Constituents of the fungi Daedalea quercina and Daedaleopsis confragosa var. tricolor

Article abstract of DOI:10.1016/S0031-9422(00)00130-8

Phytochemical examination of solvent extracts of the wood-rotting fungi Daedalea quercina and Daedaleopsis confragosa var. tricolor led to the isolation of five new triterpene derivatives and some known fungal constituents. All structures were identified

Full text of DOI:10.1016/S0031-9422(00)00130-8

Phytochemistry 54 (2000) 757±762
www.elsevier.com/locate/phytochem
Constituents of the fungi Daedalea quercina and Daedaleopsis
confragosa var. tricolor
Joachim Rosecke, Wilfried A. Konig*
Institut fur Organische Chemie, Universitat Hamburg, Martin-Luther-King-Platz 6, D-20146 Hamburg, Germany
È
È
Received 10 February 2000; received in revised form 3 April 2000
Abstract
Phytochemical examination of solvent extracts of the wood-rotting fungi Daedalea quercina and Daedaleopsis confragosa var.
tricolor led to the isolation of ®ve new triterpene derivatives and some known fungal constituents. All structures were identi®ed
by one- and two-dimensional NMR spectroscopy and mass spectrometry. From Daedalea quercina, the new natural products 16-
O-acetylpolyporenic acid C, 16a-acetoxy-24-methylene-3-oxolanost-8-en-21-oic acid, (+)-24-methylene-3,23-dioxolanost-8-en-26-
oic acid, (+)-3b,12b-dihydroxy-24-methyl-23-oxolanost-8-en-26-oic acid and 12b,23-epoxy-3a,23-dihydroxy-24-methyllanost-8-
en-26-oic acid could be isolated. From Daedaleopsis confragosa var. tricolor, the compounds 3a-carboxyacetoxyquercinic acid,
3a-carboxyacetoxy-24-methylene-23-oxolanost-8-en-26-oic acid and 5a,8a-epidioxyergosta-6,22-dien-3b-ol were identi®ed. These
are the ®rst described triterpene derivatives isolated from this fungus. 7 2000 Elsevier Science Ltd. All rights reserved.
Keywords: Daedalea quercina; Daedaleopsis confragosa var. tricolor; Polyporaceae; Basidiomycete; Triterpene derivatives
1. Introduction
only some unusual fatty acids have been described ear-
lier (Dembitsky et al., 1993). In this article, we describe
the isolation and characterization of 12 compounds,
®ve of which are new natural products.
Little e€orts have been made in the past to identify
compounds from the brown-rot fungus Daedalea quer-
cina (L.) Pers. [syn. Trametes quercina (L.) Pilat],
which grows on weak or dead stumps of deciduous
trees, mainly oak trees, and from the white-rot species
Daedaleopsis confragosa var. tricolor (Bulliard) Bon-
darzew [syn. Lenzites tricolor (Bulliard)], which grows
on various deciduous trees. Only ergosterol, ergosta-7,
22-dien-3b-ol, ergosterol peroxide (5a,8a-epidioxyer-
gosta-6,22-dien-3b-ol) and 3a-carboxyacetoxyquercinic
acid (3a-carboxyacetoxy-24-methyl-23-oxolanost-8-en-
26-oic acid) have been isolated from fruiting bodies of
D. quercina (Adam et al., 1967a; Turner and Aldridge,
1983). Ergosterol peroxide is believed to be an artefact
(Adam et al., 1967b). From D. confragosa var. tricolor
2. Results and discussion
Fresh fruiting bodies of both species were cut in
small pieces and crushed in liquid nitrogen. The
resulting powders were extracted with n-hexane. After
removing the solvent, D. quercina was ®rst extracted
with methanol and the obtained solid extract ®nally
extracted with dichloromethane. D. confragosa var. tri-
color was directly extracted with dichloromethane.
