OXIDATION OF FATS AND OILS IN INSTANT NOODLES STORED

UNDER VARIOUS CONDITIONS

NAOHIRO GOTOH1, AI IWASAWA1, HIROYUKI WATANABE2,

REIKO OSATO2 and SHUN WADA1,3
1
Department of Food Science and Technology
Tokyo University of Marine Science and Technology
Minato-ku, Tokyo 108-8477, Japan
2
Department of Nutrition
Food Science and Culinary Arts
Toita Women’s College
Minato-ku, Tokyo, Japan

Submitted for Publication August 24, 2006

Revised Received and Accepted November 27, 2006

ABSTRACT

Changes in the peroxide value (PV) in instant noodles stored under

various conditions were carefully analyzed to examine the oxidation of fat and
oils in instant noodles and to evaluate the importance of the maximal
30 mequiv/kg PV standard value in the Food Sanitation Law in Japan from a
food safety point of view. The changes in PV in instant noodles stored at 40 to
60C gradually increased and then rapidly increased after exceeding approxi-
mately 30 mequiv/kg, regardless of the oxidation temperature. This finding
indicates that the PV standard value is meaningful for suppressing the
oxidation-induced formation of toxic compounds. The relationships among PV,
acid value (AV, amount of free fatty acid) and p-anisidine value (AnV, amount
of secondary oxidized products in the oil) in oxidized instant noodles were
investigated. There was no significant relationship between PV and AV, and AV
and AnV. In contrast, there was a significant relationship between PV and AnV.
These results indicate that PV measurements are useful for predicting the
formation of toxic compounds, particularly the secondary oxidized products of
the oil, and measurements of both PV and AV are very important to accurately
determine food deterioration level.

3
Corresponding author. TEL: +81-3-5463-0605; FAX: +81-3-5463-0626; EMAIL: wada@s.kaiyodai.
ac.jp

Journal of Food Lipids 14 (2007) 350–365. All Rights Reserved.

350 © 2007, The Author(s)
Journal compilation © 2007, Blackwell Publishing
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 351

PRACTICAL APPLICATIONS

In this study, changes in peroxide value (PV) in oxidized instant noodles

under various conditions were investigated. The results showed that a sudden
increase in PV occurred after PV exceeded 30 mequiv/kg in instant noodles.
The oxidized fats and oils are toxic. Thus, suppressing the formation of
oxidized compounds from fats and oils in food is an important matter in terms
of toxicity. Acid value (AV) is used as a factor for monitoring the degree of
oxidation in instant noodles in the Codex Alimentarius Commission
(CODEX); however, AV does not directly indicate the amount of oxidized
compound. Therefore, PV, which is deeply related to the formation of oxidized
product must also be used. The results obtained in this experiment propose that
PV must be adopted as the standard factor in the evaluation of oxidation of
instant noodles, in CODEX, and the upper limit will be determined as
30 mequiv/kg.

INTRODUCTION

It is well established that oxidized fats and oils are toxic (Crampton et al.
1951; Kaneda and Ishii 1952; Kaunitz et al. 1955; Andrews et al. 1960;
Artman 1969; Iwaoka and Perkins 1976; Matsuo 1976; Tovar and Kaneda
1977; Frankel et al. 1984; Lamboni et al. 1998). Fats and oils in foods are
oxidized during processing, circulation and preservation. This reaction causes
a deterioration in taste, flavor, odor, color, texture and appearance, and a
decrease in the nutritional value of the foods (Frankel 1998; Kamal-Eldin
2003). Instant noodles are a fried food, and therefore contain a lot of fat and
oil. Approximately 40 years ago, Japan had a food poisoning epidemic caused
by the degradation of fats and oils in instant noodles (Inagaki 1966). Many
people who ate the degraded instant noodles developed acute symptoms such
as diarrhea, nausea, emesis, abdominal pain, fatigue and headache. Before and
after the incidents, many studies reported the formation of toxic compounds
in oxidized fats and oils. For example, 4-hydroxy-trans-2-nonenal is a strong
toxic compound in oxidized fats and oils in heated cooking oil (Seppanen and
Csallany 2004). These toxicities have been evaluated by general toxicity tests
such as acute toxicity, subacute and chronic toxicity tests. Oxidized fats and
oils are also reported to be neurotoxic (Gotoh et al. 2006). Consequently, the
control of the oxidation of fats and oils in food is important to prevent against
the deterioration of foods and protect human health.
Most industrialized countries set food standards. There are also food
standards for instant noodles. Instant noodles are a food invented in Japan, and
the Ministry of Health and Welfare (currently the Ministry of Health, Labor,
352 N. GOTOH ET AL.

