Phytochemistry 58 (2001) 799–810

www.elsevier.com/locate/phytochem

Chemical study of the essential oil of Cyperus rotundus

Mesmin Mekem Sonwa, Wilfried A. König*
Institut für Organische Chemie, Universität Hamburg, D-20146 Hamburg Germany

Received 4 April 2001; received in revised form 25 June 2001

Abstract
Minor constituents of the essential oil of Cyperus rotundus have been investigated. The three new sesquiterpene hydrocarbons
(
)-isorotundene, (
)-cypera-2,4(15)-diene, (
)-norrotundene and the ketone (+)-cyperadione were isolated and their structures
elucidated. The absolute configuration of (
)-rotundene was derived by chemical correlation and enantioselective gas chromato-
graphy. # 2001 Elsevier Science Ltd. All rights reserved.
Keywords: Cyperus rotundus; Cyperaceae; Sesquiterpenes; Norsesquiterpene; (
)-Isorotundene; (
)-Cypera-2,4(15)-diene; (
)-Norrotundene; (+)-
Cyperadione

1. Introduction (17) by comparison with a spectral library established

under identical experimental conditions (Joulain and
Cyperus rotundus is a cosmopolitan sedge belonging to König, 1998), cyperotundone (18) (Hikino et al., 1966),
the family of the Cyperaceae. It is encountered in tropi- mustakone (19) (Nyasse et al., 1988), cyperol (20)
cal, subtropical and temperate regions. C. rotundus is a (Hikino et al., 1967), isocyperol (21) (Hikino et al.,
traditional medicinal plant appearing among Indian, 1967) and a-cyperone (22) (Howe and McQuillin, 1955;
Chinese and Japanese natural drugs used against spasms Haaksma et al., 1992) (Fig. 1). However, several minor
and stomach disorders (Dassanayake and Fosberg, compounds could not be identified by their mass spectra
1985). The essential oil as well as solvent extracts of the and had to be isolated for further investigations. Con-
rhizomes of C. rotundus have been subject to numerous sequently, the essential oil was first separated by silica
studies (Ohira et al., 1998 and cited literature) resulting chromatography into a hydrocarbon fraction and a
in the isolation of many terpenoids. Here we describe more polar oxygenated fraction. The hydrocarbon frac-
the isolation and structure elucidation of new patch- tion was further separated by column chromatography,
oulane and rotundane sesquiterpenes from the essential TLC over silver nitrate precoated silica and preparative
oil of this plant. GC. This resulted in the isolation and characterization
of isorotundene (23), cypera-2,4(15)-diene (24) and nor-
rotundene (25). From the oxygenated fraction cyper-
2. Results and discussion adione (26), a patchoulane type sesquiterpene, was
isolated.
The analysis of the essential oil of Cyperus rotundus by
GC and GC–MS allowed the identification of cyprotene 2.1. (
)-Isorotundene (23)
(1), cypera-2,4-diene (2), a-copaene (3), cyperene (4), a-
selinene (5), rotundene (6), valencene (7), ylanga-2,4- The unknown compound 23 was assigned the mole-
diene (8), g-gurjunene (9), trans-calamenene (10), d-cadi- cular formula C15H24 ([M]+at m/z=204). The 13C NMR
nene (11), g-calacorene (12), epi-a-selinene (13), a-muur- spectrum showed 15 carbon signals, two of which were
olene (14), g-muurolene (15), cadalene (16), nootkatene olefinic ( 107.9 and 151.08). Through the DEPT tech-
nique two methyl groups, seven methylene groups, four
* Corresponding author. Tel.: +49-40-42838-2824; fax: +49-40-
methine and two quaternary carbon atoms were identi-
42838-2893. fied. The compound has four unsaturations and only
E-mail address: wkoenig@chemie.uni-hamburg.de (W.A. König). one double bond, therefore the presence of three rings
0031-9422/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0031-9422(01)00301-6
800 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

Fig. 1. Sesquiterpenes from the essential oil of Cyperus rotundus.

was concluded. The 1H NMR and 13C NMR signal H-7 also couples with the methylene group CH2-8 (
assignments, achieved by 1H–1H-COSY and HMQC, 1.05 and 1.71) which again couples with CH2-9 ( 1.44
correlations, afforded three substructures. The methine and 1.72) to give the second substructure–CH–CH2–
proton CH-5 ( 1.99) couples with the methine proton CH–CH2–CH2–. The methylene group CH2-11 ( 2.14
CH-4 ( 1.90) which is vicinal to methyl group CH3-15 and  2.27) and the olefinic proton at d 5.85 couple with
( 0.86). The latter methine proton also couples with the each other and give the third substructure–CH2–
methylene group CH2-3 ( 1.22 and 1.68) itself coupled C¼CH2. The proton-carbon long-range coupling corre-
to another methylene group CH2-2 ( 1.46 and 1.52) lations derived from the HMBC spectrum allowed to
which further couples with the methine proton CH-1 ( identify the skeleton of the compound. Long-range
1.81) giving the partial structure–CH–CH(CH3)–CH2– coupling correlations were found between the protons
CH2–CH–. The methine proton  1.99 also couples with H-11, H-13 and carbon C-7 as well as between the
a methylene group CH2-6 ( 1.00 and 1.70) itself cou- methine proton H-7 and the carbons C-11, C-12 and C-
pled to the methine proton CH-7 ( 2.52). The proton 13, indicating a bond connecting C-7 and C-12. The
 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810 801

