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: hbb38160@wave.odn.ne.jp * 2 E-mail: ochiai@toua-u.ac.jp

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)