and Welfare) in Japan set the standards in the Food Sanitation Law to protect
against food poisoning and to control the quality of instant noodles after the
food poisoning epidemic. In the law, peroxide value (PV) and acid value (AV)
were chosen as useful indices of the deterioration of fat and oil to control food
safety and quality, and standard values of PV and AV were set at not more than
30 and 3 mequiv/kg, respectively (Specifications and Standards for Foods
Food Additives in Japan). After setting these values, there have been no
reported cases of food poisoning caused by instant noodles in Japan. Recently,
there was a movement to set a standard for instant noodles in the Codex
Alimentarius Commission because instant noodles are now a universal food
(ALINORM 2004a,b, 2005). It is an important worldwide movement to
protect against food poisoning caused by the oxidation of fats and oils. These
standard values for instant noodles in the Food Sanitation Law in Japan were
set approximately 40 years ago and ambiguous data were used to determine the
standard values. In particular, PV, relating to the amount of lipid hydropero-
xide, is a very important index from a food safety point of view. Thus, in this
study, the changes in PV in instant noodles stored under various conditions
were carefully monitored to examine the oxidation of fat and oils in instant
noodles and to validate the existing standard values.

MATERIALS AND METHODS

Materials
All the reagents were obtained from Wako Pure Chemical Industries, Ltd.
(Osaka, Japan). Bag-type instant noodles manufactured by company A having
the same lot number were purchased from a shop in Tokyo. The instant noodles
were the fried type and each package of noodles weighed 92 g. The wrapper of
the instant noodles was made from oriented polypropylene and polyethylene
terephthalate.

Oxidation of Fats and Oils in Instant Noodles

The instant noodles were stored under the conditions shown in Table 1 and
used at various times to measure the oxidation degree of fats and oils. Condition
A (23C, 75%) was the average temperature and humidity of Naha City in the
Okinawa prefecture, Japan. This city was chosen as a representative hot city in
Japan. Condition B (30C, 75%) was selected to gauge the temperature effect on
the oxidation of fats and oils to compare with the effects of condition A.
Similarly, condition C (30C, 65%) was selected to observe the effect of
humidity on the oxidation of fats and oils. Condition D (23C, 75%, 2,000 lx)
was the same condition as condition A with the addition of a fluorescent lamp.
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 353

TABLE 1.
STORAGE CONDITIONS ADOPTED IN THIS EXPERIMENT

Condition Temperature (C) Humidity (%) Light Remarks

A 23 75 Dark
B 30 75 Dark
C 30 65 Dark
D 23 75 Fluorescent lamp (2,000 lx)
E* Approximately 20 Not controlled Fluorescent lamp (700 lx)
F 60 Not controlled Dark
G 50 Not controlled Dark
H 40 Not controlled Dark
I† Atmospheric temperature Not controlled Sunlight
J† Atmospheric temperature Not controlled Sunlight Without pouch

* Stored under fluorescent lamp in an air-conditioned office room. Therefore, the fluorescent lamps
were frequently turned on and off.
† Stored on the roof of the building under sunlight in summer. The package of instant noodles was
covered with an acrylic plate to protect from crows during storage. Temperature and humidity
alternated between 32 to 45C and 35 to 70%, respectively.

Condition D was used to determine the effect of light on the oxidation of fats and
oils. Condition E (approximately 20C, humidity not controlled) was considered
to be the standard display conditions of a retail shop. Conditions F (60C), G
(50C) and H (40C) were used as conditions of oxidation acceleration. Condi-
tions I (sunlight) and J (sunlight without pouch) corresponded to the display
conditions outside the shop during the summer. The instant noodles that caused
the food poisoning epidemic approximately 40 years ago were primarily stored
under sunlight conditions in summer. Under condition J, the packaging was
removed and replaced with a polyethylene film bag, because approximately 40
years ago, the instant noodle packaging contained a large transparent circle of
film in the center to make the noodles visible to the consumer.

