FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fragr. J. 2001; 16: 263–273

DOI: 10.1002/ffj.991

Germacradienols in the essential oils of the Myrtaceae

Charles P. Cornwell,1∗ Narsimha Reddy,2 David N. Leach3 and S. Grant Wyllie4
1 Australian Botanical Products, 39 Melverton Drive, Hallam, Victoria 3803, Australia
2 School of Science, University of Western Sydney, Nepean Kingswood, New South Wales 2747, Australia
3 Australian Tea Tree Oil Research Institute, Southern Cross University, Lismore, New South Wales 2480, Australia
4 Centre for Biostructural and Biomolecular Research, University of Western Sydney, Hawkesbury, Richmond, New South Wales

2753, Australia
Received 8 May 2000
Revised 10 January 2001
Accepted 12 January 2001

ABSTRACT: Two germacradienols, 4-ˇ-hydroxygermacra-1(10),5-diene (ˇ-germacrenol) and 6-˛-hydroxyger-

macra-1(10), 4-diene (kunzeaol) were identified in the solvent extracts of members of the family Myrtaceae. Acid
solvolysis associated with steam distillation or leaf ageing causes these germacradienols to rearrange to a mixture
of (C)-υ-cadinene, ˛-cadinol, -cadinol, ˛-muurolol, -muurolol and associated sesquiterpenoids. Copyright 
2001 John Wiley & Sons, Ltd.
KEY WORDS: cadinenes; cadinols; muurolols; sesquiterpenes; hydroxygermacradiene; germacradienol; germa-
crenol; acid solvolysis; carbocation; Myrtaceae; Corymbia; Eucalyptus; Kunzea; Leptospermum

Introduction
Germacradienols are found in the extracts of a variety
of plant species1 – 5 and are potentially the precursors
of the cadinene and skeletally related sesquiterpenoids
observed in the steam distillates of other species.1 A
similar situation has been proposed for germacrene D.6,7
In this work, two germacradienols have been isolated
and identified in the solvent extracts of members of the
family Myrtaceae. These sesquiterpenes are the direct
precursors of (C)-υ-cadinene, the cadinols and muurolols
and other related sesquiterpenes observed in the steam
distillates or (in some cases) the solvent extracts of
the mature growth. The carbocationic nature of the
common intermediate(s) was demonstrated using 18 O-
incorporation experiments analogous to the formation
of terpinen-4-ol from sabinene hydrate in Melaleuca
alternifolia.8,9 These germacrene-derivatives have not
been previously reported in the oils from the Myrtaceae.
Experimental
Materials and Samples
All solvents and reagents were AR quality. Column chro-
matography was performed using 70–230 mesh silica
*Correspondence to: C. P. Cornwell, Australian Botanical Products,
39 Melverton Drive, Hallam, Victoria 3803, Australia.
Contract/grant sponsor: University of Western Sydney Hawkesbury
Research and Development, Australia.
gel (Merck Silica Gel 60). Unless otherwise specified, all
plant samples were collected from Mt. Annan Botanic
Gardens, Mt. Annan, NSW, Australia. These samples
are designated by the Gardens’ unique five- or six-digit
Accession No. Samples collected were put into low-
density polyethylene plastic bags, immediately placed
onto dry ice and kept frozen (10 ° C) until analysed.
Samples for bulk distillations or extractions were kept
in paper bags protected from direct sunlight. Thujopsis
dolabrata (Accession No. 15405) was collected from
the Mt. Tomah Botanic Gardens, NSW. Angelica root
oil was purchased from a local distributor.
Nuclear Magnetic Resonance Spectroscopy
All spectra were obtained at 303 K in CDCl3 using
a Varian UNITYplus-300 Spectrometer (300.4 MHz 1 H
channel, 75.5 MHz 13 C channel). Standard protocols
were employed in obtaining all one- and two-dimensio-
nal spectra.
Gas Chromatography
High resolution gas chromatography carried out on a
Hewlett-Packard 5890 Series II chromatograph fitted
with a capillary column (below). The columns were ter-
minated at a Hewlett-Packard HP 5971A Mass Selective
Detector, with the ion source in EI mode, using an ion-
ization energy of 70 eV. The scan rate was 0.9 scans/s.
GC–MS data was processed using Hewlett-Packard

Copyright  2001 John Wiley & Sons, Ltd.

