New quinone sulfates from the crinoids Tropiometra afra macrodiscus and Oxycomanthus japonicus

Article abstract of DOI:10.1248/cpb.50.1609

Quinone pigments in the crinoids were investigated. New quinone sulfates 1 and 3 were isolated from Tropiometra afra macrodiscus and Oxycomanthus japonicus, respectively. The structures 1 and 3 were determined to be 1,6,8-trihydroxy-3-propylanthraquinone-

Full text of DOI:10.1248/cpb.50.1609

December 2002
Notes
Chem. Pharm. Bull. 50(12) 1609—1612 (2002)
1609
New Quinone Sulfates from the Crinoids Tropiometra afra macrodiscus
and Oxycomanthus japonicus
Daizo TAKAHASHI,a Takashi MAOKA,*,a Miyuki TSUSHIMA,a Kazuyoshi FUJITANI,a Mutsuo KOZUKA,a
b
Takao MATSUNO,a and Tetsuro SHINGU
a Kyoto Pharmaceutical University; Misasagi, Yamashina, Kyoto 607–8412, Japan: and b Faculty of Pharmaceutical
Sciences, Kobe Gakuin University; Nishi-ku, Kobe 651–2113, Japan. Received May 29, 2002: accepted August 26, 2002
Quinone pigments in the crinoids were investigated. New quinone sulfates 1 and 3 were isolated from Tropi-
ometra afra macrodiscus and Oxycomanthus japonicus, respectively. The structures 1 and 3 were determined to be
1,6,8-trihydroxy-3-propylanthraquinone-2-carboxylic acid 6-O-sulfate (ptilometric acid 6-O-sulfate) and 2-bu-
tanoyl-3,6,8-trihydroxy-1,4-naphtoquinone 6-O-sulfate, respectively, by chemical and spectroscopic analysis.
Ptilometric acid 6-O-sulfate (1) showed an antifeedant activity on fish.
Key words ptilometric acid 6-O-sulfate; crinoid; quinone sulfate; 2-butanoyl-3,6,8-trihydroxy-1,4-naphtoquinone 6-O-sulfate;
Tropiometra afra macrodiscus; Oxycomanthus japonicus
using sulfation shifts in 1H-NMR7) and 13C-NMR.11) Because
Biologists have been attracted by the bright external color
patterns of crinoids, leading to the isolation and structural
elucidation of many kinds of pigments from crinoids. The
color is mainly caused by quinones and carotenoids. Various
reports on the quinone pigments of crinoids have been pub-
lished.1) At present, the following four groups of polyketide-
derived pigments, linear and angular naphthopyrones,2—4)
4-acylanthraquinones,5—7) 3-alkylanthraquinones,6,8) and dia-
metric bianthrones (bianthraquinones and phenanthroperyl-
enequinones)9,10) are known to occur in crinoids. Some of the
quinone pigments in crinoid are present in sulfate form.1,7)
This prompted us to search for water-soluble quinone pig-
ments in crinoids. This paper reports the isolation and struc-
tural elucidation of new quinone sulfates from Tropiometra
afra macrodiscus and Oxycomanthus japonicus by chemical
and spectroscopic analysis. Furthermore, antifeedant activity
of anthraquinone sulfate on fish is reported.
1
no H- or 13C-NMR data were available for ptilometric acid
1
(2), we carried out complete H- and 13C-NMR assignments
of 2 in DMSO-d6 solution using two dimensional (2D)-NMR
experiments. Double quantum filtered correlation spectroscopy
(DQF-COSY) and nuclear Overhauser spectroscopy (NOESY)
experiments revealed the assignment of aromatic protons at
H-4, H-5, H-7, protons of the propyl chain (H-1Ј, H-2Ј, and
H-3Ј), and a hydrogen bonded phenolic proton at 8-OH. The
assignment of all proton-attached carbons was made from a
13C–1H COSY spectrum. 13C signals of quaternary carbons at
C-2, C-3, C-6, C-8, C-10, C-11, C-12, and C-14 were unam-
biguously assigned by correlation of proton signals in a
13C–1H long range COSY experiment. Furthermore, the re-
maining seven carbon signals (d 132.4, 167.2, 180.8, 189.3)
were assigned to C-13, C-1, 2-COOH, and C-9, respectively,
by comparison with polyhydroxyanthraquinones.12) The as-
1
signment of H- and 13C-NMR data for 1 was carried out in
Structure of a New Anthraquinone Sulfate from T. afra
macrodiscus The acetone extract of T. afra macrodiscus was
partitioned with Et2O–n-hexane (1 : 1) and water to remove
fat and carotenoids from the aqueous acetone solution. The
purple-red colored aqueous layer was extracted with EtOAc
to remove free anthraquinones. The aqueous layer was evap-
orated, and the dark purple residue was subjected succes-
sively to column chromatography on ODS and preparative
HPLC on ODS to yield a water-soluble yellow pigment 1.
