食用油としてのツバキ油の効果と抗酸化性 · -shi, fukuoka prefecture in nov....
TRANSCRIPT
食用油としてのツバキ油の効果と抗酸化性
誌名誌名 日本食品保蔵科学会誌
ISSNISSN 13441213
著者著者時枝, 久子落合, 為一
巻/号巻/号 31巻5号
掲載ページ掲載ページ p. 253-260
発行年月発行年月 2005年9月
農林水産省 農林水産技術会議事務局筑波産学連携支援センターTsukuba Business-Academia Cooperation Support Center, Agriculture, Forestry and Fisheries Research CouncilSecretariat
( 25 ) Food Preservation Science VOL. 31 NO. 5 2005 (Article) 253
Studies of Camellia Oil as Edible Oil and Its Antioxidative Activity
TOKIEDA Hisako*l and OCHIAI Tameichi*2
* 1 Department 01 Home Economics.めushuWomen's University
1 -1. Jiyugaoka Yahatanishi. Kitaわlushu.Fukuo知 807-8586
* 2 Depar加 ent01 Engineering. Universiη01 East Asia
2 -1. lchinomiyaGakuen-cho. Shimonoseki-City. Yamaguchi 751 -8503
The antioxidative ability and sensory characteristics of camellia oil were studied. to elucidate its
potential as an edible oil. The measurement of its peroxide value CPOV) showed that camellia oil was
stable for more than 3 months at 400
C. The filtrate containing antioxidative substances was
fractionated using column chromatography. and the fractions active toward Folin-Ciocalteu reagent
were collected and isolated in two materials. HPLC showed that the first material was essentially
composed of one compound and the second of three compounds. The NMR spectra of the two
materials were similar to each other. The peaks in the aromatic region were superimposable on those
of kaempferol. and the presence of a kaempferol moiety was strongly suspected. F AB-MS spectra
showed that the molecular weights of the four compounds regularly increased by 100. Detailed
structures of these compounds are now under investigation. Sensory evaluation of camellia oil in terms
of its color. flavor. taste, and total evaluation was carried out Although dressings prepared using
without heat treatment were not preferred, fried spaghetti and fried potato using heated camellia oil
were preferred over those prepared from salad oil with a significant di妊erence.
Camellia oil Ctsubaki oi!) is produced by extraction
from seeds of Camellia japonica LINNE, and has
been used traditionally as a high-grade hair oil for
women in Japan. Although the main use of this oil
has been in cosmetics, it also has potential use as an
edible oil or medicinal oil for treating skin diseases').
Most carboxylic acid moieties of camellia oil are
composed of c.s-monounsaturated acids Coleic acid,
>80%), which is less easily oxidized in air than
polyunsaturated carboxylic acids. It has also been
recently reported that monounsaturated carboxylic
acids decrease LDL concentration in the blood,
which causes aortosclerosis. From a dietary
perspective , the intake of monounsaturated
carboxylic acids should be 1. 5 times that of
saturated or polyunsaturated carboxylic acids. Thus,
camellia oil, which is rich in monounsaturated
carboxylic acids. has a potential for edible use and
is a potential oil resource. Few studies of camellia
oil as an edible oil. however. have been reported to
date2). In this paper, we describe the preservability,
cookability and sensory properties of camellia oil.
* 1 E-mail: [email protected] * 2 E-mail: [email protected]
CReceived Jan. 24, 2005 ; Accepted Sep. 5, 2005)
Preliminary experiments showed that camellia oil
is less easily oxidized in air than other oils that contain
monounsaturated carboxylic acid moieties; hence, the
presence of antioxidants in this oil was suspected. It
was reported that buds and leaves of Camellia japonica
contain antioxidants, such as cammelliatannins3)").
However, the chemical structures of antioxidants in
seeds of this plant have not yet been determined.
In this paper, we describe the separation and
structural properties of antioxidants from seeds of
Camellia japonica.