proton H-11 also correlated with the carbons C-1, C-9, order to correlate it with (+)-g-gurjunene (9). Rotun-
C-10 and C-14 while the methyl group H-14 correlates dene (6) was submitted to ozonolysis followed by
with the carbons C-1, C-9, C-10 and C-11. From these reduction with dimethyl sulfide under nitrogen
observations it was deduced that C-10 is connected to (McMurry and Bosch, 1987). The desired aldehyde was
C-11, C-14 to C-10, and C-10 to C-1. Long-range cor- not stable and was oxydised directly the to ketoacid 28
relations were observed between the protons H-6, H-5, which could be isolated. The acid 28 was then dec-
H-4 and carbon C-1 as well as between H-1 and the arboxylated (Fristad et al., 1983) to give ketone 29.
carbons C-2, C-4, C-5, C-6 and C-10 indicating a bond Subsequently 29 was allowed to react with methylene-
between C-1 and C-5. The combination of all these triphenylphosphine in dimethyl sulfoxide (Greenwald et
informations led to a rotundane skeleton with an exo- al., 1963) to yield the hydrocarbon 30 which was
cyclic double bond. The relative configuration at the hydrogenated (Harmon et al., 1969) to give compound
chiral centres C-1, C-4, C-5, C-7 and C-10 was con- 31 (Fig. 4). Compound 31 was finally compared by
cluded from the NOESY diagram. The correlations enantioselective gas chromatography with the fully
observed between protons H-1, H-4 and H-5 indicate hydrogenated products of (+)-g-gurjunene (5) and
that these protons are on the same side of the molecule. proved to have the same retention time with the major
Furthermore, the protons H-1 and H-5 show spatial hydrogenation product (Fig. 5). The co-injection was
interactions with the proton H-11a, which is only pos- made in a column which separates (+)- and (
)-g-gur-
sible if H-1 and H-5 are located at the same side as the junene (Fig. 6). Unfortunately, the small quantity of
C-11-C-12 bridge. Therefore, compound 23, which was (
)-g-gurjunene (which we isolated from the liverwort
named isorotundene, has the relative configuration as Marchantia polymorpha) was insufficient for hydro-
shown in Fig. 2. genation and comparison with 31. However, it is very
To verify the rotundane skeleton a partial synthesis of probable that the used column also would have sepa-
23 by oxymercuration-demercuration (Brown and Geo- rated the hydrogenation products of the two enantio-
ghegam, 1970) of rotundene (6) to isorotundenol (27) mers. Therefore, it is very likely that rotundene has the
and subsequent dehydration (Greenwood et al., 1968) to absolute configuration as shown for 23.
isorotundene (23) and rotundene (6) was performed
(Fig. 3). The synthesized and the isolated compounds 23 2.2. (
)-Cypera-2,4(15)-diene (24)
have identical spectroscopic data. Although rotundene
(6) has been known for a long time its stereochemistry The isolation of cypera-2,4(15)-diene (24) was
was not known as yet. The relative configuration of achieved by a combination of column chromatography
rotundene could not be derived from its NOESY dia- and preparative GC. The compound was assigned the
gram because of the overlapping of all the important molecular formula C15H22 ([M]+ at m/z=202). The 13C
signals. However, the fact that isorotundene (23) could NMR spectrum combined with the DEPT technique
be obtained from rotundene as shown in Fig. 3, proves revealed three methyl groups, four methylene groups,
that rotundene and isorotundene have the same relative one being olefinic ( 102.84), five methine groups, two of
configuration. which are olefinic ( 133.26 and 139.17) and three
The identity of the relative configuration of rotundene quaternary carbon atoms, one of which is olefinic
(6) and isorotundene (23) still leaves the question about (d 148.94).
the absolute configuration open. In order to determine The proton NMRspectrum displayed some signals
the absolute configuration of the two compounds, a common to patchoulane sesquiterpenes, namely the two
chemical transformation of rotundene was performed in

Fig. 2. Relative configuration of isorotundene with relevant NOEs. Fig. 3. Preparation of isorotundene (23) from rotundene (6).
802 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

Fig. 4. Chemical conversion of the rotundane to a guaiane skeleton.

geminal methyl groups H-12 and H-13 (d 0.99 and 1.01) All these informations from the 13C NMR and DEPT
which are coupled to each other (W-coupling), the as well as the 1H–1H-COSY led us to propose structure
methyl group at  0.77, appearing as a doublet and 24 for the compound. Its relative configuration resulted
coupling with the methine proton H-10 ( 2.12). The from the NOESY diagram which shows NOE correla-
vinylic proton H-2 (d 6.17) couples with another vinylic tions between H-5, H-9 and H-14 at one side and
proton H-3 ( 5.69), itself coupled to the olefinic meth- between H-13 and H-10 at the other. The observed cor-
ylene group H-15 ( 4.98) and also to the methine pro- relations indicate that the methine proton H-10 is on the
ton H-5 ( 3.03) giving the substructure –CH¼CH– same side with the bridge C-1–C-7, while H-5, H-9 and
C(CH)¼CH2. Moreover, the coupling constant between H-14 are located at the opposite side of the molecule
H-2 and H-3 (J=5.60 Hz) indicates that they belong to (Fig. 7). 24 was before found as a minor product during
a five-membered ring. The methine proton H-5 also the reduction of cyperotundone (18) to the correspond-
couples with the methylene group CH2-6 ( 1.68 and ing alcohol and subsequent dehydration to 2 (Fig. 8)
2.10) which further couples with the methine proton H- (Mekem Sonwa et al., 1997).
7 (d 1.85) itself coupled to the methylene group CH2-8
(d 1.51 and 1.86) which again couples with the methyl- 2.3. (
)-Norrotundene (25)
ene group CH2-9 (d 1.03 and 1.29) itself coupled to the
methine proton CH-10 (d 2.12). From these informa- The isolation of 25 was performed by the combina-
tions the substructure CH–CH2–CH–CH2–CH2–CH– is tion of many separation methods including CC at low
derived. temperature (
20 C), CC over silver nitrate precoated
 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810 803

