Volatile constituents in the liverwort Tritomaria polita

Article abstract of DOI:10.1016/S0031-9422(03)00298-X

The essential oil of the liverwort Tritomaria polita, collected in Oetztal/Tyrol (Austria), was investigated by chromatographic and spectroscopic methods. Several new compounds were isolated by preparative gas chromatography (GC) and their structures inve

Full text of DOI:10.1016/S0031-9422(03)00298-X

Phytochemistry 64 (2003) 637–644
www.elsevier.com/locate/phytochem
Volatile constituents in the liverwort Tritomaria polita
Adewale Martins Adioa, Claudia Paulb, Wilfried A. Koniga,*, Hermann Muhlec
aInstitut fur Organische Chemie, Universitat Hamburg, Martin-Luther-King-Platz6, D-20146 Hamburg, Germany
¨
¨
bThetis-IBN GmbH, Notkestrasse 85, D-22607 Hamburg, Germany
¨
Abteilung Systemqtische Botanik und Okologie, Universitat Ulm, D-89081 Ulm, Germany
¨
c
Received 7 January 2003; received in revised form 24 April 2003
Dedicated to the memory of Professor Jeffrey B. Harborne
Abstract
The essential oil of the liverwort Tritomaria polita, collected in Otztal/Tyrol (Austria), was investigated by chromatographic and
spectroscopic methods. Several new compounds were isolated by preparative gas chromatography (GC) and their structures investigated
by mass spectrometry (MS) and NMRtechniques. In addition to known constituents, the sesquiterpenoids (+)-eudesma-3,11-dien-8-one,
(+)-eudesma-3,7(11)-dien-8-one, (+)-6,11-epoxy-eudesmane, (-)-6,7-seco-eudesm-7(11)-en-6-al, (+)-6b-hydroxy-eudesm-11-ene, (-)-6a-
hydroxy-eudesm-11-ene, (+)-6,11-epoxy-isodaucane could be identified as natural compounds for the first time.
# 2003 Elsevier Ltd. All rights reserved.
Keywords: Tritomaria polita; Liverwort; Sesquiterpene hydrocarbons; Oxygenated sesquiterpenes; seco-Eudesmane
1. Introduction
known constituents were identified by mass spectro-
metry in conjunction with their gas chromatographic
retention indices by comparison with a spectral library
established under identical experimental conditions
(Joulain and Konig, 1998; MassFinder software and data
bank, Hochmuth et al., 2002). Sesquiterpenoids with
unknown mass spectra were selected for isolation by pre-
parative GC and subjected to NMRspectroscopic inves-
tigation. The known sesquiterpene hydrocarbons were
b-elemene, aromadendrene, allo-aromadendrene (1.5%),
4,5-di-epi-aristolochene, a-amorphene, eremophilene,
(+)-a-selinene (5%) (Williams et al., 1995), d-amorphene
(Melching et al., 1997), selina-3,7(11)-diene identified
(relative concentrations of major compounds in par-
entheses, others below 1%).
Tritomaria polita (a liverwort), previously taxo-
nomically separated and listed as Saccobacis polita
(Nees) Buch (Frahm and Frey, 1992), occurs at eleva-
tions above 1500 m in the Bavarian and Austrian Alps.
In contrast to Tritomaria quinquedentata, which has
been investigated before by several research groups
(Tori et al., 1993; Warmers and Konig, 1999) the species
Tritomaria polita, to our knowledge, was not. We now
report that in the essential oil of T. polita monoterpenes
and sesquiterpene hydrocarbons are present only as
minor constituents, but major peaks of oxygenated ses-
quiterpenes with an eudesmane backbone can be detected
by GC–MS. This may be an indication of the relationship
to T. quinquedentata, where the oxygenated eudesmanes
7-epi-isojunenol and 7-epi-junenol together with a-seli-
nene are major constituents (Warmers and Konig, 1999).
The optical rotation of the new compounds was
determined by polarimetric measurements. Enantio-
selective GC using cyclodextrin phases in conjunction
with chemical conversions was employed to assign the
absolute configuration.
2. Results and discussion
(+)-Eudesma-3,11-dien-8-one (1), was isolated for
the first time as a natural product. The 1H NMR
(CDCl3) and the positive optical rotation of 1 were
consistent with the reported oxidation product of ent-
8b-hydroxy-eudesma-3,11-diene (Kondo et al., 1990).
Its relative configuration was derived from the NOESY
spectrum (Fig. 1). Treatment of (+)-1 with activated
basic alumina for two days at room temperature afforded
The essential oil of T. polita was prepared by hydro-
distillation and analysed by GC and GC–MS. All
* Corresponding author. Tel.: +49-40-42838-2824; fax: +49-40-
42838-2893.
E-mail address: wkoenig@chemie.uni-hamburg.de (W.A. Konig).
0031-9422/03/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0031-9422(03)00298-X

638
A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
form solution. Compound (+)-1 was also sensitive to
heat. Isolation by prep. GC always resulted in a mixture
with (+)-2 (8%). The instability of (+)-1 could be due
to formation of a resonance stabilized allylic carbonium
ion by loss of hydrogen from the a-position to the car-
bonyl group.
Compound 2, an a,b-unsaturated ketone with positive
optical rotation, was also present in the essential oil of
T. polita. It showed the identical mass spectrum and
same retention times on both achiral and chiral capil-
lary GC columns with the product formed from 1. The
enantiomer of 2 has been reported as a constituent of
the liverwort Bazzania fauriana (Toyota and Asakawa,
1988) and the racemic compound, resulting from the
oxymercuration–demercuration reaction of germacrone
was reported by Tsankova and Ognyanov (1980). The
Fig. 1. NOE correlations observed for (+)-1.
