This article was downloaded by: [181.66.156.

48]
On: 28 May 2015, At: 20:26
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK

Agricultural and Biological Chemistry

Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/tbbb19

Antioxidative Effect of the Constituents

of Rosemary (Rosmarinus officinalis L.)
and Their Derivatives
a a a
Reiko Inatani , Nobuji Nakatani & Hidetsugu Fuwa
a
Department of Food and Nutrition, Faculty of Science of Living,
Osaka City University, Sumiyoshi-ku, Osaka 558, Japan
Published online: 09 Sep 2014.

To cite this article: Reiko Inatani, Nobuji Nakatani & Hidetsugu Fuwa (1983) Antioxidative Effect
of the Constituents of Rosemary (Rosmarinus officinalis L.) and Their Derivatives, Agricultural and
Biological Chemistry, 47:3, 521-528

To link to this article: http://dx.doi.org/10.1080/00021369.1983.10865682

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the
Content) contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever
or howsoever caused arising directly or indirectly in connection with, in relation to or
arising out of the use of the Content.

This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &
Conditions of access and use can be found at http://www.tandfonline.com/page/terms-
and-conditions
 Agric. Bioi. Chem., 47 (3),521-528,1983 521

Antioxidative Effect of the Constituents of Rosemary

(Rosmarinus officinalis L.) and Their Derivatives t

Reiko INATANI, Nobuji NAKATANI* and Hidetsugu FuwA

Department of Food and Nutrition, Faculty of Science of Living,
Osaka City University, Sumiyoshi-ku, Osaka 558, Japan
Received August 9, 1982

Antioxidative activity was measured on extracts of rosemary (Rosmarinus officinalis L.), one of
the herb spices belongs to the Family Labiatae, using different solvents. Strong activity was
observed in the weakly acidic fraction of the n-hexane extract, which was further fractionated and
purified to afford several active compounds. An ordorless and colorless compound, named
rosmanol, showed high antioxidative activity in both lard and linoleic acid, and particularly in lard,
Downloaded by [181.66.156.48] at 20:26 28 May 2015

was about four times more active than synthetic antioxidants, such as BRT and BRA.
Furthermore, the antioxidant activities of the derivatives of rosmanol and carnosol were measured
by the ferric thiocyanate method and the TBA method, to observe the correlation between chemical
structure and activity as an antioxidant.

Various efforts have been made to preserve
the quality of foods. In particular to depress
rancidity of fats and oils, synthetic antioxi-
dants such as tert- butyl-4-hydroxyanisol (BHA)
and tert-butyl-4-hydroxytoluene (BHT), and
natural antioxidants such as tocopherols, are
widely used. However, the use of BHA and
BHT as food additives is restricted in several
countries, because these compounds seem to
cause an undesirable effect on the enzymes of
liver and lung. Consequently, we have directed
our attention towards edible plants as re-
sources of safer and more effective natural
antioxidants, and have especially investigated
the anti oxidative effect and chemical structure
of the components of spices.
Spices are generally used to flavor and sea-
son foods. In addition some spices have been
applied for their antiseptic activity and have
been utilized in the storage of meat from early
times. Chipault et al. l ,2) and Palitzsch et al. 3 ,4)
measured the antioxidative effect of many
kinds of spices, and reported that the spices
belonging to the Family Labiatae have an
especially strong antioxidative activity. In
1969, Brieskorn et al. S ) presumed that the
antioxidative effect of rosemary was based on
carnosic acid and carnosol. However, Chang
et al. 6 ) have recently suggested that other
antioxidants exist in rosemary and sage, but
they have not yet reported the isolation and
structural determination of the active com-
ponents. Very recently we found a new anti-
oxidative compound, named rosmanol, from
rosemary and reported its structure. 7 ) We de-
scribe here the antioxidative effect of rosma-
nol (1), carnosol (2) and their derivatives.
Three methods were adopted for the anti-
oxidative assay. The ferric thiocyanate and
thiobarbituric acid methods represent respec-
tively the amount of hydroperoxide of linoleic
acid at an early stage, and the production of
carbonyl compounds degraded from the per-
oxide at a later stage in an aqueous ethanolic
solution. The third method was used the active
oxygen method, which indicates the peroxide
value in an oil system.