Both dichloromethane extracts and the n-hexane
extract of D. quercina were repeatedly chromato-
graphed on silica gel with step-gradients of petroleum
ether±ethyl acetate and chloroform±methanol.
* Corresponding author. Tel.: +49-404-283-82824; fax: +49-404-
283-828293.
E-mail address: wkoenig@chemie.uni-hamburg.de (W.A. Konig).
0031-9422/00/$ - see front matter 7 2000 Elsevier Science Ltd. All rights reserved.
PII: S0031-9422(00)00130-8

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J. Rosecke, W.A. Konig / Phytochemistry 54 (2000) 757±762
È È
2.1. Daedalea quercina
was 3b,12b-dihydroxy-24-methyl-23-oxolanost-8-en-26-
oic acid for which we propose the name quercinic acid
C.
Six compounds were identi®ed by one- and two-
dimensional NMR spectroscopy, mass spectrometry
and comparison with literature data as polyporenic
acid C (1), 16a-hydroxy-24-methylene-3-oxolanost-8-
en-21-oic acid (2), 16-O-acetylpolyporenic acid C (3),
16a-acetoxy-24-methylene-3-oxolanost-8-en-21-oic acid
(4), 3a-carboxyacetoxyquercinic acid (5) and ergosterol
peroxide (6). To our knowledge, compounds 3 and 4
are new natural products. In addition, compounds 1
and 2 are reported here for the ®rst time as constitu-
ents of D. quercina, and the NMR data of compounds
2 and 4 were not reported before. Compound 1 was
previously isolated from various fungi, e.g. Fomitopsis
pinicola, and converted to its 16-O-acetyl derivative
(Keller et al., 1996). Compound 2 was isolated pre-
viously from the closely related fungus Daedalea dick-
insii (Inouye et al., 1970). The NMR data of
compound 5 was described earlier (Chairul et al.,
1990). Some of the earlier assignments appeared to be
incorrect.
Compound 9 showed two ole®nic ꢀd 133.84, 134.85)
and two methine carbon signals ꢀd 75.79, 79.73) as
well as one carboxylic carbon at d 177.36. In addition,
a quaternary carbon signal at d 109.96 was found
which corresponds to a semi-acetalic carbon atom. The
latter showed long-range couplings with the hydrogen
atoms at carbons 25 ꢀd 2.22), 24 ꢀd 2.12), 20 ꢀd 2.24)
and with the hydroxylic methine proton at carbon 12
ꢀd 4.34) (HMBC). This led to the conclusion that for-
mation of a cyclic acetal had taken place involving the
oxygen at position 12 and the carbonyl group in pos-
ition 23 of quercinic acid C (8). Comparison of the
chemical shifts of the carbons in ring A of the triter-
pene skeleton with the data of many previously ident-
i®ed compounds indicated a 3a-con®guration, so that
this compound was not a simple artefact of compound
8. Analysis of all spectral data led to the conclusion
that compound 9 was 12b,23-epoxy-3a,23-dihydroxy-
24-methyllanost-8-en-26-oic acid for which we propose
the name 3a-oxepanoquercinic acid C.
Compound
7
showed four ole®nic carbons ꢀd
From the n-hexane extract, a 15:1 mixture of (E)-
(10) and (Z )-methyl 4-methoxycinnamate (11) was iso-
lated. The identi®cation was carried out by comparison
of the GC-MS and NMR spectra with literature data
(10: van Heerden et al., 1996, 11: Jacobsen et al.,
1994). Compound 10 was previously isolated from the
fungus Lentinus lepideus (Wat and Towers, 1977). A
reference sample of methyl (E )-4-methoxycinnamate
was prepared by treatment of authentic (E )-4-methoxy-
cinnamic acid with diazomethane. The spectroscopic
data corresponded to those of compound 10.