(+)-eudesma-3,7(11)-diene-8-one
(2)
and
(+)-
3,4,4aR,7,8,8aR - hexahydro - 5,8a - dimethylnaphthalen-
2(1H)-one (3), (Schemes 1 and 2). The latter is assumed
to be a degradation product since it exhibited a mole-
cular ion at m/z 178 corresponding to C12H18O. A
1
comparison of the H NMRspectral data (C 6D6) of
(+)-3 with 2 showed the disappearance of the signals at
d1.85 (s, 3H, CH3-12), 4.77 (s, 1H, H-13a) and 4.97 (s,
1H, H-13b) and thus revealed the loss of an isopropenyl
group. This was confirmed by the 1H NMRspectrum in
CDCl3. It also showed the disappearance of the signals
at d 1.78 (s, 3H, CH3-12), 4.77 (s, 1H) and 4.96 (s, 1H)
1H NMRdata (CDCl ) of (+)-2 are in agreement with
3
the reported data. The 13C NMRspectral data (C D6)
6
of (+)-1 and (+)-2 are reported for the first time
(Table 1). The absolute configuration of (+)-1 and (+)-
2 were deduced by chemical correlation with (+)-a-
selinene (5) as shown in Schemes 2 and 3. Reduction of
(+)-1 with lithium aluminium hydride gave a sesqui-
terpene alcohol whose GC–MS data were identical to
the isolated ent-8a-hydroxy-eudesma-3,11-diene (4). All
spectral data and the direction of optical rotation were
in agreement with a compound described by Kondo et
al. (1990) as a constituent of the liverwort Bazzania
spiralis. The absolute configuration of (+)-4 from T.
polita was elaborated by comparison of its hydrogena-
tion products with that of the hydrogenation products
of authentic (+)-a-selinene (5) by capillary GC with
different cyclodextrin derived chiral stationary phases
corresponding to the isopropenyl residue. The GC–MS
1
and the H NMRspectrum (CDCl ) of (+)-3 was con-
3
sistent with the enantiomeric reaction product obtained
by Vial et al. (1989) by degradation of podocarpic acid.
In addition, the mass spectrometric investigation of a
sample of (+)-3 kept in chloroform solution at room
temperature for 24 h showed a compound with a mole-
cular ion at m/z 288. This suggests further degradation
of (+)-3 in the presence of traces of HCl in the chloro-
Scheme 1.
Scheme 2.

A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
639
Table 1
13C NMRspectral data of compounds (+)-1 and (+)-2 (125.7 MHz,
C6D6); d (ppm)a
Table 2
13C NMRspectra data for compounds (+)- 6, (À)-8, (+)-10, (À)-11
and (+)12 (125.7 MHz, C6D6), d (ppm)a
Carbon
(+)-1
(+)-2
Carbon
(À)-8
(+)-6
(+)-10
(À)-11
(+)-12
1
2
37.3(t)
23.0(t)
122.1(d)
133.5(s)
45.6(d)
30.5(t)
59.1(d)
207.3(s)
55.7(t)
36.9(s)
144.3(s)
20.9(q)
113.2(t)
16.9(q)
21.3(q)
36.8(t)
22.8(t)
122.5(d)
133.9(s)
43.8(d)
29.4(t)
145.1(s)
200.6(s)
56.4(t)
33.4(s)
131.2(s)
22.7(q)
23.8(q)
17.2(q)
21.0(q)
1
2
37.3(t)
21.7(t)
34.5(t)
27.6(d)
64.2(d)
205.3(d)
125.2(d)
22.1(t)
43.9(t)
36.4(s)
131.2(s)
25.9(q)
17.6(q)
20.0(q)
20.9(q)
41.8(t)
22.2(t)
36.6(t)
33.8(d)
55.5(d)
78.0(d)
40.1(d)
18.9(t)
39.9(t)
33.1(s)
83.5(s)
31.9(q)
25.1(q)
18.2(q)
20.7(q)
44.0(t)
22.2(t)
38.0(t)
33.4(d)
53.7(d)
70.5(d)
46.3(d)
22.1(t)
38.8(t)
34.0(s)
146.5(s)
23.9(q)
113.2(t)
21.8(q)
22.8(q)
42.7(t)
21.4(t)
37.6(t)
30.0(d)
55.3(d)
66.6(d)
51.2(d)
22.5(t)
44.2(t)
34.0(s)
147.8(s)
23.0(q)
111.9(t)
20.6(q)
20.0(q)
42.6(s)
41.9(t)
26.4(t)
54.2(d)
62.8(d)
82.7(d)
41.7(d)
38.6(t)
23.5(t)
41.2(t)
81.0(s)
32.2(q)
23.8(q)
20.3(q)
28.1(q)
3
3
4
4
5
5
6
6
7
7
8
8
9
9
10
11
12
13
14
15
10
11
12
13
14
15
a
a
All assignments were confirmed by HMBC and HMQC.
All assignments were confirmed by HMBC and HMQC.
(Scheme 3). The co-existence of (+)-1, (+)-2 and (+)-4
suggested a structural and stereochemical relationship.