Experimental
1. Materials
Camellia oil was produced at Jinoshima, Munakata
-shi, Fukuoka prefecture in Nov. 1999. The seeds of
Camellia japonica L. were gathered at the same
place in Nov. 2003. Other oils were purchased from
local markets: olive oil. Ajinomoto Co., Ltd.; corn
oil. Ajinomoto Co., Ltd.; soybean oil. Wako Co.,
Ltd.; and salad oil, Nisshin Oil Products Co., Ltd.
254 Food Preservation Science VOL. 31 NO.5 2005 ( 26 )
2. Analyses of carboxylic acids from oils
Carboxylic acids from camellia oil were analyzed
by gas chromatography according to a standard
method under the following conditions: column.
FF AP on Chromosorb W (A W). 80-100 mesh. 2皿
x 2 m; temperature. 230t on injection into a 2000
C
column; and FID detector.
3. Measurement of peroxide value (POV)
The POV of each oil was measured at 40t
according to a standard method5).
4. Sensory evaluation
Sensory evaluation was performed for color. flavor.
taste. fattiness and overall acceptance using paired
preference. ranking and rating tests. The panel
comprised 15 to 20 female students aged between
19 and 22.
AX505HA mass spectrometer. and UV spectra were
recorded on a Shimazu UV吋 isiblespectrophotometer
UV1600. 1H-and 13C-NMR spectra were recorded
on a Bruker BioSpin A VNCE 500 (500 MHz)
spectrometer at room temperature. unless otherwise
mentioned. Chemical shifts were based on TMS.
HPLC was performed using a Shimazu LC-6 A
Shim-pack VP-ODS column (4.6 x 250m皿)with H,O/
MeOH/formic acid (55.8/43.9/0.3 voI) as mobile
phase at 300
C.
6. Isolation of antioxidants
The separation procedure for antioxidants is
shown in Fig. 1. Seeds (1. Okg) of Camellia japonica
L.. which were collected at ]inoshima. Munakata-shi.
Fukuoka prefecture in November 2003. were
homogenized in 1 R of methanol. The debris in the
5. Determination of chemical structures of homogenate was filtered off. and the filtrate was
antioxidants in Camellia japonica seeds concentrated to 100mR using a rotary evaporator.
The resulting solution was then extracted with 250 F AB-MS profiles were recorded on a ]EOL ]MS-
n-hexane layer (camellia oiI)
Evaporation to dryness
n-hexane
acetone
Separation by column chromatography
‘一一一一一一一 acetone
active toward Folin-Ciocalteu reagent
active toward Folin-Ciocalteu reagent
骨一一一一_ethanol/H,O
active toward Folin-Ciocalteu reagent
-一一一一一 methanol/H,O
active toward Folin-Ciocalteu reagent
Flavone 1 and Fraction B
Fig. 1 Separation procedure used in this study
( 27) (ArticleJ Antioxidative Activity of Camellia Oil 255
me of n-hexane. Subsequently, 100me of methanol was
added to the methanolic layer and the solution was
twice extracted with 50me of n-hexane. 300me of
acetone was then added to the methanolic layer
The resulting precipitates were filtered off, and the
filtrate was evaporated to dryness. The residue was
chromatographed on Wako Gel C-200 (2. Ocm x 20cm)
with acetone/MeOH (90/10) as developing solvent,
and fractions that reacted with Folin-Ciocalteu
reagent were collected. After the solvent was
removed, the residue was chromatographed on
Diaion HP-20 (2. Ocm x 20cm) with aqueous methanol
as developing solvent. The concentration of
methanol was increased stepwise ciム O→10→20→30
→40→50→60→70→80→90→100%) . Fractions that
reacted with Folin-Ciocalteu reagent were collected;
they were found to comprise four main compounds
by HLPC. They were further chromatographed on
Sephadex LH-20 (1. 0 x 30 cm) with 30% aqueous
ethanol. Fractions rich in the compound with the
shortest HPLC retention time were collected and
further purified using Diaion HP-20SS (1. Ocm x 30cm) .
Subsequently, the compound was isolated at more
than 90% purity by HPLC as a pale yellow solid
(ll mg) and named Flavone I.
The other three compounds could not be
separated by the above procedure; thus, the
mixture of these compounds was designated
Fraction B.