Extraction of Fats and Oils from Instant Noodles

Extraction of fats and oils from unoxidized and oxidized instant noodles
was performed according to the method of the Japan Agricultural Standards
(Instant Noodles 1981). The noodles were crushed into small pieces with a
hammer in a transparent polyethylene bag and the pieces were sifted with a
10-mesh sieve. The sifted pieces were placed in a 300-mL Erlenmeyer flask
with a rubber stopper, mixed with petroleum ether at approximately three
times the volume of the sifted pieces, shaken slowly and allowed to sit for 2 h.
The mixture was then filtered and moved to a separating funnel. The extracted
residue was mixed with approximately half the volume of petroleum ether
used for the first extraction in a 300-mL Erlenmeyer flask, shaken slowly,
354 N. GOTOH ET AL.

filtered and combined in a separating funnel with the first extraction. The
petroleum ether was mixed with about a half volume of distilled water in the
funnel, shaken strongly and allowed to stand still until the liquid interface was
clear. Then, the petroleum ether layer was moved to another 300-mL Erlenm-
eyer flask with a rubber stopper and dried with anhydrous sodium sulfate. The
petroleum ether was filtered again and evaporated under vacuum at 40C to
obtain the fats and oils.

Analysis of the Fatty Acid Composition

The fatty acid composition of extracted fats and oils was analyzed by gas
chromatography. Methyl esterification of the fats and oils was performed using
the following procedure. Approximately 10 mg of the extracted fats and oils
and 1 mL of 0.5-M sodium hydroxide methanol solution (NaOH-MeOH) were
placed in a 10-mL screw-capped tube and mixed. Two milliliters of 14% boron
trifluoride methanol solution (BF3-MeOH) was added to the mixture and
heated at 100C for 20 s. The heated tube was cooled to 40C under air, and
1-mL hexane was added to the solution. Saturated sodium chloride solution
(3 mL) was added to the mixture and mixed vigorously. The tube was left to
stand for a few minutes to allow the hexane and water layers to separate. The
hexane layer was subjected to a gas chromatography-flame ionization detector
system (GC14B, Shimadzu, Tokyo, Japan) equipped with a capillary column
(Omegawax320, 30 m ¥ 0.25 mm ID, Supelco, Bellefonte, PA) and a Chro-
matopac integrator (C-R6A, Shimadzu). The temperature of the injection port
and detector was 250 and 260C, respectively. The initial column temperature
was 175C, and increased to 225C at the rate of 1C/min. Helium was used as the
carrier gas at a flow rate of 32 cm/s. The fatty acid species was identified using
the retention time of a fatty acid methyl ester standard solution (Supelco 37
Component FAME Mix, Sigma-Aldrich Japan, Tokyo, Japan).

Analysis of PV
The PV of the extracted fats and oils was determined using the potenti-
ometry method (Hara and Totani 1988; Hara et al. 1997) adopted by the
Standard Methods for the Analysis of Fats, Oils and Related Materials estab-
lished by JOCS (2004). We did not use the universal method for the determi-
nation of PV in fats and oils because it requires the experimenter to determine
the titration end point at which the liquid color changes, and the method is
strongly affected by the original color of fat and oil. In contrast, the potenti-
ometry method is not affected by the original color of the fats and oils.
The extracted fats and oils were placed in a 300-mL Erlenmeyer flask
with a glass stopper and the weight of the sample was measured. Twenty-five
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 355

milliliters of degassed 2,2,4-trimethyl pentane and acetic acid mixture (2:3 by

volume) was poured into the 300-mL Erlenmeyer flask containing a magnetic
stir bar and stirred vigorously to completely dissolve the fats and oils. Satu-
rated potassium iodine aqueous solution (0.1 mL) was added to the solution
and shaken slowly on ice. Then, 100 mL distilled water was added to the
flask, shaken steadily and analyzed using an automatic potentiometric titrator
(AT-510, Kyoto Electronic Manufacturing Co., Ltd., Kyoto, Japan) equipped
with a platinum combination electrode and a burette. The prepared sample
solution was stirred with a magnetic stirrer and titrated with 0.01 mol/L
sodium thiosulfate aqueous solution. The end point of the titration was defined
as the point the electric charge per certain volume of sodium thiosulfate
aqueous solution became maximum and the point was determined by the
machine automatically. The PV was calculated as milliequivalents (mequiv) of
peroxide/kg of sample using the equation:

PV (mequiv ) = ( A − B ) × F × 10 C ,

where A is the amount of sodium thiosulfate aqueous solution (in mL) used in
the main experiment; B is the amount of sodium thiosulfate aqueous solution
(in mL) used in the blank test; C is the sample weight (g); and F is the factor
of 0.01 mol/L sodium thiosulfate solution. Respective values were measured
three times and indicated as means ⫾ SD.

