Delineating the microbial and physicalchemical changes during storage of ozone

treated wheat our

Man Li, Jing Peng, Ke-Xue Zhu , Xiao-Na Guo, Miao Zhang, Wei Peng, Hui-Ming Zhou
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, Jiangsu Province, PR China
a b s t r a c t a r t i c l e i n f o
Article history:
Received 5 March 2013
Accepted 12 June 2013
Editor Proof Receive Date 10 July 2013
Keywords:
Wheat our
Ozone treatment
Storage
Microbial change
Physicochemical property
Shelf-life
In this work, effects of ozone treatment on the inactivation of microorganisms and physicochemical changes
in wheat our during the storage were evaluated. Total plate count (TPC) in wheat our was found to be
slightly reduced immediately after ozone treatment, but then signicantly (P b 0.05) decreased in the rst
few days' of storage. Ozone had more impact on pH value and a
w
than fatty acid value and water content
of wheat our. PPO activity in ozone treated our was signicantly inhibited and got slightly lower during
storage. Ozone treated our presented reduced pasting temperature and increased peak viscosity during
storage, and no obvious changes were detected in the major volatile compounds as shown by the results of
GCMS. In addition, fresh noodles were made to evaluate the shelf-life extending effect as induced by ozone
and noodles made from treated our with 4 days' storage presented a better microbial and color stability.
Industrial relevance: This paper presents a new discovery of the non-immediately lethal effect of moderate
ozone treatment on the microorganisms in wheat our, which also indicates a potential for ozone as the alter-
native to chemical oxidants in our and eliminates the potential residue of hazardous chemicals. Some phys-
icochemical properties and the sensory qualities can be also improved. The discovery of this phenomenon
would provide a new concept for low-bacteria wheat our production and have important consequences
for the application of ozone in food industry.
2013 Elsevier Ltd. All rights reserved.
1. Introduction
Wheat is one of the most important cereal grains in the world, with
a production of nearly 600 million metric tons (MMT) in 2000 (USDA/
NASS, 2001). It is used for food (67%), feed (20%), and seed (7%)
(Sayaslan, Seib, & Chung, 2006). The most common utilization of
wheat is to be ground into ours with different rening degrees.
Wheat our is unique in its ability to produce a diverse array of food
products, including staple foods such as bread, pasta, and Asian noodles
(de Vasconcelos et al., 2013; Liyana-Pathirana & Shahidi, 2005). It also
provides the principal source of energy, protein and dietary ber for a
major portion of the world population (Abdel-Aal & Hucl, 2002).
Traditional our production process may include cleaning, con-
ditioning, breaking and sieving, etc. (Berghofer, Hocking, Miskelly, &
Jansson, 2003; Fistes & Tanovic, 2006). The exposed environment of
wheat growth and harvest imparts a lot of impurities to wheat grains
including microorganisms and bran specks, these microorganisms will
get further growth and reproduction during the conditioning and sub-
sequent processes, thus inducing a high bacteria quantity in the newly
produced our (Berghofer et al., 2003). Due to the limited water con-
tent (b14%), wheat our itself is a long term storage product because
it is not an ideal environment for microbial growth. However, as the
case may be, it is processed into food products, water is added and
sometimes partly or even totally reserved in them; thus the microor-
ganisms will proliferate immediately and lead to the degeneration of
the nal product. The high microorganism quantity in wheat our has
been recognized as one of the most important reasons leading to the
short shelf-life of such wheat-based food products.
Wheat our has such a peculiar matrix with quite small spaces
between our particles, lower water content and poor thermal con-
ductivity, thus it is difcult for the application of traditional steriliza-
tion methods such as thermal treatment, ultraviolet and microwave
irradiation (Li et al., 2012). Therefore, how to effectively inactivate the
bacteria cells in wheat our remains a concern in this industry, which
seriously affects the microbiological shelf-life of the end-products.
