ARTICLE IN PRESS

LWT 41 (2008) 561–568

www.elsevier.com/locate/lwt

Oxidative stability of fried dough from rice flour containing

rice bran powder during storage
Chatchawan Chotimarkorna,, Nattiga Silalaib
a
Faculty of Technology and Management, Prince of Songkla University, Surat Thani Campus, P.O. Box 8, Khunthalae, Muang, Surat Thani 84100, Thailand
b
Department of Food and Nutrition Sciences, University College Cork, Cork, Ireland
Received 9 March 2007; received in revised form 28 April 2007; accepted 3 May 2007

Abstract

The aim of this study was to evaluate the efficiency of rice bran powder on lipid peroxidation inhibition of fried dough from rice flour
during storage. Rice flour dough containing rice bran powder at 5, 10 and 15 g rice bran powder/100 g mixed rice flour were fried in
soybean oil at 160 1C for 1 min and stored in dark at 60 1C for 10 days. Lipid peroxidation of fried dough was determined by change of
fatty acid compositions, oxygen absorption in vial headspace, lipid hydroperoxides, TBA values including decreasing tocopherol and
gamma-oryzanol contents during storage. Polyunsaturated fatty acid decreased rapidly in fried dough without rice bran powder, while an
increase of oxygen absorption in vial headspace, lipid hydroperoxide and TBA values were significantly lower (pp0.05) in fried dough
containing rice bran powder. In addition, tocopherol degradation was significantly lower in fried dough containing rice bran powder
during storage (pp0.05). However, there was no significant difference in gamma-oryzanol contents among fried dough during storage
(p40.05). These results demonstrated that rice bran powder shows the antioxidative effects on fried dough during storage.
r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Keywords: Fried dough; Rice bran powder; Antioxidant

1. Introduction hydroxyanisole (BHA) and butylated hydroxytoluene

Fried dough base products are popular snacks in South
East Asia and Thailand. During the process of fried dough
under deep fat frying, dough is completely surrounded by
the frying oils. Frying oils are repeatedly used at elevated
temperature in the presence of atmospheric oxygen.
Heating frying oils in the presence of oxygen led to
conversion of frying oil in volatile and non-volatile
products (Giese, 1996). Oxidative stability is a major
course of the deterioration of deep fat fried food during
storage because thermal and oxidative reactions occur in
oil during frying and also absorb in deep fat fried products
(Dobarganes, Márquez-Ruiz, & Velasco, 2000). Oxidative
stability of deep fat fried food can be improved by addition
of antioxidants to fried food or frying oil. The most
commonly used synthetic antioxidants are butylated
Corresponding author. Fax: +66 77 355 453.
E-mail address: chotimarkorn.c@hotmail.com (C. Chotimarkorn).
(BHT). However, BHA and BHT are quite volatile and
easy decomposed at high temperature (Hamama & Nawar,
1991). Recently, natural antioxidants were applied for deep
fat fried food other than synthetic antioxidants such as
tocopherols (Houhoula, Oreopoulou, & Tzia, 2003; Jaswir,
Che Man, & Kitts, 2000) due to some adverse effects from
synthetic antioxidants (Ito et al., 1986; Williams, Iatro-
poulos, & Whysner, 1999). Rice bran that are by products
of rice milling is considered a valuable domestic oil in many
countries. Rice bran powder is high nutrition (Saunders,
1990), offers benefits like lowering of blood cholesterol
(Kahlon, Chow, & Sayre, 1994), and decreases the
incidence of arthrosclerosis disease (Saunders, 1985) and
had laxative effect (Saunders, 1990). The unsaponifiable
matters of rice bran comprise sterols, higher alcohol
and gamma-oryzanol about 2% (w/w) (Nicolosi, Rogers,
Ausman, & Orthoefer, 1994). Gamma-oryzanol is ubiqui-
tous components of primary plant cell walls as well as crop
bran, and it also has some bioavailabilities (Tanaka, 1971;