Results and Discussion
resulting peroxides are often used as an indicator of
the degree of deterioration of oils. A peroxide value
(POV) of 30 meq/kg is a critical value that indicates
the limit of deterioration. Four types of oil; namely,
camellia, olive, soybean and salad oils, were stored
at 40t, and the POV of each oil was periodically
measured. At 40t, oxidation was found to proceed
four times as fast as that at room temperature. The
results for this test are shown in Fig. 2. The
increase in the POV of camellia oil was low
compared with those in the POVs of the other oils.
Even after three months, the POV of this oil was
as low as 25 meq/kg, compared with its initial value
of 3. 8 meq/kg. For salad oil, no change in POV was
observed for the first 18 days, after which POV
gradually increased. After 70 days, the POV of
salad oil was 195 meq/ kg, before it rapidly
increased to 1,324 meq/kg after 90 days. The POV
of soybean oil also increased more rapidly than
those of the other oils. After 2 months, the POV of
soybean oil was 100 times as high as the initial
value of 5.85 meq/kg and peaked at 1,585 meq/kg
after 85 days. These findings show that camellia oil
is stable against 'auto-oxidation. and has significant
preservative qualities.
3. Isolation and structural determination of
chemical antioxidants in C. japonica L.seeds
The above discussion suggests that camellia oil is
stable against auto-oxidation. which we suspected to
be due to the presence of antioxidants. Since the
Folin-Ciocaltue test for camellia oil showed that the
1. Carboxylic acid composition oil contained only small amounts of antioxidants. it
The carboxylic acid composition of camellia oil was thought that it would be easier to directly
was determined by gas chromatography according
to a standard method2) (Table 1). The data
obtained were essentially identical to those 16
previously reported6) with no special features
observed in this sample.
2. Stability against oxidation
In general, oils are easily auto-oxidized during
storage to yield peroxides. and the amounts of the
Table 1 Carboxylic acid composition of camellia oil
Carboxylic acid Content
palmitic acid 6-9%
stearic acid 2-3%
oleic acid 83-86%
linolic acid 4-6%
others trace amount
u
u
m
8
6
4
(NO
H
×品』
¥σug)〉
O仏
nU
2
0
40 80 100 60 20 Reaction time (days)
Fig. 2 Changes in peroxide values of camellia oil and other
vegetable oils at 40t
• : camellia oil,ム:olive oil, 0: soybean oil, 企 :salad oil
of kaempferol (3, 4', 5, 7-tetrahydroxyflavone)
(Table 2). In addition, the13C NMR
supported the presence of
(Fig.5).
The peaks in Region 2 were complex, and their
interpretation requires further study . Since
methylene and methyne hydrogen atoms in sugars
peaked in this region, we suspected the presence of
a compound with a sugarlike structure. The peaks
in Region 1 (σ= 1. 18, D,J = 6. 1 Hz;σ= 1. 09. D.] =
6.4 Hz) indicated the presence of two types of
methyl groups connected to a methyne group.
The 'H-NMR spectrum of the mixture of the four
antioxidants was similar to that of Flavone 1, where
Regions 1 and 2 were also essentially identical.
However, we could not determine a structure that
was compatible with the mass and NMR spectra.
The precise structures of Flavone 1 and the other
three compounds are now under investigation and
will be reported in the future.
4. Sensory Evaluation
( 1 ) Non-preheated Camellia Oil Sensory
evaluations of French dressing and mayonnaise
prepared from camellia oil and salad oil in terms of
color, flavor, taste, and total evaluation were carried
out by a paired test. Non-preheated Camellia oil was
evaluated by a panel of 25 female students aged
between 19 and 22. The results are shown in Table
3. Since non-preheated camellia oil is deep yellow
with a characteristic flavor, most of the students in
the panel preferred the French dressing prepared
from salad oil in terms of all the items evaluated. In
particular, significant di丘erences were found in
isolate the antioxidants from C. japonica 1. seeds.
Antioxidants in fresh seeds of C. japonica 1. were
extracted as described in the experimental section.