133.31, 135.16, 125.46, 148.05), one acid ꢀd 178.31)
and two ketone carbon signals ꢀd 201.83, 217.90). The
chemical shifts of the keto function at d 201.83, the
exocyclic methylene group at d 125.46 and observed
long-range correlations indicated a lanost-8-ene struc-
ture with a 23-oxo-24-methylene functionalized side-
chain and an acid function in position 26 which is
common to previously identi®ed triterpenic acids from
this species. Analysis of the NMR spectra (DEPT,
HMQC, HMBC) and their comparison with the data
of other isolated metabolites indicated the structure
24-methylene-3,23-dioxolanost-8-en-26-oic acid for
which we propose the name quercinic acid B.
Compound 8 showed two ole®nic ꢀd 133.33, 136.74)
and two hydroxylic methine carbons ꢀd 71.57, 74.64)
as well as one ketone ꢀd 213.31) and one carboxylic
function ꢀd 178.04). The hydroxylic methine proton at
d 71.57 showed long-range coupling with the quater-
nary carbons 13 and 14 ꢀd 52.04, 49.36) as well as with
a methylene group at d 34.62. Further analysis left car-
bon 12 as the only possible position for this hydroxy
function. The other hydroxyl group could be assigned
to carbon 3 as the proton showed long-range couplings
with carbons 2, 4, 5 and with the methyl carbons 28
and 29. Futher analysis of the NMR data indicated
presence of a 24-methyl-23-oxo side chain with an acid
function in position 26. In the NOESY spectrum, the
hydroxylic methine proton ꢀd 4.39) only showed a cor-
relation with the methyl protons at carbon 30 ꢀd 0.94)
indicating a b-con®guration. The hydroxylic methine
proton at carbon 3 ꢀd 3.63) only showed a correlation
with the protons connected to carbon 28 ꢀd 0.92) also
indicating a b-con®guration. Therefore, this compound
2.2. Daedaleopsis confragosa var. tricolor
From the dichloromethane extract, compounds 5
and 6 were isolated. These compounds are the ®rst
triterpene derivatives isolated from this species and
were identi®ed by comparison of their spectral data
with literature data. Furthermore, a mixture of com-
pound 5 and a very similar compound was obtained.
The latter one showed di€erent carbon shifts for the
side chain compared with compound 5. The additional
presence of an ole®nic methylene carbon at d 125.34
and a quaternary carbon at d 148.05 as well as the
up®eld shift of the carbonyl carbon to d 201.54 indi-
cated that this compound was 3a-carboxyacetoxy-24-
methylene-23-oxolanost-8-en-26-oic acid (12). This was
con®rmed by analysis of the two-dimensional NMR
spectra. This compound has been isolated previously
from an unidenti®ed Indonesian Ganoderma species
(Chairul et al., 1990) but some NMR assignments
appeared to be incorrect.

J. Rosecke, W.A. Konig / Phytochemistry 54 (2000) 757±762
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759
3.2. Extraction
3.2.1. Daedalea quercina
One kilogram of fresh fruiting bodies were crushed
in liquid nitrogen. The resulting powder was extracted
twice with n-hexane (2 Â 2 l, 6 days each) and then
with methanol (2 l, 6 days). The methanol extract (5.3
g of a green solid) was extracted with 250 ml of
dichloromethane (1 day under stirring) yielding 4.8 g
of a pale green solid.
3.2.2. Daedaleopsis confragosa var. tricolor
750 g of fresh fruiting bodies were crushed in liquid
nitrogen. The resulting powder was extracted twice
with n-hexane (2 Â 2 l, 6 days each), with dichloro-
methane (2 l, 6 days), yielding 6.7 g of a pale green
solid, and ®nally with methanol (2 l, 6 days).
