FLAVOUR AND FRAGRANCE JOURNAL, VOL.

8,39-41(1993)

A Novel Sesquiterpene, 1,2-Epoxyfurano-l0(15)-germacren-6-one,

from the Resin of Commiphora holtziana Engl.*
Iain S. Cavanagh, Michael D. Cole," Simon Gibbons, Alexander I. Gray, Gordon J. Provant and Peter
G. Waterman
Phytochemistry Research Laboratories, Department of Pharmaceutical Sciences, University of Strathclyde, Glasgow GI I X W,
Scotland, UK
'Forensic Science Unit, Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow GI IXW, Scotland, UK

Three known and one novel furanogermacrenes have been isolated from the resinous exudate of Commiphora
h o l t z i a ~Engl. The structures of the known compounds were determined by comparison of 'H-and "C-NMR
spectra with those already published. The structure of the novel compound was determined as 1.2epoxyfurano-
10(15)-gemacren-6-one (IV),using spectroscopic techniques. Some previous '%-NMR assignments for the known
compounds were corrected or clarified.
KEY WORDS Comiphora holtziana Engl. Buneraceae Resin exudate Furanogermacrene NMR spectroscopy
1,2-Epoxyfurano-lO(1S)-germacrend-one

INTRODUCTION Extraction and Isolation of Sesquiterpenes

Resin (5Og) was extracted at 100mgml-' in
The genus Commiphora (Burseraceae) forms, with
Acacia (Leguminosae), the dominant flora over EtOH at room temperature for two days. The
much of northern and Kenya, southern Ethiopia EtOH was removed under vacuum, yielding a
and Somalia.' The resins from the African Bursera- yellow oil (25 8). Compounds I-III were isolated
ceae are important items of commerce, as glues, tick from 12.5g of the oil, after vacuum liquid chroma-
repellents, medicinals and perfume^.'^ The chemi- tography (silica gel, 230-4000 mesh, 6 cm x 6.5 an
i.d.) using petroleum ether:EtOAc mixtures of in-
cal constituents of the resins are still not well creasing polarity, and PTLC on Si gel plates (layer
documented.' As part of our continued investiga-
tion of the chemistry of this genus, we have exa- thickness 1 mm) developing with toluene: EtOAc:
HOAc 93:6:1 v/v/v finally yielding I (68mg), 11
mined a sample of the resin of C. holtziana Engl. We (164mg) and III(12 mg). Traces of a further com-
wish to report the isolation from this sample, of pound, later identified as IVYwere also observed
three known furanogermacrenes I-III that we had during the separation process.
isolated previously from other material of this
species,' and the isolation and structural determi-
nation of a fourth, 1,2-epoxyfurano-l0(15>germa-
men-bone (IV). The opportunity has been taken to
correct and clarify assignments in the 13C-NMR
spectra originally reported for I-III.5
I4 k
I LI
EXPERIMENTAL

Plant Material Me0

The resin used in this study was a commercial
sample obtained in Wajir, Kenya, in 1984. m Iv
Part 16 in the series 'Chemistry of the Burseraceae'. For Part 15 see Phytochemistry, 31,2065 (1992).
t Present address: The Rowett Research Institute, Bucksburn, Aberdeen AB2 9SB, Scotland, UK.
0882-5734/93/010039-03S06.5O Received I0 August 1992
Q 1993 by JohnWiley Br Sons, Ltd Accepted 2 October 1992
40 I. S. CAVANAGH ETAL

The second batch of oil (12.5 g) was subjected to presence of an a,/l-unsaturated ketone (1671 an-'),
vacuum liquid chromatography (silica gel, 3 cm x and the presence of an epoxide was suggested by
4cm i.d.) using 3 x 10ml aliquots of petroleum bands at 1245,905 and 798 an-'.
ether: EtOAC mixtures of increasing polarity The 'H-and "C-NMR spectra (Tables 1 and 2)
(100-92 % petroleum ether in 1 % steps). The sol- indicated that IV was a furanogermacrene similar
vent was removed from the fractions enriched in IV to I-III with two methyl, four methine, four methy-
and a solution of the residue prepared at lene and five quaternary carbons. A 2H singlet at 6
40mgml-' in EtOH. 2 0 0 4 aliquots of this sol- 4.94 (6, 110.6) indicated the presence of an exo-
ution were subjected to semi-preparative HPLC on cyclic methylene. The HMBC spectrum6 showed
a silica gel column (Spherisorb 5, 25 cm x 10 mm these protons to have a ' 5 coupling to a quaternary
i.d.), using an eluant of 5 % EtOAc in iso-octane,
with a flow rate of 3 ml min- and detection at
254nm. The components of the fractions further
Table 1. 'H-NMR data for compound IV
enriched in IV were finally separated by PTLC
(silica gel, 1 mm layer thickness) in to1uene:EtOAc: Proton
HOAc 97:2: 1 using multiple development, yielding no.
N as a viscous oil (11.5 mg).
H-1 2.96 brs
1,2-Epoxyfuruno-10( 15)-germacren-6-one (IV). H-2 1.92 dt (10.2,23)
IR data of IV were obtained from a thin film from H-3 1.08 m
CHC13on KCl: vmar (cm-') 2958,2930,1733,1671, H-3 2.10 m
1543, 1382, 1245, 987, 905. NMR data were re- H-4 2.15 m
corded in CDCl, using a Bruker 400MHz spec- H-5 2.32 dd (11.9, 13.9)
H-5 295 ddd (11.9. 1.4, 3.2)
trometer (Tables 1 and 2). MS (rel. int.) m/z 246 H-9 3.47 d (14.3)
(15) (calculated for C,,H,80,246.1228, found H-9 4.18 d (14.3)
246.1256), 230 (4). H-12 7.03 q (1.2)
Me-13 2.13 d (1.2)
Me-14 1.12 d (6.8)
RESULTS AND DISCUSSION
H-15
H-15 1 4.94 s

