92
T. Brukwicki et al. / Journal of Molecular Structure 832 (2007) 90–95
completed after 155 h. The solution was filtrated through a
column with a small amount of asbestos and the column
was washed by three portions of water (2 ml). The solution
was concentrated to about 15 ml on the rotary evaporator
under a reduced pressure and the rest was transferred into
a mortar. Pellets of potassium hydroxide (8 g) were cau-
tiously added and all was mixed with 12 g of diatomaceous
earth. The mixture was placed in a column with basic dia-
tomaceous earth (prepared of 5 g of diatomaceous earth
and 4 ml of 20% aqueous solution of potassium hydrox-
ide). The alkaloids were eluted with 300 ml of petroleum
ether (40–60ꢁ), 100 ml of diethyl ether and 50 ml of methy-
lene chloride. From the petroleum ether fraction 0.696 g of
chromatographically pure colourless crystals of 13a-
hydroxysparteine were obtained (92%). Mp. 145ꢁ. EI-MS
All solutions were concentrated and controlled by TLC.
The petroleum ether fraction containing chromatographi-
cally pure fraction of alkaloids was dissolved in a small
amount of diethyl ether and filtrated through a column
with 10 g of silica gel. The alkaloids were eluted with dieth-
yl ether (50 ml) and methylene chloride (50 ml). From
diethyl ether fraction, colourless crystals of 13b-hydroxys-
parteine were obtained (0.076 g, 85%). Mp. 124 ꢁC. EI-MS
+
Æ
m/z (%): 250 [M ] (45), 251 (10), 209 (32), 153 (34), 137
(100), 134 (21), 133 (19), 126 (15), 121 (15), 114 (13). IR
ꢀ1
(KBr pellets, cm ) 3342 (OH), 2819, 2795, 2772, 2679,
1
3
1
2664 (Bohlmann band); C and H NMR – Tables 1
and 2, respectively.
3. Results and discussion
+
Æ
m/z (%): 250 (30) [M ], 251 (7.5), 252 (0.87), 233 (11),
1
3.1. C NMR and H NMR spectra
3
1
2
1
1
09 (21), 153 (11), 152 (22), 150 (16), 138 (15), 137 (100),
36 (19), 134 (11), 126 (12), 122 (16), 114 (13), 113 (22),
ꢀ1
13
1
08 (13), 98 (84). IR (KBr pellets, cm ) 3364 (OH),
The C and H NMR signals in the spectra of 13a-
1
3
2
828, 2798, 2777, 2753, 2681 (Bohlmann band); C and
hydroxysparteine (5) and 13b-hydroxysparteine (6) were
1
13
H NMR – Tables 1 and 2, respectively.
3b-Hydroxylupanine was obtained from 13-oxolupa-
assigned using 2D techniques. Tentative results of the
C
1
NMR spectra were starting-points for HSQC spectra anal-
1
nine [7] by NaBH4 reduction according to the method
reported previously [8] and characterized in Ref. [5].
ysis and the assignment of H signals was verified by means
1
1
of H– H COSY. The results are shown in Tables 1 and 2,
respectively. The data for 13a- (3) and 13b-hydroxylupa-
1
3
2
.2.2. 13b-Hydroxysparteine
nine (4) [5] were included into the same tables. The
C
PtO (0.050 g) was suspended in 10 ml of 1 N HCl and
the suspension was stirred by a magnetic stirrer. Gas
hydrogen was bubbled into the suspension. After 30 min,
3b-hydroxylupanine (0.090 g) (dissolved in 5 ml of 1 N
HCl) was added to the suspension. The progress of the
reaction was controlled by TLC (silica gel, acetone–metha-
NMR spectra of 13a-hydroxysparteine (5) are in accor-
dance with the results of Bohlmann (Bohlmann inter-
changed signals for C7 and C9; the only difference, ca
2
1
3
1
1
0.6 ppm, is for carbon atom C13) [9], the C and H
1
NMR spectra for 13b-hydroxysparteine (6) and H NMR
for 5 have been recorded, as far as we know, for the first
time.
nol–methanol/NH , 5–0.5–0.5). The reaction was complet-
3
ed in 150 h. The reaction mixture was filtrated through a
column with 5 g Al O and alkalized by KOH to pH 14.
The solution was extracted by petroleum ether (100 ml),
diethyl ether (100 ml) and methylene chloride (100 ml).
A comparison of the chemical shifts of carbon atoms of
compounds 5 and 6 with those of the appropriate carbon
atoms of sparteine (1) indicates the differences involving
the appropriate atoms forming ring A can be neglected.
2
3
Table 1
1
3
C NMR chemical shifts of 13a-hydroxysparteine (5), 13b-hydroxysparteine (6), 13a- hydroxylupanine (3) and 13b-hydroxylupanine (4), (CDCl3, ppm
from TMS)
0
C atom 13a-Hydroxysparteine D = d
5
ꢀ d
1
13a-Hydroxylupanine 13b-Hydroxysparteine D = d
6
ꢀ d
1
13b-Hydroxylupanine Sparteine
(
5)
(3)
(6)
(4)
(1)
2
3
4
5
6
7
8
9
0
1
2
3
4
5
7
56.12
25.77
24.69
29.28
66.41
32.98
27.43
35.48
61.63
57.25
41.54
65.01
32.76
49.13
53.18
ꢀ0.16
171.19
33.09
19.78
27.46
60.86
32.40
26.62
34.30
46.75
57.11
40.17
64.46
31.73
49.16
52.43
56.27
25.92
24.75
29.40
66.45
33.12
27.48
35.81
61.66
61.86
43.57
69.34
35.24
52.83
52.61
ꢀ0.01
ꢀ0.04
ꢀ0.05
ꢀ0.01
ꢀ0.10
ꢀ0.08
ꢀ0.26
ꢀ0.42
ꢀ0.35
ꢀ2.60
8.79
171.25
32.42
19.50
27.31
60.57
32.24
26.62
34.35
46.71
61.17
41.56
68.97
33.84
52.78
51.49
56.28
25.96
24.80
29.41
66.55
33.20
27.74
36.23
62.01
64.46
34.78
24.95
26.06
55.44
53.65
ꢀ0.19
ꢀ0.11
ꢀ0.13
ꢀ0.14
ꢀ0.22
ꢀ0.31
ꢀ0.75
ꢀ0.38
ꢀ7.21
6.76
1
1
1
1
1
1
1
40.06
6.70
44.39
9.18
ꢀ6.31
ꢀ2.66
ꢀ0.47
ꢀ1.04