and ethanol
from Carlo Erba (Italy). 2,4,6-tris-2,4,6-tripyridyl-2-triazine (TPTZ),
1,1 diphenyl-2-picryl-hydrazyl (DPPH) and 6-hydroxy-2,5,7,8-tetramethylchroman-2 carboxylic acid (Trolox) were purchased from
Sigma-Aldrich (Steinheim, Germany).
2.2. Thermal treatments of meat samples
Fresh samples from chicken breast, pork tenderloin, beef llet and
sea bream (Sparus aurata) llets were purchased from a local market.
Proximate composition of the meats determined by AOAC (1996)
ofcial procedure is reported in Table 1. Minced meat samples were
shaped into cylinders, having a diameter of 5 cm and thickness of
1 cm, by spreading them into aluminum caps. Using this procedure,
any variation in the rate of heat transfer during cooking, which
would further affect the rate of antioxidant/pro-oxidant degradation/
formation, was limited.
Meat samples were put into a preheated and air-ventilated oven
(REX, Italy) at 180 C and cooked for 5, 10, 15 and 20 min. After
cooking, samples were immediately cooled in an ice bath and stored
at 20 C before freeze drying.
Freeze dried meat samples were ground to powder form using a
mortar and were consecutively passed through a sieve (Endecotts Test
Sieve, London, U.K.) having 50 mesh size (297 m). All raw and
cooked meat powders were stored at 20 C prior to analysis and all
analyses were performed within 3 days.
2.3. Preparation of meat samples for QUENCHER procedure
Freeze dried meat samples prepared as described above could be
directly used for the QUENCHER procedure. Due to the high
antioxidant activity (the values of absorbance were below the linear
response range of the radical discoloration/color formation) a
preliminary dilution was necessary. Dilution was performed by
mixing the freeze dried meat samples with cellulose at a ratio
between 1:1 to 1:10 (w:w) in a tube depending on the TAC values,
following by rigorously shaking or squeezing in a mortar to achieve
better homogeneity. Preliminary tests showed that a 1:5 (w:w) solidstate dilution with cellulose was suitable for both ABTS and DPPH

Table 1
Proximate composition of the raw meat samples.
Meat

Water

Protein

Lipid

Ash

Chicken (breast)
Pork (tenderloin)
Beef (tenderloin)
Fish (Sea bream, llet)

73.3 0.47
73.2 0.21
73.1 0.28
67.4 1.12

23.1 0.37
22.1 0.39
20.0 0.33
19.7 0.25

2.4 0.21
3.6 0.33
5.9 0.52
10.6 0.51

1.2 0.09
1.1 0.11
1.0 0.12
1.4 0.06

2.4.1. ABTS + solution

ABTS solution was prepared by adding 5 mL of deionized water to
38.41 mg of ABTS. Potassium persulfate solution was prepared by
mixing 5 mL of deionized water with 6.615 mg potassium persulfate.
A total of 10 mL of stock solution of ABTS + was prepared by reacting
5 mL of each solution described above which resulted in nal
concentrations of 7 mmol/L ABTS and 2.45 mmol/L potassium
persulfate. The stock solution of ABTS + was allowed to stand in the
dark at room temperature for 1216 h before use (Re et al., 1999). The
working solution of ABTS + having absorbance of 0.750.80 at
734 nm was prepared daily by diluting the 10 mL of stock ABTS +
solution with approximately 800 mL of a water/ethanol (50:50, v/v)
mixture.
2.4.2. DPPH solution
Stock solution of DPPH was prepared daily by dissolving 40 mg of
DPPH in 100 mL of ethanol. Then the ethanolic solution of DPPH was
further diluted with 100 mL of deionized water to obtain a DPPH
stock solution in ethanol/water mixture (50:50, v/v). The working
solution of DPPH having an absorbance value of 0.750.80 at 525 nm
was prepared by diluting 200 mL of stock DPPH solution with
approximately 800 mL of water/ethanol (50:50, v/v) mixture (BrandWilliams, Cuvelier, & Berset, 1995).
2.4.3. FRAP solution
FRAP solution was prepared by diluting an aqueous solution of
10 mM TPTZ and 20 mM ferric chloride in 300 mM sodium acetate
buffer (pH 3.6) at a ratio of 1:1:10 (v:v:v) as described by Benzie and
Strain (1996).
2.5. Measurement of Trolox Equivalent TAC of meats by direct
QUENCHER method
Ten (1.0) mg of powdered meat sample (diluted at a ratio of 1:5
(w:w) with cellulose for the ABTS and DPPH assays) was weighed into
a centrifuge tube. The reaction was started by adding 10 mL of
ABTS +, DPPH or FRAP working solution. The tube was shaken
rigorously for 1 min and placed on an orbital shaker in the dark. The
mixture was shaken at 300400 rpm at room temperature on the
orbital shaker until centrifugation to facilitate the surface reaction
between the solid meat particles and the ABTS +, DPPH or FRAP
solution.
15, 30, 60, 120 and 180 min after introduction of the radical
solution to the solid meat samples, centrifugation was performed at
9200 g for 2 min. The clear supernatant (2 mL) was transferred into a
cuvette and the absorbance measured at 734 nm (for ABTS assay),
525 nm (for DPPH assay) or 593 nm (for FRAP assay) at room
temperature. Measurements at ve different times (15, 30, 60, 120,
180 min) were needed to estimate the time required to reach the
plateau (i.e. the end point of the reaction).
The inhibition percentage of the ABTS + or DPPH radicals was
calculated using the following equation (Eq. (1)):