Ozone has been used for years in disinfecting water for drinking
purpose due to its potential oxidizing capacity. This powerful oxidant
has made it possible to destroy microorganisms and organic impuri-
ties. Recognized by the US as GRAS (Generally Recognized as Safe)
in 1997, this product has been used more and more often in the
food industry (Guzel-Seydim, Greene, & Seydim, 2004). It may have
many advantages in the food industry, such as decomposing rapidly
into molecular oxygen, and leaving no residue as well as can be gen-
erated on site (Sandhu, Manthey, & Simsek, 2011). The current use of
ozone in the food industry includes food surface hygiene, sanitation
of food plant equipment, reuse of waste water, and as an alternative
Innovative Food Science and Emerging Technologies 20 (2013) 223229
Corresponding authors. Tel./fax: +86 510 85919389.
E-mail addresses: kxzhu@jiangnan.edu.cn (K.-X. Zhu), hmzhou@jiangnan.edu.cn
(H.-M. Zhou).
1466-8564/$ see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.ifset.2013.06.004
Contents lists available at ScienceDirect
Innovative Food Science and Emerging Technologies
j our nal homepage: www. el sevi er . com/ l ocat e/ i f set
to methyl bromide as a fumigant in grain and our storage (Graham,
1997). There have been numerous application areas of ozone in the
industry but limited research has been performed on wheat and
wheat-based products (Sandhu et al., 2011).
The present study was conducted to determine the effect of ozone
treatment on the mortality of microorganisms and physicochemical
properties in wheat our, and the most important was to investigate
their changes during storage, aiming to provide a new concept for
low-bacteria wheat our production and prolong the shelf-life of
end our products.
2. Materials and methods
2.1. Materials and proximate analysis
High protein white wheat our was manufactured by China Oil &
Foodstuffs Corporation (used for ramen, fresh noodles and dumplings),
the contents of moisture, ash, and protein in which were 13.5%, 0.53%,
and 12.6%, respectively. All drugs and reagents used were of analytical
grade.
Moisture, protein, and ash contents of wheat our were deter-
mined according to approved methods of AACC (2000). Water activity
was determined using a Novasina Thermoconstanter model LabSwift-a
w
(Lucerne, Switzerland) according to the manufacturer's instructions.
Particle size distribution was analyzed using a laser particle size tes-
ter (Model S3500, Microtrac, USA).
2.2. Ozone treatment
The ozone gas treatment was conducted using an ozone generator
(Model OJ-8003k-A, Guangzhou, China) with a production of 5 g/h.
Wheat our was placed in an automatic revolving evaporation bottle
to ensure a homogeneous contact between our and O
3
during the
reaction and treated for 5 min, 10 min, 15 min, 20 min and 30 min,
respectively. The airow rate was set at 5 L/min. Two silicone hoses
were connected to the bottle with one for importing the ozone gas and
the other for exhausting the waste gas to the outside. For each treating
time, two single our samples were exposed for the next analysis.
2.3. Determination of TPC
Flour samples were analyzed periodically during the storage time.
Total plate count (TPC) was examined according to GB/T 4789-2008
(Code of National Standard of China, 2008). The sample (25 g) was
put into 225 mL 0.85% aseptic physiological saline and the mixture
was shaken in a stomacher bag using a stomacher machine (Lab-
blender 400, Seward Laboratory) for 60 s. Serial dilutions were then
prepared using 0.85% aseptic physiological saline and 1 mL of the
appropriate dilutions were poured and plated onto sterile plate count
agar (PCA) plates. The plates were then incubated at 37 C for 48
2 h before enumeration.
2.4. Determination of pH value
According to Li, Zhu, Guo, Peng, and Zhou (2011), the our sample
(10 g) was homogenized in 90 mL of distilled water in a stomacher
bag using a stomacher machine (Lab-blender 400, Seward Laboratory)
for 60 s to get a uniform mixture. The pH value was then measured
using the pH meter (FE20, Mettler Toledo, USA).