Four furanogermacrene sesquiterpenoids were

isolated from the resin of C.holttiuna. Three of the
compounds I-III have previously been reported as Table 2. 13C-NMR data for compounds I-IV
constituents of the resin of C. holtzianu? and were
identified by comparison of 'H and NMR Chemical shift (a,)
spectra with those reported when they were origin- Carbon
ally isolated from C. m ~ l m o l .In
~ that previous
study some "C NMR assignments were ambigu- no. I I1 m Iv
~~

ous. These have now been fully characterized using 1 133.6 d 132.8 d 135.2 d 59.9 d
the Heteronuclear Multiple Quantum Coherence 2 74.7 d 73.9 d 132.3 d 61.7 d
(HMQC) and Heteronuclear Multiple Bond Co- 3 37.1 t 31.9 t 88.5 d 40.2 t
herence (HMBC) techniques. These proton-de- 4 25.4 d 30.8 d 38.2 d 29.5 d
5 50.6 t 79.0 d 48.6 t 53.6 t
tected two-dimensional procedures6 give cross- 6 202.8 s 195.6 s 204.1 s 200.9 s
peaks indicating 1-bond (HMQC) and 2-bond (.'!) 7 119.8 s 123.3 s 117.9 s 123.6 s
and 3-bond ( , J ) (HMBC) heteronuclear (C-H) 8 151.6 s 154.4 s 152.0 s 157.3 s
couplings; in this study the HMBC experiment was 9 38.4 t 38.2 t 34.0 t 32.8 t
optimized for J = 7-Hz. Corrrection of previous 10 132.7 s 135.3 s 142.7 s 141.7 s
11 126.1 s 121.3 s 129.0 s 118.3 s
assignments was only required for C-14 and C-15 in 12 137.5 d 138.1 d 138.1 d 138.7 d
I and 11, C-5 and C-9 in I and C-7 and C-11 in II; 13 8.4 q 8.8 q 9.4 q 10.3 q
ambiguities relating to C-3 and C-9 methylenes in I 14 22.1 q 17.4 q 19.1 q 24.4 q
and II and C-1 and C-2 olefins and C-5 and C-9 15 18.2 q 18.9 q 115.9 t 110.6 t
OMe 55.6 q 55.9 q 56.8 q
methylenes in III were also resolved. Ac 170.3 s
The structure of IV was determined using spec- 20.8 q
troscooic methods. The IR sm t ru m indicated the
 1,2-EPOXYFURANO-10(15)-GERMACREN-6-ONE 41

carbon at 141.7ppm and ' J coupling to an oxy- ring then the only one is left to cover the require-
methine carbon (6, 59.9) and a methylene carbon ments for the two oxymethine carbons. This can
( ~ 5 ~ 3 2 .on
8 ) which the two non-equivalent protons only be achieved by the presence of an epoxide ring
formed an AB quartet showing no additional coup- and allows the complete assignment of the struc-
ling. The carbon at 6, 59.9 was directly bonded ture of N.The appreciable non-equivalence of the
(HMQC) to a resonance at 6 2.96. This proton two H-9 protons is presumed to be due to the close
coupled ('H-lH COSY) to a 1H doublet of triplets proximity of the exocyclic methylene. As a NOESY
at 6 1.92 (Sc61.7), that was also coupled with two spectrum reveals no interaction between H-1 and
nonequivalent methylene protons which in turn H-2 they are presumed to be trans.
coupled to a methine which could be linked directly
to a methyl and the final methylene (6 2.32, 2.95).
This allowed assignment of the partial structure Acknowledgements-1.S.C. was supported by a summer student-
R-CH,-C-(=CHz)-CH-(-O-)-CH- ship from the Isaac Sclaar Trust and S.G. by a studentshipfrom
the Science and Engineering Research Council. NMR spectra
(-O-)-CHz-CH-(CH3)-CHz-R. were obtained from the Strathclyde University NMR labora-
Further connectivities were assigned from the tory.
HMBC spectrum. This revealed the 6 2.32 proton
coupling to a carbonyl (6, 200.9) and quaternary
carbon at 6, 123.6. The latter also showed long-
range interactions with the protons of a vinylic REFERENCES
methyl and an olefinic proton (6 7.03). The olefinic
methine exhibited a further ' J coupling to an 1. A. Maradafu, Phytochemistry, 21,677 (1982).
2 G. J. Provan, A. I. Gray and P. G. Waterman, Flavour
oxygen-bearing quaternary carbon at 6,157.3. This Fragr. J., 2, 109 (1987).
allowed the formulation of a furan ring and with 3. G. Usher, Dictionary of Plants Used by Man, Constable,
the observation of an interaction between the 6, London (1984).
157.3 and the protons of the isolated methylene AB 4. J. F. Carroll, A. Maradafu and J. D. Warthen, Entomol. Exp.
quartet completed the connection of all carbons in Appl., 53, 111 (1989).
5. C. H. Brieskorn and P. Noble, Tetrahedron Lett., 21, 1511
lV.High-resolution electron impact MS gave an (1980).
empirical formula C1,H1,03. As one oxygen is 6. M. A. Quader, A. I. Gray, P. G.Waterman, C. Lavaud, G.
required for the carbooyl and another for the furan Massiot and I. H. Sadler, Tetrahedron, 47,3611 (1991).