0023-6438/$30.00 r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2007.05.005
 ARTICLE IN PRESS
562 C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568
Tanaka & Koto, 1975; Xu & Godber, 1999) such as
inhibition of tumor promotion (Yasukawa, Akihisa,
Kimura, Tamura, & Takido, 1998), reduction of serum
cholesterol levels (Guardiola, Codony, Addis, Rafecas, &
Boatella, 1996) and antioxidant properties in several oil
models (Nyström, Mäkinen, Lampi, & Piironen, 2005; Xu,
Hua, & Godber, 2001). Thus, rice bran powder has
potential for use in different food products. However, no
researches to expose the suitability of rice bran powder
available for fried products have been conducted.
Soy bean oil contains a high amount of essential fatty acids
(Wang, Wang, & Johnson, 2003), however, this oil is
susceptible to lipid peroxidation during deep fat frying due
to the high contents of polyunsaturated fatty acids (Sebedio,
Catte, Boudier, Prevost, & Grandgirard, 1996). Addition of
rice bran powder as the source of natural antioxidant in fried
dough might increase the oxidative stability during storage. In
this present study was performed to evaluate the efficiency of
rice bran powder on oxidative stability of fried dough from
rice flour during storage.
2. Materials and methods
2.1. Materials
Rice flour was purchased from Thai Wah Food Products
Co., Ltd. (Bangkok, Thailand). Rice bran powder was
obtained by milling rice grain in a local grinding mill
followed by sieving to separate grain from bran. Rice bran
was ground, then passed through to 177–297 mm sieve and
heated at 100 1C for 30 min to inactivate endogenous
lipases (Juliano, 1985). Gamma-oryzanol (Oryza Oil & Fat
Chemical, Co., Ltd., Aichi Pref, Japan) was kindly given by
Prof. Dr. Hideki Ushio, Department of Food Science and
Technology, Tokyo University of Marine Science and
Technology, Tokyo, Japan. Alpha, beta, gamma and delta
tocopherol standard was purchased from Eisai Co., Ltd.
(Tokyo, Japan). Fully refined and free synthetic antiox-
idants soybean oil was obtained from a local refinery
(Kamolkij Group of Companies, Bangkok, Thailand).
Cumene hydroperoxide (80%) was purchased from Sigma
Chemical Co. (St. Louis, MO, USA). 2-Thobarbituric acid
was purchased from Fluka (Buchs, Switzerland). Xylenol
orange tetrasodium salt was purchased from Riedel
deHaen (Seelze, Germany). HPLC grade methanol, acet-
ronitrile and dichloromethane were purchased from BDH
(Poole, UK). The analytical grades of chloroform,
1-propanol and 1-butanol were purchased from Merck
(Darmstadt, Germany). Others chemicals and organic
solvents used in this study were analytical grade without
further purification.
2.2. Methods
2.2.1. Frying of flour dough
Flour dough which was prepared from rice flour (70.0 g)
and distilled water (30.0 ml) was mixed in the food mixer
model LBM901 LB Family Home Appliance Co., Ltd.
(Hefei, China) in presence of rice bran powder at 5, 10 and
15 g of rice bran powder/100 g mixed rice flour. The rice
flour dough was passed pasta roller at 0.2 cm thickness.
Flat flour dough was cut into squares (1.5  1.5 cm) and
then fried. One batch of flat square shaped flour dough
(200 g) was fried in a 1500 ml of soybean oil at 160 1C for
1 min in a temperature controller electronic oil bath. Fried
dough (50.070.25 g) was placed in 500 ml brown glass
bottle and tightly sealed with plastic caps and then stored
in an hot air incubator at 6070.5 1C for 10 days.
2.2.2. Lipid extraction of fried dough from rice flour
Total lipid content of fried dough containing rice bran
powder was extracted according to method reported by
Folch, Lees, and Sloane-Stanley (1957) by using chloro-
form and methanol (2:1, v/v). The mixture of organic
solvent from extracts was completely evaporated by using
rotary evaporator N-N Series, Eyela (Tokyo, Japan) at
37 1C.
2.2.3. Determination of fatty acid composition
Accurate weight of oil (50 mg) was saponified by 1 M
NaOH in methanol and subsequently methylated with 14%
of boron trifluoride (BF3) in methanol obtaining free fatty
acid methyl esters (AOCS, 1998). The result of fatty acid
methyl esters was analyzed with a Shimadzu GC 14B gas
chromatography, equipped with a SUPELCOWAX-10
fused silica open tubular capillary column (0.25 mm
i.d.  30 m, 0.25 mm film thicknesses, Supelco, Tokyo,
Japan) and flame ionization detector. The column oven
was heal at 150 1C for 1 min and raised to 240 1C at the rate
of 1 1C per min. Helium was used as a carrier gas with the
column inlet pressure of 2 kg/cm2. The fatty acid methyl
esters were identified with reference standard GLC-68A
from Nu Check Prep (Elysain, NW, USA). Peak areas of
fatty acid methyl esters were normalized as a percentage of
total methyl esters.
2.2.4. Determination of lipid hydroperoxide
Lipid hydroperoxides were analyzed according to the
method reported by Nourooz-Zadeh, Tajaddini-Sarmadi,
& Wolff (1995). Oil samples were prepared for assay by
dissolving 100 mg of oil in 1.0 ml of n-propanol. An aliquot
of the oil solution (1.0 ml) was then mixed with xylenol
orange working solution (9.0 ml) in a centrifuge tube. This
mixture solution was incubated at room temperature for
30 min and then centrifuged at 1000g for 5 min at 5 1C. The
supernatant was used for measurement of absorbance at
560 nm with UV–vis spectrophotometer, model Lambda
EZ201 UV/vis spectrophotometer (Perkin Elmer, USA).
Results were given as mmol of cumene hydroperoxide
equiv/g of rice flour fried dough.
2.2.5. Determination of thiobarbituric acid value
Accurate weight of oil (100.0 mg) was dissolved in
1-butanol and determined according to the method of
 ARTICLE IN PRESS
C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568
AOCS Cd 19-90 (AOCS, 1995). Absorption was measured
at 530 nm with a UV–vis spectrophotometer (model