264 C. P. CORNWELL ET AL.
Chemstation software (G1034C, Version C.01.05, 
1989–1993). Capillary columns; SGE Cydex-B and
J&W CyclodexB: 0.25 µm ð 25 m (SGE) or 30 m ð
0.25 mm (J&W); initial temperature, 70–90 ° C; initial
time, 10–15 min; programme rate, 2 ° C/min to 125 ° C,
then at 3 ° C/min to 200 ° C or 220 ° C, hold for 10 min;
J&W DB5-HT: 0.1 µm ð 30 m ð 0.25 mm; 60 ° C for
5 min, then at 3 ° C/min to 170 ° C, then at 8 ° C/min to
240 ° C, hold for 5 min. Injector and transfer line temper-
atures: 220–260 ° C; He at 32 cm/s; 1 : 60 split. Kováts
indices on commercial ˇ-cyclodextrin columns: ()-υ-
cadinene, 1570; (C)-υ-cadinene, 1578; ˇ-germacrenol,
1705; ˛-muurolol, 1788; -muurolol, 1791; -cadinol,
1796; ˛-cadinol, 1817; kunzeaol, 1874.
Small-scale Hydrodistillation
Samples of young or mature leaves (0.05–0.4 g) were
heated at 100–110 ° C for 1 h in sealed auto sampler vials
with 100–300 µl water [or 95% H2 18 O (Novachem)]
for 18 O-runs. After cooling, CH2 Cl2 (100–400 µl) was
added and the organic layer was then analysed by
GC–MS, as described above. No correction for the
isotopic abundance of the water was applied during
18
O runs.
Small-scale Solvent Extractions
Samples of young or mature leaves (0.05–0.4 g) were
frozen with liquid N2 , partially crushed and then extrac-
ted with CH2 Cl2 or MeOH (100–400 µl) at RT in sealed
autosampler vials for 36 h, followed by the addition of
an equivalent amount of water (CH2 Cl2 extracts only).
Each sample was then analysed by GC–MS as described
above.
Hydrodistillation
Bulk hydrodistillations were carried out using a Cock-
ing–Middleton apparatus.10
Solvent Extraction/High pH Hydrodistillation
Bulk samples of leaves were extracted using CH2 Cl2
by allowing solvent to reflux through a leaf bed in
a 4 ð 20 cm glass column. K3 PO4 and K2 HPO4 were
added to prevent acid degradation of the extracts by
any plant acids that might be extracted. When finished,
the CH2 Cl2 was evaporated and the resulting mixture
of crude plant extract and residual phosphate salts was
hydrodistilled after addition of water.
Copyright  2001 John Wiley & Sons, Ltd.
Enantiomeric Composition of d-Cadinene
The enantiomeric composition of υ-cadinene was deter-
mined using a commercial J&W ˇ-cyclodextrin column
and a heptakis(2,6-di-O-methyl-3-O-pentyl)-ˇ-cyclodex-
trin (50% in OV1701, w/w) column,11,12 the ()-υ-cad-
inene in Angelica root oil11 and the mixture of (C)- and
()-υ-cadinene from Hiba oil (Thujopsis dolabrata)12
providing appropriate standards.
Isolation of b-Germacrenol
ˇ-Germacrenol was the major compound in the sol-
vent extracts/high pH hydrodistillates of Corymbia mac-
ulata (891338). NMR experiments were performed with-
out further purification. The oil yield from an ordinary
hydrodistillation of Corymbia maculata (891338) was
0.7% fresh weight (1.2% dry weight).
Isolation of Kunzeaol
Kunzeaol was isolated by column chromatography of the
solvent extracts/high pH hydrodistillates from Kunzea
sp. (933330), using a ethyl acetate/petroleum spirit gra-
dient (up to 20%). The oil yield from the solvent extrac-
tion/high pH hydrodistillation of Kunzea sp. (933330)
was 0.8% fresh weight and from an ordinary hydrodis-
tillation, 0.5% fresh weight.
Results and Discussion
The sesquiterpene region of the chromatograms from the
hydrodistillates of Corymbia maculata (891338) display
fractions rich in (C)-υ-cadinene, cadinols and muurolols
(Figure 1). In this species and in many other members of
the Myrtaceae (Table 1) these compounds are associated
with the acid-solvolysis of a compound that was identi-
fied as germacrenol1 (Figure 1) by establishing the basic
skeleton using HMBC, HMQC and DQ-COSY experi-
ments and by its characteristic mass spectrum, which
exhibits a large base peak at m/z 81 (Figure 2)13,14 and
by comparison of the 13 C- and 1 H-NMR data (Table 2)
with previous reports.1,4,15,16 This compound has not
been previously reported in the Myrtaceae, although
reports of the presence of its breakdown products are
not uncommon.17 The basic assignments of the confor-
mation and skeleton of germacrenol are detailed below.
The basic skeleton of the molecule was established
using HMBC and DQ-COSY. The coupling constant
of 15 Hz between the vinylic protons on C5 and C6
indicates that they are trans to each other. The trans-
orientation of the vinylic proton on C1 and the methyl
group on C10 (C14) was determined using a NOESY
Flavour Fragr. J. 2001; 16: 263–273
 GERMACRADIENOLS IN ESSENTIAL OILS OF MYRTACEAE 265