The new tricyclic compound (+)-6 exhibited a mole-
cular ion at m/z 222 and an elemental composition of
C15H26O. The 13C NMRspectrum revealed the presence
proton at H-6. Information from 2D 1H–1H-COSY,
HMBC, HMQC spectra in addition to the 13C NMR
and DEPT (Table 2) suggested structure 6 with an
eudesmane backbone. Its relative configuration resulted
from spatial interactions of H-14 with the protons of
H-6, H-7 and H-15 as observed from the NOESY spec-
trum (Fig. 2). Treatment of 6 with acidic ion exchange
resin (Amberlyst) for 2 h at room temperature gave an
unknown hydrocarbon (m/z 204, base peak 108) (4%)
and (+)-7 (7%), (-)-d-selinene (9) (24%) (proved by
comparison with an authentic reference compond by
enantioselective GC) and compounds (+)-10 (33%),
(À)-11 (19%) and (À)-8 (2.1%), which were also present
in the essential oil of T. polita (Scheme 4). Selcia-5,
7(11)-diene(7) with unspecified configuration was pre-
viously mentioned as a constituent of Olibanum essen-
tial oil by Maupetit (1985), but no spectrocsopic data
were given. Scheme 4 shows a possible biogenetic rela-
tionship between compounds (+)-6, (À)-8, (+)-10 and
(À)-11. Thus (À)-8 may be a direct precursor of (À)-11
as similar treatment of (À)-8 with acidic ion exchange
resin for 2 hours gave predominantly (À)-11 (75%) and
(À)-d-selinene (9) (13%). Interestingly, no trace of (+)-
10 was observed among the reaction products of (À)-8.
This suggests that (+)-10 is formed directly by acid
hydrolysis of (+)-6 via a cationic intermediate. The
formation of the isomeric alcohols (+)-10 and (À)-11
confirmed the C-O-linkage at C-6 in (+)-6. Scheme 4
also suggests that (À)-8 may be formed directly from
1
of 15 carbon resonances. H NMRand HMQC spec-
troscopic analysis indicated a total of 26 protons
1
directly attached to the carbon skeleton. The H NMR
spectrum showed signals of a doublet and three singlets
for methyl groups at d 1.09 (d, 3H, J=5.9 Hz), 0.58 (s,
3H), 1.30 (s, 3H) and 1.31 (s, 3H), respectively. The
strongly downfield shifted signal at d 4.28 (dd, 1H,
J=7.4, 8.7 Hz) was assigned to an oxygenated methine
Scheme 3.
Fig. 2. NOE correlations observed for (+)-6.

640
A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
Table 3
Important 1H-13
eudesm-7(11)-ene-6-al (8)
C
long-range HMBC couplings of (À)-6,7-seco-
Carbon
Protons
C-4
C-5
C-6
C-7
C-11
H-2a, H-2b, H-3a, H-5, H-15.
H-1a, H-3a, H-4, H-6, H-9a, H-9b, H-14, H-15.
H-4/5, H-15.
H-8a, H-8b, H-9a, H-9b, H-12, H-13.
H-8a, H-8b, H-12, H-13.
HMBC and HMQC, are listed in Table 2. The 2D
1H–1H-COSY, HMBC (Table 3) and HMQC spectra
confirmed the structure of (À)-8. In the NOESY spec-
trum of (À)-8, spatial interactions between the aldehyde
proton and the methyl protons H-14 and H-15 were
observed. This indicates that H-14 and H-15 are on the
same side of the molecule. Treatment of (À)-8 with acidic
ion exchange resin afforded (À)-d-selinene (9) (13%) and
(À)-11 (75%) (Scheme 4). The latter, which is isomeric to
(+)-10 was also isolated. The correlation with (À)-9 sug-
gests a-orientation for the methyl group at C-10. Hence,
the methyl group at C-4 should also be in the a-orienta-
tion in agreement with the NOESY spectrum.
Scheme 4.
(+)-6 via a concerted mechanism involving the opening of
the oxetane ring in the absence of protons ([2+2] cyclor-
eversion). From the co-occurrence of (+)-6 and (À)-8 it
cannot be excluded that the latter is an artefact formed
during the isolation procedure. In analogy to reactions
described by Coxon et al. (1972) it is also possible that one
of the isomeric alcohols (+)-10 or (À)-11 is a precursor of
(À)-8. The presence of (À)-d-selinene (9) in the acid hydro-
lysis products confirmed the a-orientation of the methyl
group at C-10.
The new sesquiterpene hydrocarbon (+)-7, an acid
hydrolysis product of (+)-6, exhibited a molecular ion
at m/z 204 and an elemental composition of C15H24.
The 1H NMRspectrum (C 6D6) showed signals of a
doublet and three singlets for methyl groups at d 1.09 (d,
3H, J=6.6 Hz), 1.07 (br.s, 3H), 1.70 (br.s, 3H) and 1.80
(br.s, 3H), respectively. The strongly downfield shifted
signal at d 6.32 (br.s, 1H) was assigned to the methine
proton H-6. (+)-7 has a mass spectrum almost identical
to (À)-d-selinene (9). The structure of (+)-7 was derived
from the 1H NMRspectral data by comparison with the
known data of (-)-(9). Its absolute configuration was
deduced from its relationship to (+)-6.
(À)-6,7-seco-Eudesma-7(11)-en-6-al (8), a trace com-
ponent isolated as an oil by preparative GC, has a new
seco-eudesmane skeleton. The mass spectrum exhibited
a molecular ion at m/z 222 and an elemental composi-
tion of C15H26O. The 1H NMRspectrum (C 6D6)
showed signals of a doublet and three singlets for
methyl groups at d 0.68 (3H, d, J=6.0 Hz), 0.81 (3H, s),
1.60 (3H, s) and 1.68 (3H, s), respectively. The singlets
of methyl groups at d 1.60 and 1.68 indicate that they
are attached to a double bond. The downfield signal at d
9.46 (1H, d, H-6, J=4.4 Hz) was assigned to the alde-
hyde proton. The 13C NMRspectral data, confirmed by
(+)-6b-Hydroxy-eudesm-11-ene (10), a sesquiterpene
alcohol with eudesmane skeleton exhibited a molecular
ion at m/z 222 and an elemental composition C15H26O.
