Molecules 2010, 15
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(C-6''', C-6''''), 60.7 (C-6''), 69.5 (C-4''''), 71.5 (C-5'''), 72.0 (C-2'''), 72.4 (C-2'''', C-5''''), 73.0 (C-3''''),
73.3 (C-3'''), 73.8 (C-2''), 75.1 (C-5''), 75.9 (C-3''), 78.8 (C-4''), 78.9 (C-4'''), 98.1 (C-1''), 101.5 (C-1''',
C-1''''), 101.9 (C-8), 105.1 (C-5), 115.0 (C-3', C-5'), 117.6 (C-10), 122.5 (C-1'), 124.5 (C-3), 130.3
(C-2', C-6'), 148.7 (C-6), 151.0 (C-9), 152.2 (C-7), 153.8 (C-2), 157.6 (C-4'), 178.0 (C-4).
Glycitein 7-O-β-maltoside (6): FABMS m/z: 631.1641 [M+Na]+ (calcd 631.1639 for C28H32O15Na);
1H-NMR (DMSO-d6): δ 3.09-3.77 (12H, m, H-2'', 2''', 3'', 3''', 4'', 4''', 5'', 5''', 6'', 6'''), 3.90 (3H, s,
OCH3), 5.07 (1H, d, J = 3.2 Hz, H-1'''), 5.19 (1H, d, J = 7.6 Hz, H-1''), 7.01 (2H, d, J = 8.0 Hz, H-3',
5'), 7.46 (1H, s, H-8), 7.52 (2H, d, J = 8.0 Hz, H-2', 6'), 7.76 (1H, s, H-5), 8.41 (1H, s, H-2); 13C-NMR
(DMSO-d6): δ 56.1 (OCH3), 60.2 (C-6'''), 60.9 (C-6''), 69.7 (C-4'''), 72.0 (C-2'''), 72.7 (C-5'''), 73.3
(C-2''), 73.5 (C-3'''), 75.1 (C-5''), 75.9 (C-3''), 79.9 (C-4''), 98.8 (C-1''), 100.6 (C-1'''), 101.0 (C-8),
105.1 (C-5), 115.2 (C-3', C-5'), 117.7 (C-10), 122.5 (C-1'), 124.5 (C-3), 130.3 (C-2', C-6'), 148.8 (C-6),
151.0 (C-9), 152.5 (C-7), 153.8 (C-2), 157.0 (C-4'), 178.0 (C-4).
Glycitein 7-O-β-maltotrioside (7): FABMS m/z: 793.2177 [M+Na]+ (calcd 793.2167 for
1
C34H42O20Na); H-NMR (DMSO-d6): δ 3.05-3.88 (18H, m, H-2'', 2''', 2'''', 3'', 3''', 3'''', 4'', 4''', 4'''', 5'',
5''', 5'''', 6'', 6''', 6''''), 3.92 (3H, s, OCH3), 5.07 (1H, d, J = 3.2 Hz, H-1''''), 5.09 (1H, d, J = 3.2 Hz, H-
1'''), 5.19 (1H, d, J = 7.6 Hz, H-1''), 7.00 (2H, d, J = 8.4 Hz, H-3', 5'), 7.45 (1H, s, H-8), 7.51 (2H, d,
J = 8.4 Hz, H-2', 6'), 7.76 (1H, s, H-5), 8.41 (1H, s, H-2); 13C-NMR (DMSO-d6): δ 56.1 (OCH3), 60.5
(C-6''''), 60.7 (C-6'', C-6'''), 69.7 (C-4''''), 71.5 (C-5'''), 72.1 (C-2'''), 72.5 (C-2'''', C-5''''), 73.0 (C-3''''),
73.5 (C-3'''), 73.8 (C-2''), 75.1 (C-5''), 75.7 (C-3''), 79.7 (C-4''), 79.9 (C-4'''), 98.1 (C-1''), 100.5, 100.6
(C-1''', C-1''''), 101.9 (C-8), 105.1 (C-5), 115.0 (C-3', C-5'), 117.6 (C-10), 122.5 (C-1'), 124.5 (C-3),
130.2 (C-2', C-6'), 148.8 (C-6), 151.0 (C-9), 152.5 (C-7), 153.8 (C-2), 157.0 (C-4'), 178.1 (C-4).
3.5. Suppressive action on IgE antibody formation
The inhibitory action of β-glycosides of glycitein or daidzein on IgE antibody formation was
examined as follows: 7S-globulin was used as the antigen (1 mg/rat), and Al(OH)3 and pertussis
vaccine were used as the adjuvants (20 mg and 0.6 mL/rat, respectively). Sensitization was made by
injection of a mixture (0.6 mL) of the antigen and the adjuvant into the paws of each rat (male,
ca. 200 g). Paw edema was measured 24 h after injection and the treated rats were divided in groups
with an equal average swelling volume. Each sample was dissolved in physiological saline containing
10% Nikkol and the solution was injected daily into the rat for 11 d starting on the day of grouping.
Hydrocortisone was used as the positive control. The amount of IgE was measured by the passive
cutaneous anaphylaxis method on the 15th day [17]. The results were expressed as average of plasma
IgE level of 7 rats administered a total of 10 mg/kg of each test compound.
3.6. DPPH radical scavenging activity
DPPH free-radical scavenging activities of β-glycosides of glycitein or daidzein were determined as
follows: DPPH was dissolved in ethanol (500 μM) [21]. The sample solutions were prepared by
dissolving each compound in ethanol. To a solution containing various concentrations of each sample
(0.1 mL) and ethanol (0.9 mL) was added DPPH solution (1 mL) at room temperature. Vitamin C was