Fig. 5. Chemical transformation of rotundene and GC comparison with the hydrogenation product of (+)-g-gurjunene.

silica, preparative TLC and GC. The compound has the should consist of three rings since it has a total of four
molecular formula C14H22 in agreement with its mass unsaturations, one being a double bond. The constitu-
spectrum ([M]+ at m/z =190). The 13C NMR and tion of 25 was derived from interpretation of the 1H
DEPT spectra revealed 14 carbon atoms, two methyl NMR spectrum and the phase-sensitive 1H–1H-COSY
groups, five methylene groups, six methine groups from spectrum together with the HMQC and the HMBC
which two are olefinic ( 129.15 and 139.10) and one diagrams.The methine proton CH-1 ( 2.20) couples
quaternary carbon atom. Therefore the compound with the methylene group CH2-2 ( 1.61 and 1.54) which
804 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

assumption is confirmed by the HMBC diagram which

shows long-range correlations between the protons H-
13 and C-1, C-10 and C-11. Proton H-1 shows long-
range connectivities to C-2, C-5, C-10 and C-11. The
protons of methylene group CH2-9 show connectivities
to C-1, C-8, C-10 and C-11, while the olefinic proton H-
11 is long-range coupled to C-1, C-7, C-9, C-10, C-12
and C-13. All these observations imply the two carbon-
carbon connections between C-10–C-1 and C-10–C-11,
leading to a rotundane skeleton for 25 (Fig. 9).
Fig. 6. Separation of (
)- and (+)-g-gurjunene by capillary gas chro- The relative stereochemistry at the chiral centres of
matography on a 25 m capillary column with heptakis(6-O-t.butyldi- the molecule could not be derived from the NOESY
methylsilyl-2,3-di-O-methyl)-b-cyclodextrin (50% in OV 1701, w/w) at
spectrum. The signals of the two methine protons H-1
100  C.
and H-4 overlap and hence it is not possible to know
which one of them is responsible for the NOE with the
proton H-5. However, using the phase-sensitive COSY
technique as well as the gradient selected HMQC
method, it was possible to calculate the scalar coupling
constant between H-4 and H-5 (J=5.85 Hz). This value
of the coupling constant corresponds (Karplus curve) to
an angle of =35 between the two protons, which
means that they are on the same side of the molecule.
Accordingly, both of them have spatial interactions
with H-1. It remains to investigate the stereochemistry
at C-7 and C-10. To solve this problem the NOESY
diagram was of no help because the protons H-1 and H-
5 are too far from the vinylic protons H-11 and H-12.
The solution came from the 1H–1H-COSY diagram
where a long-range coupling correlation is found
between proton H-1 and one of the H-9 protons. This is
a case of 4J coupling through a s-bond which requires a
Fig. 7. Stereostructure of cypera-2,4(15)-diene (24) and important particular geometry (W arrangement of the two pro-
NOEs. tons) for the molecule, and this is only possible if the H-
1 proton and the C-11–C12 bridge are on the same side
further couples with another methylene group CH2-3 ( of the molecule. Moreover, the axial–axial coupling
1.42 and 1.87) which couples again with the methine between H-5 and H-6a is only possible for this stereo-
proton H-4 ( 2.10) itself coupled to the methyl group chemistry of the molecule (Fig. 10). Norrotundene is
CH3-14 ( 0.90) and the methine proton CH-5 ( 1.80) probably biogenetically related to rotundene and should
which further couples with the methine proton H-1 ( have the same absolute configuration.
2.20) and gives rise to a five-ring partial structure a:
C1H–CH2–CH2–CH(CH3)–CH–C1H–. 2.4. Cyperadione (26)
The methine proton H-5 also couples with methylene
group CH2-6 ( 0.95 and 1.66) which further couples Cyperadione (26) was isolated from the oxygenated
with the methine proton H-7 ( 2.56), itself coupled to fraction of C. rotundus oil by preparative GC. The
the methylene group H-8 ( 1.12 and 1.91) which cou- compound has the molecular formula C15H24O2 ([M]+
ples again with the methylene group H-9 ( 1.52 and at m/z=236). The 13C NMR- and the DEPTspectrum
1.83) to give the substructure b: CH–CH2–CH–CH2– indicate that it has four methyl groups, two methine
CH2–. Moreover, the vinylic protons H-11 and H-12 ( protons and four quaternary carbon atoms, two of
6.02) are coupled to the methine proton H-7, indicating which being carbonyl carbons ( 208.50 and 220.25).
a partial structure c: –CH¼CH–CH–. Observing that From the 1H NMR and the 1H 1H-COSY spectra, three
the substructures a and b have C-5 in common and that substructures were assigned which helped to derive the
b and c have C-7 in common, the three partial structures full structure of the product. The methyl group CH3-14
can be connected to give a new substructure A (Fig. 9). ( 0.78) couples with the methine proton CH-10 ( 2.10)
It may be assumed that the only tertiary methyl group which also couples with the methylene group CH2-9
of the molecule CH3-13 is linked to the only quaternary ( 1.10 and 1.68) itself coupled to the methylene group
carbon and thus yielding substructure B: CH3–C. This CH2-8 ( 1.45 and 2.01) which further couples with the
 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810 805

Fig. 8. Formation of cypera-2,4(15)-diene (24) form cyperotundone (18).

