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Studies on Cause of Color Reversion of Edible Soybean Oil and its Prevention
a

Mamoru Komoda , Noriji nuki & Ichiro Harada

a

Sugiyama Chemical Research Institute, Mitaka-shi

Hohnen Oil Co., Ltd., Shimizu Plant, Shimizu-shi

Published online: 09 Sep 2014.
To cite this article: Mamoru Komoda, Noriji nuki & Ichiro Harada (1966) Studies on Cause of Color Reversion of Edible Soybean Oil and its Prevention, Agricultural and Biological Chemistry, 30:9, 906-912
To link to this article: http://dx.doi.org/10.1080/00021369.1966.10858698

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[Agr. BioI. Chem. Vol. 30, No.9, p.

906~912,

1966]

Studies on Cause of Color Reversion of Edible Soybean

Oil and its Prevention
Part I. Relation between the Moisture of Soybeans
and the Quantity of Tocopherol in them
By Mamoru

KOMODA,

Noriji

ONUKI

and Ichiro

HARADA*

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Sugiyama Chemical Research Institute, Mitaka-shi;

*Hohnen Oil Co., Ltd., Shimizu Plant, Shimizu-shi;
Received March 4, 1966
Properties and minor constituents were examined for soybean oils obtained from soybeans with various moisture contents. Yields of crude oils by extraction process of experimental scale anotocopherol content in the extracted oil were found to be correlated to the.
, .
moisture content in soybeans (moisture range, 8_18%). When moistened raw soybeans
(moisture 18%) are dried (moisture 8%), the tocopherol content in the extracted crude oil
from the dried beans increased. This phenomenon can be observed only in the case of
drying whole beans and not in the case of drying crushed raw soybeans. When soybeans
with ordinary moisture (moisture 13%, tocopherol 1.25 mg/g) are dried to reduce the
moisture to 1.9%, the tocopherol content in the extracted crude oil increases (1.90 mg/g),
but it decreases (0.33 mg/g) again when the dried soybeans are moistened (moisture 18%).
..'

INTRODUCTION

Soybean oils just after deodorization are

quite pale but become darker during storage
in the air. The degree of color change depends on the following conditions: nature of
raw soybeans, refining and deodorization condition of oil, duration of storage, temperature,
intensity of light and contact surface with the
air during storage, etc. With some oils it
takes only several hours to reach the maximum
of color reversion, while others take several
months. In both cases, the color becomes
paler after it reaches the peak.
We call the phenomenon of the color of the
oil getting darker during storage "the color
reversion of oil." As to the coloring of oil
during storage, it is hitherto known that the
oxidized-polymerized unsaturated acids affect
the coloration, but this coloring appears only
in the rancid oils and is clearly distinguished
This research has been financed by a grant made by the
United States Department of Agriculture under P. L. 480.

from what we call the color reversion, which

occurs at a very early stage of oxidation.
As to cottonseed oil, the coloration by gossypol was studied.1.2) Swift3 ) proved that the
phenomenon of refined cottonseed oil becoming reddish is due to the 2,7,8-trimethyl-2(4',8',12'-trimethyltridecyl-I-)-chroman-5,6-quinone derived by the oxidation of 2,7,8-trimethyl-2-(4', 8', 12'-trimethyltridecyl)-6-chro
manol (r-tocopherol) in the oil.
There are a few papers which treat the
color reversion of refined and deodorized soybean oil. It is reported by Beal4 ) that phos
phorus content of refined and deodorized
soybean oil is closely related to the color and
,
the oxidation stability of oil.
We recognized that the color reversioR.,is
<

1) J. M. Dechariy, R. P. Kupperman, F. H. Thurber and

R. T. O'conner, J. Am. Chern. Soc., 31, 420 (1954).
.
.
2) V. P. Rzhekin, MasloboinoZhirotJaya Pro., 19, No. 28
(1954).
<
3) C. E. Swift, G. E. Mann and G. S. Fisher, Oil & Soap,
21, 317 (1944).