Analyses of Acid Value and p-Anisidine Value

The extracted fats and oils were also subjected to AV and p-anisidine
value (AnV) analyses. AV was measured using the method of the Japan
Agricultural Standards (Instant Noodles 1981). Briefly, approximately 3 g of
the extracted fats and oils was mixed with 80 mL of mixed 1:1 (v/v) solution
of ethyl ether and ethyl alcohol in a 200-mL Erlenmeyer flask. Phenolphthalein
was used as an indicator and the titration was performed with N/20 potassium
hydroxide (KOH) ethyl alcohol solution. AV was calculated by the equation:

AV = ( A [mL ] × 2.806 × f ) B [g ],

where A is the volume of N/20 KOH ethyl alcohol solution; f is the factor of
N/20 KOH ethyl alcohol solution; and B is the weight of the extracted fats and
oils.
AnV is the factor that represents the amount of secondary oxidized
products of fat and oils. Consequently, AnV should be strongly related to the
toxicity of degraded fats and oils. AnV was determined by the method of the
Standard Methods for the Analysis of Fats, Oils, and Related Materials estab-
356 N. GOTOH ET AL.

lished by Japan Oil Chemists’ Society (p-ANISIDINE VALUE 2.5.3. 1996).

Briefly, approximately 0.5 g of sample was placed into a 25-mL Erlenmeyer
flask, weighed and dissolved with 2,2,4-trimethylpentane. The absorbances of
both samples were measured at 350 nm with a spectrophotometer. p-Anisidine
was dissolved in 99.7% (weight %) acetic acid and the concentration was
adjusted to 0.25 g/100 mL. The p-anisidine–acetic acid solution (1 mL) was
mixed with 5 mL sample solution or 2,2,4-trimethylpentane in a screw-capped
test tube and mixed vigorously. The solution was allowed to stand for 10 min
at room temperature and the absorbance at 350 nm was measured with a
spectrophotometer. AnV was calculated by the equation:

AnV = 25 × (1.2 × A − B ) C ,

where A is the absorbance of sample solution reacted with p-anisidine-acetic

acid solution; B is the absorbance of the sample solution; and C is the sample
weight. Respective values were measured three times and indicated as
means ⫾ SD.

Random Sampling and Statistical Analyses

Thirty-two samples, corresponding to one-fifth of the total sample
number (156), were randomly selected using a random number table and
subjected to AV and AnV analyses. The correlations between the two factors
(PV versus AnV, PV versus AV, and AnV versus AV) were assessed using
Pearson’s product-moment coefficient of correlation. The relationship
between two factors was considered significant when P was less than 0.05.

RESULTS

Fatty Acid Composition of Fats and Oils in Instant Noodles

The fatty acid composition of fats and oils extracted from instant noodles
is shown in Table 2. The original fats and oils were comprised mainly of oleic
acid (43.5%), palmitic acid (29.8%), stearic acid (11.9%) and linoleic acid
(9.5%).

Changes in PV in Instant Noodles

Changes in the PV under conditions A–E are shown in Fig. 1. The PV of
instant noodles stored under condition A changed minimally compared to
instant noodles stored under the other conditions, and the PV reached to
3.3 mequiv/kg at 72 weeks. Condition B also suppressed the increase in PV
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 357

TABLE 2.
FATTY ACID COMPOSITION OF FAT AND OIL EXTRACTED
FROM INSTANT NOODLES

Fatty acids Area % of gas

chromatography
chromatogram*

14:0 1.50
16:0 29.78
16:1(n-9) 1.94
17:0 0.35
17:1 0.30
18:0 11.89
18:1(n-9) 43.53
18:2(n-6) 9.45
18:3(n-3) 0.34
20:1(n-9) 0.68
20:2(n-6) 0.23
Total 100

* [(Area for each peak)/(Total area)] ¥ 100.