Abundance H OH
1100000 H OH

1000000 H OH
OH
900000 H
OH H H
800000 H H H
700000

600000

500000

400000 H

300000

200000

100000

0
Time--> 31.00 32.00 33.00 34.00 35.00 36.00 37.00 38.00 39.00 40.00
Abundance

1.2e+07
H OH
1.1e+07

1e+07
9000000
H
8000000
OH
7000000
OH H
6000000 H

5000000

4000000

3000000

2000000

1000000

0
Time--> 31.00 32.00 33.00 34.00 35.00 36.00 37.00 38.00 39.00 40.00

Figure 1. Sesquiterpene region of the small scale hydrodistillate (upper trace) and solvent extract (lower trace) of
Corymbia maculata (891338)

experiment; a clear NOESY crosspeak was observed
between the protons on C9 and the vinylic proton on
C1. NOE-difference spectroscopy also confirmed this,
and showed that irradiation of the methyl signal (of C14)
generated only weak NOE signals between the protons
on C1 and C14 and between the protons on C14 and C9.
While this evidence does not preclude a cis-orientation
of the C1–C10 double bond, the interpretation of the
results does favour the trans-orientation.
One of the acid-solvolysis byproducts of germacrenol
is (C)-υcadinene, as determined using enantioselec-
tive gas-chromatography.11,12 This establishes the stere-
ochemistry at C7 of germacrenol. NOE-difference spec-
troscopy did not provide conclusive evidence about the
stereochemistry at C4. The presence of cubebol (and
only traces of epicubebol) in the solvent extracts of C.
maculata (Figure 1), present as a possible byproduct of
the biosynthesis of germacrenol, lends support to assign-
ing ˇ- or S-stereochemistry about C4, as does compari-
son with previous NMR data1,4,15,16 (Table 2). However
in the absence of conclusive evidence this assignment
must remain tentative.
The solvent extracted oil from an unidentified Kun-
zea species (933330) was found to contain one major
product whose peak on a ˇ-cyclodextrin column dis-
played a large amount of ‘fronting’, caused by thermal
degradation of the compound on the column. Small-scale
hydrodistillation of the leaves yielded a cadinol-rich frac-
tion, not unlike that produced by germacrenol (Figure 3)
and the presence of this compound in the Myrtaceae is
largely confined to Kunzea (Table 3). The mass spec-
trum of the peak is shown in Figure 4, a large m/z D 84
being characteristic of this compound. This behaviour
suggested that the compound, named ‘kunzeaol’ for con-
venience, was a germacrene-derivative. Kunzeaol was
isolated from solvent extracts of the Kunzea species
and a full set of one and two-dimensional NMR exper-
iments were performed, identifying the compound as 6-
hydroxygermacra-1(10),4-diene (Table 4). The 1 H-NMR
spectra showed some similarities to a previous report,15
although no previous reports detailing information about
the 13 C-spectra of this compound have been found. The
13
C- and 1 H-spectra of kunzeaol display signal splitting,
indicating that the molecule has two stable conformers;

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
266 C. P. CORNWELL ET AL.