3.3. Isolation
3.3.1. Daedalea quercina
The dichloromethane extract was submitted to ¯ash
column chromatography on silica gel with a stepwise
gradient of petroleum ether±ethyl acetate (from 39:1 to
0:1, v/v) and was rechromatographed several times on
silica gel using stepwise gradients of petroleum ether±
ethyl acetate (from 39:1 to 0:1, v/v) and chloroform-
methanol (from 49:1 to 9:1, v/v). The following yields
were obtained: 1 (see below), 2 (15 mg), 3, 4 (see
below), 5 (30 mg), 6 (55 mg), 7 (14 mg), 8 (10 mg), 9
(50 mg). compounds 3 and 4 were obtained only as a
mixture and 1 was only obtained admixed with 2, but
because of the existence of similar reference com-
pounds, further separation was not necessary. The n-
hexane extract was chromatographed in the same man-
ner and yielded 5 mg of a 15:1 mixture of 10 and 11.
3.3.2. Daedaleopsis confragosa var. tricolor
The dichloromethane extract was chromatographed
in the same manner as described for the extracts of D.
quercina. Yields: 5 (5 mg), a 3:1 mixture of 5 and 12
(100 mg) and 6 (10 mg).
3.4. TLC
3. Experimental
Silica gel 60F254 (Merck); eluent chloroform-2-pro-
panol 9:1, v/v and petroleum ether±ethyl acetate, 1:1,
v/v; detection by spraying with sulphuric acid (10% in
ethanol).
3.1. Plant material
Fruiting bodies of Daedalea quercina were collected
in October 1998 near Salzgitter in the Harz Mountains
and fruiting bodies of Daedaleopsis confragosa var. tri-
color in September 1999 in the Sachsenwald near Ham-
burg (both in northern Germany) (Gerhardt, 1996).
3.5. Column chromatography
Various column sizes with silica gel 0.063±0.040 mm
(Macherey±Nagel).

760
J. Rosecke, W.A. Konig / Phytochemistry 54 (2000) 757±762
È È
3.6. Gas chromatography
3.9. Polarimetry
Carlo Erba HRGC 5300 Mega Series instrument
equipped with a fused silica capillary (25 m) coated
with CPSil-5CB (Chrompack), split injector and FID.
Temperature program: 110±2308C, 38C/min. Carrier
gas: hydrogen.
Perkin±Elmer 241, l ˆ 1 dm, l ˆ 589 nm.
3.10. EI-MS
70 eV, VG Analytical 70-250S, exact mass measure-
ment at resolution 10,000, direct probe sample intro-
duction.
3.7. GC-MS
3.11. Compound 1, polyporenic acid C, 16a-hydroxy-24-
methylene-3-oxolanosta-7,9(11)-dien-21-oic acid
VG Analytical VG 70-250S mass spectrometer (EI,
70 eV) coupled to a Hewlett-Packard HP 5890 gas
chromatograph. Carrier gas: helium.
Obtained only as a mixture with 2. 13C-NMR
(100.61 MHz, pyridine-d5): see Table 1.
3.8. NMR spectroscopy
3.12. Compound 2, 16a-hydroxy-24-methylene-3-
oxolanost-8-en-21-oic acid
Bruker WM 400 at 400.16 (1H) and 100.61 MHz
(13C) and Bruker DRX 500 at 500.13 (1H) and 125.76
MHz (13C). All NMR shifts are relative to TMS.