Finally, two fractions that were reactive toward
Folin-Ciocalteu reagent were obtained. The first
fraction was essentially composed of a single
compound (>90% by HPLC). The UV spectrum of
the first fraction showed three peaks at tlmax =
352.0run (log.:: = 4.0), 268.5run (log.:: = 4.2), and 215run
(s) (log.:: = 4. 2) , which are typical of flavenoid
compounds (Fig. 3). We thus named the first fraction
Flavone 1. The second fraction was composed of
three compounds and named Fraction B.
The mixture of the four compounds showed four
[M-Hr ion peaks at m/z 425, 525, 625, and 725 in
the F AB-mass spectrum. This shows that the
molecular weights of the four compounds regularly
increased by 100. Since the order of elution from
the Sephadex LH-20 column showed that Flavone 1
had the highest molecular weight, we estimated the
molecular weight of Flavone 1 to be 726. This
molecular weight was then used to calculate the ε's
of Flavone 1.
The 'H-NMR spectrum of Flavone 1 consisted of
three regions.
① Region 1,σ1.0-σ 1.2
② Region 2,σ3.0-σ5.5
③ Region 3,σ6.0-σ8.3
Regions 1, 2, and 3 corresponded to a methyl
group region, methylene or methyne substituted by
heteroatom region, and an aromatic hydrogen
region, respectively (Fig. 4). The peaks in Region 3
could be almost completely superimposed on those
( 28 )
spectrum
kaempferol moiety a
2005 NO.5 VOL. 31 Food Preservation Science 256
a
。凶
h山田-[
出向。.司-[・
-[NHdJ[
。∞∞HH
B 。命的。
。。∞.∞
A
日l1'l 小3
HPLC charts of A: Flavone 1 and B: Fraction B
Cコひ司
l1'l ,.....,
o ,.....,
ばコo l1'l ~司
o ,.....,
Fig.3
l1'l o
257 Antioxidative Activity of Camellia Oil CArticleJ ( 29 )
non
H,O
11 if if
司法叫。的
一
NN.的出
目。.N
o ppm 2 4 6 8
OH
NMR spectrum of Flavone 1 Fig.4
and I Flavone of data spectral Table 2 IH NMR
kaempferol O HO
OH
O OH
Flavone 1
6.17
6.38
6.90
8.07
σ=6.18 (C6・H;D)
6.38 (C 8 -H ; D)
6.89 (C3ぺH,C 5 ' -H: DD)
8.07 (C 2ぺH.C 6ぺH: DD)
Kaempferol
Chemical structure of kaempferol Fig.5
Table 3 Number of panelists who chose French dressing and mayonnaise prepared
using camellia oil and salad oil without pre-heat treatment
Mayonnaise French dressing Sensory evaluation
MS MT FS FT
15 10 15 10 Color
* 7 18 15 10 Flavor
*** 2 23
*** 3 22 Taste
*** 2 23 ハU
M
一一Total evaluation
FT or FS: French dressing prepared using camellia oil or salad oil
MT or MS: Mayonnaise prepared using camellia oil or salad oil
significance: * * *p<O.OOl, * *p<O.Ol. *p<O.05
number of panelists : 25. by paired preference testing
258 Food Preservation Science VOL. 31 NO.5 2005 ( 30 )
terms of taste (p<O.OO1) and total evaluation (p<
0.001) ; the results for mayonnaise were not
significantly di妊erent.
Since the deep yellow color and characteristic
flavor of camellia oil seemed to render the oil less
favorable, we performed a sensory evaluation of
French dressing with ingredients, such as onion,
goma-miso, and plumvinegar. by ranking. Three oils,
namely, camellia oil. salad oil. and olive oil. were
used. The results are summarized in Table 4.
Although no significant differences were observed
for all the items evaluated, camellia oil dressing
ranked the highest among the three dressings in
terms of taste and total evaluation.