Table 1. Germacradienols in the small-scale hydrodistillates (Hyd) and solvent extracts (DCM or MeOH) of the
Myrtaceaea

Eucalyptus, section Adnataria υ-cad ˇ-gol ˛-mol -mol -cdol ˛-cdol kuzol
E. microtheca 831264 Hyd 7.94 2.08 6.21 7.47 15.11
DCM 4.58 1.70 1.37 4.28 5.55 10.23
E. pruinosa ssp. pruinosa 841760 Hyd 13.56
DCM 0.64 1.52
Eucalyptus, section Bisectaria
E. blaxellii 832140 Hyd
DCM 0.18
Corymbia
C. trachyphloia ssp. carnarvonica 892010 Hyd 0.33
DCM 0.22
C. maculata 861790 Hyd 9.19 1.19 2.04 4.77 8.63 13.77
DCM 4.13 31.22 0.55 1.39 3.60 4.36
C. petalophylla 892019 Hyd 5.67 0.46 0.66 2.16 0.62
DCM 5.07 0.23 0.98 1.06 2.32 0.95
C. maculata 891338 Hyd 3.76 1.09 2.60 4.72 7.37
DCM 3.23 18.23 0.33 0.88 2.31 2.54 0.52
C. scabrida 891868 Hyd 15.34 0.16 0.91 1.60 5.94 1.97
DCM 14.11 0.47 1.04 1.51 5.09 2.05
C. clarksoniana ssp. clarksoniana 904971 Hyd 0.14
DCM 0.06
Kunzea
K. preissiana 252702 Hyd 8.92 2.12 7.91 10.52
MeOH 0.32 2.53 27.00
K. sp. 933330 Hyd 10.34 2.28 6.22 5.21 18.11
MeOH 1.02 4.10 33.76
K. sp. 904934 Hyd 0.62 0.56 0.49 1.48
MeOH 0.44 0.45 4.36
Leptospermum
L. epacridoideum 932473 Hyd 0.40
MeOH 0.12 1.90
L. spectabile 770048 Hyd 6.16 1.27 2.97 6.20 11.35
MeOH 2.75 7.61 0.66 1.82 3.56 5.41
L. Amboinese 910574 Hyd 2.86 0.63 1.47 1.71 2.59
MeOH 0.58 0.28 0.16
a
Abbreviations: υ-cad, (C)-υ-cadinene; ˇ-gol, ˇ-germacrenol; ˛-mol, ˛-muurolol; -mol, -muurolol; -cdol, -cadinol; ˛-cdol, ˛-cadinol; kuzol, kunzeaol.

Scan 1670 (37.007 min): 2201001.D

Abundance
81

2000000

1800000

1600000

1400000

1200000

1000000
43
800000

600000
123
400000 67 93 105 161
55 109
200000 207
133
147 165 179 189 222
38
0
m/z-->
40 60 80 100 120 140 160 180 200 220

Figure 2. Mass spectrum of ˇ-germacrenol isolated from Corymbia maculata (891338)

similar properties have been reported for the C6-epimer.5
These features and the identification of the stereochem-
istry of kunzeaol are detailed below.
The 1 H-spectrum displayed mostly broad ill-defined
peaks. The 13 C-spectrum showed two sharp signals,
one around 70 ppm, the other around 16 ppm; all other
signals were broad. The DEPT spectra showed that there
were four CH3 -signals, seven CH2 -signals, five aliphatic
CH-signals and four vinylic CH-signals. Four vinylic
quaternary carbons were identified in the 13 C-spectrum.