Lambda EZ201 UV/vis spectrophotometer (Perkin Elmer,
USA)) and calculated by using a formula TBA
value ¼ (50  (AB))/m; where A ¼ absorbance of the test
solution; B ¼ absorbance of the reagent blank and
m ¼ weight of the oil in test portion, g.
2.2.6. Determination of oxygen absorption in the vial
headspace during autoxidation
Oxygen absorption was determined according to method
reported by Ohshima, Fujita, and Koizumi (1993). In brief,
fried dough from rice flour was ground and accurately
weighed of fried dough 0.5 g into 40 ml volume of glass
vials, which were sealed, with teflon-lined septa. After
sealing, 0.1 ml portions of air in the headspaces of the glass
vials were collected with gastight microsyringe and
analyzed on the Shimadzu gas chromatography, model
GC3BT, equipped with a glass column (3 mm i.d.  1.7 m)
packed with molecular sieve 5A (149–177 mm, Nihon
Chromato Co., Ltd., Tokyo, Japan) and a thermal
conductivity detector. The ratio of N2 and O2 in the
headspace air was determined at the every 24 h.
2.2.7. Determination of gamma-oryzanol content in fried
dough from rice flour
Gamma-oryzanol contents in the fried dough was
measured by using reverse phase high performance liquid
chromatography (RP-HPLC) according to the method
reported by Rogers et al. (1993) with some modification.
An oil samples (100 mg) were extracted from fried dough
during storage and dissolved in 1.0 ml of n-propanol before
filtering through a syringe filter with PTFE (0.2 mm;
Acrodisc syringe filter). The HPLC consisted of an Agilent
1100 series (Palo Alto, CA, USA) including auto sampler
and column oven equipped with Hypersil ODS (4.0 
250 mm, 5 mm, Agilent Technologies, Palo Alto, CA,
USA), and variable wavelength UV–vis detector (model
G1379A) at 330 nm by using a mixture of methanol:ace-
tonitrile:dichloromethane: acetic acid (50:44:3:3 v/v/v/v) as
a mobile phase at a flow rate of 1.0 ml/min. The content of
gamma-oryzanol was calculated from peak area of gamma-
oryzanol.
2.2.8. Determination of total tocopherol and tocopherol
isomer contents
Tocopherol isomer content of fried dough was determined
by RP-HPLC according to method reported by Xu, Hua, and
Godber (2001). An oil samples (100 mg) were extracted from
fried dough containing rice bran powder during storage and
dissolved in 1.0 ml of n-propanol. Tocopherol isomers were
separated in HPLC Agilent 1100 series equipped with a
Mightysil RP-18 GP column (4.6  250 mm, 3 mm, Kanto
Chemical Co., Inc., Tokyo, Japan) and FLD G1321A
fluorescence detector operating with excitation and emission
wavelength 290 and 330 nm, respectively. The mobile phase
was methanol:acetonitrile:dichloromethane (50:44:6, v/v/v)
563
with a flow rate of 1 ml/min. Peak areas of each tocopherol
isomer were used for calculated total tocopherol and
tocopherol isomer contents.
2.2.9. Statistical analysis
The experimental data were subjected to a one-way
analysis of variance for a completely random design to
determine the least significant difference at the level of 0.05.
The data values were expressed as mean7SD (n ¼ 3).
3. Results and discussion
The main fatty acid compositions of soybean oil were
palmitic acid (11.49%), stearic acid (5.34%), oleic acid
(24.66%), linoleic acid (49.88%) and linolenic acid
(6.87%), while rice bran powder was palmitic acid
(20.93%), oleic acid (44.11%) and linoleic acid (27.82%).
The tocopherol content of soybean oil and rice bran
powder was 375.51712.75 mg/g oil and 185.23710.34 mg/g
rice bran. The gamma-oryzanol content was not found in
soybean oil while rice bran power was 1425.757160.98 mg/g
rice bran. After frying at 160 1C 1 min in soybean oil, the lipid
contents of fried dough was found in range of 325.667
32.18 mg oil/g fried dough in all treatment of studied and
there was no significant different (p40.05) of initial lipid
level. Fatty acid compositions of fried dough containing rice
bran powder were showed in Table 1. As the results, the fatty
acid composition of fried dough without rice bran powder
was similar profile to frying oil. Fried dough containing rice
bran powder had higher amount of monounsaturated and
saturated fatty acid, while it had lower amounts of
polyunsaturated fatty acid than fried dough without rice
bran powder. The lower level of polyunsaturated fatty acid in
fried dough containing higher amount of rice bran powder
would be expected to show improved the oxidative stability of
fried products. Since the oxidizability of polyunsaturated
fatty acid indicate that these values were linearly related to the
number of doubly allylic positions presence in the molecule
(Cosgrove, Church, & Pryor, 1987). However, there was no
significant difference (p40.05) in saturated, monounsatu-
rated and polyunsaturated fatty acid among groups of fried
dough before storage at 60 1C.
Changes of fatty acid compositions of fried dough
during storage at 60 1C in dark area were showed in
Table 2. In fried dough without rice bran powder, relative
content of polyunsaturated fatty decreased rapidly to
77.96%, while relative content of saturated fatty acid
increased rapidly to 172.06% after 10 days of storage.
There was a slight decrease of relative contents of
polyunsaturated fatty acid in fried dough containing 5,
10 and 15 g rice bran powder/100 g mixed rice flour to
89.17%, 91.93% and 95.68%, respectively, at the end of
storage. On the other hand, relative contents of saturated
fatty acid of fried dough containing 5, 10 and 15 g rice bran
powder/100 g mixed rice flour increased slightly to
140.09%, 125.67% and 107.78%, respectively, at the
same time. Thus, the combination of low-level initial
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Table 1
Fatty acid composition of fried dough from rice flour containing rice bran powder before storage at 60 1C