150
Condition A
Peroxide Value [meq/kg]

Condition B
Condition C
Condition D
120 Condition E

90

60

30

0
0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60

Storage Period [Weeks]

FIG. 1. CHANGES IN PEROXIDE VALUE IN INSTANT NOODLES STORED UNDER

CONDITIONS A–E

in the range of 2 to 4 mequiv/kg for the first 68 weeks, but then the value
increased to 9 mequiv/kg. Condition C suppressed the oxidation of fats and
oils in the range of 2 to 5 mequiv/kg for the first 44 weeks, but then PV
suddenly increased and eventually reached 18 mequiv/kg at 84 weeks. PV,
however, remained under 30 mequiv/kg, which was set as the standard value in
358 N. GOTOH ET AL.

480
450
420
390
Peroxide value [meq/kg]

360 Condition H
Condition F Condition G
330
300
270
240
210
180
150
120
90
60
30
0
0 4 8 12 16 20 24 28 32 36 40 44 48
Storage period [Weeks]

FIG. 2. CHANGES IN PEROXIDE VALUE IN INSTANT NOODLES STORED UNDER

CONDITIONS F–H

the Food Sanitation Law in Japan. In contrast, PV began to increase immedi-

ately in instant noodles exposed to fluorescent light (conditions D and E). The
increases fluctuated, but steadily increased. The rates of the PV increase were
similar between groups in spite of the differences in light power (condition D:
2,000 lx, condition E: 700 lx). The PVs in both conditions D and E exceeded
30 mequiv/kg at 16 and 24 weeks, respectively.
The changes in PV of noodles stored under high temperatures (conditions
F–H) are shown in Fig. 2. PV increased markedly during storage under all
conditions. The maximum values of PV (condition F, 432 mequiv/kg; con-
dition G, 311 mequiv/kg; and condition H, 174 mequiv/kg) decreased with
increases in storage temperature. Furthermore, the induction period of the
increase in PV decreased with an increase in storage temperature. Interest-
ingly, the sudden increase began after PV reached approximately 30 mequiv/
kg, regardless of the oxidation temperature.
The changes in PV under conditions I and J are shown in Fig. 3. There
was no induction period and the oxidation rate was fairly high compared
to other conditions. Furthermore, the oxidation rate under condition J was
approximately three times higher than that under condition I.

Changes in AV and AnV in Instant Noodles

The relationship between PV and AV, AV and AnV, and PV and AnV were
evaluated with extracted fats and oils from instant noodles randomly selected
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 359

450
420
390
360
Peroxide Value [meq/kg]

330 Condition J
300
270
240
210
180
150 Condition I
120
90
60
30
0
0 20 40 60 81 92 101 110 120 132 140 150 160 170 180
Storage Period [hours]

FIG. 3. CHANGES IN PEROXIDE VALUE IN INSTANT NOODLES STORED UNDER

CONDITIONS I AND J

using a random number table (Fig. 4). The coefficients of correlation for
respective relations were calculated with Pearson’s product-moment coeffi-
cient of correlation. There was a significant relationship between PV and AnV.
In contrast, there was no relationship between PV and AV, or between AV
and AnV.

DISCUSSION

The deterioration of fats and oils in foods leads to the formation of many
types of degradation products, such as lipid hydroperoxide, aldehyde, ketone,
alcohol, free fatty acids, polymerized oil, cyclic fatty acid, etc. (Frankel 1998;
Kamal-Eldin 2003). These compounds not only cause an unpleasant smell, but
also food toxicity. These products, with the exception of free fatty acids, form
via the production formation of lipid hydroperoxide (Frankel 1998). There-
fore, lipid hydroperoxide production must be suppressed to protect against the
formation of toxic compounds. The food poisoning epidemic caused by the
degradation of the fats and oils in instant noodles 40 years ago indicated that
oxidized fats and oils are toxic to both animals and humans. Therefore, PV and
AV are now used to monitor the deterioration of fats and oils in food in Japan.
The concepts behind measuring PV and AV are different. PV is measured to
360 N. GOTOH ET AL.

