FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fragr. J. 2001; 16: 253–256

DOI: 10.1002/ffj.994

Constituents of the essential oil of six Helichrysum

species from Madagascar
Jean-François Cavalli,1,2 Lalasoa Ranarivelo,3 Michel Ratsimbason,3 Antoine-François Bernardini2 and
Joseph Casanova1∗
1 Université
de Corse, Equipe Chimie et Biomasse, UMR-CNRS 6134, Route des Sanguinaires, 20 000 Ajaccio, France
2 Université
de Corse, Equipe Chimie des Produits Naturels, UMR-CNRS 6134, BP 52, 20 250 Corte, France
3 Centre National d’Application des Recherches Pharmaceutiques, BP 702, Tananarive 101, Madagascar

Received 14 July 2000

Revised 12 January 2001
Accepted 16 January 2001

ABSTRACT: The essential oils of six Helichrysum species from Madagascar were investigated and 46
components were identified by GC, GC–MS and 13 C-NMR spectrometry. The main constituents were 1,8-
cineole for H. gymnocephalum (59.7%) and H. bracteiferum (27.3%); ˇ-pinene for H. selaginifolium (38.2%);
(E)-caryophyllene for H. cordifolium (55.6%), H. faradifani (34.6%) and H. hypnoides (34.0%). Copyright 
2001 John Wiley & Sons, Ltd.
KEY WORDS: Helichrysum gymnocephalum; H. bracteiferum; H. selaginifolium; H. cordifolium; H. faradifani ;
H. hypnoides; Asteraceae; essential oil composition; Madagascar; GC–MS; 13 C-NMR.

Introduction deposited at the Herbarium of the Centre National

The genus Helichrysum (Asteraceae), to which more
than 400 species belong, is widespread all around the
world (Europe, Africa, Australia, North America). In
Madagascar 115 species, mostly endemic, are currently
known1,2 and some are used in folk medicine. Essen-
tial oils and extracts are obtained from the whole plant
or from different parts of the plant. They are used
in perfumery and aromatherapy. Some of them exhibit
an antibacterial activity.3,4,5 The chemical composition
of Helichrysum oils has been recently reviewed by
Lawrence.6
As part of our ongoing work on the composi-
tion of the essential oils from Madagascar, we inves-
tigated six Helichrysum species: H. gymnocephalum
(DC) H. Humbert, H. bracteiferum (DC) H. Humbert,
H. selaginifolium Vig. et Humb., H. cordifolium DC.,
H. faradifani Sc. Ell., and H. hypnoides (DC) Vig. et
Humb. To our knowledge, the oil composition of three
of these is reported for the first time.
Experimental
Plant Material and Oil Isolation
d’Application des Recherches Pharmaceutiques
(CNARP; Department of Botany). The fresh aerial parts
of the plants, cut into small pieces, were water-distillated
using a Clevenger-type apparatus. For each species, the
locality of harvest, the date of harvest, the voucher spec-
imen number and essential oil yield are as follows:
H. gymnocephalum, Anjozorobe, March 1997, HE 2830,
0.41%; H. bracteiferum, Behenjy, February 1999, HE
1075, 0.16%; H. selaginifolium, Anjozorobe, May 1995,
HE 2831, 0.01%; H. cordifolium, Fandriana, June 1997,
HE 2793, 0.03%; H. faradifani, Anjozorobe, May 1995,
HE 2792, 0.07%; H. hypnoides, Anjozorobe, May 1995,
HE 2833, 0.012%.
Analytical GO
GC analyses were carried out using a Perkin-Elmer
Autosystem GC apparatus equipped with FID detec-
tor and fused-silica capillary columns (50 m ð 0.22 mm
i.d., film thickness 0.25 µm), BP-1 (polydimethylsilox-
ane) and BP-20 (polyethyleneglycol), as previously
reported.7

Helichrysum plants were harvested in different locali- GC–MS

ties in Antananarivo province. Voucher specimens were
*Correspondence to: J. Casanova, Université de Corse, Equipe Chimie
et Biomasse, UMR-CNRS 6134, Route des Sanguinaires, 20 000
Ajaccio, France. E-mail: casanova@vignola.univ-corse.fr
Samples were analysed with a Perkin Elmer Turbo mass
detector, directly coupled to a Perkin Elmer Autosys-
tem XL equipped with fused-silica capillary columns
(50 m ð 0.22 mm i.d., film thickness 0.25 µm) BP-1

Copyright  2001 John Wiley & Sons, Ltd.