Fatty acid composition (%) Rice bran powder (g)/100 g mixed rice flour

0 5 10 15

Myristic acid (C14:0) 0.1270.02 0.2170.01 0.2570.02 0.3070.01

Palmitic acid (C16:0) 11.5270.53 11.9470.66 12.1270.75 12.3670.54
Palmitoleic acid (C16:1) 0.1370.03 0.2170.02 0.2270.01 0.3470.02
Stearic acid (C18:0) 5.4370.66 5.2170.34 4.9270.66 4.7770.64
Oleic acid (C18:1) 24.8271.08 25.7371.45 26.4271.65 26.8471.37
Linolenic acid (C18:2) 50.6471.05 49.5371.22 49.0271.34 48.2371.55
Linolenic acid (C18:3) 6.9570.54 6.6170.44 6.4270.35 6.2170.27
Arachidic acid (C20:0) 0.1370.01 0.1570.03 0.1670.02 0.1870.02
Saturated fatty acid 17.1171.32 17.5171.14 17.4571.90 17.6171.22
Monounsaturated fatty acid 24.9571.66 25.9471.34 26.6471.55 27.2870.99
Polyunsaturated fatty acid 57.4971.21 56.0471.24 55.5472.20 54.3372.41

Values are mean7SD (n ¼ 3).