Inhibitionsample % = Absblank Abssample = Absblank 100

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A. Serpen et al. / Meat Science 90 (2012) 6065

where, Absblank is the initial absorbance of the ABTS + or DPPH

radical solution without sample and Abssample is the absorbance of the
ABTS + or DPPH radical solution having sample after t min (15, 30,
60, 120, 180 min) of incubation.
Trolox was used as a standard reference to convert the inhibition
capability of each sample to the trolox equivalent antioxidant capacity
(TEAC). First, standard trolox solutions in methanol were prepared at
a concentration range between 0 and 600 g/mL. A 0.1 mL of each
trolox solution was added to 9.9 ml of ABTS + or DPPH radical
solution (024 M trolox in radical solution). After 30 min of
incubation at room temperature, (enough time to reach a stable
value) 2 mL of the radical solution was transferred into a cuvette and
the absorbance measured at 734 nm (for ABTS assay) or 525 nm (for
DPPH assay).
Standard calibration curves were constructed by plotting percentage inhibition (Eq. (2)) against concentration of trolox at 734 nm and
525 nm for the ABTS and DPPH assays, respectively (Fig. 1a and b).
Inhibitiontrolox % = Absblank Abstrolox = Absblank 100

where, Absblank is the absorbance of the ABTS + or DPPH radical

solution without trolox (only methanol) and Abstrolox is the
absorbance of the ABTS + or DPPH radical solution with trolox
after 30 min of incubation.
The ratio between % inhibition of the sample and the slope of the
trolox calibration curve was dened as the TEAC, which was used to
indicate the scavenging free radical capability of the samples on a dry
basis by the following equation (Eq. (3)).

TEAC

mmol Trolox Eq:

kg meat d:w:


=

% inhibitionsample
 10
sm

where, s represents the slope of trolox calibration curve for ABTS

(sABTS = 4.0895) and for DPPH (sDPPH = 3.0069) and m is the sample
amount in mg dry basis, 10 is the conversion factor to obtain TEAC
values in mmol Trolox Eq./kg of meat (d.w).
Since color formation is monitored in the FRAP assay, no inhibition
percentage was calculated as in the DPPH or ABTS assays. A calibration
curve was constructed at room temperature by plotting concentration
of trolox (020 M in radical solution) against the absorbance at
593 nm (Fig. 1c). TEAP values of the samples were calculated by
(Eq. (4))
TEAP



mmol Trolox Eq:
Abs593nm n
=
 10
kg meat d:w:
sm

where, Abs593nm represents the absorbance of the FRAP solution with

the sample at 593 nm after t min (15, 30 60, 120, 180 min) of
incubation. n and s represent the intercept (0.04) and the slope
(0.0444) of the trolox calibration curve of the FRAP assay, respectively
and m indicates the sample amount in mg (dry basis), 10 is the
conversion factor to obtain TEAC values in mmol Trolox Eq./kg of meat
(d.w).
2.6. Statistical analysis
The analytical data are reported as mean standard deviation of
triplicate independent measurements and were subjected to ANOVA,
the signicance of mean differences was determined by Duncans'
posthoc test and t test using SPSS version 14.0 (2005).
3. Results and discussion
Most natural antioxidant or neo-formed antioxidants upon
processing are multifunctional, and in complex heterogeneous foods
such as meat and meat products, their activity cannot be evaluated by
a single method (Perez-Jimnez & Saura-Calixto, 2005). Two or more