254 J.-F. CAVALLI ET AL.
(polydimethylsiloxane) and Rtx-WAX (60 m ð 0.25 mm
i.d., film thickness 0.25 µm, polyethyleneglycol). Ion
source temperature, 180 ° C; energy ionization, 70 eV;
electron ionization mass spectra were acquired oven the
mass range 35–350 Da; Split, 1/80. Other GC conditions
were as described for GC.
13
C-NMR
NMR spectra were recorded on a Bruker AC 200 Fourier
Transform spectrometer operating at 50.323 MHz for
13
C, equipped with a 10 mm probe (200 mg of the oil
in 2 ml CDCl3 ). Other parameters were as previously
reported.7
Component Identification
Identification of the components was based: on (a) their
GC retention indices (RI) on polar and apolar columns,
determined relative to the retention time of a series of
n-alkanes, with linear interpolation with those of authen-
tic compounds or literature data,8 (b) computer matching
with commercial mass spectral libraries9,10 and compar-
ison of spectra with those of our own library or literature
data;8,11 (c) 13 C-NMR, following the methodology first
reported by Formácek and Kubeczka12 and improved and
computerized in our laboratory.13
Results and Discussion
Analysis of the six samples of Helichrysum oils by GC,
GC–MS and 13 C-NMR spectroscopy allowed the iden-
tification of 46 components: 14 monoterpene hydrocar-
bons, eight oxygenated monoterpenes, 16 sesquiterpene
hydrocarbons, seven oxygenated sesquiterpenes and one
phenyl propanoid (Table 1).
Helichrysum gymnocephalum produces a monoter-
pene rich oil (83.2%) characterized by the preemi-
nence of 1,8-cineole (nearly 60% of the oil) while
the 10 identified sesquiterpenes represented only 8.8%
of the whole composition. The main components of
H. selaginifolium oil are ˇ-pinene (38.2%), ˛-pinene
(16.3%), caryophyllene and its oxide (respectively 7.5%
and 9.3%), while 1,8-cineole accounted for only 7.1%.
1,8-Cineole (27.3%), ˇ-pinene (11.9%) and ˛-pinene
(5.9%) are the major monoterpenes of H. bracteiferum
oil, while two sesquiterpene hydrocarbons, ˛-humulene
and (E)-caryophyllene (10.1% and 7.1%) are present in
appreciable amounts.
The compositions of our samples are very close
to these reported respectively by Möllenbeck et al.14
for the two former and by Ramanoelina et al.15 for
Copyright  2001 John Wiley & Sons, Ltd.
the latter, at least for the major components. How-
ever, several differences occurred in the qualitative
and quantitative analysis of the minor components.
Indeed, several sesquiterpene hydrocarbons (such as
ˇ-elemene, germacrene D and ˛-muurolene), as well as
oxygenated sesquiterpenes (such as spathulenol, glob-
ulol, humulene-6,7-epoxide, T-cadinol) and the phenyl
propanoid eugenyl acetate, not previously reported, are
present in our samples at moderate to appreciable
amounts. A special mention should be made of humu-
lene epoxide, which was present in the three sam-
ples (0.9–1.1%) and was identified by comparison of
its carbon chemical shifts with those reported in the
literature.16 Conversely, the composition of our sam-
ple of H. gymnocephalum differed from that reported by
De Medici et al.17 (1,8-cineole, 15–17%) or by Theron
et al.18 (1,8-cineole, 14–20%).
To our knowledge, the composition of the essential
oils obtained from the three other species has not been
reported previously in the literature. In contrast to the
samples previously investigated, as well as other oils
from Helichrysum species, the sesquiterpene hydrocar-
bon (E)-caryophyllene is the major constituent of the
three oils.
H. cordifolium produced a (E)-caryophyllene-rich oil
(55.6%). Besides caryophyllene, the two biologically
related sesquiterpenes, ˛-humulene and caryophyllene
oxide, are the main constituents (12.1 and 7.1% respec-
tively). We noted the presence, in the oil, of humulene
epoxide (1.4%) as well as several sesquiterpenes bearing
the bisabolane or the bicyclo[4.4.0]decane skeletons.
(E)-Caryophyllene represented around the third of
the composition of H. faradifani and H. hypnoides oils
(34–35%). These oils differ by the second major com-
ponent, which is linalool (16.1%) for the former and
1,8-cineole (13.4%) for the latter.
Our samples of Helichrysum oil were characterized by
an appreciable to high content of 1,8-cineole, ˇ-pinene or
(E)-caryophyllene. To our knowledge, none of the com-
positions reported in the literature for other Helichrysum
oils exhibited high contents of 1,8-cineole or ˇ-pinene,
even though these compounds have been identified in
H. splendidum oil (8.6% and 10.2%, respectively).19
Conversely, (E)-caryophyllene was recently reported as
a major component (38.5%) of H. heldreichii essen-
tial oil from Greece20 and was present in H. stoechas
subsp. barrelieri oil from Greece21 (15.6% associated
with ˇ-elemene, 13.1%) and H. odoratissimum oils from
Cameroon22 (13.8 and 5.1%).
Our results confirmed that Helichrysum species exhib-
ited a very important chemical variability. The compo-
sition of the samples investigated in this study differed
drastically from those reported for other Helichrysum
oils. Indeed, below are the major components of essen-
tial oils whose compositions are reported in the literature,
as well as the origin when available:
Flavour Fragr. J. 2001; 16: 253–256
 ESSENTIAL OIL OF HELICHRYSUM 255