1H-NMR (400.13 MHz, pyridine-d5): d 0.98 (3H, d,
J ˆ 3:1 Hz), 0.99 (3H, d, J ˆ 3:1 Hz), 1.00 (3H, s, H-
Table 1
13C shifts of identi®ed compounds (pyr = pyridine-d5)
C
1 (pyr)
2 (pyr)
3 (CDCl3)
4 (CDCl3)
5 (CDCl3)
6 (CDCl3)
7 (CDCl3)
8 (pyr)
9 (CDCl3)
12 (CDCl3)
1
36.38 t
34.54 t
214.79 s
47.08 s
50.64 d
23.47 t
120.31 d
142.43 s
144.35 s
37.13 s
117.24 d
35.85 t
44.63 s
48.96 s
43.95 t
75.98 d
57.20 d
17.20 q
21.96 q
48.12 d
178.24 s
31.04 t
32.83 t
155.64 s
33.73 d
21.63 q
21.48 q
21.63 q
25.95 q
25.24 q
106.64 t
35.72 t
34.29 t
215.83 s
46.95 s
50.90 d
19.22 t
26.27 t
133.00 s
135.26 s
36.69 s
20.51 t
29.25 t
45.82 s
48.48 s
43.31 t
76.17 d
56.93 d
17.44 q
18.22 q
48.27 d
178.37 s
31.18 t
32.83 t
155.69 s
33.73 d
21.63 q
21.48 q
20.93 q
26.00 q
25.00 q
106.64 t
36.70 t
34.75 t
216.47 s
47.48 s
50.67 d
23.66 t
121.16 d
141.29 s
144.48 s
37.28 s
116.51 d
35.43 t
43.82 s
48.55 s
40.92 t
78.71 d
53.06 d
17.11 q
22.00 q
46.07 d
180.53 s
29.94 t
31.66 t
154.82 s
34.02 d
21.85 q
21.90 q
22.42 q
25.33 q
25.65 q
106.85 t
170.79 s
21.33 q
36.07 t
34.50 t
217.48 s
47.42 s
51.18 d
19.30 t
26.30 t
133.35 s
134.63 s
36.95 s
20.58 t
28.95 t
45.12 s
48.16 s
40.28 t
78.98 d
52.91 d
17.38 q
18.63 q
46.20 d
180.77 s
30.05 t
31.66 t
154.82 s
34.02 d
21.85 q
21.90 q
21.27 q
26.19 q
24.85 q
106.85 t
170.79 s
21.33 q
30.70 t
23.23 t
80.50 d
36.86 s
45.36 d
18.01 t
25.97 t
134.17 s
134.49 s
36.86 s
20.93 t
30.89 t
44.59 s
50.03 s
30.82 t
28.37 t
50.26 d
15.80 q
18.97 q
32.68 d
13.45 q
48.76 t
213.53 s
48.13 d
40.79 d
181.41 s
14.26 q
21.78 q
27.64 q
24.29 q
19.82 q
166.74 s
40.78 t
171.49 s
34.69 t
30.08 t
66.46 d
36.90 t
82.16 s
135.42 d
130.73 d
79.43 s
51.08 d
36.96 s
23.40 t
39.74 t
44.56 s
51.68 d
20.63 t
28.64 t
56.20 d
12.87 q
18.17 q
39.74 d
20.88 q
135.20 d
132.30 d
42.77 d
33.06 d
19.64 q
19.95 q
17.56 q
36.10 t
34.63 t
217.90 s
47.43 s
51.28 d
19.44 t
26.35 t
135.16 s
133.31 s
36.94 s
21.08 t
30.90 t
44.69 s
50.06 s
28.40 t
21.08 t
50.72 d
15.94 q
19.73 q
33.96 d
18.71 q
44.69 t
201.83 s
148.05 s
40.19 d
178.31 s
15.89 q
21.32 q
26.20 q
24.41 q
125.46 t
30.55 t
26.49 t
74.64 d
37.76 s
44.18 d
18.17 t
25.83 t
133.33 s
136.74 s
36.89 s
34.62 t
71.57 d
52.04 s
49.36 s
31.27 t
25.61 t
50.38 d
10.44 q
19.01 q
29.91 d
22.55 q
48.21 t
213.31 s
48.46 d
41.58 d
178.04 s
14.41 q
22.21 q
28.68 q
23.82 q
13.26 q
31.79 t
23.18 t
79.73 d
36.78 s
45.21 d
17.81 t
25.58 t
134.85 s
133.84 s
36.78 s
29.45 t