Compound 1 showed UV–vis absorption maxima at 225,
262, and 435 nm, indicating a hydroxyanthraquinone chro-
mophore and IR absorption maxima at 1620 and 1481 cmϪ1,
which are characteristic absorption bands of anthraquinone.8)
Furthermore, 1 showed strong IR absorptions at 1249 and
1057 cmϪ1, suggesting the presence of a sulfate group.7) Neg-
ative ion FAB-MS gave a quasi-molecular ion at m/z 443,
corresponding to the formula [C18H14O10SNaϪH]Ϫ. The char-
acteristic fragment ions at m/z 421 [MϪNa]Ϫ and 341
[MϪSO3Na]Ϫ were compatible with the presence of a
sodium sulfate group. Furthermore, the presence of sulfur
and sodium atoms in 1 was confirmed by inductively coupled
plasma (ICP) spectra. Acid hydrolysis of 1 gave ptilometric
acid (2). Therefore, 1 was determined to be a sodium sulfate
of ptilometric acid (2).
the same manner described above. All proton-attached car-
bons were assigned by 13C–1H COSY. Quaternary carbons at
C-6, C-8, C-10, C-11, and C-14 were also assigned by
13C–1H long range COSY. The remaining seven carbon sig-
nals at d 125.2, 133.8, 134.0, 153.6, 168.8, 182.5, and 187.2
were assigned to C-2, C-12, C-13, C-3, C-1, 2-COOH, and
C-9, respectively, by comparison with polyhydroxyan-
1
thraquinones12) and ptilometric acid (2). The total H- and
13C-NMR assignments for 1 and 2 are compiled in Table 1.
It is known that, in the 13C-NMR spectra of phenol com-
pounds, the introduction of O-sulfate, an electron withdraw-
ing group, results in an increased electron density of the car-
bon carrying the sulfate group and a decreased electron den-
sity of the carbons ortho and para, thus resulting in an up-
field shift for the former carbons and a downfield shift for the
latter carbons.11) Sulfate 1 showed an upfield shift of 6.6 ppm
at C-6, and downfield shifts of 4.9 ppm at C-7 and 0.8 ppm at
C-5 compared with 2. These results indicated that the hy-
droxy group at C-6 in 1 was present as the sulfate ester. Fur-
thermore, about 0.4 ppm downfield shifts of H-5 and H-7 in 1
relative to 2 agreed with the 6-O-sulfate structure.7) There-
fore, a new anthraquinone sulfate 1, isolated from T. afra
macrodiscus, was determined to be a sodium salt of ptilomet-
ric acid 6-O-sulfate (1,6,8-trihydroxy-3-propylanthraquinone-
The position of the sulfate group in 1 was determined by
* To whom correspondence should be addressed. e-mail: maoka@mbox.kyoto-inet.or.jp
© 2002 Pharmaceutical Society of Japan

1610
Vol. 50, No. 12
Table 1. 1H- (500 MHz) and 13C-NMR (125 MHz) Data for Ptilometric Acid 6-O-Sulfate (1) and Ptilometric Acid (2) in DMSO-d6
Ptilometric acid 6-O-sulfate (1)
Ptilometric acid (2)
Carbon No.
13C mult.
1H mult.
J (Hz)
13C mult.
1H mult.