The 1H NMR(C 6D6) spectrum showed signals of a
doublet and two singlets for methyl groups at d 1.31 (d,
3H, J=6.3 Hz), 0.82 (s, 3H) and 1.63 (s, 3H), respec-
tively. The olefinic carbon signals at 113.2 (t) and 146.5
(s), suggested an exomethylene double bond, which was
1
confirmed by two signals in the H NMRspectrum at d
4.81 (s, 1H) and 4.88 (s, 1H). The signal at d 70.5 was
assigned to a secondary carbon with the hydroxyl
group. All this information from 13C NMRand DEPT
spectroscopic (Table 2) as well as from 2D 1H–1H-
COSY, HMQC, and HMBC analysis were consistent
with the assigned structure (+)-10. Its relative con-
figuration was derived from the NOESY spectrum
(Fig. 3). Rigorous catalytic hydrogenation of (+)-10
resulted in a simultaneous hydrogenation and dehy-
dration to give two fully saturated diastereoisomeric
eudesmanes (molecular mass 208), which were com-
pared with the fully hydrogenated products of authentic
(+)-a-selinene (5). Enantioselective GC showed that
one of the hydrogenation products of (+)-10 gave
identical retention times with a hydrogenation product
Fig. 3. NOE correlations observed for (+)-10.

A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
641
Considering the major sesquiterpenes constituents in
the investigated Tritomaria species, the inclusion of T.
polita into the Tritomaria family appears chemotax-
onomically justified as eudesmane type structures are
predominant in both T. polita and T. quinquedentata.
(+)-a-Selinene as a major constituent in both species
may serve as a precursor for the oxygenated secondary
metabolites formed, however, the site of oxygenation is
different. No sesquiterpene lactones are present in T.
polita but quite abundant in T. quinquedentata.
Fig. 4. NOE correlations observed for (À)-11.
3. Experimental
Fig. 5. NOE correlations observed for (+)-12.
3.1. General experimental procedures
3.1.1. Gas chromatography
of (+)-5. This is possible if an intermediate with a C-6-
C-7 double bond is formed, which generates two dia-
stereoismeric products upon hydrogenation. Thus the
absolute configuration at the stereogenic centers C-5
and C-10 could be assigned and was in agreement with
the NOESY spectrum. The b-orientation of the
hydroxyl group at C-6 and of the isopropenyl group at
C-7 was derived from the spatial interactions of H-6 and
H-7 with a-H-14. Interestingly, compound (À)-11, a
sesquiterpene alcohol with a molecular ion at m/z 222
and an elemental composition of C15H26O showed
Orion Micromat 412 double column instrument with
25 m fused silica capillaries with polysiloxane CPSil-5
and polysiloxane CPSil-19 (Chrompack); Carlo Erba
Fractovap 2150 or 4160 gas chromatographs with 25 m
fused silica capillaries with octakis(2,6-di-O-methyl-3-O-
pentyl)-g-cyclodextrin, heptakis(2,6-di-O-methyl-3-O-pen-
tyl)-b-cyclodextrin or heptakis(6-O-tert-butyldimethylsi-
lyl-2,3-di-O-methyl)-b-cyclodextrin in OV 1701 (50%,
w/w), split injection; split ratio approx. 1:30; FID; car-
rier gas 0.5 bar H2; injector and detector temperatures
ꢀ
were 200 and 250 C, respectively.
identical GC- and mass spectra as well as 1H NMR, 13
C
NMR( Table 2), 2D 1H–1H-COSY, HMBC and HMQC
spectral data as (+)-10. (À)-11 also gave similar results
when subjected to catalytic hydrogenation, but in this
case the different absolute configuration at C-7 was
clearly shown by the NOESY spectrum (Fig. 4), which
indicated spatial interactions between H-5, H-6 and
H-7, thus suggesting that H-7 is in b-orientation with
respect to H-5. The structures of the two isomeric alco-
hols (+)-10 and (À)-11 are in agreement with the
correlations shown in Scheme 4.
(+)-12, a new tricyclic skeleton with isodaucane back-
bone was isolated with the hydrocarbon fraction of the
liverwort. (+)-12 exhibited a molecular ion at m/z 222 and
an elemental composition of C15H26O. The 1H NMR
showed signals of a doublet and three singlets for methyl
groups at d 1.21 (d, 3H, J=6.6 Hz), 0.85 (s, 3H), 1.17 (s,
3H) and 1.30 (s, 3H), respectively. A signal at d 3.22 (t, 1H,
J=9.8 Hz) was assigned to the oxygenated methine H-6
which coupled with an adjacent proton H-5 at d 1.99 (t,
1H, J=9.5 Hz). The 2D 1H–1H-COSY, HMQC and
HMBC spectra in addition to the 13C NMR (Table 2)
confirmed the structure. Its relative configuration resulted
from the NOESY spectrum (Fig. 5). No visible changes
were observed in the GC and GC–MS of (+)-12 after
treatment with acidic ion exchange resin for 2–8 h. This
suggests that (+)-12 is a relatively stable compound. Its
absolute configuration remains undetermined.
