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College of Pharmaceutical Sciences, Matsuyama University, 4-2 Bunkyo-cho, Matsuyama, Ehime 790-8578, Japan
Division of Food Additives, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku,
Tokyo 158-8501, Japan
2
Received November 26, 2008; Accepted February 9, 2009; Online Publication, May 7, 2009
[doi:10.1271/bbb.80832]
presence of a wide array of compounds, including monoterpenes, such as 1,8-cineole, sesquiterpenephloroglucinol derivatives, avonoids, tannins, and
related polyphenols.3,713)
As part of studies evaluating ocial food additives
conducted by research groups of the Ministry of Health,
Labour and Welfare of Japan since 2003, eucalyptus
leaf extract was found to include gallic acid, quercetin
3-O--D-glucuronide, kaempferol 3-O--D-glucuronide,
globuluside, cryptmeridiol, 4-epi-cryptmeridiol, 3,13dihydroxyurs-11-en-28-oic acid, -eudesmol, and macrocarpal I, of which the rst three were the main
constituents.14) However, -diketones, dened in the
ocial list as the main constituents, were not detected.
Thus the standardized markers in eucalyptus leaf extract
are currently undened.
This paper describes a re-examination of the constituents of the commercial food additive eucalyptus leaf
extract based on a detailed characterization of the
antioxidant polyphenols of the raw material (E. globulus
leaves). It also proposes a new standard, including a
characteristic marker ingredient, for the quality of this
antioxidant food additive.
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Results
HPLC analysis of eucalyptus leaf extract
Two commercial eucalyptus leaf extracts products
(eucalyptus A and B) used as antioxidant food additives
were analyzed. Figure 1 shows their HPLC chromatograms at 270 nm. Eucalyptus A showed an HPLC prole
34
2
10
20
30
40
50
min
1062
Y. AMAKURA et al.
OH
HO
OH
HO
HO
COOH
O
H2 C
O
O
OH
HO
HO
HO
HO
OH
O
CH 2
OH
4: R=OH
5: R=H
OH
OH
6: R=OH
7: R=H
O
R
HOOC
OH
OH
O
OH
OH
HO
HO
OH
HO
OH
OH
HO
R
O
O
O
OH
HO
HO
HO COOH
OH
OH
HO
OH
OH
HO
OH
OH
HO
OH
OH
HO
HO
HO
HO
OH
OH
O
O
HO
OH
OH
HOH2 C
OH
OH
OH
HO
OH
HO
OH
HO
HO
O
OH
CH2 OH
H 2C
O
O
OH
O
OH
HO
OH
HO
OH
OH
OH
10
HO
OH
HO
HO
O
HO
HO
O
O
O
O
H2 C
HO
HO
O
O
HO
O
OH
OH
HO
OH
HO
OH
O
OH
OH
OH
OH
14
OH
HO
H 3C
CH2OH
O
HO
OH
OH
OH
12: R=R'=H
13: R=H, R'=G
15: R=R'=G
OH
HO
OH
OH
OH
HO
O
H2C
O
O
O
C
O
OH
OH
O
OH
OH
OH
11
OH
OH
OH
OH
H 3CO
OH
OH
HO
R'OH2C
RO
O
C
OH
O
HO
OCH 3
Galloyl (G) =
C
O
OH
OH
18
16: R=H
17: R=CH3
1063
0h
1h
8h
10
20
30
40
50
min
s.f.
ori.
a'
b'
c'
A B
a b
a'
c'
1064
Y. AMAKURA et al.
E. globulus (fresh leaves)
Chloroform extract
EtOH extract
50% aq. EtOH extract
Food additives
Eucalyptus A
Eucalyptus B
0
1000
2000
3000
4000
5000
mol TE/g
Total polyphenolsb
(mg/g)
Eucalyptus A
Eucalyptus B
19.0
390.0
360
29
162.7
23.5
19.1
34
237
548
477.0
44.1
24.7
30
357
408
>100 mg/ml
>100 mg/ml
>100 mg/ml
29
Eucalyptus Oil A
Eucalyptus Oil B
Eucalyptus Oil C
a
b
Discussion
Besides the previously reported major constituents
(1, 4, and 5) of the food additive eucalyptus leaf extract,
compounds 2 and 3 were newly detected as other major
components of eucalyptus A, which showed a potent
antioxidative eect in this study. Oenothein B (2) is a
unique hydrolyzable tannin dimer with a macrocyclic
structure that is known to exhibit diverse biological
activities, such as anti-tumor, antiviral, and a restoration
eect of antibiotics toward methicillin-resistant Staphylococcus aureus (MRSA) as well as antioxidant
Acknowledgments
The authors thank Ms. Mie Tokuhara of Matsuyama
University, for technical assistance. Thanks are also
extended to the Japan Food Additives Association for
providing eucalyptus leaf extracts, and to Nagaoka
Perfumery Co., Ltd., for donating dried leaves of
E. globulus. This work was supported by a Health and
Labour Sciences Research Grant from the Ministry of
Health, Labour and Welfare of Japan.
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References
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