t This study was presented at the Annual Meeting of the Agricultural Chemical Society of Japan, Tokyo, April
1982.
* Address correspondence to this author.
 522 R. INATANI, N. NAKATANI and H. FUWA
OH
2 3
CH30~OCH3
OH
4 5
MATERIALS AND METHODS
Downloaded by [181.66.156.48] at 20:26 28 May 2015
Extraction of rosemary. Dried and ground (20-60
mesh) leaves of rosemary (500 g) from France were extract-
ed with n-hexane (1200ml) under stirring at room tem-
perature. This procedure was repeated three times. The
combined extracts were freed from the solvent to yield up
to 19.7 g from the n-hexane extract. The residual leaves
were successively extracted with CH 2Ci2 four times
(1500ml each) and then EtOH twice (1500ml each) to give
36.8 g and 60.7 g, respectively. Both the hexane and the
CH2Cl2 extracts were separately steam distilled to give the
volatile and the non-volatile fractions. Each non-volatile
fraction was fractionated into the basic, the strongly
acidic, the weakly acidic and the neutral fractions in the
usual way.8) The EtOH extract was divided into the
benzene soluble and insoluble fractions.
Chromatographic fractionation of the weakly acidic com-
ponents. The weakly acidic components of the n-hexane
extract were subjected to chromatography on a silica gel
(Merck Kiselgel 60) column with benzene-acetone as the
elution solvent system to separate into twelve fractions.
The antioxidative activity of these fractions was measured
by the ferric thiocyanate method and thiobarbituric acid
(TBA) method. The antioxidative activity was also as-
sayed by the active oxygen method (AOM) on five groups,
e.g. groups I, II, III, IV and V comprising the chromatog-
raphed fraction Nos. 1-5, 6-9, 10, 11 and 12,
respectively.
Antioxidative assay.
1. Ferric thiocyanate method. The methods of Mitsuda et
al. 9 ) and Osawa et al.lD) were slightly modified. Briefly, a
mixture of a 2ml sample in 99.5% ethanol, 2.052ml of
2.51 % linoleic acid in 99.5% ethanol, 4ml of 0.05 M
phosphate buffer (pH 7.0) and l.948 ml of water was
placed in a vial (4)=38, h=75mm) with a screw cap and
placed in a oven at 40C in the dark. To 0.1 ml of this
sample solution was added 9.7 ml of 75% ethanol and
0.1 ml of 30% ammonium thiocyanate ..Precisely 3 min
OH
CH31A-rr---<C-O
W'l::::::../H
OH
after the addition of O.l ml of 2 x 10- 2 M ferrous chloride
in 3.5% hydrochloric acid to the reaction mixture, the
absorbance of the red color developed was measured at
500nm.
2. Thiobarbituric acid method (TBA).!!) To 2ml of the
sample solution, which was prepared and incubated as
discribed above, was added 2 ml of 20% trichloroacetic
acid aq. solution and 1 ml of 0.67% thiobarbituric acid aq.
solution. This mixture was placed in a boiling water bath
for 10 min and, after cooling, was centrifugated at 3000
rpm for 10 min. The absorbance of the supernatant was
measured at 532 nm.
3. Active oxygen method (AOM). The antioxidative
effect in lard was determined by the Tentative Method Cd
12-57, of the American Oil Chemists' Society (A.O.C.S.).
Lard (20 g) and the sample in a test tube (25 x 200 mm)
were kept at 98C in an oil bath under airation (2 ml/sec),
and the peroxide values were measured by the A.O.C.S.
Official Method Cd 8-53.
Samples for antioxidative assay.
1. Rosmanol (1). Rosmanol was obtained from fraction
Nos. 8 and 9 by repeated column chromatography on
silica gel as previously reported,7) mp 241C.
2. Carnosol (2). Carnosol was isolated from fraction
Nos. 6 and 7 by recrystallization from acetic acid, mp
233C. 7 )
3. Rosmadial (3). Rosmadial was obtained from frac-
tion No.4 by repeated chromatography and recrystallized
from benzene, mp 225C.!2)
4. Flavone I (5-hydroxy-7,4'-dimethoxy flavone) (4).
The fraction Nos. 3 and 4 were rechromatographed on a
silica gel column using dichloromethane as the elution
solvent to afford pale yellow crystals of flavone I, which
were recrystallized from benzene, mp 172C. MS m/z 298
(M+, base), 269, 255,166,135,132.
5. Flavone II (5,4'-dihydroxy-7-methoxy flavone) (5).
The fraction No.8 was triturated with a large amount of
acetone, and the mixture was filtered. The insoluble matter
was recrystallized from DMF to give pale yellow needles,
mp 273C. MS m/z 284 (M+, base), 255, 241, 167, 166,
 Antioxidative Effect of the Constituents of Rosemary 523