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
 GERMACRADIENOLS IN ESSENTIAL OILS OF MYRTACEAE 267

Table 2. 13 C- and 1 H-NMR spectral data for ˇ-germacrenol from the MeCl2 extracts of
Corymbia maculata (891338)a
13 C 1H 13 Cb 1 Hc 13 Cd 1 Hd 1 Hc 1 Hc
(ppm) (υ) ˇ-OH ˇ-OH ˛-OH ˛-OH ˛-OH ˛-OH
CDCl3 CDCl3 CDCl3 CDCl3 CDCl3 CDCl3 CDCl3 C6 H6
C1 128.8 4.90 128.9 4.95 129.0 4.94 4.95 4.97
C2 23.6 1.90 23.7 1.95 (˛) 23.7 1.91 1.96 1.96
2.48 2.50 (ˇ) 2.49 2.51 2.67
C3 39.6 1.60 39.6 1.54 41.3 1.59 1.52 1.35
1.64 1.65 1.50
C4 73.0 73.1 73.1
C5 140.0 5.21e 140.1 5.25 140.1 5.24 5.17 5.06
C6 125.7 5.14e 125.8 5.17 125.8 5.19 5.25 5.30
C7 52.8 1.95 52.8 2.02 52.9 2.00 2.02 1.96
C8 25.9 1.38 26.0 1.39 39.7 1.39 1.41 1.35
C9 41.2 2.21 41.3 2.25 26.0 2.25 2.26 2.21
C10 132.5 132.6 132.5
C11 33.0 1.38e 33.0 1.39 30.0 1.39 1.41 1.35
C12 20.6 0.81 20.6 0.82 20.6 0.83 0.84 0.95
C13 18.9 0.79 19.0 0.78 19.0 0.79 0.80 0.91
C14 16.7 1.50 16.7 1.54 16.1 1.54 1.55 1.59
C15 30.7 1.15 30.7 1.19 30.8 1.20 1.21 1.12
a values from this work are in bold. Spectra were obtained at 303 K.
b
Nagahama et al.1
c
Bohlmann et al.4,15
d
Izac et al.16
e
J5,6 D 15 Hz, J6,7 D 9 Hz, J11,12/13 D 8 Hz.
Initially it was thought that the extra signals were due to
an impurity in the sample. However, closer inspection
indicated that most of the signals were split in a ratio
of 4 : 3, the two narrow signals (about six times the
width at the base line as the chloroform signals) had
approximately the same integration as a split pair of
broad signals (their widths could be a high as 20 times
that of a chloroform peak). HMQC indicated that signal
splitting was also occurring in the 1 H-dimension.
In the HMBC experiment, the signals that were not
split allowed the basic skeleton to be established, since
both peaks from a split signal would correlate with the
unsplit signals. For instance, the single signal from the
proton on C6 would correlate with the split signals in
the carbon dimension from C5 and C8 and to the weaker
split signals from C4, C7 and C11. Similarly, the unsplit
signal from C15 correlated with the split signals from
C3, C4 and C5. Similar results were observed in two-
dimensional proton experiments, such as DQ-COSY and
TOCSY. This confirmed that the signal splitting was real,
coming from a single molecule and was therefore due
to two stable conformers of the same molecule, the data
in Table 4 showing the minor peaks (¾40% of the total
signal) in brackets.
The conformation of the C4 to C5 double bond is
trans, assigned on the basis that (C)-υ-cadinene and
the cadinols/muurolols are acid-solvolysis byproducts.
These reactions do not involve either C4 or C5, so the
original conformation about these two atoms will be pre-
served. The configuration of the C1–C10 double bond
is also trans, as determined by NOE-difference spec-
troscopy. Irradiation of the C1 signal at υ4.85 resulted
Copyright  2001 John Wiley & Sons, Ltd.
in enhancement of the signals for C9, thereby confirming
the trans-orientation of the double bond. The conforma-
tion about C6 was assigned the S- or ˇ-configuration
due to the preferential formation of cadinanes and
muurolanes during acid solvolysis. This can only occur
if the carbocation localized on C6 is attacked on the ˇ-
face by the electrons forming the new bond between C1
and C6. Such a conformation could be best adopted by
a molecule with a ˇ-hydroxyl group on C6.
Both germacrenol and kunzeaol produce oils rich in
(C)-υ-cadinene, cadinols and muurolols during distil-
lation. 13 C-NMR (Table 3),18 – 23 mass spectral13,20 and
Kováts indices on a DB-5 column13 allowed the identifi-
cation of the major sesquiterpene alcohols as ˛-cadinol,
-cadinol and -muurolol. ˛-Muurolol was identified
using the latter two methods. The similarity in the prod-
ucts produced by these two hydroxygermacradienes on
degradation indicates that they come from the same
carbocation, namely the germacradien-6-yl carbocation
(carbocations II and IV; Scheme 1).
The preferential formation of (C)-υ-cadinene, the
cadinols and the muurolols during the acid solvolysis
of germacrenol and kunzeaol implies that the reaction is
stereospecific about C6, the carbocation having a geome-
try such that the proton on C6 will be anti to the proton
on C7 (Scheme 1, carbocations I, II, III and IV). No
products from the alternative C6 conformers (such as
carbocation VII; Scheme I), in which the proton on C6
is syn to the proton on C7, were observed in the product
mix.
The preferential formation of cadinols over muurolols
is a result of the transition state preferentially adopting
Flavour Fragr. J. 2001; 16: 263–273
268 C. P. CORNWELL ET AL.