3.1.2. Preparative GC
Modified Varian 1400 and 2800 instruments, equip-
ped with stainless steel columns (1.85 m  4.3 mm) with
10% polydimethylsiloxane SE-30 on Chromosorb W-
HP or with 2.5% octakis(2,6-di-O-methyl-3-O-pentyl)-
g-cyclodextrin in OV-1701 (50%, w/w) on Chromosorb
G-HP or with 6% heptakis(6-O-tert-butyldimethylsilyl-
2,3-di-O-methyl)-b-cyclodextrin in SE-52 (50%, w/w)
on Chromosorb W-HP; FID; helium as carrier gas at a
flow rate of 240 ml/min.; injector and detector tem-
ꢀ
peratures were 200 and 250 C, respectively (Hardt and
Konig, 1994).
3.1.3. GC–MS
Electron impact (70 eV) and chemical ionization
(NH3) GC–MS were carried out on a Hewlett Packard
HP 5890 gas chromatograph coupled with a VG Ana-
lytical 70-250S magnetic field mass spectrometer.
3.1.4. NMR spectroscopy
NMRspectra were recorded with either a Bruker
WM 400 or a Bruker WM 500 instrument in C6D6 and/
or CDCl3 using TMS as internal standard.
3.1.5. Polarimetry
Measurements were performed with a polarimeter 341
(Perkin-Elmer) at 589 nm at 20 ꢀC. Due to the very

642
A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
small amounts of isolated compounds only the direction
of optical rotation is given to avoid inaccuracies.
CDCl3): d 0.79 (3H, s), 1.25 (1H, br. s), 1.42 (1H, dd,
J=6.0, 12.9 Hz), 1.57 (1H, dt, J=6.9, 12.6 Hz), 1.68
(3H, s), 1.72 (1H, dd, 12.9, 25.9 Hz), 1.78 (3H, s), 1.99–
2.11 (2H, m), 2.16 (1H, ddd, 3.8, 6.6, 13.7 Hz), 2.22 (1H,
dd, J=13.9, 29.6 Hz), 2.49 (1H, br. d, J=13.9 Hz), 3.03
(1H, dd, J=6.3, 12.3 Hz), 4.77 (1H, s), 4.96 (1H, s), 5.42
(1H, br. s). 13C NMR(125.7 MHz, C 6D6) data see
Table 2; MS (EI, 70eV), m/z (rel. int.) 218 [M+] (45),
203 (12), 200 (12), 190 (3), 175 (9), 159 (14), 147 (22),
121 (33), 108 (100), 93 (74), 83 (30), 77 (38), 67 (26), 53
(37), 41 (74). HRMS calc. for C15H22O [M]+: m/z
218.1671, found: 218.1672.
3.1.6. Thin layer chromatography
Thin layer chromatography was effected using glass
or aluminum supported plates of silica 60 F254 (Merck).
An ethanolic solution of sulfuric acid (10%) and anis-
aldehyde was used as spray reagent. The solvent system
used was n-hexane:ethylacetate (3:1, v/v).
3.1.7. Reactions
Hydrogenation reactions were performed by bubbling
hydrogen gas through a stirred solution of ca. 1 mg of
sample in 1 ml n-hexane and 0.5 mg Pd/C at room
temperature for 1 h. The reaction mixture was filtered
and the reaction products were analyzed by GC–MS
and by GC on several capillary columns with cyclo-
dextrin derivatives.
Dehydration was carried out with ca. 1 mg of sample
in 0.5 ml of pyridine and 1 drop of phosphoryl chloride
under ice cooling. After 1 h of stirring at room temp. the
reaction was quenched by adding a few drops of water
and the mixture was extracted three times with n-hexane.
The organic phase was washed several times with water
and dried over Na2SO4.
3.5. (+)-Eudesma-3,7(11)-dien-8-one (2)
Colourless oil; RICPSIL 5=1745 ; Rf=0.82; sense of
optical rotation (chloroform): (+); 1H NMR(500
MHz, C6D6): d 0.76 (s, 3H, CH3-14), 1.12–1.17 (m, 2H,
H-1), 1.48 (s, 3H, CH3-12), 1.56 (br.s, 3H, CH3-15),
1.74–1.83 (m, 1H, H-2a), 1.87–1.96 (m, 3H, H-6a, H-2b,
H-5), 2.01 (d, 1H, H-9a, J=16.3 Hz), 2.20 (s, 3H, CH3-
13), 2.30 (d, 1H, H-9b, J =16.2 Hz), 2.57 (dd, 1H, H-6b,
J=3.8, 14.5 Hz), 5.29–5.34 (m, 1H, H-3). 1H NMR(500
MHz, CDCI3) d 0.85 (s, 3H), 1.39–1.45 (m, 2H), 1.70
(br.s, 3H), 1.84 (s, 3H), 1.94–2.07 (m, 3H), 2.09 (s, 3H),
2.12–2.29 (m, 3H), 2.80 (dd, 1H, J=3.8, 14.2 Hz), 5.42
3.2. Plant material and essential oil
(br.s, 1H). 13C NMR(125.7 MHz, C D6) see Table 2.
6
MS (EI, 70eV), m/z (rel. int.): 218 [M+] (100), 203 (22),
189 (4), 175 (17), 161 (19), 147 (30), 135 (17), 121 (55),
108 (47), 98 (20), 93 (46), 91 (44), 83 (40), 77 (33), 67
(32), 53 (35), 41 (73). HRMS calcd for C15H22O [M]+:
m/z 218.1671, found: 218.1667.