J (Hz)
1
2
3
4
5
6
7
8
9
10
11
12
168.8 s
125.2 s
153.6 s
117.4 d
109.7 d
159.1 s
112.8 d
163.2 s
187.2 sa)
182.5 s
112.5 s
133.8 s
134.0 s
115.9 s
182.5 sa)
37.1 t
167.2 s
130.4 s
147.6 s
118.6 d
108.9 d
165.7 s
107.9 d
164.4 s
189.3 sa)
180.8 s
113.8 s
135.0 s
132.4 s
113.8 s
180.8 sa)
35.2 t
7.23 s
7.45 d
7.52 s
7.10 d
2.3
2.3
1.8
1.8
7.04 d
6.57 d
13
14
2-COOH
1Ј
2Ј
3.16 t
1.58 tq
0.94 t
7.5
7.5
7.5
2.65 t
1.63 tq
0.93 t
7.3
7.3
7.3
23.9 t
14.1 q
23.2 t
13.7 q
3Ј
8-OH
13.3 br s
12.0 br s
a) Assignments may be interchangeable within the same column.
Table 2. 1H- (300 MHz) and 13C-NMR (75 MHz) Data for 2-Butanoyl-
3,6,8-trihydroxy-1,4-naphthoquinone 6-O-Sulfate (3) in D2O, and H-NMR
(300 MHz) Data for 2-Butanoyl-3,6,8-trihydroxy-1,4-naphtoquinone (4) in
Acetone-d6
0.73, 1.45, 2.64) and two aromatic protons with an AX spin
system (d 6.93, 7.23). The coupling constant (Jϭ2.3 Hz) be-
tween the aromatic protons indicated meta-orientation. The
13C-NMR spectra, including a distortionless enhancement by
polarization transfer (DEPT) experiment, showed 14 carbon
signals due to one methyl, two methylenes, two methines and
nine quaternary carbons. All proton-attached carbons were
assigned by a 13C–1H COSY experiment. Of the nine quater-
nary carbons, three were ascribed to carbonyl carbons (d
184.4ϫ2C, 186.5) and three to phenol substituted carbons (d
172.2, 160.9, 154.5) by their chemical shifts. These spectral
data strongly suggested that 3 was a trihydroxy-1,4-naphtho-
quinone substituted with a butanoyl group.13) Acid hydrolysis
of 3 gave a free naphthoquinone (4), which showed a molec-
ular ion peak at 276.0649 compatible with the formula
C14H12O6. An electron impart (EI)-MS fragment ion at m/z
206 [MϪCOC3H6] indicated the presence of a butanoyl
group in 4. The structure of 4 for 2-butanoyl-3,6,8-trihy-
droxy-1,4-naphthoquinone was postulated by UV–vis, IR,
1H-NMR, and EI-MS spectral data comparison with those of
flaviolin (3,6,8-trihydroxy-1,4-naphthoquinone).14) The posi-
tion of the sulfate group in 3 was determined from sulfation
1
3
4
Carbon No.
13C mult. 1H mult.
J (Hz)
1H mult.
J (Hz)
1
2
3
186.5 sa)
115.9 s
172.2 s
184.4 s
4
5
6
7
8
113.5 d
154.5 s
111.5 d
160.9 s
112.4 s
132.3 s
184.4 sa)
45.5 t
7.23 d
6.93 d
2.3
2.3
6.97 d
6.56 d
1.4
1.4
9
10
1Ј
2Ј
3Ј
4Ј
2.64 t
1.45 tq
0.73 t
7.5
7.5
7.5
2.86 t
1.58 tq
0.86 t
7.5
7.5
7.5
18.0 t
12.9 q
a) Assignments may be interchangeable within the same column.
