J Nat Med (2011) 65:191–193
193
The antioxidant activity of 1–7 has been studied with
superoxide dismutase (SOD) assay kit. Vitamin C was used
as a positive control (IC50 66.2 lM). Among these, com-
pounds 2, 5, and 6 exhibited significant SOD-like activity,
IC50 3.6, 46.0, and 21.2 lM, respectively.
Acid hydrolysis of compound 1
A solution of 1 in 5% H
SO –dioxane (1:1) was heated at
4
2
100°C for 2 h. The reaction mixture was diluted with H
O
2
and then neutralized with Amberlite IRA-35 and evapo-
rated in vacuo to dryness. The identification and the D or
L configuration of rhamnose was determined by using RI
detection (Shimadzu RID-10A) and chiral detection (Sho-
Experimental
dex OR-1) by HPLC (Shodex RSpak NH P-50 4D,
2
General
CH CN–H O–H PO , 95:5:1, 1 mL/min, 47°C), by com-
3
2
3
4
parison with an authentic sugar (10 mmol of L-rham). The
Optical rotation was taken on a JASCO DIP-1000 polarimeter.
IR spectra were measured on a JASCO FT/IR-5300 instru-
ment. NMR spectra were recorded on a Varian UNITY 600
spectrometer. The chemical shifts are given in d (ppm) in
C D N solution, using tetramethylsilane (TMS) as an internal
sugar portion gave the following peak of L-(?)-Rham at
4.60 min.
Superoxide dismutase-like activity
5
5
standard.NMRexperimentsincludedCOSY,HMQC,HMBC,
and ROESY. Coupling constants (J values) are given in Hz.
HR-FAB-MS were measured on a JEOL JMS-700 MS station.
SOD-like activity was determined according to the method
of Ukeda et al. [11] using an SOD Assay Kit-WST (Dojindo
Lab., Kumamoto). A test sample was dissolved in DMSO to
obtain a final DMSO concentration of 0.8% (v/v).
Material
Acknowledgments The authors are grateful to Ms. Y. Watanabe,
MIC Co. Ltd., Mr. L. Bergeron, L.B. Maple Treat Inc., and
Dr. T. Ishiguro, BioPharm Toyono, for supplying the material of
A. saccharum.
A. saccharum was collected at St. Roberts, Quebec, Canada,
in May 2003. A voucher specimen (TB 5429) is deposited at
the Herbarium of the Department of Pharmacognosy,
Tokushima Bunri University, Tokushima, Japan.
References
Extraction and isolation
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extracted withEtOHatroomtemperature for1.5 months. The
EtOH extract was partitioned between EtOAc and H O. The
2
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EtOAc soluble-portion (54.0 g) was subjected to silica gel
column chromatography with hexane–isopropyl ether–
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1
57.8 mg) from fraction 6, and compound 2 (14.6 mg) from
fraction 7, respectively. Fraction 9 (2.25 g) was subjected to
silica gel column chromatography with isopropyl ether–
6. Achenbach H, Benirschke G (1997) Joannesialactone and other
compounds from Joannesia Princeps. Phytochemistry 45:
MeOH–H O(25:3:0.1)andpurifiedbyHPLC(ODS, 40–50%
2
1
49–157
0
MeOH) to afford cleomiscosin C (5, 85 mg) and 5 -dem-
ethylaquillochin (8, 4.3 mg). Fraction 10 (4.99 g) was also
subjected to silica gel column chromatography with isopropyl
7
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Cleomiscosin D, a coumarino-lignan from seeds of Cleome
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apelta. Fitoterapia 71:341–342
ether–MeOH–AcOEt–H O (6:2:4:1) and purified by HPLC
2
(
ODS, 30–60% MeOH) to afford compound 1 (22 mg),
koaburside (3, 38.7 mg), icariside E (4, 138.6 mg), cle-
9
. Deyama T, Ikawa T, Nishibe S (1985) The constituents of
Eucommia ulmoides Oliv. II. Isolation and structures of three new
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4
omiscosin C (5, 37.3 mg), and cleomiscosin D (6, 13.4 mg).
1
1
0. Yoshikawa K, Matsumoto K, Arihara S (1999) New lanostanoid
glycosides from the fruit body of Laetiporus versisporus. J Nat
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1. Ukeda H, Kawana D, Maeda S, Sawamura M (1999) Spectro-
photometric assay for superoxide dismutase based on the
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xanthine oxidase. Biosci Biotechnol Biochem 63:485–488
Compound 1
Amorphous solid. [a] -40.1° (c 1.7, MeOH). FT-IR (dry
D
-
1
film) cm : 3400 (OH), 1760 (C=O), 1050 (OH). HR-FAB-MS
m/z: 549.2006 (calculated for C H O : 549.1973).
27 34 12
123