Tritomaria polita was collected on wet rocks in July
2001 close to Obergurgl in the Otztal, Tyrol/Austria at
an elevation of approx. 2300 m. A voucher specimen is
kept at the Abteilung fur Systematische Botanik und
Okologie, University of Ulm. The essential oil was pre-
pared by hydrodistillation (2 h) of aqueous homo-
genates of partially air-dried plants using n-hexane as
collection solvent. Because of the greatly differing
weight, the plant material was not weighed.
3.6. (+)-3,4,4aR,7,8,8aR-Hexahydro-5,8a-dimethyl-
naphthalen-2(1H)-one (3)
Colourless oil; RICPSIL 5=1461; sense of optical rota-
1
tion (chloroform): (+); H NMR(500 MHz, C D6), d
6
3.3. Isolation of single constituents of the essential oils
0.63 (s, 3H), 1.04 (dd, 1H, J=6.0, 12.6 Hz), 1.15–1.22
(m, 3H), 1.46 (br.s, 3H), 1.76 (d, 1H, J=13.5 Hz), 1.77–
1.85 (m, 4H), 2.07 (dd, 1H, J=2.0, 13.9 Hz), 2.29 (dt,
All isolations were carried out using SE-30- and/or
SE-52-columns combined with at least one cyclodextrin
phase column.
1
1H, J=2.5, 14.8 Hz), 5.25 (br.s, 1H). H NMR(500
MHz, CDCI3), d 0.78 (s, 3H), 1.41 (dd, 1H, J=5.67,
12.6 Hz), 1.67 (s, 3H), 2.02–2.10 (m, 3H), 2.13–2.22 (m,
3H), 2.28–2.39 (m, 3H), 2.44–2.50 (m, 1H), 5.41 (br.s,
1H). MS (EI, 70eV) m/z (rel. int.): 178 [M+] (45), 163
(100), 150 (47), 145 (10), 135 (18), 121 (40), 107 (41), 105
(37), 95 (40), 93 (61), 91 (42), 81 (82), 79 (63), 77 (39), 67
(37), 53 (38), 41 (63).
3.4. (+)-Eudesma-3,11-dien-8-one (1)
Colourless oil; RICPSIL 5=1668; Rf=0.81; sense of
optical rotation (chloroform): (+); 1H NMR(500
MHz, C6D6): d 0.67 (s, 3H, CH3-14), 1.06 (dd, 1H, H-
1a, J=6.0, 12.6 Hz), 1.23 (dt, 1H, H-1b, J=6.6,12.6
Hz), 1.50 (s, 3H, CH3-15), 1.59 (dd, 1H, H-6a, J=12.6,
25.5 Hz), 1.78–1.83 (m, 2H, H-9a, H-2a), 1.85 (s, 3H,
CH3-12), 1.87–1.93 (m, 2H, H-6b, H-2b), 2.02 (br.d, 1H,
H-5, J=12.9 Hz), 2.08 (d, 1H, H-9b, J=13.2 Hz), 2.68
(dd, 1H, H-7, J=6.3, 12.6 Hz), 4.77 (s, 1H, H-13a), 4.97
3.7. (+)-8ꢀ-Hydroxy-eudesma-3,11-diene (4)
Colourless oil; RICPSIL 5=1669; Rf=0.77; sense of
1
optical rotation (benzene): (+); H NMR(500 MHz,
C6D6): d=1.09 (dd, 1H, H-9a, J=3.4, 13.9 Hz), 1.20–
1.28 (m, 1H, H-1a), 1.31 (s, 3H, CH3-14), 1.38 (dd, 1H,
1
(s, 1H, H-13b), 5.26 (s, 1H, H-3). H NMR(500 MHz,

A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
643
H-1b, J=6.6, 12.6 Hz), 1.55 (s, 3H, CH3-12), 1.57–1.59
(m, 1H, H-6a), 1.62–1.70 (m, 1H, H-6b), 1.64 (s, 3H,
CH3-15), 1.81–1.88 (br.d, 2H, H-5, H-7, J=11.4 Hz),
1.92 (dd, 1H, H-9b, J=2.2, 13.9 Hz), 1.95–1.99 (m,1H,
H-2a), 2.09–2.16 (m, 1H, H-2b), 3.90 (br.d, 1H, H-8,
J=2.5 Hz), 4.79 (s, 1H, H-13a), 4.85 (s, 1H, H-13b),
5.35 (br.s, 1H, H-3). 1H NMR(500 MHz, CDCl 3) d 1.04
(s, 3H), 1.23-1.28 (m, 1H), 1.32 (dd, 2H, J=4.1, 13.6
Hz), 1.41 (dd, 1H, J=6.3, 12.3 Hz), 1.61 (m, 1H), 1.66
(3H, br.s), 1.81 (s, 3H), 1.90 (dd, 1H, J=2.5, 14.5 Hz),
1.96–2.00 (m, 2H), 2.06–2.12 (m, 1H), 2.12–2.17 (m,
1H), 4.05 (br.d, 1H, J=2.52 Hz), 4.87 (s, 1H), 5.02 (s,
H-1a), 1.12–1.17 (m, 1H, H-1b), 1.20–1.32 (m, 4H, H-2,
H-9), 1.42–1.47 (m, 1H, H-3b), 1.60 (s, 3H, CH3-13),
1.61–1.65 (m, 2H, H-4, H-5), 1.68 (s, 3H, CH3-12), 1.85–
1.94 (m, 1H, H-8a), 2.06–2.13 (m, 1H, H-8b), 5.16 (dt,
1H, H-7, J=1.6, 7.3 Hz), 9.46 (d, 1H, H-6, J=4.4 Hz).