2.5. Determination of fatty acid value
To determine the impact of ozone exposure on the fat components
in wheat our, fatty acid value was evaluated during the storage based
on the method described by Rose, Ogden, Dunn, and Pike (2008). 10 g
of our was added to a 150 mL Erlenmeyer ask, and 50 mL of absolute
ethanol was added. The asks were placed on an orbital shaker (Model
SHA-B, Ronghua Instrument Company, Shanghai, China) for 10 min at
140 rpm. After shaking, the mixture was decanted through lter paper
(Whatman No.1) into a 50 mL colorimetric cylinder. Then 25 mL of
the ltrate was transferred into another 150 mL Erlenmeyer and
20 mL double-distilled water was added to precipitate alcohol solu-
ble proteins in the system. After that the mixture was titrated using
0.1 M KOHethanol solution with phenolphthalein as an indicator.
The nal results were expressed in terms of milligrams of KOH
required to neutralize the free fatty acid contained in 100 g wheat
our.
2.6. Particle size analysis
The particle size distributions of ozone treated or untreated wheat
our were measuredby the Laser Particle Size Analyzer (S3500, Microtrac
Inc., USA). The measurements were triplicates and data were tted by
Origin (version 8.0).
2.7. Color measurement of wheat our and noodle sheets
A Chroma Meter (Konica Minolta CR-100, Japan) equipped with a
D65 illuminant, using the CIE 1976 L*, a* and b* color scale was used
to measure the color of the wheat our samples and fresh noodle
sheets. L* is a measurement of brightness (0100), a* represents the
redgreen coordinates ( is green while + is red) while b* measures
the blueyellow coordinates (is blue with + indicating yellowness)
of a product. Flour samples were equally weighed into the matched
powder testing box for measuring while noodle sheet samples were
cut into pieces of 5 cm in diameter and measured within 5 min.
2.8. Viscosity analysis
Pasting properties of control wheat our, 15 min ozone exposed
our (ozone 0 day) and 15 min treated samples stored for 4 days
(ozone 4 day) were determined with a Rapid Visco Analyser (RVA,
Model Super-3, Newport Scientic, Warriewood, Australia), according
to AACC method 76-21 (AACC, 2000). Suspensions were made using
pure deionized water (25 g) and the mixed our (3.48 g) and manually
homogenized using the plastic paddle right before the RVA test, and
then the tests were conducted in a programmed heating and cooling
cycle.
2.9. Polyphenol oxidase (PPO) activity analysis
The PPO for the test was extracted from wheat our according to Li
et al. (2012). 2 g of wheat our was incubated in 10 mL of extraction
buffer (0.1 M phosphatecitric acid buffer, pH 6.0) in a 15 mL centri-
fuge tube and shaken at 4 C for 12 h, followed by centrifuging at
10,000 rpm for 15 min and the supernatant was used as the crude ex-
tracts of PPO. Polyphenol oxidase activity was determinedby measuring
the absorbance at 420 nm (A 420) using a spectrophotometer (Unocal
UV-2800, USA). The PPO activity was calculated as the difference in
the absorbance of sample and control and expressed as 420/ming
our.
2.10. Volatile compound analysis
According to Li et al. (2011), volatile compounds of our samples
were extracted and collected by headspace solid phase microextraction
(HS-SPME). Two gram of sample was hermetically sealed in a 15 mL
vial. A well conditioned SPME ber (CARPDMS) (Supelco, Bellefonte,
PA, USA) was used to extract the volatiles at 55 C for 30 min. After-
wards, the HS-SPME ber was inserted into the injection port of the
gas chromatographymass spectrometry (GCMS) system for thermal
desorption.