75.79 d
52.65 s
49.18 s
30.94 t
23.62 t
42.99 d
11.79 q
18.86 q
28.07 d
19.65 q
40.74 t
109.96 q
50.61 d
42.25 d
177.36 q
13.68 q
21.82 q
27.58 q
24.71 q
13.01 q
30.69 t
23.22 t
80.47 d
36.85 s
45.35 d
18.00 t
25.97 t
134.17 s
134.47 s
36.83 s
20.94 t
30.88 t
44.58 s
50.02 s
30.81 t
28.45 t
50.72 d
15.81 q
18.96 q
34.05 d
19.67 q
44.66 t
201.54 q
148.05 s
40.31 d
179.76 s
15.84 q
21.78 q
27.65 q
24.28 q
125.34 t
166.74 s
40.78 t
171.49 s
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
1'
2'
3'

J. Rosecke, W.A. Konig / Phytochemistry 54 (2000) 757±762
È È
761
29), 1.05 (3H, s, H-18), 1.13 (3H, s, H-19), 1.14 (3H, s,
H-28), 1.46 (3H, s, H-30), 4.53 (1H, dd, J ˆ 7:6, 6:6
Hz, H-16), 4.85 (1H, s, H-31a), 4.98 (1H, s, H-31b).
13C-NMR (100.61 MHz, pyridine-d5): see Table 1.
(8), 467 [M
(13), 325 (87), 151 (100), 69 (94).
Me]+ (23), 449 [M
Me
H2O]+
3.18. Compound 8, quercinic acid C, 3b,12b-dihydroxy-
24-methyl-23-oxolanost-8-en-26-oic acid
3.13. Compound 3, 16-O-acetylpolyporenic acid C, 16a-
acetoxy-24-methylene-3-oxolanosta-7,9(11)-dien-21-oic
acid
Amorphous light brown solid, ‰a20 ‡ 86:0Š (CH2Cl2,
c0.05). 1H-NMR (400.13 MHz, pyridine-d5): d 0.92
(3H, s, H-28), 0.94 (3H, s, H-30), 1.05 (3H, s, H-18),
1.08 (3H, s, H-19), 1.15 (3H, d, J ˆ 6:9 Hz, H-31),
1.23 (3H, s, H-29), 1.34 (3H, d, J ˆ 7:2 Hz, H-27),
1.35 (3H, d, J ˆ 7:3 Hz, H-21), 2.57 (1H, dd, J ˆ 17:3,
10.1 Hz, H-20), 2.86 (1H, dd, J ˆ 17:6, 6.9 Hz, H-22a),
3.17 (2H, m, H-24, H-25), 3.63 (1H, br. s, H-3), 4.39
(1H, t, J ˆ 7:9 Hz, H-12). 13C-NMR (100.61 MHz,
pyridine-d5): see Table 1. EI-MS m/z (rel. int.): 502
Obtained only as a mixture with 4. 13C-NMR
(100.61 MHz, CDCl3): see Table 1.
3.14. Compound 4, 16a-acetoxy-24-methylene-3-
oxolanost-8-en-21-oic acid
Obtained only as a mixture with 3. 13C-NMR
(100.61 MHz, CDCl3): see Table 1.
[M]+ (7), 484 [M
Me]+ (100), 451 [M
309 (31), 69 (74).
H2O]+ (11), 469 [M
2 H2O
Me]+ (47), 325 (38),
H2O
3.15. Compound 5, 3a-carboxyacetoxyquercinic acid,
3a-carboxyacetoxy-24-methyl-23-oxolanost-8-en-26-oic
acid
3.19. Compound 9, 3a-oxepanoquercinic acid C, 12b,23-
epoxy-3a,23-dihydroxy-24-methyllanost-8-en-26-oic acid
1H-NMR (400.13 MHz, CDCl3): d 0.74 (3H, s, H-
18), 0.87 (3H, d, J ˆ 7:6 Hz, H-21), 0.89 (3H, s, H-29),
0.93 (3H, s, H-30), 0.94 (3H, s, H-28), 1.00 (3H, s, H-
19), 1.09 (3H, d, J ˆ 7:1 Hz, H-31), 1.21 (3H, d, J ˆ
6:9 Hz, H-27), 3.45 (2H, s, H-2'), 4.76 (1H, br. s, H-3).