Table 2
Saturated, monounsaturated and polyunsaturated fatty acid composition of fried dough from rice flour containing rice bran powder after storage at 60 1C

Rice bran powder Storage time (days) Fatty acid composition (%)
(g)/100 g mixed rice
flour Saturated fatty acid Monosaturated fatty acid Polysaturated fatty acid

0 0 17.1171.32 (100.00) 24.9571.66 (100.00) 57.4971.21 (100.00)

2 17.9971.01 (105.13) 25.3471.22 (101.56) 56.3771.32 (97.54)
4 19.6871.22 (115.02) 25.9871.45 (104.12) 54.1170.96 (94.12)
6 23.9871.03 (140.15) 26.2571.05 (105.21) 49.5371.42 (86.15)
8 26.3270.98 (153.83) 26.7870.98 (107.33) 46.6571.31 (81.14)
10 29.4471.13 (172.06) 27.4171.64 (109.86) 44.8272.01 (77.96)
5 0 17.5171.14 (100.00) 25.9471.34 (100.00) 56.0471.24 (100.00)
2 17.8170.77 (101.71) 26.1170.85 (100.65) 55.5770.99 (99.16)
4 18.7771.07 (107.20) 26.2370.65 (101.12) 54.4971.01 (97.23)
6 22.1371.03 (126.38) 26.3370.76 (103.30) 51.0370.65 (91.06)
8 23.4170.78 (133.70) 27.0070.98 (104.89) 49.0870.77 (87.58)
10 24.5370.97 (140.09) 27.1970.87 (104.82) 47.9771.12 (89.17)
10 0 17.4571.90 (100.00) 26.6471.55 (100.00) 55.5472.20 (100.00)
2 17.9970.99 (103.09) 26.7171.12 (100.26) 54.9370.67 (98.90)
4 18.5471.22 (106.25) 26.7770.99 (100.49) 54.3270.66 (97.80)
6 20.2170.65 (115.52) 26.8171.01 (100.64) 52.6170.96 (94.72)
8 21.4471.11 (122.87) 26.9070.88 (100.98) 51.2970.87 (92.35)
10 21.9370.97 (122.87) 26.9571.01 (101.16) 51.0670.88 (91.93)
15 0 17.6171.22 (100.00) 27.2870.99 (100.00) 54.9372.41 (100.00)
2 17.6271.01 (100.06) 27.1870.76 (96.63) 54.8970.87 (99.93)
4 17.7370.87 (100.68) 27.3571.02 (100.26) 54.6570.97 (99.49)
6 17.9770.77 (102.04) 27.7670.65 (101.76) 54.0371.01 (98.36)
8 18.5671.02 (105.39) 28.2170.47 (103.41) 53.0270.87 (96.52)
10 18.9870.78 (107.78) 28.6271.02 (104.91) 52.5671.24 (95.68)

Number in parenthesis is relative % of saturated, monounsaturated and polyunsaturated fatty acid base on the initial saturated, monounsaturated and
polyunsaturated fatty acid content before storage.
Values are mean7SD (n ¼ 3).

polyunsaturated fatty acid and their markedly slower rate
of decreasing polyunsaturated fatty acid relative content
during storage probably provided protection of fried
dough against lipid oxidation during storage. These results
were consistent with several previous reports showing that
contents of polyunsaturated fatty acid decrease rapidly,
whereas contents of saturated fatty acid increased during
lipid peroxidation (Warner & Mounts, 1993; Tyagi &
Vasishtha, 1996; Chung, Lee, & Choe, 2004).
Antioxidant effect of tocopherol was mainly by giving
hydrogen atom to a lipid peroxyl radical to forming lipid
hydroperoxide and tocopheryl radicals (Kamal-Eldin &
Appelqvist, 1996). It was important to understand the
interaction of rice bran powder with tocopherols, the main
 ARTICLE IN PRESS
C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568 565