224 M. Li et al. / Innovative Food Science and Emerging Technologies 20 (2013) 223229
GCMS consists of a Trace GC and a Trace MS (Finnigan Trace
GCMS, Finnigan, USA) equipped with a DB-WAX column (30 m
0.25 mm 0.25 m, J&W Scientic, Folsom, CA, USA). The injector
port was heated to 230 C, an initial temperature was set at 40 C
for 3 min and increased to 80 C at 5 C min
1
, then to 230 C at
10 C min
1
and held for 7 min. The carrier gas was helium at a ow
rate of 0.8 mL min
1
. The mass spectra were obtained using a mass
selective detector working in an electron impact mode at 70 Ev, with
an ion source temperature of 200 C and a scan range of 33450 amu.
Compounds were identied by comparing their mass spectra with
those contained in the NIST05 (National Institute of Standards and
Technology, Gaithersburg) library and/or Wiley library. The results are
reported as relative abundance expressed as total area counts.
2.11. Preparation of fresh noodles
The noodle formula consisted of 100 g of our and 34 mL of distilled
water. Noodle dough was formed using a Kitchen Aid Mixer (Kitchen
Aid, St. Joseph, MI). The prepared dough was placed to rest in a plastic
bag for 20 min. Then, the dough crumbles were passed through a
small noodle machine (Model JMTD-168/140, Beijing, China) for 68
times with the roller gap reduced gradually, to get dough sheets. The
dimensions of the resultant noodle strands were 1 mm in width and
0.9 mm in thickness.
2.12. Statistical analysis
The data obtained in this study were expressed as the mean of at
least three replicate determinations, SPSS statistical software (version
16.0, SPSS Inc., Chicago, IL, USA) was used to verify signicant differences
of evaluated parameters among samples with or without ozone treat-
ment and during storage by one-way-analysis of variance (ANOVA).
P b 0.05 was considered to be signicant.
3. Results and discussion
3.1. Lethal effects on microorganisms during storage
As a strong oxidizing agent, ozone has long been used in food indus-
try; it has non-thermal, lethal effects on microorganisms due to cell
membrane permeabilization and damage (Patil, Bourke, Frias, Tiwari,
& Cullen, 2009). Fig. 1 shows the total plate count (TPC) changes in
ozone treated (untreated) wheat our during the storage period. With
the increase of treating time, TPC in wheat our decreased signicantly
but not absolutely; however, most interestingly, although microorgan-
isms in wheat our could not be mostly lethal even when exposed for
30 min, during the storage, TPC decreased rapidly with the increase of
storage time, especially for samples treated for more than 10 min, TPC
of 15 min treated samples was reduced from 8933 CFU/g of the control
to 7350 CFU/g at 0 day and then to 1500 CFU/g after 4 days and kept
stable during the next storage time. Although there was no difference
in logarithm, the decrease in TPC was signicant. This was mainly due
to the lower logarithmic base in wheat our as compared to other per-
ishable foods. These results indicated that ozone treatment may not
result in an immediately lethal effect on the microorganisms in wheat
our, that might be due to organic compounds such as starch and pro-
teins in wheat our competing with microorganisms to react with
ozone, which thus reduced its sterilizing effect, just like the situation
reported by Jaeger, Schulz, Karapetkov and Knorr (2009) for the pro-
tective effect of milk constituents on Lactobacillus rhamnosus during
PEF inactivation. However, it caused irreparable damage on the liv-
ing cells thus they were nally dissolved and put to death. In the
present study, the cells injured by ozone seemed to be susceptible
to the low-moisture and low water activity in wheat our and there-
fore decreased signicantly during the storage. The discovery of this
phenomenon would have important consequences for the applica-
tion of ozone in our production industry. In addition, microorgan-
isms in ozone treated our may have the potential to be further
decreased while combined with unsuitable storage conditions such
as cold temperature, which is being conducted in our present work.
However, the underlying causes are still not clear at present, and fur-
ther clarication of mechanism of the phenomenon is still needed
from a molecular level study.