121, 118.
6. Diacetyl carnosol (6). To a solution of carnosol (2)
(70 mg) in pyridine (0.5 ml), acetic anhydride (0.5 ml) was
added and the mixture was allowed to stand at 38C
overnight. The reaction mixture was worked up in
the usual manner to afford diacetyl carnosol (87 mg),
crystallized from n-hexane as colorless prisms, mp
158C.
7. Methyl derivatives (7, 8, 9 and 10) of carnosol.I 2 ) To a
cooled solution at 5C of carnosol (2) (1.32 g) in dry
acetone (50ml) were added methyl iodide (IOml) and
anhydrous K 2C0 3 (3 g). This mixture was stirred for 6 hr
at 7 - 10C. After filtration of this reaction mixture, the
filtrate was concentrated in vacuo. Ether and H 20 were
added to the residue and the mixture was shaken. The
organic layer was dried over anhydrous MgS04 and
concentrated to dryness. The product (1.4 g) was purified
by chromatography on a silica gel column using n-hexane-
Downloaded by [181.66.156.48] at 20:26 28 May 2015
Nos. 32~45 and 54-75 gave compounds 13 and 14,
respectively. Compound 13: mp 180C (isopropyl ether),
UV Amax (EtOH) nm (loge): 209 (4.43),227 sh. (4.12), 287
(3.38). IR Vmax (Nujol) cm- I : 3400, 1745, 1250, 1220, 1200,
1170,1158,1110,1025. NMR (CDCI3 ).5: 0.95 (3H, s), 1.04
(3H, s), 1.21 (3H, d, J=7.2Hz), 1.25 (3H, d, J=7.2Hz),
1.4-2.3 (5H, m), 2.15 (lH, s), 2.8-3.5 (3H), 3.80 (3H, s),
4.57 (lH, d, J= 3.6 Hz), 4.75 (lH, br d, J=3.6Hz), 5.98
(1H, s), 7.08 (lH, s). MS m/z 360 (M+, base), 316, 301,
287, 273, 259, 245. Compound 14: mp 196C (isopropyl
ether). UV Amax (EtOH) nm (loge): 208 (4.51), 226 sh
(4.18),289 (3.47). IR Vmax (Nujol) cm- I : 3500, 3450,1760,
1250, 1220, 1172, 1120, 1078, 1048, 1000. NMR (CDCI3 )
15: 0.93 (3H, s), 1.02 (3H, s), 1.21 (6H, d, J= 6.6 Hz),
1.4-2.5 (2H), 2.22 (lH, s), 3.0-3.5 (3H), 3.74 (3H, s),
4.55 (lH, d, J=3.6Hz), 4.75 (lH, br d, J=3.6Hz), 6.12
(1H, s, D 20 exchangeable), 6.87 (IH, s). MS (m/z): 360
(M+, base), 316, 301, 287, 273, 259, 245.