Fig. 9. Important proton-carbon long-range correlations in the HMBC spectrum of norrotundene (25).

group. This is confirmed by the HMBC diagram where

correlations are observed between the protons H-6 and
carbonyl carbon C-7. From all this information sub-
structure CH3–CH–CH2–CH2–CH–CH2–CO can be
derived. The two methyl groups at  0.98 and 1.28
appearing as singlets are coupled to each other (W cou-
pling), what indicates that they are linked to the same
quaternary carbon atom giving the substructure CH3–
C–CH3. The deshielded methyl group CH3-15 at  2.18
Fig. 10. Relative configuration of norrotundene (25). Note the W- is probably adjacent to a carbonyl function (this is also
arrangement between H-1 and H-9e, and the axial-axial disposition confirmed by the HMBC spectrum) and shows a long-
between H-5 and H-6a. These two geometrical patterns which are range coupling correlation to the methylene group CH2-
supported by the observed coupling correlations are only possible for 3 ( 2.35 and 2.44) which further couples with the meth-
the given stereostructure of the molecule.
ylene group CH2-2 ( 1.50 and 2.11) to give the partial
structure CH3–CO–CH2–CH2–. Additional structural
methine proton CH-7 ( 1.90) itself coupled to the informations were drawn from the HMBC diagram. The
methylene group CH2-6 ( 1.96 and 2.48). It should be methylene protons H-2 are long-range correlated to
pointed out that the signals of this methylene group carbons C-1, C-3, C-4, C-5, C-10 and C-11 while the
appearing at lower field and showing no other coupling methine proton H-10 shows connectivities to C-1, C-2,
correlation to a proton may be adjacent to a carbonyl C-5, C-9, C-11 and C-14. From these observations the
806 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

three carbon–carbon connections C-1–C-2, C-1–C-10 The biosynthesis of cyperene and rotundene may
and C-1–C-5 were deduced. Moreover, the methyl pro- involve two successive cyclisation steps of farnesyl di-
tons H-12 and H-13 are long-range correlated to C-1, C- phosphate FDP leading via a cyclodecadienyl to a
7 and C-11, establishing the two connections C-11–C-1 cationic azulane type intermediate. At this stage two
and C-11–C-7. All these considerations led to structure pathways a and b may take place: Path a proceeds by a
26 (Fig. 11), with a skeleton related to the patchoulane 1,2- proton shift followed by a deprotonation leading to
skeleton of cyperene (4). To confirm structure 26 we a-guaiene from which cyperene is formed by subsequent
performed an ozonolysis of cyperene (4) (Fig. 12). The protonation, cyclisation and deprotonation. Path b
reaction product displayed the same spectral data as the proceeds by ring formation followed by a deprotonation
isolated compound. yielding rotundene or isorotundene (Fig. 13).

2.5. Biogenesis of patchoulane and rotundane type

sesquiterpenes 3. Experimental

Patchoulane and rotundane sesquiterpenes are char- 3.1. GC-MS

acteristic for the genus Cyperus. Cyperene (4) is always
the major hydrocarbon component in Cyperus species. Electron impact (70 eV) GC–MS measurements were
Rotundene (6) also is always present as a major sesqui- carried out on a Hewlett-Packard HP 5890 gas chro-
terpene hydrocarbon component. These two compounds matograph equipped with a 25 m polydimethylsiloxane
appear to be of some chemotaxonomic importance. We (Chrompack CP Sil 5 CB) capillary column and coupled
have previously reported the isolation of cyprotene (1), to a VG Analytical VG 70-250S mass spectrometer.
a norsesquiterpene biogenetically related to cyperene Helium was used as carrier gas.
from the essential oil of Cyperus alopecuroides. Here, we
described a new norsesquiterpene, norrotundene (25), 3.2. NMR spectroscopy
which most likely has a biogenetic relationship to
rotundene. Besides 1 compound 25 was also identified in NMR spectra were recorded using a Bruker DRX 500
C. papyrus. It, therefore, seems to be a peculiarity of (1H: 500 MHz, 13C: 125 MHz) and a Bruker DRX 400
Cyperus species to produce norsesquiterpenes. (1H: 400 MHz, 13C: 100 MHz). Tetramethylsilane
(TMS) was used as reference signal (=0.00).

3.3. Capillary GC

Orion Micromat 412 double column instrument with

25 m fused silica capillaries with CPSil 5 and CPSil 19
(Chrompack); Carlo Erba Fractovap 2150, 4160 instru-
ments with 25 m fused silica capillaries with hepta-
kis(2,6-di-O-methyl-3-O-pentyl)-b- (2,6-Me-3-Pe-b-CD)
and - g - cyclodextrin, respectively, and heptakis(6 - O -
t.butyldimethylsilyl-2,3-di-O-methyl)-b-cyclodextrin (6T-
2,3-Me-b-CD), all in polysiloxane OV1701 (50 wt%);
Fig. 11. Long-range correlations from the HMBC spectrum of cyper- split injection, flame ionization detection; carrier gas 0.5
adione (26). bar hydrogen.