antioxidants in soybean oil during storage of fried dough at
60 1C. The decrease of total tocopherol and tocopherol
isomer contents of fried dough containing 5, 10 and 15 g
rice bran powder/100 g mixed rice flour during autoxida-
tion was done to test the effect of fried dough against
autoxidation during storage. Table 3 showed the initial
total tocopherol contents of fried dough after frying; the
initial tocopherol contents were increased when increasing
the amount of rice bran powder. The lower initial total
tocopherol content were 112.6572.45 mg/g sample in fried
dough without rice bran powder, while the amounts of
initial total tocopherol contents which ranged from
120.0971.99 to 133.1272.11 mg/g fried dough increased
when increasing amount of rice bran powder in fried
dough. The increase of initial tocopherol contents of fried
dough containing rice bran corresponding to previous
report showing that rice bran is the source of tocopherols
and tocotrienols (Aguilar-Garcia, Gavino, Baragaño-Mos-
queda, Hevia, & Gavino, 2007). Consequently, fried dough
containing rice bran powder 5, 10 and 15 g rice bran
powder/100 g mixed rice flour had a much higher initial
tocopherol contents comparing with fried dough without
rice bran powder. The important tocopherol as antiox-
idants suggests that fried dough containing higher level
would be expected to exhibit greater oxidative stability.
Furthermore, tocopherol isomer content of fried dough
was also important since the previous report showed that
the different forms of tocopherols exhibit the different
antioxidant effects and the order of antioxidant effect of
tocopherol was d4g4b4a during accelerated temperature
(Madhavi, Singhal, & Kulkarni, 1996). The a, b, g and d
tocopherol isomer contents of fried dough and fried dough
containing with rice bran powder was also shown in Table
3. After frying, fried dough composed g and d forms which
accounted higher than 90% of total tocopherol contents in
fried products. The increase of rice bran powder in fried
dough caused the higher level of initial g and d forms
between 1.07 to 1.18 folds when comparing to fried dough
without rice bran powder. Decreasing total tocopherol
content of fried dough during storage was done by
measuring the residual amount of total tocopherol during
storage. Among the groups of fried dough, total tocopher-
ols in fried dough without rice bran powder degraded
substantially faster than in that containing rice bran
powder. The degradation rates of total tocopherol in fried
dough without rice bran powder were 2.13 folds, which was
faster than that in fried dough containing 15 g rice bran
powder/100 g mixed rice flour during 10 day storage. After
10-day storage, the residual concentration of total toco-
pherols was reduced to 25.41% of initial content, while the

Table 3
Decreasing tocopherol isomer contents, total tocopherol of fried dough from rice flour containing rice bran powder during storage at 60 1C for 10 day

Rice bran Storage time Tocopherol content (mg/g sample)

powder (g)/100 g (days)
mixed rice a-Toc b-Toc g-Toc d-Toc Total

0 0 5.6470.41 2.32710.11 67.6071.32 37.1870.98 112.6572.45 (100)

2 1.22710.21 2.0170.05 65.5871.54 34.5171.23 95.6271.65 (84.88)
4 0 0.9770.07 43.4471.66 27.1572.31 71.5671.11 (63.54)
6 0 0 35.3272.19 24.8671.88 60.1871.23 (53.46)
8 0 0 30.4471.09 23.3270.98 53.7671.22 (47.76)
10 0 0 11.6771.08 17.3572.01 28.6270.89 (25.41)

5 0 6.0170.33 2.4170.09 72.1172.01 39.6571.23 120.0971.99 (100)

2 2.0970.21 2.0770.05 68.6671.09 38.8371.43 111.6571.02 (92.97)
4 0 1.1270.04 65.1171.45 37.7772.03 103.4272.33 (86.12)
6 0 0 58.8772.01 32.8171.22 91.6871.98 (76.34)
8 0 0 45.7772.33 30.0071.42 75.7772.33 (63.09)
10 0 0 31.7671.99 24.4370.98 56.1971.45 (46.79)
10 0 6.3970.32 2.5570.02 76.4671.88 42.1571.43 127.7372.43 (100)
2 1.9870.33 2.3270.04 71.6572.10 40.7272.22 116.6771.89 (93.34)
4 0 1.7770.09 68.2372.12 39.8171.34 109.8172.65 (85.97)
6 0 0 59.8771.87 36.2472.56 96.117l.67 (75.24)
8 0 0 47.5472.33 32.5471.43 79.9972.15 (62.62)
10 0 0 30.6371.67 25.5671.09 56.1971.41 (43.99)
15 0 6.6670.22 2.6670.03 79.8771.65 43.9371.67 133.1272.11 (100)
2 2.0170.13 2.3270.04 76.6672.01 40.6372.22 121.6272.31 (91.09)
4 0.9870.04 1.9970.07 73.3171.98 39.2671.87 115.5471.99 (86.79
6 0 0.9870.08 69.8972.01 39.0971.55 109.9670.98 (82.60)
8 0 0 61.7771.99 32.3671.88 94.1371.31 (70.71)
10 0 0 55.3372.23 31.1172.09 86.5772.31 (65.03)

Number in parenthesis is relative % of residual total tocopherol content.