3.2. Changes in pH value and free fatty acid value during storage
Due to the strong oxidizing properties, ozone treatment may cause
some physico-chemical quality changes in wheat our as a result of
the oxidation reactions of starch, protein and fat components (Li et al.,
2012; Sandhu, Manthey, & Simsek, 2012). Fig. 2 shows the free fatty
acid (a) and pH value (b) changes in our samples with or without
ozone treatment during storage. As can be seen, fatty acid value did
not increase immediately after ozone treatment as expected, instead
they got decreased slightly but not signicantly, and thenkept relatively
stable during the whole storage time, expect for samples exposed for
20 min and 30 min in ozone gas. Fatty acid values of the 5 min,
10 min and 15 min treated ours were even lower than that of the con-
trol at the end of storage. This unexpected result indicated that changes
in other components seemed to induce a protective effect on the small
amount of fat present in wheat our. Speculation concerning the mech-
anism underlying this phenomenon may assert that ozone induced a
lethal effect on the microorganisms and common insect (pest) larvae
which existed in wheat our, as well as an inhibiting effect on some hy-
drolases, which were considered as the leading cause of fat oxidation
and decomposition (Tilton, Brower, & Cogburn, 1974; Wang, Wang, &
Wen, 2007). Thus ozone treatment at a decent level could slow the
0
2000
4000
6000
8000
10000
12000
14000
0 2 4 6 10 40
Storage time/day
T
P
C
/
C
F
U
/
g
0min
5min
10min
15min
20min
30min
Fig. 1. TPC changes in wheat our exposed to different times in ozone during storage time.
225 M. Li et al. / Innovative Food Science and Emerging Technologies 20 (2013) 223229
increase of fatty acidvalue. Inaddition, it was reportedthat the free fatty
acid may also participate in the oxidation reaction and therefore result
in a lower extractable fatty acid value (Olpin, 2004). However, excessive
ozone treatment would be liable to directly affect the fat components in
our, leading to more decomposition and higher fatty acid values.
In contrast, changes in pH value induced by ozone treatment were
more signicant and decreased with the increase of treating time
(from 6.04 for the control to 5.78 and 5.34 for 15 min and 30 min
samples, respectively). In addition, pH value of ozone treated samples
further decreased during the storage, with the 20 and 30 min samples
changing more rapidly than others, especially after 4 days. Changes in
pH value might be due to the variation in amino acid composition and
starch hydroxyl groups as affected by ozone treatment. This indicated
that ozone treatment generally showed more signicant inuence on
the water-soluble acid than alcohol-soluble acid in wheat our.
3.3. Water content and water activity changes during storage
Water content and water activity (a
w
) changes after ozone treat-
ment and during the storage were investigated; samples exposed
for 15 min in ozone were selected for the analysis due to their better
microbial and storage stability. As shown in Fig. 3, water content of
our samples slightly and not signicantly decreased initially after
ozone treatment, due to the ventilation during the exposure. More
remarkably, signicant decrease was observed in a
w
of the ozone
treated samples, this was mainly due to the more compact interaction
of water and other inner components caused by ozone treatment.
In addition, during the 35 days' storage, no signicant changes were
detected in water content while the a
w
kept progressively decreasing
with the increase of storage time, especially on the rst few days,
with ozone treated samples kept more lower than the control during
the whole period. These changes in a
w
may account for at least part of
the reasons of microbial decrease in ozone treated our during the
storage time. However, as decrease of a
w
during storage was also
detected in control our samples, the microbial inhibiting effect of
reducing a
w
was recognized as negligible.