Fig. 12. Preparation of cyperadione by ozonolysis of cyperene.

 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810 807

Fig. 13. Proposed biogenetic pathway for cyperene, rotundene and isorotundene.

3.4. Column chromatography at low temperature an oven. An ethanolic solution of sulfuric acid (10%)
was used as a spray reagent.
Prefractionations of essential oils were performed on
a silica gel column equipped with a condenser and con- 3.6. Essential oil
nected to a Kryoflex KF 40 cooling system from Mess-
geräte-Werk Lauda at
20  C. Ethanol was used as The essential oil of C. rotundus was a gift of K.-D.
cooling liquid. Protzen, Paul Kaders GmbH, Hamburg.

3.5. Thin layer chromatography 3.7. Isolation of compound 23

Thin layer chromatography was effected using glass A preliminary separation of the essential oil was per-
and aluminum plates of silica 60 F254 (Merck). Silver formed by column chromatography at low temperature
nitrate coated plates were prepared by immersing the (
20  C). This resulted in a fractionation of the oil into
plates into an ethanol–water (4:1) solution of silver a hydrocarbon and an oygenated fraction. One gram of
nitrate (5 mg AgNO3 in 50 ml of solvent). After 30 min the hydrocarbon fraction was then separated by TLC
the plates were removed from the solution and dried in using petroleum ether as eluting solvent. The aim of this
808 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

separation was to eliminate the major hydrocarbon 3.9. Preparation of isorotudene (23) from rotundene
(cyperene) from the sample before any other operation. (6)
Three bands were obtained, the first two containing
mainly cyperene and a-copaene. Only the third fraction 3.9.1. Hydration of rotundene by oxymercuration-
(Rf=0.45) was interesting because it contained the demercuration
unknown products. This fraction was further separated In a 50 ml flask, fitted with a magnetic stirrer, 48 mg
by preparative GC (column 6T-2,3-Me-b-CD, tempera- of mercuric acetate were placed. To this, 3 ml of water
ture programme: from 110 to 180  C, heating rate 2 / were added followed by 3 ml of THF. After addition
min). Fraction 5 of the chromatogram contained (+)- of 30 mg (0.148 mmol) of rotundene the reaction mix-
ylanga-2,4(15)-diene (8). Fraction 8 was analysed by ture was stirred for 15 min at room temp. to complete
GC–MS and three major products were present: a-seli- the oxymercuration. 3 ml of 3.0 M sodium hydroxyde
nene (5), valencene (7), and an unknown product. This were added followed by 3 ml of a solution of 0.50 M
sample was further separated by preparative TLC on sodium borohydride in 3.0 M sodium hydroxide.
silver nitrate precoated plates, using petroleum ether as Reduction of the mercurial complex is almost instan-
eluting solvent. Two bands were obtained, the second of taneous. The mercury is allowed to precipitate. Sodium
which (Rf=0.35) was taken up in petroleum ether and chloride was added to saturate the water layer. The
purified by preparative GC. Spectroscopic analysis upper layer of THF was removed and purified by pre-
resulted in the structure of isorotundene 23. parative GC, yielding 26 mg (80%) of isorotundenol
(27).
3.8. (
)- Isorotundene (23)
3.9.2. Isorotundenol C15H25OH (27)
1
H NMR (500 MHz, C6D6): see Table 1. 13C NMR 1
H NMR (500 MHz, C6D6):  0.75 (3H, d, J=6.61
(100 MHz, C6D6):  16.10 (q, C-15), 25.20 (t, C-2), 27.53 Hz), 0.76 (3H, s), 1.02 (3H, s), 1.08-1.54 (12H, m), 1.57–
(t, C-8), 29.28 (t, C-9), 31.48 (t, C-3), 32.28 (q, C-14), 1.65 (2H, m), 1.75 (1H, m), 2.15 (1H, m). MS (EI, 70
33.15 (t, C-6), 34.38 (s, C-10), 38.25 (d, C-4), 38.64 (d, eV), m/z (rel. int.) : 222 (5), 204 (19), 189 (27), 175 (7),
C-7), 41.52 (d, C-5), 44.36 (t, C-11), 56.19 (t, C-1), 161 (14), 147 (12), 133 (13), 121 (33), 108 (100), 93 (57),
107.98 (t, C-13), 151.08 (s, C-12). MS (EI, 70 eV), m/z 81 (39), 67 (24), 55 27), 41 (35).
(rel. int.) : 204 (33) [M]+, 189 (84), 175 (25), 161 (64),
144 (45), 133 (41), 119 (54 ), 108 (100), 93 (91), 79 (71), 3.9.3. Dehydration of isorotundenol
67 (46), 55 (51), 41 (82). Phosphoryl chloride (0.5 ml) was added to a pyridine
solution (1 ml) of isorotundenol (10 mg). The mixture
was stirred for 10 h and pyridine was then removed by
Table 1 distillation at low pressure. The remaining residue was
1
H NMR spectral data for compounds 23 and 25a taken up in hexane and separation by prep. GC yielded
H 23 25
3 mg of isorotundene and 3 mg of rotundene.

