2394 Bull. Korean Chem. Soc. 2010, Vol. 31, No. 8
Notes
FAB-MS m/z 547.1 [M + Na]+; HR-FAB-MS m/z 547.1068
[M + Na]+ (calcd for C23H24O14Na, 547.1064).1H NMR (400
MHz, C5D5N) δ 4.19-4.77 (1H, t, J = 8.0 Hz, H-2''; 1H, dd, J =
2.8, 8.0 Hz, H-3''; 1H, d, J = 2.8 Hz, H-4''; 1H, t, J = 6.0 Hz,
H-5''; 1H, dd, J = 6.0, 10.8 Hz, H-6''a; 1H, dd, J = 6.4, 10.8 Hz,
H-6''b), 3.97 (6H, s, OCH3), 6.41 (1H, d, J = 7.6 Hz, H-1''), 6.90
(1H, d, J = 2.0 Hz, H-6), 8.17 (2H, s, H-2',6'). 13C NMR (100
MHz, C5D5N) δ 57.3 (3',5'-OCH3), 62.4 (C-6''), 70.3 (C-4''), 73.9
(C-2''), 75.3 (C-3''), 78.1 (C-5''), 100.4 (C-6), 104.6 (C-1''),
105.7 (C-4a), 108.8 (C-2'), 108.8 (C-6'), 121.5 (C-1'), 127.5
(C-8), 135.5 (C-3), 141.2 (C-4'), 149.2 (C-5'), 149.3 (C-3'),
146.8 (C-8a), 155.2 (C-5), 155.7 (C-7), 157.9 (C-2), 179.6 (C-4).
Determination of sugar component. The monosaccharide
subunit of 1 was obtained by acid hydrolysis. Compound 1
(4 mg) in 10% HCl-dioxane (1:1, 1 mL) was heated at 80 oC
for 4 h in a water bath. The reaction mixtures were neutralized
with Ag2CO3, filtered, and then extracted with EtOAc. After
concentration, each H2O layer (monosaccharide portion) was
evaporated in vacuo to give residue, which was subjected to a
silica gel column chromatography (CHCl3-MeOH-H2O (55:45:
10) to yield D-galactose. The sugar was compared with authentic
sample by TLC. The Rf value for the above sugar was 0.19.
Cytotoxic activity assay. The cancer cell lines were main-
tained in RPMI 1640, which included l-glutamine with 10% FBS
and 2% penicillin-streptomycin. Cells were cultured at 37 oC
in a 5% CO2 incubator. Cytotoxic activity was measured using
a modified MTT assay.18 Viable cells were seeded in the growth
medium (100 µL) into 96-well microtiter plates (1 × 104 cells per
well) and incubated at 37 oC in a 5% CO2 incubator. The test
sample was dissolved in DMSO and adjusted to final sample
concentrations ranging from 5.0 to 150 µM by diluting with
the growth medium. Each sample was prepared in triplicate.
The final DMSO concentration was adjusted to < 0.1%. After
standing for 24 h, 10 µL of the test sample was added to each
well. The same volume of DMSO was added to the control wells.
On removing medium after 48 h of the test sample treatment,
MTT (5 mg/mL, 10 µL) was also added to the each well. After
4 h incubation, the plates were removed, and the resulting
formazan crystals were dissolved in DMSO (150 µL). The OD
was measured at 570 nm. The IC50 value was defined as the
concentration of sample that reduced absorbance by 50% rela-
tive to the vehicle-treated control.
Acknowledgments. This research was supported by research
grants from Catholic University of Daegu in 2010.
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