( 2) Preheated Camellia oil French dressing and
mayonnaise were prepared from camellia oil
preheated between 90-100t for 10 min and then
subjected to sensory evaluation using the paired
test. The results are summarized in Table 5. For
French dressing, the flavor of the camellia dressing
was preferred over salad oil with a significant
difference (p< 0.005). For the other items evaluated,
no significant di任'erenceswere observed. Mayonnaise
prepared from camellia oil was preferred over
French dressing in terms of color (p<0.005) and
taste (p<O.OO1) with significant di妊erences.
( 3) Sensory evaluation of fried spaghetti and
fried potato Since the above results showed that
preheating increased the number of the panelists
who prefered camellia oil, we used the oil in
cooking fried spaghetti and fried potato. The results
of the sensory evaluation of preheated camellia oil
and salad oil by the paired test are summarized in
Table 6. For fried spaghetti, camellia oil was
significantly preferred over salad oil; in terms of
fattiness for fried potato, camellia oil was preferred
over salad oil in terms of all the items evaluated 0. e.,color, flavor, taste, and total evaluation p<O.Ol ;
fattiness p < 0.05) with significant differences
Sensory evaluation by scoring also showed the
preference for camellia oil (Table 7).
( 4) Increase in POV during preheating The
POV of camellia oil was measured after preheating
under various conditions. The results are shown in
Table 8. Too much preheating promoted auto-
oxidation. Although preheating at 1000
C for only 10
Table 4 Rank sum of French dressings prepared using camellia oil, olive oil and salad oil
French Onion Plum vinegar Goma miso dressing
Senalsuoaruy 。 Camellia Olive Salad Camellia Olive Salad Camellia Olive evalUauon
Color 36 29 25 24 40 28 31 23
Flavor 31 27 32 31 30 29 26 29
Taste 37 26 27 31 33 26 31 25
Total evaluation 39 25 26 27 34 29 31 25
number of panelists : 15, by rank testing.
Table 5 Number of panelists who chose French dressing and mayonnaise prepared using camellia oil and salad oil with pre-heat treatment at 90-100
o
C for 10 min
French dressing Mayonnaise Sensory evaluation
FT FS MT
* Color 10 15 19
* Flavor 18 7 17
*** Taste 17 8 21
* Total evaluation 16 9 19
FT or FS: French dressing prepared using preheated camellia oil or salad oil MT or MS: Mayonniase prepared using preheated camellia oil or salad oil
significance: * * *p<O.001, * *p<O.01, *p<O.05 number of panelists : 25 by paired preference testing
MS
6
8
4
6
Salad
36
34
33
34
259 Antioxidative Activity of Camellia Oil (ArticleJ ( 31 )
Number of panelists who chose pre-heat-treated camellia oil and salad oil Table 6
Salad oil
qJaaτnLntnL
Fried potato
Camellia oil
..
‘ .
.
e
・
--------
4aτquFDAHvrD
。ゐ
ηLqLnLηL
Salad oil
弓
t
ハUP
O
凋仏
τβb
唱
EA
唱
EA
Fried spaghetti
Camellia oil
ηLAUdqJRdqJ
唱』
A
噌
EA
唱
Ei
九_______________ Cooking
Sensゅryevaluation ____________
Color
Flavor
Taste
Fattiness
Total evaluation
Significance: **p<O.Ol, *p<O.05 number of panelists・19-27,by paired preference testing
was strongly suggested. Sensory evaluation of
camellia oil showed that dressings prepared from
this oil when not heat-treated were not preferable.
although fried spaghetti and fried potato cooked
using camellia oil were found to be preferable with
significant differences.
Table 7 Sensory scores of French fries prepared using
camellia oil and salad oil
Average
References
1) FUKATSU. T. : Touyousyokubutu (The
University Publication Station. Tokyo).
(1993)
2) TOKIEDA. H.. T AKIZA WA. K. and IKEDA. R.:
States of Utilization of Camellia Oil Producer on ¥
the Solitary Island in Kyushu Area as a Cooking
Oil and Preference to Oil, ; The Japan Association
for the Integrated Study of Dietary Habits. 10 (2).
30-36 (1999)
3) YOSHIDA. T.. CHOU. T.. MARUY AMA. Y. and OKUDA.