1
2-carboxylic acid 6-O-sulfate).
shifts in the H-NMR.7) The protons at H-5 and H-7 in 3
Structure of a New Naphthoquinone Sulfate from O. were shifted downfield by 0.26 and 0.37 ppm, respectively,
japonicus In the same manner as described above, a water- relative to those of 4. These data indicated that the hydroxy
soluble reddish pigment 3 was isolated from the acetone ex- group at C-6 was present as the sulfate ester. Therefore, the
tract of O. japonicus. Compound 3 showed UV–vis absorp- structure of 3 was determined to be a sodium salt of 2-bu-
tion maxima at 249, 295, and 374 nm, and an IR absorption tanoyl-3,6,8-trihydroxy-1,4-naphthoquinone 6-O-sulfate.
maximum at 1630 cmϪ1, characteristic of hydrogen-bonded
Furthermore, the sodium sulfate of 1,3,6,8-tetrahydroxy-
quinones.1) Furthermore, 3 showed strong IR absorptions at anthraquinone (5) was also obtained. A negative ion FAB-
1275 and 1057 cmϪ1, suggesting the presence of a sulfate MS gave a quasi-molecular ion at m/z 373, compatible with
group.7) A negative ion FAB-MS gave a quasi-molecular ion the structure of a sodium sulfate of 5. However, the position
at m/z 378, corresponding to the formula [C14H11O9SNa]Ϫ. of the sulfate group could not be determined because of the
The characteristic fragment ions at m/z 355 [MϪNa]Ϫ and small sample size.
275 [MϪSO3Na]Ϫ were compatible with the presence of a
In addition to these quinone sulfates, the following five
1
1
sodium sulfate group. H-NMR spectra, including a H–1H known quinones, 1,3,6,8-tetrahydroxyanthraquinone (5), 2-
COSY experiment, revealed the presence of a propyl chain (d acetylemodin (6), flaviolin (7), rhodolamprometrin (8), and

December 2002
1611
Table 3. The Inhibitory Effect of Ptilometric Acid 6-O-Sulfate (1) and Ptilometric Acid (2) on Feeding by Fish
Concentration of
test substance
in diet
Species of fish
Poecilia reticulata (nϭ6) Oplegnathus fasciatus (nϭ24) Parapristipomatriline atum (nϭ10)
Test substance
Ptilometric acid (2)
Ptilometric acid 6-O-sulfate (1)
Ptilometric acid 6-O-sulfate (1)
1.0%
0.2%
1.0%
Ϫ
Ϯ
ϩ
Ϫ
ϩ
ϩ
Ϫ
Ϯ
ϩ
ϩ, Substantial preference for control diet; Ϫ, no statistically significant preference for either diet; Ϯ, marginal preference for control diet.
with MeOH. Crystallization from aqueous AcOH afforded ptilometric acid
(2) (2 mg).
Ptilometric Acid 6-O-Sulfate (1): Reddish crystal. UV–vis: (30% MeOH)
nm (e) 225 (11270), 262 (11240), 435 (4660). IR cmϪ1: 3442, 2962, 1620,
1481, 1374, 1249, 1057. Negative ion FAB-MS: m/z 443 [MϪH]Ϫ, 421
1
[MϪNa]Ϫ, 341 [MϪSO3Na]Ϫ, 297 [MϪSO3NaϪCO2]Ϫ. H- and 13C-NMR
(DMSO-d6): Table 1. DQF-COSY correlations: between H-5 to H-7, H-1Ј to
H-3Ј. NOESY correlations: H-4 to H-1Ј, H-2Ј, and H-3Ј, H-1Ј to H-2Ј, H-2Ј
to H-3Ј, OH-8 to H-7. 13C–1H long range COSY correlations: C-6 to H-7, C-
8 to H-7, C-10 to H-4 and H-5, C-11 to H-7 and H-5, C-14 to H-4, C-2Ј to
H-3Ј. ICP-optical emission spectrum: 589.592 nm (Na), 180.731 nm (S).
Ptilometric Acid (2): Reddish crystal. UV–vis: (EtOH) nm (e) 228
(34500), 277 (29240), 313 (9700) and 444 (14860). IR cmϪ1: 3418, 2965,
1715, 1674, 1626. EI-MS m/z: 342 [M]ϩ, 324 [MϪH2O]ϩ, 298 [MϪCO2]ϩ,
270 [MϪCO2ϪCH2CH2]ϩ. High resolution (HR)-EI-MS: m/z 342.0731
(C18H14O7 Calcd for 342.0737). 1H- and 13C-NMR (DMSO-d6): Table 1.