Abundance H OH
H OH
1800000 H OH

1600000
H
1400000 H
H
1200000 H

1000000 H OH

800000

600000
H
400000

200000

Time--> 0
32.00 34.00 36.00 38.00 40.00 42.00 44.00 46.00

Abundance

6000000
OH
5500000

5000000

4500000
H
4000000

3500000

3000000

2500000 OH

2000000
β-caryophyllene caryophyllene oxide
1500000

1000000
α-gurjunene α-humulene
500000
0
Time-->
32.00 34.00 36.00 38.00 40.00 42.00 44.00 46.00

Figure 3. Sesquiterpene region of the small-scale hydrodistillate (upper trace) and solvent extract (lower trace) of
Kunzea sp. (933330)

Table 3. 13 C-NMR data for the cadinols

13 C-NMR
˛-cadinol -muurolol -cadinol

(ppm) CDCl3 CDCl3 b CDCl3 CDCl3 c CDCl3 CDCl3 d

CH3 15.0 15.2 15.3 15.4 15.1 15.1
20.6 20.9 21.5 21.6 21.3 21.4
21.4 21.2 23.5 23.6 23.7 26.0
23.7 23.9 29.2 29.3 28.6 28.4
CH2 21.8 21.9 19.3 19.3 19.8 19.6
22.6 22.6 20.9 20.9 22.5 22.5
30.8 31.1 31.2 31.2 30.9 30.8
42.1 42.4 34.6 34.6 40.3 40.2
CH 25.9 26.0 26.6 26.6 23.4 23.6
39.7 40.0 34.5 34.5 37.7 37.5
46.6 46.8 43.9 43.9 46.7 46.5
49.9 50.1 46.0 46.1 47.9 47.9
CH 122.2 122.5 124.8 124.8 122.6 122.5
COH 72.2 72.2 72.2 72.3 70.5 70.4
C 134.8 134.9 133.3 133.4 134.1 134.0
a
Values from this work are in bold. Spectra were obtained at 303 K.
b
Bottini et al.,18 Sanz and Marco19 and Chalchat et al.20
c
Asakawa et al.21
d
Connolly et al.22

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
 GERMACRADIENOLS IN ESSENTIAL OILS OF MYRTACEAE 269

Scan 2157 (46.948 min): 0601044.D

Abundance 84

650000

600000
550000

500000

450000

400000

350000 41
55
300000
250000 69
109
200000
95 121
150000
39 107
100000
137 161
50000
151 179 189 204 222
167
m/z-->0
40 60 80 100 120 140 160 180 200 220