acetone (7: I) as the eluent. The fraction Nos. 14-18,

22-27 and 37-50 afforded compounds 9, 7 and 8,
respectively.I2) Compound 7: colorless prisms, mp
149 _150C (n-hexane). Compound 8: colorless crystals,
mp 69C (isopropyl ether). Compound 9: colorless oil, M+
372. Compound 10 was obtained according to the pro-
~dure reported previously,12) mp 123C (n-hexane).
8. Triacetyl rosmanol (11) and Dimethyl rosmanol (12).
Acetylation and methylation of rosmanol were carried out
as previously reported. 7 ) Compound 11: colorless prisms,
mp 218C (benzene-n-hexane). Compound 12: colorless
needles, mp 226C (benzene).
9. Monomethyl rosmanol (13) and (14). To a solution of
rosmanol (100 mg) in dry acetone (20 ml) were added
methyl iodide (2ml) and anhydrous K 2C0 3 (1 g) at 5C
under stirring, the mixture being allowed to stand for 3 hr
at 5 _10C. After filtration of the reaction mixture, the
filtrate was worked up in the usual manner, followed by
purification using column chromatography. The fraction
Analytical method. Melting points were measured with a
Yanagimoto micro melting point apparatus and were
uncorrected. UV and visible absorption spectra were
determined on a Hitachi 220 spectrophotometer and IR
spectra were recorded by a Jasco IR-S. IH-NMR (60
MHz) spectra were run on a Hitachi R600 and MS were
obtained on a Shimadzu GCMS-7000S. Column chroma-
tography was performed using Merck silica gel 60 (70 - 230
mesh) and TLC was run using silica gel GF-254.
RESULTS AND DISCUSSION
Antioxidative effect from extracts of rosemary
The antioxidative activity of each fraction of
the n-hexane, dichloromethane and ethanol
extracts was measured by the active oxygen
method (AOM) when added at a concen-

Hexane ext. Dichloromethane ext. Ethano 1 ext.

{ SA fro (non V.) JNeut. fro (non V.)

Cant. / Volatile fro Cont./lVolatile fro Cant.
50 " 50 \ 50 \

>-
o
"-
..... WA fro (non V.).

fr. (non V.)

.......... WA fr. (non V.)

SA fr. (non V.)

"" Benzene sol. fro

insol. fro

25 50 5 25 50 25 50 hr

FIG. 1. Antioxidative Effect from Extracts of Rosemary with Different Solvents. (AOM)
Cont., control (without additives); SA fr., strongly acidic fraction (0.01%); WA fr., weakly acidic fraction
(0.01 %); Neut. fr., neutral fraction (0.01 %); non V., non-volatile fraction.
 524 R. INATANI, N. NAKATANI and H. FUWA

tration of 0.01 % into fresh lard. As shown in
Fig. 1, the volatile fraction, the strongly acidic
and neutral fractions of non-volatile com-
ponents from the n-hexane and dichloro-
methane extracts had no activity, and neither
had the ethanol extract. On the other hand, the
weakly acidic fraction of the n-hexane extract
showed a strong antioxidative activity and that
of the dichloromethane extract showed a slight
activity.
Antioxidative effect of fractionated weakly aci-
dic components n-hexane extract
The groups I, II, III, IV and V were ob-
tained by chromatography from the non-
Downloaded by [181.66.156.48] at 20:26 28 May 2015
extract. As shown in Fig. 2, the antioxidative
activities of groups I, II and III were higher
than that of a-tocopherol when measured by
AOM. In particular a remarkable activity was
observed in group II. In addition, the activity
of the fractionated weakly acidic components
(fraction Nos. 1 ~ 12) was examined by the
ferric thiocyanate method, as shown in Fig. 3.
Fraction No. 1 had no activity, but the other
fractions were effective. Especially, strong ac-
tivity was observed in the fraction Nos. 2, 3, 4,
6, 8, 9, 11 and 12. Also similar results were
obtained when the TBA values were measured
on the 7th day (Fig. 4). This weakly acidic
fraction may have contained several com-

volatile weakly acidic fraction of n-hexane ponents.