Values are mean7SD (n ¼ 3).
 ARTICLE IN PRESS
566 C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568
residual tocopherol concentration occurred between
46.79% and 65.03% of fried dough containing 5, 10 and
15 g rice bran powder/100 g mixed rice flour. These results
showed that the degradation rate of total tocopherol and
tocopherol isomer content of fried dough containing rice
bran powder tended to be slower than in fried dough
without rice bran powder at the same time of storage.
Tocopherol isomer degradation was found faster in order
of a4b4g4d when comparing to the same fried dough
groups at the same incubation time. This effectiveness of
different forms of tocopherol was in agreement of
previously observation in autoxidation of purified fish oil
triacylglycerol (Kulas & Ackman, 2001). In addition, the
tocopherol degradation during autoxidation was consistent
with previous reports of tocopherol decreasing during
autoxidation of refined tuna oil (Chotimarkorn, Ohshima,
& Ushio, 2005) and fried products in soybean oil blending
with sesame oil during storage (Chung, Lee, & Choe, 2006).
Therefore, the addition of rice bran powder for fried dough
showed the higher initial tocopherol contents of g and d
forms as well as the slower tocopherol degradation than
that without rice bran powder during storage. As a
consequence, it would be expected to exhibit higher
oxidative stability of fried dough containing rice bran
powder.
Gamma-oryzanol was a mixture of phytosteryl and
campesteryl ferulate (Xu & Godber, 1999). It has been
found that gamma-oryzanol exhibits antioxidant proper-
ties in many types of vitro model systems (Nyström et al.,
2005; Xu et al., 2001). Gamma-oryzanol contents of fried
dough containing 5, 10 and 15 g rice bran powder/100 g
mixed rice flour were 46.2772.88, 102.8574.32 and
187.5074.45 mg/g fried dough, respectively. Gamma-
oryzanol was not found in fried dough without rice bran
powder. Gamma-oryzanol contents of fried dough tended
to be stable during 10 days of storage as showed in Fig. 1.
From this result, it is clear that gamma-oryzanol was not
decreased during storage fried dough. The antioxidant
activity of gamma-oryzanol and phytosterols also was
studied by other researchers showing that the antioxidative
effects of them were lower than of tocopherol. What’s
more, the antioxidant effect of gamma-oryzanol was also
not significantly different at the lower concentration (Xu
et al., 2001). Furthermore, the combination of higher level
of initial tocopherol, slowly rate of tocopherol degradation
and the stability of gamma-oryzanol in fried dough
containing of rice bran powder during storage probably
exposed synergistic antioxidant effect and protected
tocopherol from degradation.
The oxygen absorption in vial headspace of fried dough
during storage at 60 1C was shown in Fig. 2. Oxygen
disappearance in the vial headspace is directly related to
lipid peroxidation because of oxygen reaction with poly-
unsaturated fatty acid to produce peroxy radicals and lipid
hydroperoxides (Jung, Yoon, & Min, 1989). From this
result, oxygen absorption in vial headspace of fried dough
containing rice bran powder was significantly lower
Fig. 1. Gamma-oryzanol contents of fried dough containing rice bran
powder during storage at 60 1C. Values are mean7SD (n ¼ 3). K: 0 g rice
bran powder/100 g mixed rice flour, J: 5 g rice bran powder/100 g mixed
rice flour, .: 10 g rice bran powder/100 g mixed rice flour, ,: 15 g rice
bran powder/100 g mixed rice flour.
(pp0.05) than that of fried dough without rice bran
powder during 10-day storage. The oxygen absorption of
fried dough containing rice bran powder increased slightly
during 4-day storage, while there was a rapid increase of
oxygen absorption in fried dough without rice bran powder
after 2-day storage. These results indicated that rice bran
powder prolonged initiation period of lipid peroxidation
during storage at 60 1C by decreasing the oxygen absorp-
tion to fried products for forming lipid hydroperoxides. At
the end of 10-day storage, the oxygen absorption of fried
dough without rice bran powder increased 2.06 folds when
comparing with fried dough containing 15 g rice bran
powder/100 g mixed rice flour.
The antioxidant effects of rice bran powder in fried
dough during storage were directly investigated by
measuring the primary and secondary lipid peroxidation
products of fried dough during autoxidation. Initial lipid
hydroperoxides and TBA value were not significantly
different (p40.05) among groups of fried dough after