350 350
(a) (b) r=0.325
300 300
r=0.791 p=0.075

PV [meq/kg]
PV [meq/kg]

250 250
p<0.01
200 200

150 150

100 100

50 50

0 0
0 20 40 60 80 0 10 20 30 40
AnV AV
80
(c)
70
60
50
AnV

40
30
20
r=0.286
10
p=0.118
0
0 10 20 30 40
AV

FIG. 4. THE PLOT (a) PV VERSUS AnV, (b) PV VERSUS AV AND (c) AnV VERSUS AV
The coefficients of correlation (r) for respective relations and P values were calculated with
Pearson’s product-moment coefficient of correlation and the results are indicated in the respective
figures. PV, peroxide value; AnV, p-anisidine value; AV, acid value.

monitor the formation of lipid hydroperoxide. In contrast, AV is measured to

monitor food quality. During food processing and storage, free fatty acids are
formed in the food by hydrolysis of the fats and oils. Free fatty acids are not
toxic compounds (Tovar and Kaneda 1977), but hydrolysis causes a reduction
in flavor and taste. The purpose of measuring AV is to check the free fatty
acid levels in food. Monitoring PV in food is very important for protecting
against food poisoning. Japan has adopted a PV standard of not more than
30 mequiv/kg in the Food Sanitation Law for instant noodles. There have been
no concrete data, however, regarding the selection of this particular value or to
indicate why the value has been sufficient to protect against food poisoning.
Thus, in the present study, representative bag-type instant noodles in Japan
were stored under several different oxidation conditions and changes in PV
were monitored.
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 361

The fatty acid composition of fats and oils contained in instant noodles is
such that the fats and oils are easily autoxidized because approximately 10%
linoleic acid is contained in the fats and oils (Table 2). Also, there is approxi-
mately 40% oleic acid in the fats and oils. Oleic acid is not easily oxidized by
autoxidation, but is easily oxidized by photoxidation (Frankel 1998). Condi-
tions A–C mildly facilitated the oxidation of fats and oils in instant noodles
(Fig. 1). The shelf life of instant noodles in Japan is 6 months, or approxi-
mately 24 weeks. The PV of all of the instant noodles stored under these
conditions did not exceed 30 mequiv/kg for 24 weeks. The oxidation induction
periods under conditions B and C, however, were shorter than that under
condition A, indicating that a slight change in temperature and humidity
affects the oxidation of fats and oils in food. Condition D is the condition in
which a 2,000-lx fluorescent lamp was added to condition A and PV increased
without an induction period. This finding indicates that light energy strongly
affected the oxidation rate of fats and oils and the antioxidants contained in
instant noodles were not adequate under the light energy conditions. The
oxidation of instant noodles maintained under condition D might be dominated
by photoxidation. Condition E was also the condition in which the noodles
were exposed to light energy in an air-conditioned room adjusted to approxi-
mately 20C. This condition was set as the representative display condition of
the shops in Japan. The light energy was approximately 700 lx, but the rate of
increase of PV was almost the same as that under condition D. In fact, there
was not much difference between the two conditions except for light energy.
One difference is that the instant noodles under condition E were left in an
office room where the fluorescent lamp was frequently turned on and off.
Therefore, this light–dark cycle might have affected the oxidation rate of the
fats and oils, and caused the fairly high rate of oxidation. Further studies are
required.
Conditions F–H were accelerated oxidation conditions without light
energy. All the conditions showed an induction period prior to the increase
in PV (Fig. 2). The oxidation under these conditions was the result of autoxi-
dation and the increase in temperature affected the rate of autoxidation. The
results indicated that both the formation and decomposition of hydroperox-
ide occur simultaneously and the decomposition reaction gradually became
dominant with the increase in storage temperature. Also, the induction
period for the increase in PV decreased with the increase in storage tem-
perature. The point of the rapid increase in PV was near 30 mequiv/kg for all
conditions (Fig. 2). This point was not recognized under conditions A–C
because the oxidation rates were fairly slow and PV did not reach
30 mequiv/kg during the storage period. These results indicate that the
hydroperoxide level of fats and oils in instant noodles under autoxidizing
conditions increases suddenly after PV reaches 30 mequiv/kg. This is a very
362 N. GOTOH ET AL.

