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19
The International Conference on Sustainable Community Development27-29 January 2011
Monitoring lipid oxidation in Chicken and Processed chicken products by TBA assay
Pornpimol Muangthai1 , Panisa Makachan1 and Jaturong Jongcharoen1 1Department of Chemistry ,Faculty of Science ,Srinakharinwirot University
114 Sukhumvit 23, Bangkok 10110, Thailand
Tel (662) 649-5000 ext 8455 , Fax (662) 259-2097
E-mail: pornpi@swu.ac.th
Abstract The lipid oxidation occurs in food systems can cause
the flavor deterioration in food. The aim of this work was to
monitor the lipid oxidation in chicken and processed chicken
from boiling, steaming and frying by analysis malondialdehyde
content. The malondialdehyde was analysed by TBA technique.
The result showed that malondi- aldehyde was found in fresh
chicken about 1.05 µg/g and the steaming chicken contained
the highest content of malondialdehyde about 5.75 µg/g.
However, the effect of heating time and heating temperature
on lipid oxidation were also studied.The result showed that the
heating time for cooking chicken breast related with the MDA
content as linearity form and the heating temperature related with
the MDA content as an exponential form. This work showed
that malondialdehyde content represent the lipid oxidation in
processed chicken products depend on the method to cook
chicken and the reaction rapidly increase as exponential with
heating temperature.
Keywords: lipid oxidation , malondialdehyde , TBA assay
1. Introduction Lipid oxidation is one of the most important reaction
for deterioration of food, which affects fatty acids, particularly
polyunsaturated fatty acids(PUFA) [1-3].The autooxidation of lipid
gives many degradation products that effect on the food quality
such as colour, aroma, flavour, texture and nutritive value. [4-6].
Animal meat , especially fish contains high content of PUFA
which highly susceptible to lipid oxidation during manipula-
tion ,processing and cooking. As a consequence of oxidative
spoilage, lipid hydroperoxides are formed and decomposed to
aldehydes, ketones, alcohols, acids or hydrocarbons [7] These
are known as secondary oxidation products which change food
quality [8].
One of the most important oxidation product is
malondialdehyde (MDA), which is thought to be a carcinogenic
initiator and mutagen. MDA has often been used as marker of
oxidative damage in biological samples [9] and foods [7]. MDA
is a three carbon dialdehyde with carbonyl groups at the C-l and
C-3 positions (Fig. 1).
Figure 1. Structure of Malondialdehyde.
There are different theories about the possible mechanisms of
MDA formation[10], through hydroperoxides formed from PUFA,
with three double bonds(triene) or more, associated with phos-
pholipids, their primary substrates in animal food. MDA was
found in natural such as cotton seed, rancidity food. [10-11].
However,there are many works that refered the MDA content in
fish such as,canned fish[12], oil fish [13] ,storage sardine [14] ,
cured and uncured frozen cooked pork[15],sauage[16],infant
milk[17], milk powder [18-19][27] etc. Factors which determine
the extent and amount of MDA formation from peroxidized PUFA
are: the degree of fatty acid unsaturation [20-21]; the presence
of metals [22]; pH [23] ;the temperature and duration of heating
time[24]. Iron catalyzes fatty acid hydroperoxide decomposi-
tion to MDA at physiological pH and temperature [22]. These
studies suggest that the degradation products of fatty acid
hydroperoxides in living and post-mortem tissues may differ from
the degradation products obtained by heat and acid treatment
during the TBA test. It was found that the type of cooking (e.g.
microwaves, roasting, etc.), time and temperature affected MDA
content [25].
The aim of this work was to study the lipid oxidation
in chicken breast and processed chicken breast products by
normal cooking method such as boiling , steaming and frying.
The MDA content was an index to evaluate the lipid oxidation
reaction which was quantitative analysed by TBA method.
The reaction between MDA with TBA (2-thiobarbituric acid
(C4H4N2O2S) reagent occurs by attack of the monoenolic form
of MDA on the active methylene groups of TBA (Fig. 2).
(C4H
4N
2O
2S)
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The International Conference on Sustainable Community Development27-29 January 2011
Figure 2 . Reaction between MDA and TBA in TBA assay.
TBA pigment absorbs the visible light at 530- 535 nm
[26]. The intensity of colour was compared with the standard
calibration curve of MDA contents to evaluate MDA concentration
in samples[27]. This work also studied the effect of heating time
and heating temperature on MDA content which was no report
in any work about the relation of MDA content with heating time
and temperature in process chicken breast product.
2. Materials and methodPart 1 Sample preparation
1.1 Chicken preparation
The fresh chicken breast samples without their skin
were purchased from the poultry section in the Mall department
store in Bangkok . Those samples were also immediately
cooled at 10oC in the ice box before analysis.