Table 1. Constituents of six essential oils collectives from Helichrysum species from Madagascar. H. gymnocephalum;
H. bracteiferum; H. selaginifolium; H. cordifolium; H. faradifani; H. hypnoides

RI
No. Compound BP-1 BP-20 A B C D E F Identification
1 ˛-Thujene 924 1023 0.5 0.6 — — — — RI, MS
2 ˛-Pinene 931 1023 4.9 5.9 16.3 1.0 2.2 5.0 RI, MS, 13 C

3 ˛-Fenchene 942 1058 — — — 0.2 — 0.9 RI, MS, 13 C

4 Camphene 944 1068 — — 0.2 0.2 0.4 0.4 RI, MS
5 Sabinene 967 1124 1.4 2.0 1.2 — — — RI, MS, 13 C

6 ˇ-Pinene 971 1111 3.3 11.9 38.2 2.0 0.5 1.7 RI, MS, 13 C

7 Myrcene 981 1160 1.0 1.0 0.3 0.3 1.5 2.1 RI, MS, 13 C

8 ˛-Terpinene 1011 1183 — 0.5 — — — — RI, MS, 13 C

9 p-Cymene 1013 1271 6.3 2.0 2.2 0.3 0.8 2.0 RI, MS, 13 C

10 Limonene 1023 1204 1.7 4.0 1.7 2.1 4.6 6.9 RI, MS, 13 C

11 1,8-Cineole 1023 1215 59.7 27.3 7.1 0.2 2.9 13.4 RI, MS, 13 C
12 (Z)-ˇ-Ocimene 1025 1232 — — — 0.3 0.3 0.2 RI, MS
13 (E)-ˇ-Ocimene 1037 1249 — — — 0.9 0.6 1.4 RI, MS, 13 C