DQF-COSY correlations: between H-5 to H-7, H-1Ј to H-3Ј. NOESY corre-
lations: H-4 to H-1Ј, H-2Ј, and H-3Ј, H-1Ј to H-2Ј, H-2Ј to H-3Ј. 13C–1H
long range COSY correlations: C-2 to H-14, C-3 to H-4 and H-1Ј, C-4 to H-
ptilometric acid (2), were isolated from O. japonicus.
Antifeedant Activity of Ptilometric Acid (2) and Its Sul-
fate (1) in Fish It has been reported that quinone pigments,
especially the sulfate forms, provide a chemical mechanism
against predatory fish for some species of crinoids.9) In this 1Ј, C-5 to H-7, C-6 to H-7, C-7 to H-5 and OH-8, C-8 to H-7 and OH-8, C-
10 to H-14 and H-5, C-12 to H-5, C-14 to H-4, C-1Ј to H-1Ј and H-3Ј, C-3Ј
to H-1Ј and H-3Ј.
Acid Hydrolysis of 1 Compound 1 (3 mg) was dissolved in 20 ml 1 N
HCl and stored at 37 °C for 4 h. Then, the pigment was extracted with
study, ptilometric acid 6-O-sulfate (1) showed concentration-
dependent antifeedant activity in fish, but ptilometric acid (2)
was ineffective (Table 3). Antifeedant activities of 2-bu-
tanoyl-3,6,8-trihydroxy-1,4-naphtoquinone (4) and its sulfate
(3) could not be measured because of the small sample size.
EtOAc and was purified by preparative TLC on silica gel G with benzene–
EtOAc–AcOH (60 : 40 : 1) to give ptilometric acid (1.5 mg). The identifica-
1
tion of ptilometric acid was based on UV–vis, IR, EI-MS, HR-EI-MS, H-,
and 13C-NMR data.
Experimental
Extraction and Isolation of Quinone Pigments from O. japonicus
Nineteen fresh specimens of O. japonicus (1100 g) were immediately im-
mersed in acetone to yield a deep red solution which was then partitioned
between n-hexane–Et2O (1 : 1) and water to remove fat and carotenoid from
the acetone solution. The water layer was extracted with EtOAc to remove
free quinines. The aqueous layer was concentrated by a rotary evaporator.
The dark red residue was subjected to column chromatography on ODS. The
yellow fraction eluted with 20% MeOH was further purified by preparative
HPLC on ODS with 25% MeOH to afford a sulfate of 5 (0.7 mg), and the
red fraction eluted with 20% MeOH was further purified by preparative
HPLC on ODS with 25% MeOH to afford a new compound 3 (2.5 mg). The
EtOAc solution was also concentrated by a rotary evaporator. The dark red
residue (1.8 g) was subjected to column chromatography on silica gel (Merck,
70—230 mesh) with mixed solvents of benzene–EtOAc–AcOH as eluents.
The yellow fraction eluted with benzene–EtOAc–AcOH (95 : 5 : 1) was fur-
ther purified by HPLC on ODS with 80% MeOH to afford 2-acetylemodin
(6, 0.6 mg). The yellow fraction eluted with benzene–EtOAc–AcOH (90 :
10 : 1) was further purified with the same HPLC system to afford 1,3,6,8-
tetrahydroanthraquinone (5, 0.7 mg) and flaviolin (7, 1.0 mg). The orange
fraction eluted with benzene–EtOAc–AcOH (80 : 20 : 1) was further purified
with the same HPLC system to afford rhodolamprometrin (8, 1.1 mg). The
orange fraction eluted with benzene–EtOAc–AcOH (60 : 40 : 1) was further
purified with the same HPLC system to afford ptilometric acid (2, 4 mg).
Identification of these five known quinones, 2-acetylemodin (6),15) 1,3,6,8-
tetrahydroxyanthraquinone (5),16) flaviolin (7),14) rhodolamprometrin (8),16)
and ptilometric acid (2) were made by UV–vis, IR, EI-MS, and 1H-NMR
data and by comparison with authentic samples14—16) on HPLC.