1 1.81 ddd (9.5, 9.5, 5.9) 2.20 dddd (13.5, 6.6, 6.4, 7.0) 3.10. Ozonolysis of rotundene
2 1.46 dd (9.5, 3.2) 1.54 m
1.52 m 1.61 m
3 1.22 dddd (13.6, 11.4, 11.4, 6.7) 1.42 dddd (13.5, 11.0, 11.0, 6.7) A solution of rotundene (50 mg) in dichloromethane
1.68 m 1.87 m (20 ml) was treated at
78 C with a stream of ozone
4 1.90 dddd (13.6, 6.9, 6.6, 6.4) 2.10 m until a persistant blue colour was observed. The solu-
5 1.99 dddd (13.2, 6.4, 6.4, 5.9) 1.80 dddd (13.2, 6.4, 6.4, 6.4) tion was then purged with nitrogen for 5 min. After
6 1.00 t (13.2) 0.95 dd (13.2, 13.2)
addition of dimethylsulfide (3 ml) the solution was
1.70 m 1.66 ddd (13.2, 9.1, 6.4)
7 2.52 m 2.56 m washed with ice-cold sodium bicarbonate solution (25
8 1.05 dd (12.9, 9.5) 1.12 ddd (13.8, 11.7, 5.1) ml), water and brine. The organic layer was dried
1.71 m 1.91 ddd (13.8, 9.4, 5.0) (NaSO4) and the solvent was evaporated. Purification of
9 1.44 m 1.52 ddd (11.7, 9.4, 6.1) the residue by prep. GC gave the keto-acid 28 (45 mg).
1.72 m 1.83 m 1
H NMR (400 MHz, CDCl3):  1.10 (3H, d, J=6.61
11 2.14 dt (17.0, 2.5, 2.5) 6.02 m
2.27 dd (17.0, 2.2) Hz), 1.30 (1H, m), 1.35 (3H, s), 1.65–1.92 (9H, m), 2.11
12 6.02 m (1H, m), 2.20 (1H, m), 2.29 (3H, s), 2.58–2.85 (2H,m).
13
13 5.85 m 1.13 s C NMR (100 MHz, CDCl3): 16.73, 25.10, 25.66,
5.85 m 27.47, 28.30, 30.07, 31.09, 31.60, 39.64, 41.70, 48.56,
14 0.80 s 0.99 d (6.6)
48.64, 50.55, 184.59, 212.01. MS (EI, 70 eV) m/z (rel.
15 0.86 d (6.6)
int): 252 (13) [M]+, 234 (7), 206 (15), 191 (11), 163 (36),
a
All the measurements were done in C6D6. The assignments were 149 (7), 133 (10), 121 (18), 107 (25), 95 (68), 81 (43), 67
established by 13C–1H HMBC and HMQC (22), 55 (28), 43 (100).
 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810 809

