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L.S.M. Velozo et al. / Phytochemistry 52 (1999) 1473±1477
3. Experimental
13C-NMR spectral data: Table 1. Negative LSIMS, m/
z (rel. int.): 475 [M-H] (58), 313 [M-163] (100).
3.1. General
3.5. Acid hydrolysis of 1
Mps were determined by an Electrothermal 9200
micro melting point apparatus and are uncorrelated,
or were measured on a Perkin Elmer 243B polarimeter.
UV and IR spectra were measured on a Shimadzu
UV-1601 and on a Perkin Elmer 599B, respectively.
1H- and 13C-NMR spectra were obtained on a Varian
Gemini 200 NMR spectrometer operating at 200 MHz
The compound (1, 100 mg) was hydrolyzed with 1
M HCl in dioxane±H2O (1 : 1, 10 ml) under re¯ux for
30 min. After diluting with H2O (90 ml), the aglycone
was extracted with CHCl3±MeOH (4 : 1) and evapor-
ated to dryness in vacuo. The residue was dissolved in
MeOH, and on concentration the solution yielded a
pale yellow compound, which on further recrystalliza-
tion gave odoratin (1a, 49 mg), mp 214±2168C [lit.
(Hayashi & Thomson, 1974; Galina & Gottlieb, 1974)
mp 213±2158C]. Odoratin 1a was analyzed by silica
gel-TLC in the above described solvent system. After
spraying with orcinol-H2SO4 it gave a yellow spot at
Rf 0:45. UV lMmaexOH nm ꢀlog e): 223 (4.33), 260 (4.19),
320 (3.90), (NaOAc): 215 (4.35), 255 (4.26), 346 (4.03);
(NaOAc + H3BO3): 213 (4.36), 259 (4.18), 323 (3.86).
for H and 50 MHz for 13C in DMSO-d6, with TMS
1
as internal standard. GC was carried out with FID,
using a glass capillary column (0.31 mm  25 m) SE-
30. EIMS was recorded at 70 eV. Negative LSIMS
was carried out using HMPA-glycerol as matrix, 35
kV anodic voltage, 8 kV accelerating voltage using Cs
ions.
Silica gel column (230±400 mesh ASTM, Merck)
was used for CC. TLC was performed on silica gel
(Merck) using the following solvent systems: (A)
CHCl3±MeOH (85 : 15) for 1, (B) CHCl3±MeOH (97 :
3) for 1a, (C) CHCl3±MeOH (98 : 2) for 1b and 1c
and (D) 1-BuOH±pyridine±H2O (6 : 4 : 3) for sugar.
Compounds 1, 1a, 1b and 1c were detected under UV
of 254 and 366 nm and by spraying with orcinol-
H2SO4. Sugar was detected by spraying with orcinol-
H2SO4.
IR nmax cm 1: 3513 (OH), 3157, 2994, 1622, 1578,
KBr
1566, 1514, 1480, 1455, 1320, 1207, 1183, 1137, 1051,
1015, 859, 762. H- and 13C-NMR spectral data: Table
1
1. EIMS (probe) 70 eV m/z (rel. int.): 314 [M]+ (100),
299 (21), 285 (5), 271 (17), 243 (11), 167 (10), 105 (13).
HRMS m/z: 314.2970 [M+]1 C17H14O6. Calcd. =
314. The aqueous layer was adjusted to pH 6 by ad-
dition of NaHCO3. After lyophilization, sugar was dis-
solved in pyridine and analyzed by silica gel-TLC in
the above described solvent system. After spraying,
glucose gave a blue spot at Rf 0:70. The con®gur-
ation of the sugar was established by capillary GC of
its TMSi ( )-2-butyl glycosides (Gerwig, Kamerling &
Vliegenthart, 1978).
3.2. Plant material
Roots of B. virgilioides Kunt were collected at Paco
do Lumiar, Maranhao, on March 1997, and identi®ed
by T.J.A.S. Rego. A voucher specimen (no. 01137) is
deposited at the Atico Seabra Hebarium.
3.6. Acetylation of 1
3.3. Extraction and isolation
Compound 1 (100 mg) was treated overnight with
Ac2O and pyridine at room temperature. Usual warm-
up gave (1b, 141 mg, TLC: Rf 0:55), mp 103±1058C,
Dried and powdered roots of B. virgilioides (1 kg)
were extracted with cold MeOH. Evaporation of sol-
vent gave a residue (20 g), a part of which (10 g) was
submitted to CC (90 Â 3 cm) on silica gel which was
eluted with CHCl3±MeOH mixtures of increasing po-
larity (up to 10% MeOH). Frs. eluted with CHCl3±
MeOH (9 : 1) yielded 1 as a homogeneous compound
(495 mg), TLC, Rf = 0.25, dark blue color with orci-
nol-H2SO4 and negative FeCl3 test.
20
D
MeOH
max
a
52.48 (DMSO, c 0.001). UV l
nm ꢀlog e):
244 (4.73), 322 (4.40), IR nmax cm 1: 2944, 1758, 1641,
1619, 1514, 1495, 1434, 1371, 1272, 1217, 1130, 1068,
1040, 906, 878, 820, 752. 1H- and 13C-NMR spectral
data: Table 1. CIMS, m/z (rel. int.): 704 [M + NH4]+
(78), 686 [M] (7).
KBr
3.7. Acetylation of 1a
3.4. Odoratin 7-O-b-D-glucopyranoside (1)
Compound 1a (100 mg) was treated as 1 to give (1c,
113 mg, TLC: Rf 0:50), mp 185±1878C [lit. (Hayashi
MeOH
Pale yellow amorphous powder from MeOH, mp
20
& Thomson, 1974; Galina & Gottlieb, 1974) mp 185±
MeOH
max
KBr
185±1888C, a
83.28 (DMSO, c 0.001). UV l
1878C]. UV l
nm ꢀlog e): 208 (4.41), 260 (4.38),
D
max
cm 1: 2957, 2849, 1770, 1655, 1618,
1515, 1494, 1436, 1372, 1272, 1198, 1183, 1126, 1015,
KBr
max
nm ꢀlog e): 219 (4.14), 262 (4.05), 319 (3.67). IR nmax
cm 1: 3484 (OH), 1624 (%$C10), 1494, 1435, 1289,
1262, 1219, 1201, 1085, 1020, 872, 810, 719. 1H- and
326 (3.90). IR l
905, 821, 601. H- and 13C-NMR spectral data: Table
1