General The UV–vis spectra were recorded in aqueous MeOH or EtOH
solution on a Shimadzu UV-240 spectrophotometer. The IR spectra were ob-
tained by a Perkin Elmer FT-IR 1600 as KBr pellets. The EI- and FAB-MS
spectra were recorded using a JMS-DX 300 mass spectrometer. FAB-MS
were obtained using glycerin as a matrix. The 13C- and 1H-NMR spectra
were measured with a Bruker ARX-500 spectrometer or Varian XL-300
spectrometer. The ICP spectra were recorded with a ICPS-8000 (Shimadzu)
in water at a concentration of 300 mg/ml. HPLC was performed on a Shi-
madzu LC-6AD instrument with a Shimadzu SPD-6AV spectrophotometer
set at 440 nm. The column used was
a Shim-Pack PREP-ODS
(20 mmϫ250 mm ID, particle size 5 mm, Shimadzu) using 20% MeOH as
the mobile phase.
Animal Material Specimens of T. afra macrodiscus and O. japonicus
were collected from the sea at Goza in Mie Prefecture in July 1992 and in
June 1993, respectively. Voucher specimens have been deposited at Kyoto
Pharmaceutical University.
Extraction and Isolation of Quinone Pigments from T. afra macrodis-
cus Seventeen fresh specimens of T. afra macrodiscus (800 g) were imme-
diately immersed in acetone to yield a deep red solution, which was then
partitioned between n-hexane–Et2O (1 : 1) and water to remove fat and
carotenoids from the acetone solution. The purple-red water layer was ex-
tracted with ethyl acetate to remove free anthraquinones. The aqueous layer
was concentrated by a rotary evaporator. The obtained dark purple residue
was subjected to column chromatography on ODS (Fuji Silysia Chemical,
Ltd., DM-1020T, 100—200 mesh). The purple-red fraction eluted with 20%
MeOH was further purified by preparative HPLC on ODS with 20% MeOH,
and was crystallized from EtOH–EtOAc–water to afford compound 1 (25
mg).
2-Butanoyl-3,6,8-trihydroxy-1,4-naphthoquinone 6-O-Sulfate (3): A red-
dish amorphous powder; UV–vis: (30% MeOH) nm (e) 249 (12740), 295
(11980), 374 (2720). IR cmϪ1: 3500—3300, 2950, 1630, 1275, 1057. Nega-
tive ion FAB-MS: m/z 378 [C14H11O9SNa]Ϫ, 355 [MϪNa]Ϫ, 275 [MϪ
SO3Na]Ϫ. 1H- and 13C-NMR (D2O): Table 2.
On the other hand, the EtOAc extract was subjected to preparative TLC on
silica gel G (Kieselgel 60, Merck) with benzene–EtOAc–AcOH (60 : 40 : 1).
A yellow band (Rf 0.30) was extracted with 2% AcOH in EtOAc and was
further chromatographed on Sephadex LH-20 (Pharmacia Fine Chemicals)

1612
Acid Hydrolysis of 3 Compound 3 (1.2 mg) was dissolved in 8 ml 1 N
Vol. 50, No. 12
thank Miss M. Hikawa, Shimadzu Co., Ltd., for the measurement of ICP
HCl and stored at 37 °C for 4 h. The pigment was extracted with EtOAc and spectrum, and Mr. T. Kawakami, Nippon Formula Feed Manufacturing Co.,
was purified by preparative HPLC on ODS with 65% MeOH to give 4
Ltd. for measurement of antifeedant activity for fish.
(0.6 mg).
2-Butanoyl-3,6,8-trihydroxy-1,4-naphthoquinone (4): An orange amor- References
phous powder; UV–vis: (1% AcOH/MeOH) nm (e) 234 (12640), 268
(14660), 300 (16180). IR cmϪ1: 3443, 2965, 1621, 1479. HR-EI-MS: m/z
276.0649 (C14H12O6, Calcd 276.0640). EI-MS m/z (rel. int. %) 276 [M]ϩ
(68), 234 [MϪC3H6]ϩ (45), 206 [MϪCOC3H6]ϩ (80), 60 (100). 1H-NMR
(acetone-d6): Table 2.