frying in soybean oil for 1 min at 160 1C. Lipid hydroper-
oxide formation and TBA values of fried dough containing
rice bran powder tended to be significantly lower (pp0.05)
than those of fried dough without rice bran powder. Lipid
hydroperoxide of fried dough without rice bran powder
increased significantly (pp0.05) from 2 days and increased
rapidly to 14.8171.01 mmol/g fried dough at the 6-day
storage (Fig. 3). Formation of lipid hydroperoxides in fried
dough without rice bran powder was very rapid with no
apparent of induction period. The addition of rice bran
powder in fried dough retarded the formation of lipid
hydroperoxide at all level of addition. Furthermore,
addition of rice bran powder in fried dough had no
significant effect on the lipid hydroperoxide formation at
day 4 of storage. Thus, induction period of lipid peroxida-
tion of fried dough containing rice bran powder was
 ARTICLE IN PRESS
C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568
Fig. 2. Oxygen absorption in vial headspace of fried dough containing rice
bran powder during storage at 60 1C. Values are mean7SD (n ¼ 3). K:
0 g rice bran powder/100 g mixed rice flour, J: 5 g rice bran powder/100 g
mixed rice flour, .: 10 g rice bran powder/100 g mixed rice flour, ,: 15 g
rice bran powder/100 g mixed rice flour.
Fig. 3. Lipid hydroperoxide contents of fried dough containing rice bran
powder during storage at 60 1C. Values are mean7SD (n ¼ 3). K: 0 g rice
bran powder/100 g mixed rice flour, J: 5 g rice bran powder/100 g mixed
rice flour, .: 10 g rice bran powder/100 g mixed rice flour, ,: 15 g rice
bran powder/100 g mixed rice flour.
extended about 4 days of this investigation. During
autoxidation, tocopherol was gradually consumed during
the incubation period, thereafter absorbable oil in fried
dough more rapidly to oxidize and the end of the induction
period can be determined by a dramatic increase in lipid
hydroperoxide. The tocopherol degradation, oxygen ab-
sorption and lipid hydroperoxide formation of the addition
of 5, 10 and 15 g rice bran powder/100 g mixed rice flour for
fried dough were probably exerted antioxidant effect of rice
bran powder on fried dough during storage.
Secondary lipid peroxidation products of fried dough
during storage were determined by examining TBA values.
567
Fig. 4. TBA value of fried dough containing rice bran powder during
storage at 60 1C. Values are mean7SD (n ¼ 3). K: 0 g rice bran powder/
100 g mixed rice flour, J: 5 g rice bran powder/100 g mixed rice flour, .:
10 g rice bran powder/100 g mixed rice flour, ,: 15 g rice bran powder/
100 g mixed rice flour.
Determination of TBA values was based on color intensity of
the reaction of TBA reagent and secondary oxidation
products, mainly malondialdehyde (MA). The formation of
TBA values during storage of fried dough was shown in
Fig. 4. TBA values of fried dough without rice bran powder
were significantly higher (pp0.05) than those of fried dough
containing rice bran powder from 4 days and it increased
rapidly between 6 and 10 days of storage as 11.6570.61 and
15.8870.59, respectively. The formation of secondary lipid
peroxidation products was supported by previously reports
(Bittner et al., 2002; Gal, Pinchuk, & Lichtenberg, 2003)
showing that after the rate of lipid hydroperoxide decom-
position exceeded to the rate of their production, the
secondary lipid peroxidation products such as aldehyde
increased rapidly. These results suggested that the lipid
hydroperoxides formation of fried dough without rice bran
powder were exceeded to their decomposition after 6-day
storage and there was lower TBA values in fried dough with
rice bran powder than in that without rice bran powder,
which may be due to the presence of higher amount of
tocopherol and gamma-oryzanol in fried dough. At the end
of 10-day storage, TBA values of fried dough without rice
bran powder increased 4.21 folds of fried dough containing
15 g rice bran powder/100 g mixed rice flour.
These investigations could be inferred that rice bran
powder showed excellent performance and it is useful for
protecting fried food against oxidation during storage
period. Moreover, rice bran powder probably acts as
potential source of natural antioxidants such as tocopherol
and gamma-oryzanol.
Acknowledgments
This work supported in part by a Grant-in-Aid from the
Faculty of Technology and Management, Prince of
 ARTICLE IN PRESS
568 C. Chotimarkorn, N. Silalai / LWT 41 (2008) 561–568
Songkla University through Contact No. T50/087, T50/089
and T50/091.
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