cone. 0.02% cone. 0.01 %

V\ 1/ ;-TOC. Cont.
\
5: Cont I 50

~
'-
0-
W
E
I
>
o
"-
"'-II
50 hr 25 50 hr

FIG. 2. Antioxidative Effect of Fractionated Weakly Acidic Components of n-Hexane Extract. (AOM)

0.0.
2.0 0'41L- Cont. / Fr. 5

~
A/Fr.3
-==-=-_Fr.4
o~!:::~~~~~=:! - Fr. 2
2 4 6

...- Fr. 7

~:~~: 8,9
6
1.0
$
10

12
BHA 11
~~~~e~8! BHT
2 4 6 day

FIG. 3. Antioxidative Effect of Fractionated Weakly Acidic Components of n-Hexane Extract. (Ferric
Thiocyanate Method)
Each fractionated sample, conc. 0.02%; BHA, BHT and a-Toc., conc. 0.01%.
 Antioxidative Effect of the Constituents of Rosemary 525

Antioxidant effect of constituents of rosemary TBA methods. From the results of measure-
From the fraction Nos. 6 and 7, carnosol (2) ment by AOM at a concentration of 0.02%
was isolated as the main compound of the shown in Fig. 5, the antioxidant activities of
weakly acidic fraction. Rosmanol (1) was ob- carnosol and rosmanol are much higher than
tained from the fraction Nos. 8 and 9, and those of (X-tocopherol, BRA and BRT. At a
rosmadial (3) from the fraction Nos. 3 and 4. lower concentration (0.01 %), carnosol showed
Furthermore, the fraction Nos. 3", 4 and 8 '" 9 an activity comparable with that of BRA
afforded flavone I (4) and flavone II (5), (0.02%). Rosmanol was more active than car-
respectively. The activities of these compounds nosol and the synthetic antioxidants. The ac-
were measured by the AOM, thiocyanate and tivity of rosmanol at a concentration of
0.005% was still stronger than that of the
0.02% concentration of BRA and BRT. By a
BHT
calculation simply based on the concentration,
BHA
rosmanol has an antioxidative activity four
'" -Toc. times more strong than that of BRA. On the
Downloaded by [181.66.156.48] at 20:26 28 May 2015

other hand, rosmadial and the two flavones

2
were less active.
3
When the activity of the sample in an aque-
4
ous ethanoIic solution was measured by the
5
ferric thiocyanate method and TBA, rosmanol
6
and carnosol showed stronger activity than (X-
7
tocopherol, but less than BRT. It seems that
8
these diterpene antioxidants are more effective
9
in oil (lard) than in aqueous ethanolic solution.
10
In addition rosmadial gave about half the
11 activity of (X-tocopherol as shown in Fig. 6.
12
cont. ..:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.;.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:'
Antioxidative effect of carnosol derivatives
o 0.5 1.0 o. D. Figure 7 shows the activity of carnosol and
FIG. 4. Antioxidative Effect on the 7th Day of Weakly its derivatives at two different concentrations
Acidic Components of n-Hexane Extract. (TBA) (0.01 and 0.02%) in an aqueous ethanolic
Each fractionated sample, conc. 0.02%; BHT, BHAand IX- solution. The fully O-substituted compounds
Toc., conc. 0.01%. (B, C and E) showed negligibly weak anti-

Carnosol Rosmanol Others (O.02%)

Cont. 0.005% Cont. , Fl avone II

50 " I 50 " Rosmadial

Flavone I

o>
Co.

0.02%
5 25 50 25 50 25