3.4. Size distribution changes
Immediately after exposure to ozone, wheat our was imparted
with poor dispersibility in water. Therefore, particle size distribution
of these samples was determined to analyze the reasons underlying
this phenomenon. The curves in Fig. 4 showthat the size distribution
of samples immediately after ozone treatment (ozone 0 day) was
partly different fromthe control, the main size peak was right shifted
and the mean volume diameter of the particles increased from
70.36 0.24 m to 75.19 0.19 m (Table 2). However, after 4 days'
storage, size distribution curve was again similar to that of the control
and the mean diameter was not signicantly different from the. Some
physical connections might be induced between the particles immedi-
ately after ozone treatment due to the oxidation of our components,
Storage time/day
m
g

K
O
H
/
1
0
0
g

f
l
o
u
r
0min
5min
10min
15min
20min
30min
30
35
40
45
50
5 55
60
0 3 6 12 35
4
4.5
5
5.5
6
6.5
7
0 2 4 10 35
Storage time/day
p
H

v
a
l
u
e
0min
5min
10min
15min
20min
30min
a
b
Fig. 2. Changes of fatty acid and pH values of ozone treated wheat our during storage; (a): Changes in fatty acid value; (b): Changes in pH value.
C
O
C
O
13.1
13.2
13.3
13.4
13.5
13.6
13.7
13.8
13.9
14
14.1
0 2 4 10 20 30 35
Time (day)
W
a
t
e
r

c
o
n
t
e
n
t

(
%
)
0.56
0.57
0.58
0.59
0.6
0.61
0.62
0.63
0.64
0.65
a
w
Water content
Water activity
Fig. 3. Changes in water content and water activity of ozone treated our during the
storage time; C: control; O: ozone treated samples.
226 M. Li et al. / Innovative Food Science and Emerging Technologies 20 (2013) 223229
thus they were more difcult to be separated by the air blower of the
Particle Size Analyzer. The connection of these our particles may also
induce an increased supercial area and make them able to oat on
the surface of water, leading to poor dispersibility. In addition, these
connections would disperse automatically after several days' storage.
3.5. Effect of ozone treatment on PPO activity, color, and viscosity
properties of wheat our
The color of wheat our is an important trait that strongly inuences
consumer acceptance of the our itself as well as its end-products (Li et
al., 2011). In this study, ozone treated (15 min) wheat our showed sig-
nicantly increased L* value and decreased b* value (Table 1), indicating
a white and bright appearance for these samples. This was mainly due to
the decomposition of the naturally existing yellow pigments caused by
ozone, showing that ozone could be used as a potential alternative to
chemical bleaching agent in wheat our such as benzoyl peroxide and
chlorine (Chittrakorn, 2004; Li et al., 2012). After four days' storage, no
signicant changes were detected in L* value while b* value presented
a slight increase and then kept relatively constant during the next days
(data not shown). Changes in wheat our color may impart a more com-
mercially acceptable appearance to wheat products such as white salted
noodles, white bread, and Chinese steamed bread.
Polyphenol oxidase (PPO) has long been considered as a predomi-
nant factor leading to the discoloration in raw Asian noodles and
other wheat products (Kruger, Hatcher, & DePauw, 1994). Darkened
wheat products were claimed to be unacceptable by consumers, thus
there has been a considerable effort to reduce levels of PPO activity in
wheat our (Fuerst, Anderson, & Morris, 2006). As shown in Table 1,
PPO activity was signicantly inhibited after exposed to ozone gas for
15 min, and it further decreased slightly after four days' storage.
Results in Table 1 also showed that swelling power of ozone treated
our increased signicantly and presented negligible changes during
the storage. In addition, ozone also induced signicant changes in the
viscosity properties of wheat our, the exposed samples generally had
a much higher and steeper RVA viscosity prole than the control
(curves not shown), with lower pasting temperature, higher peak vis-
cosity, through viscosity and nal viscosity. The higher peak viscosities
and lower pasting temperatures indicated larger water binding capaci-
ties of ozone treated our samples (Wood, 2009), which were much
easier for gelatinization. These results implied complicated changes in
the starch component in wheat our as affected by ozone treatment.