13C-NMR (100.61 MHz, CDCl3): see Table 1. EI-MS
Amorphous white solid. 1H-NMR (400.13 MHz,
CDCl3): d 0.76 (3H, s, H-18), 0.84 (3H, s, H-29), 0.89
(3H, s, H-28), 0.98 (3H, d, J ˆ 8:2 Hz, H-21), 0.99
(3H, s, H-19), 0.99 (3H, s, H-30), 1.10 (3H, d, J ˆ 6:7
Hz, H-31), 1.24 (3H, d, J = 7.1 Hz, H-27), 2.65 (1H,
m, H-17), 4.34 (1H, t, J ˆ 8:1 Hz, H-12), 4.74 (1H, br.
s, H-3). 13C-NMR (100.61 MHz, CDCl3): see Table 1.
m/z (rel. int.): 528 [M
CO2]+ (9), 513 [M
CO2
Me]+ (13), 453 (80), 435 (21), 309 (52), 153 (71), 43
(100).
3.16. Compound 6, ergosterol peroxide, 5a,8a-
epidioxyergosta-6,22-dien-3b-ol
3.20. Compound 10, methyl (E )-4-methoxycinnamate
Obtained only as a mixture with 11. 1H-NMR
(400.13 MHz, CDCl3): d 3.80 (3H, s, OMe), 3.85 (3H,
s, COOMe), 6.32 (1H, d, J ˆ 16:2 Hz, H-8), 6.92 (2H,
dt, J ˆ 9:1, 1.6 Hz, H-3, H-5), 7.49 (2H, dt, J ˆ 9:1,
1.6 Hz, H-2, H-6), 7.66 (1H, d, J ˆ 16:2 Hz, H-7). 13C-
NMR (100.61 MHz, CDCl3): d 51.61 (q, COOMe),
55.41 (q, OMe), 114.36 (d, C-3, C-5), 115.31 (d, C-8),
127.16 (s, C-1), 129.75 (d, C-2, C-6), 144.56 (d, C-7),
153.74 (s, C-4), 161.43 (s, C-9). EI-MS m/z (rel. int.):
1
Waxy brown solid, ‰a20 20:0Š (CHCl3, c 0.2). H-
NMR (400.13 MHz, CDCl3): d 0.82 (3H, d, J ˆ 12:6
Hz, H-26), 0.82 (3H, s, H-18), 0.83 (3H, d, J ˆ 17:1
Hz, H-27), 0.88 (3H, s, H-19), 0.91 (3H, d, J ˆ 6:6 Hz,
H-21), 1.00 (3H, d, J ˆ 6:6 Hz, H-28), 3.97 (1H, m, H-
3), 5.14 (1H, dd, J ˆ 15:3, 8.2 Hz, H-23), 5.22 (1H, dd,
J ˆ 15:2, 7.6 Hz, H-22), 6.24 (1H, d, J ˆ 8:7 Hz, H-7),
6.51 (1H, d, J ˆ 8:7 Hz, H-6). 13C-NMR (100.61
MHz, CDCl3): see Table 1. EI-MS m/z (rel. int.): 396
[M O2]+ (30), 363 (15), 81 (41), 69 (100).
192 [M]+ (72), 161 [M
OMe]+ (100), 133 [M
COOMe]+ (38), 118 (13), 89 (18).