important point for setting the standards for PV for instant noodles. Oxidized
fats and oils with a PV value of 100 mequiv/kg induce neurotoxicity in rats
(Gotoh et al. 2006). The PV reached 100 mequiv/kg quickly after exceeding
30 mequiv/kg under all conditions (Fig. 2). Consequently, suppressing the
formation of lipid hydroperoxide to under 30 mequiv/kg in instant noodles is
extremely important for protecting against food poisoning. In essence, the
standard value of PV for instant noodles set in Food Sanitation Law in
Japan coincides exactly with the value observed in the present experi-
ments. Thus, the established value set by the Food Sanitation Law of Japan
(JETRO 2004) is effective because a PV of 30 mequiv/kg appears to be a
threshold, which, once broken, quickly leads to the neurotoxic values of
100 mequiv/kg.
Under conditions I and J, fats and oils in instant noodles oxidized very
quickly (Fig. 3). These rapid oxidation rates are thought to be due to the
combination of mild temperature (32–45C) and strong UV energy. The oxi-
dation rate under condition J was approximately three times higher than that
under condition I. The instant noodles maintained under condition J were
surrounded by a transparent polyethylene film instead of the original oriented
polypropylene and polyethylene terephthalate packaging. This result indicates
that the original film protects against the oxidation of fats and oils by cutting
UV light. There was also no induction period for the increase in PV under
conditions I and J, like under conditions D and E. The antioxidants in instant
noodles might not be adequate to protect against the formation of hydroper-
oxide of fats and oils induced by photoxidation.
The relationships among PV, AV and AnV in oxidized fats and oils were
investigated to determine whether it is necessary to measure both PV and AV
to monitor the deterioration of fats and oils in food. There was a highly
significant relationship between PV and AnV (r = 0.791, P < 0.01 [1.18 ¥
10-7]). The secondary oxidized products of fats and oils were the cause of the
toxicity of the deteriorated fats and oils via the formation of hydroperoxide.
AnV indicates the amount of secondary oxidized products. Consequently, it is
reasonable that both factors represent food deterioration. In contrast, AV was
not significantly related with PV (r = 0.325, P = 0.075) or AnV (r = 0.286,
P = 0.118). This finding is reasonable because AV represents the hydrolysis of
free fatty acid, rather than oxidation to hydroperoxide. In fact, in a study of
instant noodle soups displayed in shops in 15 different countries, there was no
significant relationship between PV and AV (Gotoh and Wada 2006). These
results indicate that it is not possible to determine the deterioration level of fats
and oils in instant noodles, or food in general, using only PV or AV. As
mentioned earlier, PV and AV are monitored for different reasons. Therefore,
measuring both PV and AV is very important to accurately determine food
deterioration level.
 OXIDATION OF FATS AND OILS UNDER VARIOUS CONDITIONS 363

CONCLUSION

In the present study, changes in PV in oxidized instant noodles under

various conditions were investigated. A sudden increase in PV occurred after
PV exceeded 30 mequiv/kg in autoxidized instant noodles. This unique
finding, however, was observed only in autoxidized instant noodles. In pho-
toxidized instant noodles, there was no oxidation induction period; therefore,
a PV standard of 30 mequiv/kg is not meaningful for these instant noodles.
Furthermore, the changes in PV depended on the antioxidant content and the
fatty acid composition of the fats and oils in the instant noodles. In the Food
Sanitation Law in Japan, the standard value of PV for instant noodles is not
more than 30 mequiv/kg and this value has served to protect against food
poisoning for a long period in Japan. From the viewpoint of food poisoning
protection, suppressing the oxidation of fats and oils in instant noodles to
100 mequiv/kg in PV is very important. Monitoring PV of fats and oils in
instant noodles to 30 mequiv/kg suppresses the formation of oxidized fats and
oils, which occur at a PV of 100 mequiv/kg. In conclusion, using the PV
standard value of 30 mequiv/kg to monitor the deterioration of fats and oils in
instant noodles is meaningful for suppression of the formation of toxic com-
pounds by oxidation, not only in instant noodles, but in other foods as well.

ACKNOWLEDGMENTS

We are deeply grateful to the Yokohama Center of IAA Center for Food
Quality, Labeling, and Consumer Service for supporting the long-term storage
tests. This research was supported in part by a Grant-in-Aid for Scientific
Research from the Ministry of Health, Labor, and Welfare.

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