3.6. Volatile compound identication
Ozone treatment may impart some unpleasant smell to wheat our
due to the unique odor itself (Li et al., 2012), which will increase with
the increasing treating time. In this study, 15 min treated samples
were claimed to be acceptable by the panelists (not measured with ob-
jective data) and this odor will become weaker after kept for several
days as ozone breaks down. To identify the main compounds associated
with the odor changes induced by ozone treatment, both ozone 0 day
and ozone 4 day samples (15 min exposed) were analyzed by GC/MS,
as compared to the control.
Volatile aldehydes and some alkanes were found to be the major
compounds in untreated wheat our (the control), nonanal and
2-nonenal contributed to the largest proportion and followed by
hexanal, tetradecanoic acid, decanal and docosane. As shown in
Table 2, after ozone treatment, more kinds of aldehydes and alkanes
were observed in the wheat our, which increased the total kinds of
volatile compounds collected in the samples. However, no obvious
changes in the types of major volatile compounds were detected in
ozone treated samples, both after 0 day and 4 days' storage. Compared
to the control, relative contents of some detected compounds in ozone
treated samples changed to a certain extent, nonanal was still found
to be the leading compound while the proportion of hexanal increased
and exceeded that of 2-nonenal. These results indicated that 15 min ex-
posure to ozone will not change the major volatile compounds in wheat
our and the slight odor changes were more likely due to the propor-
tion variations of the detected components.
3.7. Microbial growth and color change in resultant fresh noodle samples
In order to ascertain the applicability of ozone treated wheat
our in extending the shelf-life of the end products, fresh noodles
made from 15 min ozone treated wheat our (0 day and 4 day stor-
age) were used to analyze the color change and microbial growth as
compared to the control, results were presented in Fig. 5a and b.
-5
0
5
10
15
20
25
0 10 20 30 40 50 60
Size(m)
%
C
h
a
n
Control
Ozone 0 day
Ozone 4 day
Fig. 4. Particle size distribution of ozone treated (untreated) samples after 0 day and 4 day storage.
Table 1
Physico-chemical property changes in ozone treated wheat our after 0 day and 4 days'
storage.
Quality parameters Samples
Control Ozone 0 day Ozone 4 day
PPO activity (A/ming) 0.034 0.001
a
0.022 0.002
b
0.020 0.00
b
L* 92.70 0.05
a
93.01 0.15
b
92.99 0.09
b
Color a* 0.13 0.01
a
0.13 0.03
b
0.01 0.02
c
b* 8.85 0.09
a
6.91 0.04
b
7.66 0.06
c
MV (m) 70.36 0.24
a
75.19 0.19
b
71.04 0.08
a
Peak viscosity 2301 0.0
a
2404 29.7
a
2725 75.0
b
RVA
viscosity
Pasting
temperature
70.6 0.64
a
67.38 0.46
b
67.43 0.60
b
Through
viscosity
1540.5 7.8
a
1655.5 57.3
a
1759.5 40.3
b
Final viscosity 2646 29.7
a
2772 70.7
a
3087 30.4
b
Swelling power (%) 8.90 0.14
a
9.35 0.17
b
9.32 0.04
b
a
Values are presented as means standard deviations.
b
Mean values followed by the same letter in the same rows are not signicantly
different at P b 0.05 level.
227 M. Li et al. / Innovative Food Science and Emerging Technologies 20 (2013) 223229
Firstly, initial microbe quantity in the control, ozone 0 day, and ozone
4 day noodle samples was 9633 CFU/g, 6000 CFU/g, and 3500 CFU/g,
respectively. These differences were mainly due to the TPC reduction
in raw materials. Then, as can be seen, control noodles deteriorated
immediately at the second day, with a TPC value over 7 lg CFU/g; in
contrast, microbial growth in noodles made from ozone treated wheat
our was signicantly inhibited, with ozone 4 day samples showing
a more remarkable microbial stability than others. This indicated that
the reduced microbe quantity in wheat our could surely extend the
microbiological shelf-life of the end products.