Figure 4. Mass spectrum of 6ˇ-hydroxygermacra-1(10),4-diene (kunzeaol)

Table 4. 13 C- and 1 H-NMR spectral data for 6-˛-hydroxygermacra-1(10),

4-diene (kunzeaol) from the MeCl2 extracts of Kunzea sp. (933330)a
13 C (ppm) 1 H (υ) 1 Hb 13 Cc 1 Hc
CDCl3 at 303 K CDCl3 CCl4 CDCl3 CDCl3
C1 121.1 (128.5) 4.85 4.90 128.7 (121.3)
C2 41.2 (35.6) 1.60 (1.58) 30.3 (25.2)
2.05 (2.30)
C3 36.9 (38.8) 2.02 37.2 (35.7)
C4 132.3 (132.2) 133.3 (133.0)
C5 131.7 (133.8) 4.90 (4.95) 4.90 133.5 (131.4) 4.95
C6 68.5 4.50 4.51 68.8 (68.6) 4.59
C7 51.9 (49.2) 0.70 (0.75) 52.2 (49.3)
C8 24.9 (29.9) 1.19 (1.25) 24.3
1.63 (1.60)
C9 24.3 2.00 41.3 (39.0)
2.25
C10 138.4 (135.3) 138.9 (135.7)
C11 31.9 (31.5) 1.59 32.1 (31.7)
C12 20.0 0.95 0.99 22.1 (21.5) 0.99
C13 20.0 0.95 0.97 21.3 (21.2) 0.97
C14 16.7 (21.7) 1.49 (1.51) 1.57 21.0 (16.5) 1.42
C15 16.2 1.35 1.44 16.5 (16.4) 1.42
a Values from this work are in bold. Values in brackets represent the shifts for the minor conformer.
b
Bohlmann et al.15
c For 6ˇ-hydroxygermacra-1(10),4-diene at 298 K; Barrero et al.5

a conformation in which C14 and the proton on C6
lie on the same face of the ‘chair’ form of the ger-
macryl carbocation (Scheme I; carbocations I and III).
The preferential formation of ˛-cadinol over its C10
epimer, -cadinol, is a direct result of the initial con-
formation adopted by the cadinyl carbocation (car-
bocation V; Scheme I). This conformation favours ˇ-
attack by water onto C10 (the site of the emerging
carbocation); unfolding of the carbocation will expose
the ˛-face of C10, thereby allowing formation of -
cadinol. The attack by water on the carbocation can
be demonstrated by distilling the leaves of C. macu-
lata in H2 18 O, the cadinols so produced showing 18 O-
incorporation, evidenced by a +2 amu shift in either
the parent ion or m/z 43–45 (Figure 5). Similar results
were obtained when Kunzea sp. was treated in a similar
fashion.
Muurolols are a result of an alternative conformation
of the germacradienyl carbocation, in which the proton
on C6 and the proton on C1 both lie on the same face
of a ‘pseudo-chair’ form (Scheme I; carbocations II and
IV). The conformation of the emerging muurolyl car-
bocation (carbocation VI; Scheme I) favours ˛-attack
by water onto C10 to yield -muurolol. Unfolding of
the carbocation will expose the ˇ-face of C10, allow-
ing the formation of ˛-muurolol. 18 O-Incorporation also
occurred for the muurolols when the leaves were distilled
in H2 18 O (Figure 6).

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
270 C. P. CORNWELL ET AL.