T.: Tannins of Theaceous Plants. n. Camellia A
and B. Two New Dimeric Hydro-lyzable Tannins
from Flower Buds of Camellia japonica L. and
Camellia sasanqua THUNB.. Chem.Phanη.Bull.. 38
(10). 2681-2686 (1990)
4) HATANO. T.. SHDA. S.. HAN. L. and OKUDA. T.:
Tannins of Theaceous Plants. III. Camelliatannins
A and B. Two New Complex Tannins from
Camellia japonica L.. Chem. Phann. Bull.. 39 (4).
876-880 (1991)
5) ]ournal of the ]apanese society for food science
and technology. Food analysis method editing
committee Shokuhin-bunsekihou (Korin. Tokyo).
p.553 (1992)
6) SINTANI. 1: Syokuhinyusinokagaku (Saiwai Syobou.
Tokyo). p.4 (1992)
Hosei
p.231
The
4.20
3.30
3.67
3.00
4.07
2.74
3.48
2.85
4.00
2.74
Sum
114
89
99
81
110
74
94
77
108
74
Sample A
Sample B
Sample A
Sample B
Sample A
Sample B
Sample A
Sample B
Sample A
Sample B
Color
Flavor
Fattiness
Taste
Total
evaluation
Sample A: camellia oil. sample B: salad oil number of panelists : 19-27 by score testing
Table 8 Amount of POV in camellia oil pre-heat-treated
at 100 or 180t
Jemperaturel 180t
AUnunU
唱
EAnr“n〈JU
57.8 meq/kg
68.8
77.2
100t
7.6 meq/kg
52.6
68.4
Time (min)
min slightly increased the POV. the POV after
preheating at 100t for 20 min reached 52.6 meq/
kg from an initial value of 6. 9 meq/kg.
The antioxidative properties of camellia oil were
studied and sensory evaluation was also carried out
determine its potential as an edible oil. POV
measurement showed that camellia oil remains
stable for more than 3 months at 40t. Separation
and structural identification of the antioxidants
present in Camellia japonica L. seeds were
attempted. and the presence of a kaempferol moiety
POV of camellia oil without pre-heat treatment : 6.9 meq/kg
Conclusions
to
260 Food Preservation Science VOL. 31 NO.5 2005 ( 32 )
食用油としての
ツバキ油の効果と抗酸化性
時枝久子判・落合為一叫
* 1 九州女子大学
(干807-8586 福岡県北九州市八幡西区自由ヶ丘 1-1)
* 2 東亜大 学
(干751-8503 山口県下関市ーノ宮学園町 2-1)
ツバキ油の食用油としての可能性を探るため,ツバキ
油の抗酸化性および、晴好性についての研究を行った。ツ
バキ油は400
Cの恒温器内で 3カ月以上酸化されず,良好
な抗酸化性を示した。抗酸化性物質を同定するため,
Camellia japonicaの種子から抗酸化物質の抽出を試みた。
ツバキの種子を粉砕物からのメタノール抽出物をカラム
クロマトグラフイ(シリカゲル, HP-20,ゲルクロマト
グラフイー)により分画し, Folin-Ciocalteu試薬に活性
な部分を追跡した。最終的にHPLC的にほぼ単一物質を
含むフラクションと 3種の物質が含まれるフラクショ
ンが得られた。前者の単一物質と思われる物質のNMR
スベクトルと,後者の 3種の混合物質のNMRスベクト
ルは極似しており,芳香族領域,糖領域,メチル基領域
に吸収を示した。芳香族領域の吸収はkaempferolの吸
収とほぼ完全に一致した。質量分析 (FAB-MS)の結果,
これら 4種の物質は分子量が100づっ異なる物質 (Mw
= 426, 526, 626, 726)であることが判明した。さらに
詳しい構造を検討中である。ツバキ油の晴好性は女子学
生をパネラーとして色,香り,味,脂っこさ,総合評価
の官能検査を行った。サラダ油との比較において,非加
熱料理の各種ソース類ではツバキ油はあまり好まれなか
った。加熱料理においてはツバキ油が有意差をもって好
まれた。
(平成17年1月24日受付,平成17年9月5日受理)