14 -Terpinene 1049 1244 — 0.6 — — — 0.3 RI, MS, 13 C

15 Terpinolene 1080 1283 — 0.2 — — 0.3 — RI, MS
16 Linalool 1085 1547 0.6 1.2 1.3 1.0 16.1 4.4 RI, MS, 13 C

17 trans-Pinocarveol 1125 1654 — — 1.0 — — — RI, MS, 13 C

18 Borneol 1150 1705 — — — — 0.6 0.3 RI, MS, 13 C

19 Terpinen-4-ol 1164 1604 2.6 1.8 — — — — RI, MS, 13 C

20 Myrtenal 1171 1632 — — 0.5 — — — RI, MS
21 ˛-Terpineol 1173 1698 1.2 1.1 1.1 — 1.9 1.3 RI, MS, 13 C

22 Myrtenol 1181 1791 — — 0.7 — — — RI, MS, 13 C

23 ˛-Copaene 1376 1488 — 0.3 0.5 — 0.6 1.8 RI, MS, 13 C

24 ˇ-Elemene 1389 1588 — 1.2 — — — — RI, MS, 13 C

25 (E)-Caryophyllene 1423 1593 1.6 7.1 7.5 55.6 34.6 34.0 RI, MS, 13 C
26 Aromadendrene 1440 1604 — 1.0 0.4 — 0.4 — RI, MS, 13 C
27 allo-Aromadendrene 1448 1638 — — 0.3 — 0.6 0.4 RI, MS
28 ˛-Humulene 1453 1665 1.8 10.1 0.9 12.1 5.4 4.7 RI, MS, 13 C

29 -Muurolene 1470 1680 0.3 0.9 — 0.3 0.5 0.5 RI, MS, 13 C

30 ˛-Curcumene 1470 1767 — — — 1.4 1.3 0.4 RI, MS, 13 C

31 -Curcumene 1473 1685 — — — 0.5 — — RI, MS, 13 C

32 Germacrene D 1479 1705 — 1.5 — — — — RI, MS, 13 C

33 ˇ-Selinene 1483 1713 — 1.2 0.2 — 0.8 0.3 RI, MS, 13 C

34 Eugenyl acetate 1486 2263 — 0.2 — — — — RI, MS
35 Bicyclogermacrene 1494 1729 — 1.3 — — — — RI, MS, 13 C

36 ˛-Muurolene 1495 1720 0.5 — — — — — RI, MS, 13 C

37 -Cadinene 1507 1752 0.8 0.5 0.3 0.7 2.0 1.2 RI, MS, 13 C
38 Calamenenea 1510 1826 0.2 0.2 — 0.2 0.5 0.3 RI, MS
39 υ-Cadinene 1515 1749 0.5 0.9 — 1.1 3.0 1.5 RI, MS, 13 C
40 Spathulenol 1571 2125 — 2.0 0.7 — — — RI, MS, 13 C

41 Caryophyllene oxide 1576 1984 1.6 0.8 9.3 7.1 4.3 5.3 RI, MS, 13 C

42 Globulol 1576 2076 — 0.4 0.2 — — — RI, MS, 13 C

43 Humulene 6,7-epoxide 1601 2044 1.1 0.9 0.9 1.4 0.5 0.7 RI, MS, 13 C

44 T-Cadinol 1629 2170 0.4 0.2 — 0.5 1.3 — RI, MS, 13 C

45 ˇ-Eudesmol 1639 2232 — 0.4 — 0.4 0.7 — RI, MS, 13 C

46 ˛-Bisabolol 1668 2215 — — — 0.4 — — RI, MS, 13 C

Totalb 92.0 91.2 93.0 90.2 89.2 91.4

a
Correct isomer not identified.
b Percentage on a BP-20 column.