14 CH3 CH3
H H
1 7

+
6 +
H H
H H
H
I II +

+ +
H , −H2O H , −H2O
H
V
OH H OH
H kunzeaol
β-germacrenol H H
H H
1
7
6 +
+
CH 3 H CH 3 H

III IV

H OH H H (+)−δ-cadinene
+

H H H
VI
α-muurolol γ-muurolene

H OH H H OH H

H H H α-cadinol H α-cadinene

τ-muurolol α- muurolene
H OH H
H

H
7
+
τ-cadinol γ-cadinene
6
H H
CH3 H
H
VII

amorphene and
H
bulgarene group

Scheme 1

The appearance of both cadinols and muurolols imp-
lies that the C1–C10 double bond can alter its orienta-
tion, although the stereospecificity about C6 indicates
that the conformation adopted by these hydroxyger-
macradienes about C6 and C7 does not change5 and
may be determined during biosynthesis when ring clo-
sure occurs. These results are in agreement with other
reports of stereospecificity about C7 in the acid-catalysed
breakdown of germacrene D in acetic acid,7 the major
products being (C)-υ-cadinene (41%), ()-1 -cadinene
(13%) and ()-˛-muurolene (16%), with traces of (C)-
-cadinene (3%), ˛-cadinyl acetate (8%) and other
acetylated products.
Acknowledgements—The authors would like to acknowledge the man-
agement and staff, especially R. Johnstone and T. Armstrong, of the
Mt. Annan Botanic Gardens (Mt. Annan, NSW) for their assistance in

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
 GERMACRADIENOLS IN ESSENTIAL OILS OF MYRTACEAE 271

Scan 1477 (33.085 min): 1101013.D (*)

Abundance
95

9000
8000
7000
121
6000
5000
4000 43
81 161 204
3000 105 99
2000 55 69
1000 39 139 149
179 189 222 224
166
m/z--> 0
40 60 80 100 120 140 160 180 200 220

#691: alpha-cadinol (*)

Abundance 95

9000
8000
43
7000
121
6000
5000
4000
71 81
3000 109 161
105 204
2000 55
137
1000 222
39 149 189 223
166 179
0
m/z-->
40 60 80 100 120 140 160 180 200 220

Scan 1450 (32.534 min): 1101013.D (*)

Abundance
161

9000
8000
7000
6000
5000
4000 81 204
43 105
3000 95
2000 79 121
55 69 134
189
1000 39 205
131 147 164
0
m/z-->
40 60 80 100 120 140 160 180 200

#693: tau-cadinol (*)

Abundance
161

9000
8000
7000
6000
5000 43
4000
81 105 204
3000 95
79 119
2000 55 134
69
1000 189
39 179 205
123 147 164
0
m/z-->
40 60 80 100 120 140 160 180 200

18
Figure 5. Mass spectra of ˛- and -cadinol showing O-incorporation (upper spectra of each pair) associated with the
hydrodistillation of the young leaves of Corymbia maculata (891338)

the collection of plant material and for granting access to their exten- References
sive collection. The authors would also like to thank the management
and staff of the Mt. Tomah Botanic Gardens for providing samples 1. Nagahama S, Tazaki M, Kobayashi H, Sumimoto M. Phytochem-
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Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 263–273
272 C. P. CORNWELL ET AL.

Scan 1461 (32.758 min): 1101013.D (*)

Abundance
161

9000
8000
7000 119
6000
105
5000 95
4000 43
3000 81 204
79
69
2000 55
1000 39 133 189
123 149
m/z-->0
40 60 80 100 120 140 160 180 200

#694: alpha-muurolol (torreyol) (*)

Abundance
161

9000
8000
7000
43 119
6000
5000 105
95
4000
3000 81
79 204
2000 55 69
1000 39 136 189
123 147 179 207
m/z-->0
40 60 80 100 120 140 160 180 200

Scan 1454 (32.616 min): 1101013.D (*)

Abundance
95

9000
8000
7000
121
6000 161
5000 43
4000 204
79 105 99
3000 69
2000 55
1000 39 133 152 181 189 224
0
m/z-->
40 60 80 100 120 140 160 180 200 220

#692: tau-muurolol (*)

Abundance
95

9000
43
8000
7000
6000 121
161
5000
4000
71
204
3000 79 105 109
58
2000
1000 39 134 149 189 222
179
177 223
0
m/z-->
40 60 80 100 120 140 160 180 200 220

Figure 6. Mass spectra of ˛- and -muurolol showing 18 O-incorporation (upper spectra of each pair) associated with
the hydrodistillation of the young leaves of Corymbia maculata (891338)

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