3.17. Compound 7, quercinic acid B, 24-methylene-3,23-
dioxolanost-8-en-26-oic acid
3.21. Preparation of methyl (E )-4-methoxycinnamate
Waxy yellow solid, ‰a20+37.5] (CHCl3, c 1.0). 1H-
NMR (400.13 MHz, CDCl3): d 0.77 (3H, s, H-18),
0.90 (3H, d, J ˆ 8:1 Hz, H-21), 0.91 (3H, s, H-30),
1.07 (3H, s, H-28), 1.10 (3H, s, H-29), 1.13 (3H, s, H-
19), 1.36 (3H, d, J ˆ 7:2 Hz, H-27), 5.95 (1H, s, H-
31a), 6.20 (1H, s, H-31b). 13C-NMR (100.61 MHz,
CDCl3): see Table 1. EI-MS m/z (rel. int.): 482 [M]+
Commercially available (E )-4-methoxycinnamic acid
was recrystallized from chloroform and 2 mg were
treated with diazomethane solution until a yellow col-
our remained, indicating an excess of diazomethane.
Finally, the solvent and the diazomethane were
removed in a stream of nitrogen yielding pure methyl
(E )-4-methoxycinnamate.

762
J. Rosecke, W.A. Konig / Phytochemistry 54 (2000) 757±762
È È
3.22. Compound 11, methyl (Z )-4-methoxycinnamate
References
Obtained only as a mixture with 10. 1H-NMR
(400.13 MHz, CDCl3): d 3.80 (3H, s, OMe), 3.85 (3H,
s, COOMe), 5.85 (1H, d, J ˆ 12:7 Hz, H-8), 6.86 (1H,
d, J ˆ 12:7 Hz, H-7), 6.90 (2H, d, J ˆ 8:7 Hz, H-3, H-
5), 7.70 (2H, d, J ˆ 8:7 Hz, H-2, H-6). EI-MS m/z (rel.
Adam, H.K., Bryce, T.A., Campbell, I.M., McCorkindale, N.J.,
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the
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Carboxyacetylquercinic acid Ð a novel triterpene conjugate from
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int.): 192 [M]+ (69), 161 [M
OMe]+ (100), 133 [M
COOMe]+ (44), 118 (21), 89 (32).
Dembitsky, V.M., Rezanka, T., Shubina, E.E., 1993. Unusual
hydroxy fatty acids from some higher fungi. Phytochemistry 34
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3.23. Compound 12, 3a-carboxyacetoxy-24-methylene-
23-oxolanost-8-en-26-oic acid
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Obtained only as a mixture with 5. 1H-NMR
(400.13 MHz, CDCl3): d 0.74 (3H, s, H-18), 0.89 (3H,
s, H-29), 0.90 (3H, d, J ˆ 7:6 Hz, H-21), 0.93 (3H, s,
H-30), 0.94 (3H, s, H-28), 1.00 (3H, s, H-19), 1.36
(3H, d, J ˆ 7:2 Hz, H-27), 3.46 (2H, s, H-2'), 4.76
(1H, br. s, H-3), 5.94 (1H, s, H-31a), 6.20 (1H, s, H-
31b). 13C-NMR (100.61 MHz, CDCl3): see Table 1.
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Keller, A.C., Maillard, M.P., Hostettmann, K., 1996. Antimicrobial
steroids from the fungus Fomitopsis pinicola. Phytochemistry 41
(4), 1041±1046.
Turner, W.B., Aldridge, D.C., 1983. Fungal metabolites II.
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Acknowledgements
van Heerden, P.S., Bezuidenhoudt, B.C.B., Ferreira, D., 1996.
Tetramethylethylene diamine/trimethylsilyl chloride mediated ad-
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The ®nancial support of the Fonds der Chemischen
Industrie is gratefully acknowledged. We also thank
Helmut and Gisela Rosecke for collecting the fruiting
bodies of Daedalea quercina.
Wat, C.-K., Towers, G.H.N., 1977. Production of methylated pheno-
lic acids by species of Lentinus (Basidiomycetes). Phytochemistry
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