FIG. 5. Antioxidative Effect of Constituents of Rosemary. (AOM)

BHA, BHT and IX-Toc., conc. 0.02%.
 526 R. INATANI, N. NAKATANI and H. FuwA

Ferric Thiocyanate Method

0.0.
2.0 "&;.><;;:'_r. _ Fl avone II
- Control

-Flavone I

1.0

__ Rosmadial

a-Toe.
::::: Rosmanol
I~~~~~~~~~~t- Carnosol
o f = - - - BHT
2 4 6 day
Downloaded by [181.66.156.48] at 20:26 28 May 2015

TBA (6th day)

BHT
a- Toe.
Carnosol
Rosmanol
Rosmadial
F1a von e :':':':':':':'.:.:.:.:.
Flavone II ~:.:~.:.:~.:.:.~:.:.~:.:.~:.:.:~.:.:~.:.:~.:.:.~:.:.;:':'~:':':~':':~':':~':':';:':'~: : :~: :'~:':'~:':~J
Control ~.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:':':':':':':::1

o 0.5 1.0 0.0.

FIG. 6. Antioxidative Effect of Constituents of Rosemary. (Ferric Thiocyanate Method and TBA)
Isolated compounds, cone. 0.02%; BHT and ex-Toe., cone. 0.01%.

oxidative activity, while the O-monosubstituted adjacent to an isopropyl group, had a higher
derivatives (D and F) were as strong as car- value than that of E'.
nosol (A) at a concentration of 0.02%. There
was no difference in the activity between the
CONCLUSION
compound with a lactone ring in the molecule
and that without it. Especially, compound F Antioxidative activity was measured for the
was particularly effective. extracts of rosemary with different solvents.
Strong activity was observed in the weakly
Antioxidative effect of rosmanol derivatives acidic fraction of the n-hexane extract, which
As shown in Fig. 8, triacetyl (B') and di- was further fractionated and purified by col-
methylrosmanol (C') showed weak activity umn chromatography on silica gel to afford
and the monomethylated compounds (D' and several active compounds. Amongst these, car-
E') were as strong as rosmanol (A') at a nosol and the new antioxidant rosmanol show-
concentration of 0.02%. However, at a 0.01 % ed remarkable activity. Rosmanol especially
concentration, the activities of D' and E' demonstrated an activity more than four times
decreased to one third that of A'. When the higher than BHA and BHT by AOM. This
effectiveness of D' was compared with that of compound is odorless and tasteless. The anti-
E', D', in which a methoxy group is located oxidative effect of several derivatives of these
 Antioxidative Effect of the Constituents of Rosemary 527

A R,R'= H (Carnoso1, 2) E R,R'=CH 3 (9)

B R,R'= Ac (6) F R= H, R'=CH 3 (10)
C R,R'= CH 3 (7)
D R,R'=H,CH 3 (8)

1.0 0.02%
A P
c :::.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.
D~
Downloaded by [181.66.156.48] at 20:26 28 May 2015

0.5 E ::::::.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:
F b
cant ..:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:
o'--.......- -.......----r- o 05 1.0 O.D.
3 5 day

0.01%
AI2J
B :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.::::::::::.:.:.:.:.:.:.:.:.:.:.:
C :.:.:....:..:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.::::3
D.:.:.:.:.:.:.:.:.:.......
0.5
E ..:.:.:.....:.:....:.:...:.:.:.:.:.:.:.:.:.:.:.:.:.::::::::::::i