ž ˛-Pinene: H. stoechas var. ˛-syncladum (59%, Por-
tugal23 ); H. odoratissimum (40.6 and 47.1%, Came-
roon,22 43.4%, Kenya,24 15.0% associated with
˛-humulene 13.0%, Zimbabwe25 ); H. stoechas subsp.
stoechas (28.3%, associated with epi-˛-bisabolol,
21.9%, Spain26 ); H. italicum (up to 29.9%, Adriatic
coast27 and 21.7%, Yugoslavia28,29 ).
ž -3-Carene: H. picardii (60.0–74.3%, Spain30 ).
ž -Terpinene: H. splendidum (14.9%, Zimbab-
we19 ).
ž Geraniol : H. taenari (50.0%, associated with cam-
phene, 18.6%, Greece21 ), H. italicum subsp. italicum
(35.6%, Greece4 ) and H. amorginum (32.1%, associ-
ated with geranyl acetate, 20.8%, Greece4 ).
ž Neryl acetate: H. italicum subsp. microphyllum
(38.6 š 15.1%, North America,31 and 28.9 š 3.5%,
Sardinia32 ), H. italicum subsp. italicum (16.6 š 2.0%,
North America,31 15.8–42.5%, Corsica33 ).
ž Carvacrol : H. doerfleri (42.5%, during anthesis, Gre-
ece20 ).

Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 253–256
256 J.-F. CAVALLI ET AL.
ž ˇ-Selinene: H. italicum subsp. microphyllum (17.1%
associated with -curcumene 13.7%, Greece20 ).
ž ˛-Curcumene: H. italicum (up to 28.6%, Adriatic
coast27 ).
ž Rosifoliol : H. italicum subsp. microphyllum (20.2 š
3.2%, associated with -curcumene 18.2 š 2.2% and
linalool (14.9 š 2.2%, Sardinia32 ).
ž Guaiol : H. italicum subsp. serotinum (9.0%, associ-
ated with nonadecane, 7.4% and nerol, 7.0%, Spain26 ).
ž ˛-Eudesmol : H. doerfleri (11.7% associated with three
isomers, before anthesis, Greece20 ).
ž Manoyl oxide: H. rupestre (33.3%, Balearic Islands5 ).
ž 2,4,6-Tris(1,1-dimethylethyl)benzoic acid : H. ambi-
guum, (48.6%, Balearic Islands5 ).
ž Hexadecanoic acid : H. orientale (17.5% associated
with nonacosane, 11%, Greece20 ).
From this literature review it can be seen that:
(a) Helichrysum oils from Mediterranean countries with
a temperate climate [continental Spain and Balearic
Islands, France (Corsica), Italy (Sardinia), Adriatic coast,
Greece], exhibited an immense chemical variability;
(b) North American oils were reported as neryl acetate-
rich oils; (c) Helichrysum oils from Africa (Cameroon,
Kenya, Zimbabwe) were reported as ˛-pinene-rich oils.
Our results confirmed the originality of Helichrysum oils
from Madagascar, whose compositions were dominated
by cineole, ˇ-pinene or (E)-caryophyllene.
References
1. Humbert H. Flore de Madagascar et des Comores, 189e. Famille:
Composées, Tome II. Museum National d’Histoire Naturelle:
Paris, 1962.
2. Koechlin J, Guillaumet JL, Morat P. Flore et Végétation de Mada-
gascar. A.R.G. Gantner Verlag: Vaduz, 1997.
3. Ramanoelina ARP, Terrom GP, Bianchini JP, Coulanges P. Arch.
Inst. Pasteur Madagascar 1987; 53: 217.
4. Chinou IB, Roussis V, Perdetzoglou D, Loukis A. Planta Med.
1996; 62: 377.
5. Roussis V, Tsoukatou M, Chinou IB, Ortiz A. Planta Med. 1998;
64: 675.
6. Lawrence BM. Perfum. Flavor. 1998; 23(5): 55.
7. Mazzoni V, Tomi F, Casanova J. Flavour Fragr. J. 1999; 14:
268.
Copyright  2001 John Wiley & Sons, Ltd.
8. Joulain D, König WA. The Atlas of Spectral Data of Sesquiterpene
Hydrocarbons. E.B.-Verlag: Hamburg, 1998.
9. National Institute of Standards and Technology. PC Version 1.5a
of The NIST/EPA/NIH Mass Spectral Library. Perkin-Elmer Cor-
poration, 1997.
10. McLafferty FW, Stauffer DB. Wiley Registry of Mass Spectral
Data 6th edn. Mass Spectrometry Library Search System Bench-
Top/PBM, version 3.10d. Palisade: Newfield, 1994.
11. McLafferty FW, Stauffer DB. The Wiley/NBS Registry of Mass
Spectral Data 4th edn. Wiley-Interscience: New York,
1988.
12. Formácek V, Kubeczka KH. Essential Oil Analysis by Capillary
Gas Chromatography and Carbon-13 NMR Spectroscopy. Wiley:
Chichester, 1982.
13. Tomi F, Bradesi P, Bighelli A, Casanova J. J. Magn. Reson. Anal.
1995; 1: 25.
14. Möllenbeck S, König T, Schreier P, Schwab W, Rajaonarivony J,
Ranarivelo L. Flavour Fragr. J. 1997; 12: 63.
15. Ramanoelina PAR, Bianchini JP, Gaydou EM. J. Essent. Oil Res.
1992; 4: 531.
16. Gatilova VP, Korchagina DV, Gatilov YV, Bagryanskaya IY,
Barkhash VA. Zh. Org. Khim. 1991; 27: 2040.
17. De Medici D, Pieretti S, Salvatore G, Nicoletti M, Rasoanaivo P.
Flavour Fragr. J. 1992; 7: 275.
18. Theron E, Holeman M, Potin-Guatier M, Pinel R. Rivista Ital.
EPPOS 1994; 13: 33.
19. Chagonda LS, Makanda C, Chalchat JC. J. Essent. Oil Res. 1999;
11: 573.
20. Roussis V, Tsoukatou M, Petrakis PV, Chinou I, Skoula M, Har-
borne JB. Biochem. Syst. Ecol. 2000; 28: 163.
21. Chinou IB, Roussis V, Perdetzoglou D, Tzakou O, Loukis A.
Planta Med. 1997; 63: 181.
22. Kuiate JR, Zollo PHA, Nguefa EH, Bessière JM, Lamaty G,
Menut C. Flavour Fragr. J. 1999; 14: 82.
23. Proença da Cunha A, Cardoso do Vale J. Boletim da Faculdade
de Farmácìa, Universidade de Coimbra 1974; 34: 5.
24. Lwande W, Hassanali A, Wanyama OB, Ngola S, Mwangi JW. J.
Essent. Oil Res. 1993; 5: 93.
25. Gundidza MG, Zwaving JH. J. Essent. Oil Res. 1993; 5: 341.
26. Tsoukatou M, Roussis V, Chinou I, Petrakis PV, Ortiz A.
J. Essent. Oil Res. 1999; 11: 511.
27. Blazevic N, Petricic J, Stanic G, Males Z. Acta Pharm. 1995; 45:
517.
28. Weyerstahl P, Marschall-Weyerstahl H, Weirauch M et al. In
Progress in Essential Oil Research, Brunke E-J (ed.). Walter de
Gruyter: Berlin, 1986; 177.
29. Leimner J, Marschall H, Meier N, Weyerstahl P. Chem. Lett.
1984; 1769.
30. Puerta R, Garcia MD, Saenz MT, Gil AM. Planta Med. 1993; 59:
94.
31. Tucker AO, Maciarello MJ, Charles DJ, Simon JE. J. Essent. Oil
Res. 1997; 9: 583.
32. Satta M, Tuberoso CIG, Angioni A, Pirisi FM, Cabras P.
J. Essent. Oil Res. 1999; 11: 711.
33. Bianchini A, Tomi P, Costa J, Bernardini AF. Flavour Fragr. J.
2001; 16: 30.
Flavour Fragr. J. 2001; 16: 253–256