Inaddition to microbial growth, color is consideredas another major
determinant of the marketability of wheat products, especially for those
who are liable to darken. Therefore, to improve shelf-life, it is also desir-
able to stop or at least slow the darkening rate (Asenstorfer, Appelbee,
& Mares, 2009). In this study, changes in L* value of noodle sheets
made from wheat our with or without exposure to ozone gas were
recorded at 12 h intervals and the results were presented in Fig. 5b.
and as can be seen, L* value of ozone treated wheat our noodles
increased signicantly, while ozone 4 day samples were slightly higher
than that of 0 day samples due to the larger water absorbing capacity.
During storage, L* value of ozone noodle sheets decreased signicantly
slower as compared to the control, showing that ozone treatment could
increase the lightness and whiteness of the nal products and inhibited
the darkening progress. This may mainly be because of the PPO activity
inhibiting effect induced by ozone, as shown in Table 1.
4. Conclusions
Microbial and physicochemical changes in ozone treated wheat
our during storage were evaluated in this study. It was found that
microorganisms slightly decreased immediately after ozone treatment
and then further decreased signicantly during storage. Ozone induced
a more signicant impact on pH value compared with fatty acid value
of wheat our, and water activity (a
w
) was found to be progressively
reduced after ozone treatment and during the storage. The L* value of
ozone treated our increased and PPO activity was shown to be lower
Table 2
Volatile compounds in ozone treated wheat our after 0 day and 4 days' storage.
Compounds Retention
time/min
Relative content (%)
Control Ozone 0 day Ozone 4 day
Hexanal 4.371 6.968 37.513 41.66
1-Propene 5.473 0.023 0.537
Heptanal 6.668 0.255
Furan 9.042 0.302 0.105 0.424
Hydroperoxide 9.322 0.196 0.457
Hexanoic acid 9.664 0.027 1.051
2-Octenal 10.77 0.641 0.934
Nonanal 12.136 51.23 40.819 28.3
2-Nonenal 13.471 9.079 11.113 14.34
2,4-Nonadienal 14.228 0.12 0.167
Decanal 14.556 6.53 5.37 4.786
Trans,trans-nona-2,4-dienal 14.716 0.411 0.533
Undecanal 16.816 0.2 0.261
2-Octenal 18.167 0.118 0.351
Dodecane 18.616 0.731 0.047
Phenol 20.656 3.926 0.074 0.106
Pentadecane 21.982 0.535 0.035
Docosane 22.002 5.83 0.107
2-Pentadecanone 25.462 0.167
Phthalic acid 25.728 0.796
Hexadecenoic acid 26.624 1.298
Tetradecanoic acid 26.878 8.675
A means not detected.
Storage time (day)
T
P
C

(
l
g
C
F
U
/
g
)
Control
Ozone 0 day
Ozone 4 day
2
3
4
5
6
7
8
0 1 2 3 4 5 6
40
50
60
70
80
90
100
0 12 24 36 48
Time (h)
L

v
a
l
u
e
Control
Ozone 0 day
Ozone 4 day
a
b
Fig. 5. Effect of ozone treatment and storage on the microbial growth and L* value changes in fresh noodles; (a): TPC changes; (b): L* value changes.
228 M. Li et al. / Innovative Food Science and Emerging Technologies 20 (2013) 223229
after 4 days' storage. In addition, fresh noodles made from ozone treat-
ed wheat our presented a longer shelf-life, with darkening rate and
microbial growth more signicantly inhibited in samples made from
4-day-stored wheat our. Overall, the results of this study signify that
ozone has an excellent potential as an alternative to chemical oxidants
in wheat our and to combat spoilage of the nal wheat products.
Acknowledgments
This work was supported by the National Key Technology R&D
Program (Grant No. 2012BAD37B04 and No. 2012BAD34B01), the
Fundamental Research Funds for the Central Universities (JUDCF11021),
and the Agricultural Key Technology R&D Program of Jiangsu Province
(BE2011377). The authors are thankful to Dr. Zhao-Feng Li for the RVA
analysis of the our samples.
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