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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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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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