Fig. 1. Calibration curve for (a) ABTS, (b) DPPH and (c) FRAP assay probes as a function
of standard trolox concentration.

radical scavenging capacity assays are required to investigate

heterogeneous samples since each assay involves different chemical
mechanism(s) and may reect different aspect(s) of their antioxidant
properties. Here three common radical probes were used, namely
ABTS, DPPH and Fe + 3 (FRAP) to assess in-vitro antioxidant activity/
power of meat in the QUENCHER procedure. Scavenging of DPPH
radical allows evaluation of the hydrogen-donating potency of
antioxidative compounds (Brand-Williams et al., 1995) while the
ABTS radical determines the single electron-transfer capabilities of

A. Serpen et al. / Meat Science 90 (2012) 6065

Antioxidant Capacity/Power,mmol Trolox Eq./kg

60
ABTS

DPPH

FRAP

50

40

30

20

10

0
0

30

60

90

120

150

180

reaction time, min

Fig. 2. Representative time antioxidant capacity/power curve of pork cooked for 15 min
at 180 C using ABTS, DPPH and Fe+ 3 (FRAP) as antioxidant assay probes.

60.0
ABTS
DPPH

50.0

TAC, mmol Trolox Eq./kg.

these (Re et al., 1999). Fe + 3 probe in FRAP assay reects the reductive
antioxidant power (Benzie & Strain, 1996).
To determine the appropriate end point for the evaluation of the
TAC of meat, the reaction with the different probes was followed for
different times. In Fig. 2 representative time-antioxidant capacity
curves obtained with the three radical solutions (ABTS, DPPH, FRAP)
for the pork samples cooked at 180 C for 15 min are reported. Similar
time-antioxidant capacity curves were obtained for all raw or cooked
meat samples studied. As shown in Fig. 2, the reaction between the
probes and the meat samples is completed within 60 min for the ABTS
and FRAP assays and within 120 min for the DPPH assay. Therefore,
60 min for the ABTS and FRAP assays and 120 min for the DPPH assay
were considered satisfactory end-points to determine the TAC of meat
by the direct QUENCHER procedure.
TAC of the samples determined by the ABTS + and DPPH radical
probes are presented in Fig. 3 and expressed in mmoles of Trolox per
kg of dry weight. The total ABTS + scavenging capacity of the raw meat
samples was in the range 25.9 1.0 mmol Trolox Eq./kg d.w. to 51.7
1.2 mmol Trolox Eq./kg (d.w). Comparing the ABTS + results of the
raw meat samples, the TAC of chicken was signicantly higher than
that of pork, beef and sh (p b 0.05), while no signicant differences
were observed when the DPPH probe was used. Considering per fresh
weight values (the average content of water in meat is 70%), the TAC
for raw meats is around 10 mmol Trolox per kg.
Different components are responsible of this TAC. It has been
demonstrated that proteins and peptides have an important antioxidant action in meats due to their ability to scavenge free radicals and
chelate prooxidative metals (Diaz & Decker, 2004; Elias, Kellerby, &
Decker, 2008; Elias, McClements, & Decker, 2007). Chan and Decker
(1994) reported that chicken meat is rich in histidine-containing
dipeptides such as carnosine and anserine, which have high
antioxidant activities. The presence of these antioxidant peptides in
chicken could account for its higher ABTS + scavenging activity.
Sacchetti et al. (2008) investigated the ABTS + scavenging
antioxidant capacities of hydrophilic and lipophilic extracts of chicken
meat by classical extraction dependent procedures. They reported the
average ABTS + scavenging antioxidant capacities of chicken meat
(breast) as 10.3 mmol Trolox Eq./kg (d.w.) and 5.3 mmol Trolox Eq./
kg (d.w.) for the hydrophilic and lipophilic extracts, respectively. This
led to a total of 15.6 mmol Trolox Eq./kg (d.w.) which is almost 70%
lower than the value obtained in present study (51.7 mmol Trolox Eq./
kg d.w.). This result, which was previously found comparing
QUENCHER with conventional extraction based procedures on other
matrixes, is likely due to the fact that the QUENCHER procedure