4) R. E. Beal, E. B. Lancaster and D. E. Brakke, 'J. Am.

Chern. Soc., 33, 619 (1956).

Downloaded by [113.161.64.163] at 20:50 02 August 2015

Studies on Cause of Color Reversion of Edible SObyean Oil and its Prevention.

Part I

907

characteristic to refined and deodorized soyFive kinds of moisture-controlled soybeans

bean oil and wrote a number of papers on were crushed and extracted with n-hexane.
this phenomenon. 5 - I2 ) We happened to find Yield of crude oil and the analytical results
that the color reversion was far slighter in the of the meal are shown in Table I.
refined and deodorized oil prepared from the
TABLE I. YIELD OF CRUDE OILS AND ANALYTICAL
crude oil obtained by extraction process from
RESULTS OF SOYBEAN MEALS
the 1963 crop of Illinois soybeans than in the
Moisture of
Soybean Meals
refined and deodorized oils hitherto manufacExtracted
Soybeans
,

Crude
Oil
tured. We knew by the reports from U.S.A.
(Xylene
Oil
(Dry Basis)
Moisture
Method)
(Dry Basis)
that the 1963 crop soybeans in Illinois had
(%)
(%)
(%)
(%)
experienced a draught in the growing and
7.8
20.2
6.99
1.67
harvesting season, which led us to deduce that
10.1
21.5
8.03
1.26
the low moisture might be a cause of the
12.1
21.8
9.95
1.05
14.8
22.4
llAO
slighter color reversion. Therefore, we have
0.72
17.6
23.2
13.30
0.50
studied1S ) the relation between the moisture
content of soybeans and the color reversion
TABLE II. PROPERTIES OF CRUDE OIL
of the refined and deodorized oils obtained
ConTotal
from them. From the experiments described Moisture
Iodine
of
Acid
jugated
Lipoid
Tocoin the previous paper,13) we found that the Soybeans Value Value Diene
pherol
(Wijs)
(%)
(%)
(%)
(mg/g)
moisture of soybeans is closely related to the
7.8
0.99
0.27
0.42
1045
130.8
color reversion of refined and deodorized oil.
0.35
0.96
10.1
1.22
129.6
1.28
Now, we have found further that the mois0.41
1.75
0.89
12.1
1.29
128.3
ture of soybeans affects the quantity of toco0.43
14.8
1.28
129.6
2.33
0.71
pherol in crude oil. The mechanism involved
0.83
17.6
1.29
127.7
0.57
2.53
in the changes of the tocopherol in soybeans
The
percent
of
the
yield
of
crude
oil
is
is not yet clear but we have found several new
proportional
to
the
moisture
content
of
soyphenomena about the behavior of tocopherol
beans.
The
crude
oil
obtained
from
soybeans
in soybeans.
with less moisture (moisture 7.8, 10.1, 12.1%)
is orange in color and transparent. The color
EXPERIMENT AND RESULT
of crude oil becomes darker and the crude oil
1) Properties of crude oils extracted from soyhas more sludge as the moisture of the raw
beans with various moisture contents.
soybeans increases (moisture 14.8, 17.6%). The
Raw soybeans used in this experiment were less the moisture of soybeans, the more is the
the 1963 crop Illinois soybeans with 12% residual oil in the meal and the darker is the
moisture. From them soybeans with the mois- extracted matter.
ture content of 7.8, 10.1, 14.8 and 17.6% were
Figs. 1 and 2 indicate the ultraviolet absorpprepared by the same process described al- tion spectrum of each extracted oil. The
ready.IS)
amount of chlorophyll which has maximum
5) M. Nakamura, S. Tomita and M. Komoda, Yukagaku, 9,
absorption
at
670
lllJL
decreases
as
the
moisture
319 (1960).
6-8) I. Harada, Y. Saratani and M. Ishikawa, ,. AgT. Chem. of soybeans decreases.
Soc. lapan, 34, 545, 551, 558 (1960).
The crude oil from soybeans with high
9i"'12) I. Harada, Y. Saratani and M. Ishikawa, ibid., 35,668,
674, 800, 807 (1960).
moisture (14.8, 17.6%) has distinct absorption
13) S. Tomita, M. Kornoda, S. Enomoto and N.Onuki Yukagaku, 13, 530 (1964).
maximum at 350 lllJL and has weak absorption

908

Mamoru KOMODA, Noriji

and tocopherol are most noticeable. Phosphatides increase and tocopherol decreases as the
moisture of soybeans increases.