3.11. Decarboxylation of keto-acid 28 under normal pressure for 15 min. The solvent was
removed under reduced pressure and petroleum ether
The keto-acid (40 mg, 0.16 mol) and silver nitrate (1 was added, since most of the catalyst remains undis-
mg) were dissolved in acetonitrile (6 ml) and water (2 solved. Filtration through a column of alumina afforded
ml) and heated to reflux. To this solution potassium the pure alkane 31. MS (EI, 70 eV), [M]+ m/z=208.
peroxosulfate (K2S2O8) (0.32 mg) in water (5 ml) was b. The hydrogenation of (+)-g-gurjunene (9) followed
slowly added. Refluxing was continued for another 5 the same procedure as that of the alkene 30 except the
min before the reaction mixture was cooled and extrac- catalyst was replaced by nickel on charcoal.
ted with a saturated sodium bicarbonate solution (10
ml, three times), dried (MgSO4) and purified by prep. 3.14. Isolation of cypera-2,4(15)-diene and
GC to give 14 mg of ketone 28. norrotundene
1
H NMR (500 MHz, CDCl3): 0.96 (3H, dd, J=6.62
Hz), 1.06 (3H, s), 1.40 (1H, m), 1.60–1.84 (7 H, m), For the above described chemical transformation, a
1.97–2.1 (2H, m), 2.13–2.24 (3H, m), 2.27 (3H,s), 2.44 (1 relatively large amount of rotundene had to be isolated
H, m), 2.69 (1H, dd, J1=13.87 Hz, J2=6.94 Hz). 13C from the essential oil of C. rotundus. For the isolation 5 g
NMR (100 MHz, CDCl3): 15.10 (q), 21.52 (q), 26.09 (t), of the hydrocarbon fraction of the oil were separated by
26.80 (t), 27.85 (q), 30.26 (t), 33.25 (t), 37.46 (t), 37.99 column chromatography over silver nitrate precoated
(d), 38.04 (d), 46.82 (d), 56.70 (d), 61.65 (d), 207.93 (s). silica using a petroleum ether–chloroform gradient for
MS, (EI, 70 eV), m/z (rel. int): 208 (5) [M]+, 190 (7), 175 elution. The eluent from the column was monitored by
(6), 161 (5), 150(8), 135 (9), 123 (27), 109 (23), 95 (100), capillary GC and 16 fractions were collected. Fractions
81 (31), 67 (25), 55 (29), 43 (52). 1–6 obtained by elution with petroleum ether contained
only known hydrocarbons. Fractions 7–10 were obtained
3.12. Reduction of ketone 29 by elution with petroleum ether–chloroform (7/3) and
contained the rotundene. It could be proved by GC–MS
In a three-necked flask sodium hydride (0.45 mmol) of fraction eighth that the sample also contained an
was washed with several portions of pentane to remove unknown minor component. The separation of this frac-
the mineral oil. The flask was then equipped with rubber tion was performed by prep. GC (column: 2,6-Me-3Pe-b-
stopper, a reflux condenser and a magnetic stirrer. The CD; temp. programme: 100–140  C, heating rate 2  C/
system was alternately evacuated and filled with nitrogen; min). The chromatogram displayed five peaks the fifth
3 ml of dimethyl sulfoxide was introduced via a syringe containing the pure unknown product which was char-
and the mixture was heated at 75–80 C for 45 min. The acterized as cypera-2,4(15)-diene (24). Fractions 11–15
resulting solution of methylsulfinyl carbanion was cooled were eluted with petroleum ether–chloroform (1/1).
in an ice-water bath, and 108 mg of methyltriphenylpho- GC–MS revealed the presence of an unknown product
sphonium bromide in 100 ml of warm dimethyl sulfoxide of molecular weight 190 among many known sesqui-
were added. The resulting dark red solution of the ylide terpene hydrocarbons. In order to isolate the unknown
was stirred at room temperature for 10 min before 7 mg of compound, the joined fractions 14 and 15 were first
the ketone 29 in two ml of dimethyl sulfoxide were added. separated by prep. TLC over silver nitrate precoated
The reaction mixture was then heated at 56 C for 16 h plates, using petroleum ether–chloroform (7/3) as elut-
after which the solution was allowed to cool and 10 ml of ing solvent. Three bands were obtained and the third
water were added. The two phases were extracted three one, which contained the norsesquiterpene as major
times with n-pentane (40 ml). The n-pentane fractions product was separated by preparative GC (column: SE
were combined and washed with 10 ml of a (1/1) water/ 30, temp. 125  C). Spectroscopic data permitted the
dimethyl sulfoxide solution and then with 30 ml of 50% characterization of the compound as norrotundene (25).
saturated sodium chloride solution. The pentane layer
was then dried over anhydrous sodium sulfate and pur- 3.14.1. (
)-Cypera-2,4(15)-diene (24)
1
ification by preparative GC gave 2 mg of the alkene 30. H NMR (400 MHz, C6D6): see Table 2. 13C NMR
MS (EI, 70 eV), m/z (rel. int): 206 (12) [M]+, 191 (18), (400 MHz, C6D6) : 17.32 (q), 19.61 (q), 26.36 (q), 26.64
177 (6), 163 (51), 149 (25), 122 (39), 107 (56), 95 (60), 81 (q), 27.70 (t), 28.48 (s), 30.16 (t), 32.58 (t), 44.96 (d), 47.66
(100), 67 (57), 55 (66), 41 (79). (d), 61.93 (s), 102.84 (t), 133.26 (d), 139.17 (d), 148.94 (s).
MS (EI, 70 eV) m/z (rel. int.): 202 (83) [M]+, 187 (24),
3.13. Hydrocarbon 31 173 (6), 173 (6), 159 (46), 145 (24), 131 (37), 118 (85), 106
(100), 91 (78), 77 (33), 69 (17), 65 (18), 55 (26), 41 (61).
a. 30 (1 mg) was dissolved in a mixture of ethanol (1
ml) and benzene (1 ml). The catalyst [(Ph)3P]3RhCl (1 3.14.2. (
)-Norrotundene (25)
1
mg) was added and the solution turned orange. A gentle H NMR (500 MHz, C6D6):see Table 1. 13C NMR
stream of hydrogen was then passed through the solution (125 MHz, C6D6) : 14.73 (q), 22.95 (t), 28.00 (t), 28.67
810 M.M. Sonwa, W.A. König / Phytochemistry 58 (2001) 799–810

Table 2 vent evaporated. The residue was then purified by prep.

1
H NMR spectral data for compounds 24 and 26a GC to give 3 mg of cyperadione (26).
H 24 26

2 6.17 d (5.6) 1.50 ddd (15.4, 11.7, 5.7)