Sulfate of 1,3,6,8-Tetrahydroxyanthraquinone (5): A yellow amorphous
powder; UV–vis: (30% MeOH) nm (e) 248, 297, 428. IR cmϪ1: 3444, 1633,
1614, 1283, 1100. Negative ion FAB-MS: m/z 373 [MϪH]Ϫ. 1H-NMR
(D2O) d: 6.77 (2H, br s, H-2, H-7), 7.06 (2H, br s, H-3, H-6). Acid hydroly-
sis of this compound with 1 N HCl at 37 °C for 4 h gave 1,3,6,8-tetrahydroan-
thraquinone (5). Identification of 5 was based on UV–vis and co-HPLC with
an authentic sample.
1) Thomson R. H., “Naturally Occurring Quinones III: Recent Ad-
vances,” Chapman and Hall, London, 1987, pp. 345—526.
2) Kent R. A., Smith I. R., Sutherland M. D., Aust. J. Chem., 23, 2325—
2335 (1970).
3) Smith I. R., Sutherland M. D., Aust. J. Chem., 24, 1487—1499 (1971).
4) Sakuma Y., Tanaka J., Higa T., Aust. J. Chem., 40, 1613—1616 (1987).
5) Singh H., Moore R. E., Scheuer P. J., Experientia, 23, 624—626
(1967).
6) Erdman T. R., Thomson R. H., J. Chem. Soc., Perkin Trans. 1, 1972,
1291—1292 (1972).
7) Rideout J. A., Sutherland M. D., Aust. J. Chem., 34, 2385—2392
(1981).
Antifeedant Activity for Fish Commercial diets, Flake Food (Tetra
Werke Co., Germany) and Nipai No. 1 (Nippon Formula Feed Manufactur-
ing Co., Ltd., Japan) were used for fresh-water fish (Poecilia reticulata) and
marine fish (Oplegnathus fasciatus, Parapristipomatriline atum), respec-
8) Matsuno T., Fujitani K., Takeda S., Yokota K., Yoshimizu S., Chem.
Pharm. Bull., 20, 1079—1082 (1972).
9) Rideout J. A., Smith N. B., Sutherland M. D., Experientia, 35, 1273—
1274 (1979).
tively, as control diets. The anthraquinone-containing diet was prepared by 10) Riccardis F. D., Iorizzi M., Minale L., Riccio R., Forges B. R., Debitus
adding ptilometric acid and its sulfate, which was dissolved in acetone and C. J., J. Org. Chem., 56, 6781—6787 (1991).
30% MeOH, to the control diet. The time required to eat each sample was 11) Barron D., Varin L., Ibrahim R. K., Harborne J. B., Williams C. A.,
recorded.
Phytochemistry, 27, 2375—2395 (1988).
12) Berger Y., Castonguay A., Org. Mag. Reson., 11, 375—377 (1978).
Acknowledgments We gratefully thank Drs. H. J. Banks (CSIRO), P. 13) Bruce F. B., Donald W. C., Maxwell J. C, Geoffrey I. F., Peter G. G.,
Brassard (Laval University), B. C. Bycroft (Nottingham University), and K. David P. K., Aust. J. Chem., 32, 769—777 (1979).
A. Francesconi (Western Australian Marine Research Laboratories) for 14) Baker P. M., Bycroft B. W., J. C. S. Chem. Commun., 1968, 71—72
kindly providing 2-acetylemodin, 1,3,6,8-tetrahydroxyanthraquinone, flavolin (1968).
and rhodolamprometrin, respectively. We also thank Mrs. H. Kogo (Tagawa 15) Banks H. J., Cameron D. W., Crassley M., J. Aust. J. Chem., 34,
Junior School, Osaka) and M. Sava (Ise High School, Mie) for identification 2385—2392 (1981).
,
of crinoids, and Dr. Y. Fujiwara and Miss K. Oda (Kyoto Pharmaceutical 16) Castonguay A., Brassard P., Can. J. Chem., 55, 1324—1332 (1976).
University) for measurement of NMR and MS spectra. The authors also 17) Francesconi K. A., Aust. J. Chem., 33, 2781—2784 (1980).