63

40.0

30.0

20.0

10.0

0.0
Chicken

Pork

Beef

Fish

Fig. 3. TAC of raw meats determined using ABTS and DPPH as antioxidant probes.

includes the contribution of compounds which are not solubilized

with the traditional extraction procedure.
The total DPPH scavenging capacity of the raw meat samples in
this were between 19.1 1.8 mmol Trolox Eq./kg (d.w.) and 31
0.9 mmol Trolox Eq./kg (d.w.). The DPPH scavenging capacity of
chicken, pork and beef was similar (p N 0.05) while that of sh was
signicantly lower (p b 0.05).
Interestingly, the TAC value measured with the two total radical
scavenging antioxidant capacity assays was signicantly different for
chicken and sh meats but not for raw pork and beef (p b 0.05) which
could be ascribed to the different afnities of the radicals to scavenge
the various antioxidant groups present in different samples. Dean,
Yamamoto, and Niki (1991) demonstrated that hydrophobic radicals
have less ability to attack macromolecules such as proteins in solution.
Therefore, it could be one of the reasons that DPPH, a relatively
hydrophobic radical, would have less interaction with polar macromolecular antioxidant compounds than more hydrophilic probes like
ABTS. In addition, DPPH is likely more selective than ABTS + in the
reaction with H-donors (Roginsky & Lissi, 2005) and this could
explain the low TAC values obtained by the DPPH compared with the
ABTS assay.
On the other hand, the TAC values of sh samples were the lowest
in both assays. Beside a lower content of blood in the sh sample the
loss of endogenous antioxidant compounds that may inhibit unsaturated lipid oxidation could be one of the reasons for the low total
radical scavenging antioxidant capacity of sh compared with beef,
pork and chicken.
FRAP of raw meat samples is presented in Fig. 4 expressed as
mmoles of Trolox Eq. per kg of meat (d.w.). Among the raw meat
samples, the highest FRAP value was in beef (4.9 0.2 mmol Trolox
Eq./kg d.w.) whereas the lowest level of 3.0 0.1 mmol Trolox Eq./kg
(d.w.) was that of the sh sample. The differences in FRAP values
between the raw beef and chicken were not statistically signicant
(p N 0.05). Similarly, there were no signicant differences between the
FRAP values of raw sh and pork. In general, reductive antioxidant
power of the raw samples was lower as compared to the ABTS + and
DPPH scavenging capacities. The FRAP method is based on the
reduction of the Fe + 3TPTZ complex to the ferrous form at low pH.
The low values of FRAP could be due to the poor ability of meat
antioxidants to reduce ferric ion to its ferrous form.
The inuence of thermal treatment on the TAC of the meat samples
was measured using the ABTS + and DPPH radical probes; Figs. 4 and

64

A. Serpen et al. / Meat Science 90 (2012) 6065

6.0

FRAP, mmol Trolox Eq./kg.

5.0

4.0

3.0

2.0

1.0

0.0
Chicken

Pork

Beef

Fish

Fig. 4. FRAP of raw meats.

5, respectively. The TAC of most meat samples increased on heating at

the beginning of the cooking period peaking after 510 min using
both assays. The maximum TAC values were clearly visible in all the
meat samples but not the sh. On increasing the heating time, TAC
levels started to decrease, although at the longest heating time a
further increase of TAC is seen, particularly using the ABTS probe.
The behavior on heating of proteins, which are the main
component of meat, helps explain this result. Thermal treatments
can alter the secondary and tertiary structure of proteins, resulting in
modications of their physical properties (Sante-Lhoutellier, Aubry, &
Gatellier, 2007; Tironi, Tomas, & Anon, 2002). Unfolding of the
proteins (i.e. denaturation) can increase their ability to scavenge
radicals by increasing the solvent exposure of antioxidant amino acids
that would normally be located in the core of the native protein
structure (Elias et al., 2007). Therefore, it is possible that mild heat
treatments could increase the antioxidant activity of some proteins
due to changes in their quaternary and tertiary structures.
On the other hand, it is well known that more severe thermal
treatments can cause thermoxidation of different components of