2.11 m
Acknowledgements
3 5.69 dd (5.6, 1.0) 2.35 ddd (16.4, 11.7, 4.4)
2.44 ddd (16.4, 12.3, 5.7) The financial support of DAAD (scholarship for M.
5 3.03 m Mekem Sonwa) and of the Fonds der Chemischen
6 1.68 m 1.96 d (19.0)
2.10 m 2.48 dd (19.0, 7.3) Industrie is gratefully acknowledged. We also thank Dr.
7 1.85 m 1.90 ddd (6.6, 3.3, 3.3) V. Sinnwell, University of Hamburg, for his support in
8 1.51 dt (14.8, 6.1) 1.45 dq (13.6, 3.2) obtaining the NMR spectra and to A. Meiners and M.
1.86 m 2.01 dddd (13.6, 12.9, 6.0, 2.7)
9 1.03 m 1.10 dddd (14.5, 12.9, 12.9, 6.6) Preusse for recording the mass spectra.
1.29 m 1.68 ddd 14.5, 6.0, 6.0)
10 2.12 dddd (13.2, 6.1, 6.1, 6.1)
12 0.99 s 1.22 s References
13 1.01 s 0.98 s
14 0.77 d (6.6) 0.78 d (6.6)
15 4.98 m 2.18 s Brown, H.C., Geoghegam jr, P.J., 1970. Solvomercuration-demer-
4.98 m curation. I. The oxymercuration-demercuration of representative
olefins in aqueous system. A convenient mild procedure for the
a
All the measurements were done in C6D6 for 24 and in CDCI3 for Markownikov hydration of the carbon–carbon double bond. Jour-
26. The assignments were established by 13C–1H HMBC and HMQC nal of Organic Chemistry 35, 1844–1850.
Dassanayake, M., Fosberg, F.R., 1985. A Revised Handbook of the
(t), 29.36 (q), 30.35 (d), 30.54 (t), 31.97 (t), 35.72 (s), Flora of Ceylon, part V. A. A. Balkema, Rotterdam, p. 181.
Fristad, W.E., Fry, M.A., Klang, J.A., 1983. Persulfate/silver ion
36.32 (d), 42.18 (d), 50.74 (d), 129.15 (d), 139.10 (d). decarboxylation of carboxylic acids. Preparation of alkanes,
MS (EI, 70 eV) m/z (rel. int.) : 190 (10) [M]+, 175 (15), alkenes, and alkohols. Journal of Organic Chemistry 48, 3575–3577.
161 (20), 148 (7), 133 (9), 119 (7), 107 (14), 94 (100), 79 Greenwald, R., Chaykovsky, M., Corey, E.J., 1963. The Wittig reac-
(50), 67(12), 55 (14), 41 (23). tion using methylsulfinyl carbanion-dimethyl sulfoxide. Journal of
the American Chemical Society 28, 1128–1129.
Greenwood, J.M., Solomon, M.D., Sutherland, J.K., Torre, A., 1968.
3.15. Isolation of cyperadione (26) A cyclisation of humulene. Journal of the Chemical Society (C),
3004–3008.
The fraction of oxygenated sesquiterpenes was sub- Haaksma, A.A., Jansen, B.J.M., de Groot, A., 1992. Lewis acid cata-
mitted to prep. TLC using petroleum ether–ethyl ace- lysed Diels-Alder reactions of S-(+)-carvone with silyloxy dienes.
tate (8/2) as eluting solvent. The second band (Rf=0.35) Total synthesis of (+)-a-cyperone. Tetrahedron 48, 3121–3130.
Harmon, R.E., Parsins, J.L., Cooke, D.W., Gupta, S.K., School-
was then taken up in diethyl ether and submitted to enberg, J., 1969. Homogeneous homocatalytic hydrogenation of
further separation by prep. GC (column: 6T-2,3-Me-b- unsaturated organic compounds. The Journal of Organic Chemistry
CD, 120–180  C, 2  C/min). 34, 3684–3685.
Hikino, H., Ito, K., Aota, K., 1966. Structure and absolute config-
3.15.1. Cyperadione (26) uration of cyperotundone. Chemical and Pharmaceutical Bulletin
1 14, 890–899.
H NMR (400 MHz, CDCl3): see Table 2. 13C NMR Hikino, H., Takemoto, T., 1967. Structure and absolute configuration
(100 MHz, CDCl3) :  16.76 (q), 21.87 (t), 22.18 (q), of cyperol and isocyperol. Chemical and Pharmaceutical Bulletin
26.42 (t), 27.05 (q), 28.10 (t), 30.03 (q), 31.32 (d), 38.71 1929–1933.
(t), 41.79 (d), 41.84 (t), 42.96 (s), 58. 21 (s), 208.50 (s), Howe, R., McQuillin, F.J., 1955. The structutre of cyperone. Part IV.
220.25 (s). MS (EI, 70 eV), m/z (rel. int.): 236 (66) [M]+, The synthesis of natural (+)-a-cyperone, its enantiomorph and
epimer. Journal of the Chemical Society, 2423–2428.
221 (89), 207 (7), 193 (25), 179 (63), 161 (17), 150 (20), Joulain, D., König, W.A., 1998. The Atlas of Spectral Data of Ses-
135 (22), 123 (22), 107 (25), 95 (19), 81 (20), 67 (23), 55 quiterpene Hydrocarbons. E. B.-Verlag, Hamburg.
(27), 43 (100). McMurry, J.E., Bosch, G.K., 1987. Synthesis of macrocyclic terpenoid
hydrocarbons by intramolecular carbonyl coupling: bicyclogerma-
3.16. Ozonolosis of cyperene (4) crene, lepidozene and casbene. Journal of Organic Chemistry 52,
4885–4893.
Mekem Sonwa, M., König, W.A., Kubeczka, K.-H., Motl, O., 1997.
Three mg of cyperene in dichloromethane (20 ml) Sesquiterpenes from the essential oil of Cyperus alopecuroides. Phy-
were treated at
78  C with a stream of ozone until a tochemistry 45, 1435–1439.
persistant blue colour was observed. The solution was Nyasse, B., Ghogumu Tih, R., Sodengam, B.L., Martin, M.T., Bodo,
B., 1988. Mandassidione and other sesquiterpenic ketones from
then purged with nitrogen for 5 mins. 1 ml of dimethyl-
Cyperus articulatus. Phytochemistry 27, 3319–3321.
sulfide was then added, and washed with 10 ml of an Ohira, S., Hasegawa, T., Hyashi, K.-I., Hoshino, T., Takaoka, D.,
ice-cold sodium bicarbonate solution, water and brine. Nozaki, H., 1998. Sesquiterpenoids from Cyperus rotundus. Phy-
The organic layer was dried over MgSO4 and the sol- tochemistry 47, 1577–1581.