0-

2)

_El

,0

"'.....
'"~

8%

Quantitative estimation of total tocopherol.

10%

-2

and Ichiro HARADA

ONUKI

10%

-4

51--

Emmerie-Engel method14 ) was usecl. for the

quantitative estimation of total tocopherol: As
crude soybean oil contains various kinds of
minor constituents such as carotenoid' pigments and sterols that interfere in the absorption spectrum of tocopherol analysis,. thes~
minor constituents were removed by Green's
method. 15 )
.
In the check test in which dla-tocophe~@
',;,}"
(E~:~ 292 m,a max.=73.2, ethyl alcohol) was
.' ,
used as a standard sample, the recovered
I","
amounts were satisfactory as shown,' in
Table III.
'.

200

300

400

500

600

700

Wave length (mp)

FIG.

1.

Absorption Curves of Crude Soybean Oils

from Dried Soybeans.

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TABLE III.

Tocopherol
Added
(mgjg)

No.

-1

0.071

4-3

3)

Soybean
Total
Oil
Tocopherol
(g)

0.071
0.143

1
2

RATE OF RECOVERY OF TOTAL TOCOPHEROl.

dl-lX-

".'

0.2
0.2

(mgjg)

0.070
0_144
0.218
0.290
0.072

Rec0very

(%)
97.9

100.7 .

100.6

'

Residual tocopherol in soybean meal. "

The tocopherol content in extracted crude

oil decreases as the moisture of soybeans

Increases.
There is some doubt that the decrease of
tocopherol may partly be attributed to' the
decrease in the solubility of tocopherol in
n-hexane due to the moisture from the soy'
beans. The following experiment was carried
out to clarify this question.

200

300

400

500

600

700

Wave length (mp)

FIG.

2.

Absorption Curves of Crude Soybean Oils

from Raw Beans and Moistened Soybeans.

maximum at 400-500 mfL characteristic to Extraction of meal first with n-hexane and the:
carotenoid pigments. The carotenoid pig- with ethyl ether.
ments are most extractable at 10-12% moisRaw soybeans (moisture 12%) and moistureture of soybeans.
14) A. Emmerie and C. Engel, Nature, 142,!873' (1938); RIC,'
Chim., 57, 1351 (1938); ibid., 58, 283 (1939). I ' ': ;'1'
As to the minor constituents of crude soy- Trav.
'i5) Vitamine E Panel, Analytical Methods Committee 1969,
.
bean oil, changes in the yield of phosphatides Analyst, 84, 356 (1959).

Studies on Cause of Color Reversion of Edible Sobyean Oil and its Prevention.

TABLE

IV.

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.~

__

Yield of
Oil

(%)

(%)

8.1
12.1
17.7

18.34
19.40
18.74

909

YIELD OF CRUDE OIL ANn TOTAL TOCOPHEROL

Extd. with n-Hexane

Moisture of Soybeans
(Xylene Method)