_F F m
~::::"---==::::::=:=--"""A cont.i:o:.::r.'.:.::::::::.:::::::::::::::::::::::::::::::.:.:::::::.:::::.:.:::::.:.:::::.:.:""'.:.:""'.:.:"'.:.:"'.:.:"'.:.:"'.:.:""'.:.
o~ .......---~---~- o 0.5 1.0
3 5 day 0.0.
(Ferric Thiocy~nate Method) (TBA, 6th day)

FIG. 7. Antioxidative Effect of Carnosol Derivatives.

diterpenes was also measured in the aqueous
ethanolic solution. The fully O-substituted de-
rivatives showed negligible activity, while the
activity of the mono-substituted ones was as
strong as that of original compounds, ros-
manol and carnosol.
Acknowledgments. We are grateful to Dr. T. Osawa,
Nagoya University, for valuable suggestions on the
measurement of antioxidant activity. We are indebted to
Dr. T. Yoshida and Mr. H. Kanno, Takasago Perfumery
Co. Ltd., for technical co-operation on AOM measure-
ments and for suppling rosemary.
REFERENCES
1) J. R. Chipault, G. R. Mizuno, J. M. Hawkins and W.
O. Lundberg, Food Res., 17, 46 (1952).
2) J. R. ChipauJt, G. R. Mizuno and W. O. Lundberg,
Food Technol., 10, 209 (1956).
3) A. Palitzsch, H. Schulze, F. Metzl und H. Bass, Die
Fleischwirtshaft, 49, 1349 (1969).
4) A. Palitzsch, H. Schulze, G. Lotter und A. Steichele,
Die Fleischwirtschaft, 54, 63 (1974).
5) C. H. Brieskorn und H. J. Domling, Z. Lebensmitt-
Untersuch, 141, 10 (1969).
6) S. S. Chang, B. O. Matijasevic, O. A. L. Hsieh and C.
L. Huang, J. Food Sci., 42, 1102 (1977).
7) R. Inatani, N. Nakatani, H. Fuwa and H. Seto,
Agric. Bioi. Chern., 46, 1661 (1982). The stereochem-
istry of the OH group on C-7 of rosmanol has been
revealed to be an ex-configuration as the structure 1
on the basis of a reexamination of the NOE analysis.
This result will be reported soon in this journal.
8) N. Nakatani, R. Inatani and H. Fuwa, Agric. Bioi.
 528 R. INATANI, N. NAKATANI and H. FUWA

A' R,R',R"= H (Rosmano1, 1)

B' R,R ',R"= Ac (11)
C' R,R'= CH 3 , R"= H (12)
D' R,R"= H, R' = CH 3 (13)
E' R = CH 3 , R',R"= H (14)

0.0.
0.02%
1.0
A' ~
B' .;.;.;.;.;.;...;.;.;.;.;.;.;.
C' .;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;
0.5
0' ~
_ E' E' ~
Downloaded by [181.66.156.48] at 20:26 28 May 2015

It:--II!I!!!~::::=::::':'~: cont ..;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;

o~~-----r-----r-- o 0.5 1.0 0.0.
3 5 day

0.0. 0.01%
1.0
-E' A' ~
-D'
B' ;...;.;.;.;.;.;.;...;........................................................;.........;.;.....
C' .;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.;.
0.5

E'.
o/~

- A' cont. .;.;.;.;.;.;.;.;.;.;.;.;.;.;.;...;.;.;.;.;.;.....;.;.....;...........;.;.;...

o 3 day
o 0.5 1.0 0.0.

(Ferric Thiocyanate Method) (TBA, 6th day)

FIG. 8. Antioxidative Effect of Rosmanol Derivatives.

Chern., 44, 2831 (1980). 11) A. Ottolenghi, Arch. Biochern. Biophys., 79, 355
9) H. Mitsuda, K. Yasumoto and K. Iwami, Eiyo to (1959).
Shokuryo, 19,210 (1967). 12) N. Nakatani and R. Inatani, Agric. BioI. Chern.,
10) T. Osawa and M. Namiki, Agric. BioI. Chern., 45, 735 47, 353 (1983).
(1981).