13C NMR(125.7 MHz, C D6): see Table 2. MS (EI,
6
70eV), m/z (rel. int.): 222 [M+] (4), 207(4), 180 (3), 161
(2), 147 (2), 137 (38), 123 (7), 109 (35), 95 (21), 81 (22),
69 (53), 55 (39), 43 (23), 41 (100). HRMS calc. for
C15H26O [M]+: m/z 222.1984, found: 222.1961.
3.11. (+)-6ꢁ-Hydroxy-eudesm-11-ene (10)
1H), 5.32 (br.s, 1H). 13C NMR(125.7MHz, C D6): d
6
18.1 (q, C-14), 21.4 (q, C-15), 22.6 (q, C-12), 23.1 (t, C-
2), 23.3 (t, C-6), 32.3 (s, C-10), 38.7 (t, C-1), 45.9 (t, C-
9), 47.6 (d, C-5), 51.3 (d, C-7), 67.1 (d, C-8), 111.9 (t, C-
13), 121.5 (d, C-3), 134.7 (s, C-4), 147.2 (s, C-11). MS
(EI, 70eV) m/z (rel. int.): 220 [M+] (33), 205 (12), 202
(31), 187 (41), 177 (17), 161 (44), 147 (34), 131 (22), 123
(23), 121 (54), 119 (49), 108 (47), 107 (78), 105 (67), 95
(31), 93 (84), 9 1(77), 81 (54), 79 (51), 77 (49), 67 (35), 55
(54), 41 (100).
Colourless oil; RICPSIL 5=1643; Rf=0.91; sense of
1
optical rotation (benzene): (+); H NMR(500 MHz,
C6D6),: d 0.82 (s, 3H, CH3-14), 0.97–1.03 (m, 1H, H-3a),
1.04–1.09 (m, 2H, H-1a, H-5), 1.15–1.21 (m, 1H, H-9a),
1.29–1.34 (m, 1H, H-1b), 1.31 (d, 3H, CH3-15, J=6.3
Hz), 1.35–1.42 (m, 2H, H-9b, H-8a), 1.44–1.53 (m, 3H,
H-8b, H-2a, H-4), 1.57–1.62 (m, 1H, H-2b), 1.63 (s, 3H,
CH3-12), 1.64–1.67 (m, 1H, H-3b), 2.42–2.47 (m, 1H, H-
7), 3.61–3.66 (m, 1H, H-6), 4.81 (s, 1H, H-13a), 4.88 (s,
1H, H-13b). 13C NMR(125.7 MHz, C 6D6): see Table 2.
MS (EI, 70eV) m/z (rel. int.): 222 [M+] (3), 207 (7), 189
(6), 180 (3), 161 (2), 149 (2), 137 (47), 123 (57), 109 (39),
95 (27), 81 (48), 69 (44), 55 (56), 41 (100). HRMS calc.
for C15H26O [M]+: m/z 222.1984, found: 222.1965.
3.8. (+)-6,11-Epoxy-eudesmane (6)
Colourless oil; RICPSIL 5=1534; Rf=0.86; sense of
1
optical rotation (benzene): (+); H NMR(500 MHz,
C6D6): d 0.58 (s, 3H, CH3-14), 0.90–1.00 (m, 1H, H-3a)
1.05–1.12 (m, 1H, H-1a), 1.09 (d, 3H, CH3-15, J=5.9
Hz), 1.14–1.21 (m, 2H, H-4, H-9a), 1.22–1.28 (m, 2H,
H-1b, H-5), 1.30 (s, 3H, CH3-13), 1.31 (s, 3H,CH3-12),
1.36–1.48 (m, 4H,H-9b, H-8a, H-2), 1.49–1.53 (m, 1H,
H-8b), 1.55–1.62 (m, 1H, H-3b), 2.55–2.60 (m, 1H, H-7),
4.28 (dd, 1H, H-6, J=7.4, 8.7 Hz). 13C NMR(125.7
MHz, C6D6): see Table 2. MS (EI, 70eV), m/z (rel. int.):
222 [M+] (4), 207 (5), 189 (3), 180 (3), 164 (34), 149 (77),
137 (40),123 (50), 109 (90), 95 (35), 81 (50), 69 (46), 55
(52), 41 (100). HRMS calcd for C15H22O [M]+: m/z
222.1984, found: 222.1971.
3.12. (À)-6ꢀ-Hydroxy-eudesm-11-ene (11)
Colourless oil; RICPSIL 5=1598; sense of optical
1
rotation (benzene): (À); H NMR(500 MHz, C D6): d
6
0.57 (dd, 1H, H-5, J=2.2, 10.7 Hz), 0.88–0.93 (m, 1H,
H-3a), 0.98 (d, 3H, CH3=15, J=6.3Hz), 1.02–1.08 (m,
2H, H-1), 1.23 (s, 3H, CH3-14), 1.26–1.31 (m, 4H, H-2,
H-9), 1.46–1.49 (m, 2H, H-8), 1.59 (s, 3H, CH3-12),
1.71–1.78 (m, 2H, H-7, H-3b), 1.89–1.95 (m, 1H, H-4),
3.92 (br.s, 1H, H-6), 4.76 (s, 1H, 13a), 4.83 (s, 1H, 13b);
MS (EI, 70eV), m/z (rel. int.): 222 [M+] (6), 207 (8), 204
(16), 189 (14), 180 (5), 161 (6), 139 (35), 137 (56), 123 (100),
109 (40), 95 (32), 81 (52), 69 (41), 55 (53), 41 (79). HRMS
calc. for C15H26O [M]+: m/z 222.1984, found: 222.1974.