Part I

A~

_ __

Total
Tocopherol
(mg/g) (A)
1.66
0.96
0.09

Extd. with Ethyl Ether

'~--''----,

Yield of
Oil
(%)
0.92
0.60
0.42

Total
Tocopherol
(mg/g) (B)
2.28
0.80
0.90

Total
Tocopherol
(A) + (B) (mg/g)
on Dried Basis
of Raw Soybeans
1.8
1.1
0.1

controlled soybeans (moisture 8, 18%) are all were dried in the sun in the form of raw beans
crushed to pass a 700fL sieve. Five hundred until the moisture decreased to about 8% and
grams of pulverized beans were extracted with the tocopherol in the crude oil from these
1500 ml of n-hexane at 40C. After extrac- dried beans was determined. The results are
tion, the meal was cooled to 25C and ex- shown in Table V.
tracted with 1500 ml of ethyl ether. The
Ethyl ether is hydrophilic and will gradually
residual solvent was completely distilled under dissolve the residual moisture in the meal, so
the reduced pressure. The yield of the crude that this solvent dissolves almost all of the
oil and the total tocopherol are shown in tocopherol in the meal. But the tocopherol
in the ethyl ether extract was small contrary
Table IV.
to
expectation.
Extraction of pulverized and then dried soybeans.
From the above experiments it may be conFour kinds of sovbeans
with
various
mois
cluded that the tocopherol is transformed to
ture contents were crushed and dried at 90C
non-reducing substance in soybeans with high
in a vacuum dryer until the moisture is remoisture level.
duced to 1-2%.
We have already described in 1) that the
The pulverized and dried soybeans were
tocopherol in extracted crude oil increases
extracted by Soxleht extractor with n-hexane
when raw soybeans are dried, crushed and
and the tocopherol in the crude oil was dethen extracted. By this experiment we have
tenllined. Among the four kinds of the above
further confirmed that the tocopherol in exsoybeans, those with 16% and 18% moisture tracted crude oil does not increase when moistened soybeans are crushed, dried and then
TABLE V. YIFLD OF CRUDE OIL AND
extracted.
TOTAL TOCOPHEROL
As
moistened
soybeans
in
this
case
are
Moisture of
crushed
and
then
dried
before
the
extraction
Soybeans
Total
Oil
,

,
of
oil,
the
moisture
in
the
crushed
beans
is
Toco(Dried
Note
Before Before
pherol
Basis)
considered to give no influence on the extracPulveriz- Extracmg
tlOn
tion of oil. Table V indicates that the toco(mg/g)
(%)
(%)
(%)
pherol in crude oil is less in quantity when
7.9
1.54
1.56
20.80
soybeans have more moisture before the pro11.2
1.63
21.37
1.17
16.0
0.27
cess of pulverizing.
22.04
2.08

17.8
8.0

7.7

1.67
1.36

2.32

21.65
21.56
21.71

0.25
1.67
1.36

18% moistened
soybeans dried
in the sun
16% moistened
soybeans dried
in the sun

4) Changes in the quantity of tocopherol during

the storage of moistened soybeans.
_

The changes of the tocopherol content in

soybeans with 15, 18 and 20% moisture, respectively, were determined during the storage

Mamoru

910

KOMODA,

Noriji

ONUKI

and Ichiro

HARADA

make clear these points.

.

1.0

0.8

""El

Changes of tocopherol content by drying moisten~d:

soybeans in the sun.

. .
~

-8

t'

..."

8'"

...
<ii
...

,\

~\.-

0.2

18%

15%

--_L---- .

\",'

E-<

20%

Storage (day)

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FIG.

3.

'.

Changes in the Quantity of Tocopherol during

the Storage of Moistened Soybeans.

of the soybeans at the room temperature and

in the diffused light as shown in Fig. 3.
The 1964 crop U.S. soybeans with 13%
moisture (Xylene method) were used in this
experiment.
The tocopherol decreased rapidly after the
soybeans were stored for one day and only
slight changes could be observed afterward.
In the case of soybeans with ordinary moisture
(13%), the quantity of tocopherol was almost
constant during the storage, while it decreased
sharply in the case of beans kept with 15%
moisture.
This result indicates that there exists a close
relation between the quantity of moisture of
beans and the quantity of tocopherol.