1.2 MDA extraction
The fresh chicken breasts without skin were
chopped and minced to reduce the sampled size. Then a por-
tion of fresh minced chicken breast was weighed 1.00 gram
in a 50 ml centrifuge tube and diluted with 30 ml of 7.5% TCA
solution(TCA analytical grade was purchased from BDH), 0.1%
(w/v) EDTA(AR grade, purchased from BDH), 0.1% (w/v) propyl
gallate (AR grade, purchased from BDH). The above mixture was
homogenised with a Polytron PT3000 blender for 1 min at 5000
rpm and filtered through filter paper (Whatman no.1). Filtrate was
centrifuged for 10 min at 6000 rpm. Supernatant was collected
to analysis in part 3.
Part 2 Study effect of heating temperature and heating
time on MDA content
The raw fresh chicken breast without skin was boiled
in the pot by varation heating temperature at 50 oC , 80 oC and
100 oC with controlled heating time at 10 minutes.Then ,boiled
chicken breasts were minced and treated as part 1.2. The fresh
chicken breast without skin was processed again by boiling
in the pot by variation heating time at 5,10,20 and 30 minutes
with controlled heating temperature at 80 oC.
Part 3 Study effect of cooking method on MDA content
The fresh chicken breast without skin was cooked
again by boiling in the pot at 100 oC, steaming at 100 oC,frying in
the fresh new soy bean oil( Kook Brand ). All cooking methods
were treated on fresh chicken breast sample as above condition
in the laboratory room until chicken ripen as normally cooking.
All ripen chicken breasts samples were treated as 1.2.
Part 4 TBA assay
This MDA determination (method A) was performed
according to Vyncke’s methodology [28]. The TEP (1,1,3,3-
tetraethoxypropane) was purchased from Merck) and used as
the MDA standard, without hydrolysis prior to the TBA reaction.
A standard curve was made from TEP standard solution diluted
in 7.5%TCA solution, at concentrations 2x10-6- 5x10-6µM.
A 5 ml sample supernatant from part 1(1.2) part 2 and
part 3 , standard or blank was transferred into a screw-capped
tube, 5 ml of 20 mM TBA solution purchased from Sigma (USA)
was added, mixture was vigorously agitated in a vortex mixer
and placed in a boiling water bath for 60 min. After cooling,
MDA-TBA complex was measured at 530 nm using an UV-VIS
spectrophotometer.
3. Results and discussion he standard calibration curve was prepared as in
figure 3 and calculated the correlation equation to be a standard
equations for analysis MDA in sample.
Figure 3. Standard MDA calibration curve
.
From the standard calibration curve showed the
regression equation as y = 191127x - 0.0582 with R² = 0.992
.This mean that the TBA method gave the high accuracy for
measurement MDA content in sample.
The results from the effect of heating time and heating
temperature were presented as in figure 4 and 5 respectively.
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The International Conference on Sustainable Community Development27-29 January 2011
Figure 4 Effect of heating time on MDA content in boiling
chicken breast
Figure 5 Effect of heating temperature on MDA content in
boiling chicken breast
From figure 4, showed the relationship between
the heating time with MDA contents in boiling chicken breast
at constant temperature trend to be a linear relation as the
regression equation Y=0.2066x + 3.8473 with R2 0.9802.
However, the the relationship between the heating temperature
with MDA contents in boiling chicken breast at constant time
trend to be a exponential relation as the regression equation
Y=9E-07x 3.4909 with R2 0.9792 in figure 5.This means that the lipid
oxidation in chicken process product trends to rapidly increase
in cooking system as increase the temperature of heating process
more than increase heating time.
The results of MDA in raw fresh chicken and
processed chicken were presented as in figure 6
Figure 6. MDA content in fresh chicken and processed chicken
From figure 6, the result showed that fresh chicken
contained a few amount of MDA content, the steaming chicken
contained a highest MDA content approximately 5.75 µg/g.Since
the steaming process took a long time in cooking ,so chicken
must contact the heat in longer time than other process. There
have to much chance to form the lipid oxidation in the system
of steaming process. In frying process could detected the MDA
content at the average level of 5 µg/g ,so the cooking chicken by
frying also induce the lipid oxidation reaction too. The oil quality
such as freshess of oil that used in frying process also effected
on the lipid oxidation as refered by other work[25]. So, this work
can support the research work of [29] and [25]. However,MDA
is very reactive,it can reacts with aminoacids, proteins, glycogen
and other constituents in food to form products[30].
Conclusion Lipid oxidation occurs in the system of processing.
The MDA contents varied depend on the way of cooking,
however in fresh chicken breast without skin always contain
a few amount of MDA. MDA was found in the all processed
chicken breast products, especially in the steaming chicken
breast.However, heating time and heating temperature that used
in cooking process also effects on the lipid oxidation reaction in
the processing. Lipid oxidation in chicken breast and processed
product from chicken breast without skin showed the MDA
contents in lower level.This work give the information about the
lipid oxidation that occurred in chicken breast product and also
applied to measure the lipid oxidation in other part of chicken
meat such as its wings.
Acknowledgement The author grateful to the research affair of Faculty
of Science for financial support in this work.
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The International Conference on Sustainable Community Development27-29 January 2011
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