Fig. 5. Effects of time at 180 C on TAC values of meats measured using ABTS as the
antioxidant probe.

muscle foods resulting in consumption of antioxidant substances and

a decrease of TAC.
The trend of TAC in the rst minutes of heating (Figs. 5 and 6) can
be explained because in the initial period, the consumption of
antioxidant substances is less marked compared to the exposure of
hidden antioxidant amino acids due to protein denaturation.
Moreover some antioxidant compounds (for example GSH) can be
released as a consequence of cell membrane destruction, thus being
more reactive towards the radical probes.
Prolongation of the heating process reduced the TAC of meats in
both radical scavenging assays (Figs. 5 and 6). This can be due to the
accumulation of oxidized proteins and the loss in functionality of
active meat peptides. Moreover, degradation of endogenous antioxidative factors such as vitamin E, vitamin C, carotenoids, ubiquinols,
polyphenols, and cellular thiols could be promoted by heating. In this
stage, the prooxidantantioxidant balance was dominated by the
prooxidant properties.
In the last stage of the heating process, the TAC of some meat
samples showed a marked increase in both assays. This could be due
to reactions between reducing sugars and free amino acids or free
amino groups in proteins, the Maillard reaction. There have been
several reports on the antioxidative properties of MRPs (Bailey & Um,
1992; Borrelli, Visconti, Mennella, Anese, & Fogliano, 2002; Serpen et
al., 2007). Since the Maillard reaction is favored at low water activity a
signicant increase in the MRPs concentration could be achieved only
at the end of the cooking time. The highest increases in TAC levels in
the nal cooking period were observed in chicken, beef and sh
samples by the ABTS assay while the net increase was only observed
in pork samples by the DPPH assay. Low reactivity of the hydrophobic
DPPH radicals against hydrophilic MRPs could explain the differences
between the two assays.
The effects of heating on the FRAP values of meats are reported in
Fig. 7. The FRAP assay gave different results showing a sharp increase of
activity for all samples, particularly beef. All the samples produced MRPs
upon thermal treatment, but with different properties depending on
their composition of proteins and carbonyl groups.The rapid increase of
FRAP values in beef in the last heating stage might be due to the
accumulation of MRPs with higher reducing properties than those of
sh, chicken and pork.
In order to detect a possible release of free iron from myoglobin to
the matrix that would account for the increase in FRAP value during
heating, the FRAP measurements were performed without adding ferric
iron to the reaction mixture, this resulted in no color development

Fig. 6. Effects of time at 180 C on TAC values of meats measured using DPPH as the
antioxidant probe.

A. Serpen et al. / Meat Science 90 (2012) 6065

Fig. 7. Effects of time at 180 C on FRAP values of the meats.

indicating that there was no detectable free ferrous iron formed from
the reduction of iron released from the heme group. Also there were no
detectable agents in the heated meat samples that could react directly
with TPTZ to form the blue chromogen. These results agree with
previous reports regarding the relatively high stability of the heme
group to thermal treatments and the binding of released iron to proteins
(Berisha, Yasushi, & Fujimoto, 2003; Kristensen & Andersen, 1997). As a
result it is concluded that the increase of FRAP values with heating is
mainly related to the development of MRPs and does not reect the loss
of endogenous antioxidative factors during the cooking of meat.
4. Conclusions
Traditional extraction-based methodologies for TAC evaluation are
only partially useful as they take into account only the soluble and
extractable hydrophilic or/and lipophilic fractions of meat. The
QUENCHER procedure which measures the antioxidant capacity of
both soluble and insoluble parts together, gives a reliable measure of
the TAC value for raw and processed meat. The QUENCHER approach
revealed that the effect of cooking on TAC is dependent on meat type
and severity of heating. The balance between phenomena occurring
during cooking such as i) denaturation and exposure of reactive sites
of proteins; ii) thermoxidation and degradation of endogenous
antioxidants; iii) formation of antioxidant MRPs might be responsible
for the overall trend of TAC observed for different cooked meats. Using
the direct QUENCHER procedure it will be possible to build a database
and compare results for the TAC of meat samples, particularly after
very different cooking procedures.
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