Soybeans with 18% moisture (total toco.

pherol 0.28 mg/g) were used as sample. They
were dried in the sun on a fine day," The
degree of moisture was determined by the
weight loss of the soybeans. This experiment
was carried out in summer and the drying
was very fast. But it was impossible to reduce
the moisture below 7% by drying in the sun.
One hundred and fifty grams of soybeans
with 18% moisture were put in a fiat glass
basin and were dried in strong sunlight for
8 hrs, the maximum atmospheric temperature
being 33C, and the decrease of moisture and
the increase of tocopherol were determined
as shown in Fig. 4.
The close relation between the moisture of
soybeans and the quantity of tocopherol as
pointed in experiment 4) has now been clear
ly observed in Fig. 4. The increase of toco
pherol is slow when the moisture of soybeans

1.5f-

.....
'"

""El

-.,...

...

5) Increase of tocopherol by drying moisteneIl.

soybeans.

In the experiment in 3) and 4), we have

observed that tocopherol in moistened soy
beans is less than in soybeans with ordinary
moisture but increases again when soybeans
are dried.
We supposed that the changes of the quanti.
ty of tocopherol might be accelerated by
sunlight and the enzyme action might be
involved in the phenomenon. The following
experiments were carried out in order to

LO

'"
0

...
<ii
...
()

E-<
0.5

18

16

14

12

10

Moisture of soybeans (%)

FIG.

4. Changes of Tocopherol Content by Drvmg

Moistened Soybeans
in the Sun and in the Dark

at 40C.

Studies on Cause of Color Reversion of Edible Sobyean Oil and its Prevention.

Part I

911

is from 18% to 15%, but it increases very

rapidly when the moisture is less than 15%.
When the moisture of beans falls below 11 %
the curve becomes flat again. Mark EB on the
curve in Fig. 4 indicates the position of the
soybeans (moisture 13%) before moistening.

must be stopped when the enzyme becomes

inactive by heating.
Therefore, the following two severe heating
conditions were chozen in this experiment.
Soybeans with 18% moisture (total tocopherol 0.28 mgjg) were placed for 7 hrs in a vacuChanges in the quantity of tocopherol by drying um dryer kept at 90C under 15-20 mmHg
and cooled under reduced pressure for a night.
moistened soybeans in the dark at 40C.
In
the
other
experiment,
the
same
soybeans
Changes in the quantity of tocopherol during the drying the moistened soybeans in the were heated at 120C for an hour. The
analysis
of
the
dried
soybeans
is
shown
in
.dark were determined in order to make clear
Table
VII.
whether the increase of tocopherol during the
drying of moistened soybeans in the sun is
TABLE VII. CHANGES OF TOCOPHEROL CONTENT BY
accelerated by sunlight.
HEAT DRYING OF MOISTENED SOYBEANS
Soybeans with 18% moisture (total tocoMoisture of
Total
Oil
Soybeans
Tocopherol
pherol 0.28 mgjg) were put in a completely
Drying
(Dried
(Xylene
in Crude
Conditions
Basis)
dark were determined in order to make clear
MethOd)
Oil
(%)
(mgjg)
(%)
the thermostat were stirred by a fan. The
90C, Vacuum,
2.6
20.8
1.52
changes of the tocopherol content by the dry7 hrs.
ing are shown in Table VI.
120C, Ordinary
9.2

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Pressure, 1 hr.