3.9. (+)-Eudesma-5,7(11)-diene (7)
Colourless oil; RICPSIL 5=1543; sense of optical
1
rotation (benzene): (+); H NMR(500 MHz, C D6): d
3.13. (+)-6,11-Epoxy-isodaucane (12)
6
1.07 (br.s, 3H), 1.09 (d, 3H, J=6.6 Hz), 1.70 (br.s, 3H),
1.80 (br.s, 3H), 6.32 (br.s, 1H). MS (EI, 70eV), m/z (rel.
int.): 204 [M+] (90), 189 (100), 175 (5), 161 (38), 149
(23), 148 (25), 147 (21), 133 (57), 119 (30), 107 (26), 105
(52), 91 (56), 77 (25), 65 (18), 55 (31), 41 (69).
Colourless oil; RICPSIL 5=1468 ; Rf=0.97; sense of
1
optical rotation (benzene): (+); H NMR(500 MHz,
C6D6): d 0.85 (s, 3H, CH3-15), 0.94 (dq, 1H, H-8a,
J=2.5,12.0 Hz), 1.17 (s, 3H, CH3-13), 1.15–1.23 (m, 1H,
H-10a), 1.21 (d, 3H, CH3-14, J=6.6 Hz), 1.22–1.27 (m,
1H, H-9a), 1.30 (s, 3H, CH3-12), 1.40–1.49 (m, 3H, H-2,
H-9b), 1.57–1.60 (m, 4H, H-7, H-3, H-10b), 1.66–1.72
(m, 1H, H-8b), 1.99 (t, 1H, H-5, J=9.5Hz), 2.35–2.40
(m, 1H, H-4), 3.22 (t, 1H, H-6, J=9.8 Hz); 13C NMR
(125.7 MHz, C6D6): see Table 2. MS (EI, 70eV), m/z
(rel. int.): 222 [M+] (4), 208 (16), 207 (100), 189 (16),
3.10. (À)-6,7-seco-Eudesm-7(11)-en-6-al (8)
Colourless oil, RICPSIL 5=1614; Rf=0.95; sense of
optical rotation (benzene): (À);1H NMR(500 MHz,
C6D6): d 0.55–0.65 (m, 1H, H-3a), 0.68 (d, 3H, CH3-15,
J=6.0 Hz), 0.81 (s, 3H, CH3-14), 0.93–1.03 (m, 1H,

644
A.M. Adio et al. / Phytochemistry 64 (2003) 637–644
164 (10), 151 (17), 149 (30), 135 (9), 123 (33), 109 (22),
107 (24), 93 (26), 81 (64), 79 (28), 67 (26), 55 (42), 43
(83), 41 (77). HRMS calc. for C15H26O [M]+: m/z
222.1984, found: 222.1974.
Hochmuth, D.H., Joulain, D., Konig, W.A., 2002. MassFinder 2.3
software and data bank. University of Hamburg.
Joulain, D., Konig, W.A., 1998. The Atlas of Spectral Data of Ses-
quiterpene Hydrocarbons. E.B.-Verlag, Hamburg.
Kondo, K., Toyota, M., Asakawa, Y., 1990. ent-Eudesmane-type ses-
quiterpenoids from Bazzania species. Phytochemistry 29, 2197–
2199.
Maupetit, P., 1985. New constituents in Olibanum resinol and essen-
tial oil. Perfumer & Flavorist 9, 19–37.
Acknowledgements
Melching, S., Bulow, N., Wihstutz, K., Jung, S., Konig, W.A., 1997.
Natural occurrence of both enantiomers of cadina-3,5-diene and d-
amorphene. Phytochemistry 44, 1291–1296.
We gratefully acknowledge financial support of
DAAD (scholarship for A.M. Adio) and the Fonds der
Chemischen Industrie. We also thank Dr. V. Sinnwell
for his support in recording the NMRspectra and
Mrs. A. Meiners and Mr. M. Preusse for GC–MS
measurements.
Tori, M., Nagai, T., Asakawa, Y., Huneck, S., Ogawa, K., 1993. Terpe-
noids from six Lophoziaceae liverworts. Phytochemistry 34, 181–190.
Toyota, M., Asakawa, Y., 1988. Sesquiterpenoids from the liverwort
Bazzania fauriana. Phytochemistry 27, 2155–2159.
Tsankova, E., Ognyanov, I., 1980. Transannular cyclisation of ger-
macrone and isogermacrone via oxymercuration-demercuration.
Tetrahedron 36, 669–671.
References
Vial, C., Thommen, W., Naf, F., 1989. 155.Structure-activity rela-
tionship in Ambergris-type woody odorants possessing a hydro-
naphthalene skeleton. Helvetica Chimica Acta 72, 1390–1399.
Warmers, U., Konig, W.A., 1999. (À)-7-epi-Isojunenol and 7-epi-
junenol, constituents of the liverwort Tritomaria quinquedentata.
Phytochemistry 52, 1519–1524.
Coxon, J.M., Garland, R.P., Hartshorn, M.P., 1972. The pyrolysis of
pinanes. VII. The pyrolysis of some hydroxypinane derivatives.
Australian Journal of Chemistry 25, 947–957.
Frahm, J.-P., Frey, W., 1992. Moosflora. Ulmer-Verlag, Stuttgart.
Hardt, I.H., Konig, W.A., 1994. Preparative enantiomer separation
with modified cyclodextrins as chiral stationary phases. Journal of
Chromatography A 666, 611–615.
Williams, H.J., Sattler, I., Moyna, G., Scott, A.I., Bell, A.A., Vinson,
S.B., 1995. Diversity in cyclic sequiterpene production by Gossypium
hirsutum. Phytochemistry 40, 1633–1636.