21.7

1.50

TABLE VI. CHANGES OF TOCOPHEROL CONTENT

BY DRYING MOISTENED SOYBEANS
IN THE DARK AT 40C

The values of increase of tocopherol are

nearly equal to those described before, in
which case moistened soybeans were dried in
Moisture of
Total
Oil
Soybeans
Tocopherol
the
sun.
It
is
expected
that
the
heating
of
Drying
(Dried
(Xylene
in Crude
Conditions
Basis)
soybeans at 90C for 7 hrs or at 120C for
Method)
Oil
(mgjg)
1 hr will make the enzyme inactive, but the
(%)
(%)
40'C Dark 7 hrs.
12.0
1.32
22.3
values of increase of tocopherol in either case
23 hu.
1.53
8.4
21.6
"
are nearly equal to those described in 3) in
47 hrs.
1.57
21.6
7.4
"
which moistened soybeans were dried in the
As shown in Fig. 4 both curves are identical sun.
when the moisture of soybeans is less than 12%
This fact shows that the enzymes in soybeans
and the increase of tocopherol in the case of concerned with the transformation of tocodrying in the dark is a little less than in the pherol are relatively resistant to heat.
case of drying in the sun, from which it can
Changes in tocopherol by repeated treatment of
be concluded that sunlight gives practically no moistening and drying of soybeans.
influence on the increase of tocopherol.
Raw soybeans (moisture 13%) and soybeans
with 18% moisture were dried in a vacuum
dryer kept at 85C for 15 hrs until the weight
The increase or decrease in the quantity of became almost constant. These dried soytocopherol in dried or moistened soybeans is beans were moistened again by the same prosupposed to be due to the biochemical re- cess described already. Table VIII shows the
action. If it is caused by enzyme action, it analysis of soybeans at each stage of drying
Changes in the quantity of tocopherol by heat
drying of soybeans.

Mamoru KOMODA, Noriji ONUKI and Ichiro HARADA

912

TABLE VIII. CHANGES IN TOCOPHEROL IN REPEATED

DRYING AND MOISTENING OF SOYBEANS

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A Raw Soybeans
B A-moistened
C A -dried
DB-dried
E C-moistened
F D-moistened

Moisture of
Soybeans
(Xylene
Method)

Oil
(Dried
Basis)

(%)

(%)

13.0
18.2
1. 9
1. 9
18.0
18.3

22.0
22.0
20.5
20.7
21.9
22.1

Total
Tocopherol
in Crude
Oil
(mgjg)
1.25
0.28
1.90
1.87
0.33

0.34

and moistening.
It was already pointed that the tocopherol
in raw soybeans with ordinary moisture (13%)
increased when they were dried, and decreased
when they were moistened.
The noticeable and interesting point in the
results of this experiment is that the quantity
of tocopherol changes according to the moisture of soybeans during the repeated drying
and moistening treatment of soybeans and
that it shows a certain fixed value at a certain
moisture level of the beans.
DISCUSSION

The tocopherol in raw soybeans increases

or decreases according to the moisture of
beans, especially in the moisture range from
II to 15%. Hitherto, nothing has been reported as to the relation between the quantity
of tocopherol and the moisture of soybeans.
M. Roche 16 ) observed that the loss of tocopherol was least when wheat germ or flour
was stored at a low temperature in the atmo16) M. Roche, W. Feldheim and B. Thomas, Ernahrungsforschung, 3, 386 (1958).

sphere of high moisture, ahd that the loss

increased when the moisture exceeded. ,12%.
M. Roche reported also that the mechanism
for the above phenomenon was not clear and
that the loss of tocopherol during the storage
of wheat germ was not caused by the oxidative enzymes. We observed that the decrease
of tocopherol in soybeans became sharp when
the moisture of beans exceeded 12%.
In the case of soybeans, the decrease of
tocopherol appeared rapidly when soybeans
were moistened, while, in the case of wheat
flour, the loss of tocopherol occurred very
slowly during a long storage, from which it is
supposed that the loss of tocopherol in both
cases is caused by different functions.
The tocopherol content in moistened soybeans is small but it increases again when the
beans are dried. Moreover, this phenomenon
is only observed in raw whole soybeans and
not in crushed and dried soybeans.
Therefore, the changes in the quantity of
tocopherol according to the moisture of the
soybeans may not be the simple oxidation of
tocopherol by air but are supposed to be due
to the biochemical reaction.
To prove this, drying of soybeans at 90C
or at 120C was attempted, but the increase
of tocopherol in each case was nearly equal
to the case when beans were dried at 40C or
dried in the sun.
From this we have learned that the enzymes
in soybeans concerned with the transformation
of tocopherol are relatively resistant to heat.
The roles of enzymes in soybeans in the transformation of tocopherol will be made cleat by
further studies.