FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fra`r[ J[ 1999^ 04] 066Ð070

Composition and Stereoanalysis of Cymbopogon

winterianus Jowitt Oil from Southern Brazil
D. Lorenzo,1 E. Dellacassa,1* L. Atti-Serafini,2 A. C. Santos,2 C. Frizzo,2 N. Paroul,2 P. Moyna,1,2 L. Mondello3
and G. Dugo3
1
Cátedra de Farmacognosia, Facultad de Quı́mica, Universidad de la República, Avda. General Flores 2124, UR-11800 Montevideo,
Uruguay
2
Instituto de Biotecnologı́a Universidade de Caxias do Sul, Rua Fco. Getúlio Vargas 1130 Caxias do Sul, RS, Brazil
3
Dipartimento Farmaco-chimico, Facoltá di Farmacia, Università di Messina, Via SS. Annunziata, 98168 Messina, Italy

Received 0 May 0888

Revised 09 December 0888
Accepted 05 December 0888

ABSTRACT] The hydrodistilled essential oil from aerial parts of Cymbopo`on winterianus Jowitt\ cultivated in
Southern Brazil\ was analysed by GCÐMS[ Thirty!one components\ representing 85) of the oil\ were char!
acterized[ Enantiomeric ratios of limonene\ linalool\ citronellal and b!citronellol were obtained by mul!
tidimensional gas chromatography\ using a developmental model set up with two GC ovens[ The enantiomeric
distributions are discussed as indicators of origin authenticity and quality of this oil[ Copyright Þ 1999 John
Wiley + Sons\ Ltd[

KEY WORDS] Cymbopo`on winterianus Jowitt^ citronella^ volatile fraction composition^ enantioselective gas
chromatography

Introduction is also one of the most important sources of natural

Citronella oils are produced from two distinct types of
citronella grass] Cymbopo`on nardus L[ Rendle\ known
as lenabatu or Ceylon citronella\ and C[ winterianus
Jowitt\ known as maha pen`iri or Java!type citronella[
The Ceylon type is the original cultivated variety and
the Java type seems to have arisen as a distinct form of
the Ceylon type[
Until the early part of this century the Ceylon type
citronella was the most widely produced oil[ Gradually
the Java type has come to dominate the market\ due to
its higher yield of oil[ It is grown commercially on a
worldwide scale in places like Haiti\ Central America\
the South Paci_c and tropical Africa[ More recently
other countries\ such as Brazil\ have contributed to the
citronella oil production[0
Java citronella oil is used directly in perfumery\ but
 Correspondence to] E[ Dellacassa\ Catedra de Farmacognosia\ Facultad de
Qu(mica\ Universidad de la Republica\ Avda[ General Flores 1013\ UR!00799
Montevideo\ Uruguay[
Contract:grant sponsor] Ministero per gli A}ari Esteri Italiano\ Italy[
Contract:grant sponsor] Direzione Generale per la Cooperazione e lo Sviluppo\
Instituto Italo!Latino!Americano[
Contract:grant sponsor] International Foundation for Science[
Contract:grant number] F:1085!1[
Contract:grant sponsor] Conselho Nacional de Desenvolvimento Cienti_co e
Tecnologico[
Contract:grant number] CNPq\ PSDT:DTI[
citronellal\ this component being present at higher con!
centration than in Ceylon oil[1\2
Before the advent of chemical sprays\ Ceylon cit!
ronella was used in combination with Virginian red
cedarwood oil in commercial insect repellents[ Probably
due to its association with insect repellency\ citronella
is usually overlooked as an aromatherapy oil ingredient[
A comprehensive survey on the composition of cit!
ronella oils has been published\3 showing considerable
di}erences in the components of the Java oils in relation
to genetic\ climatic and geographical origins\ as well as
on the stage of development of the plants used for the
oil distillation[
Given the wide use of citronella oils and their frequent
use as a basis of blends with other oils\ as well as their
variability\ chemical evaluation of their monoterpenoid
constituents is of fundamental interest[ Furthermore\
the enantiomeric distribution of the di}erent mono!
terpenic components has been reported as criteria for
genuinity in citronella oils[4Ð7
As part of our research on Brazilian essential oils\ we
report here the results relative to the composition and
enantiomeric distribution of four monoterpenes "lim!
onene\ linalool\ citronella and b!citronellol# of Cym!
bopo`on winterianus Jowitt cultivated in the South of
Brazil[

Copyright Þ 1999 John Wiley + Sons\ Ltd[

067 D[ LORENZO ET AL[
Experimental
Materials
The aerial parts of Java citronella "C[ winterianus Jow!
itt# were harvested from a small!scale experimental area
cultivated at the Prefeitura de Santo Antonio da
Patrulha in Rio Grande do Sul State "Brazil#\ in April
0886[ The essential oil was obtained from the aerial
parts after 1 h of hydrodistillation in the pilot plant of
the Instituto de Biotecnolog(a\ Universidade de Caxias
do Sul[ The average yield was 9[63 "w:w#[
GC Analysis
The composition of the oil was carried out by GC on a
Shimadzu 03 B gas chromatograph equipped with a
FID and a Shimadzu data processor software EZ!
Chrom\ using two capillary columns[ The _rst was a
SE!41 "Mega\ Legnano\ Italy# cross!linked fused!silica
capillary column "14 m×9[21 mm i[d[#\ coated with 4)
phenyl!polymethylsiloxane "9[39Ð9[34 mm phase thick!
ness#^ column temperature\ 59>C "7 min# to 079>C at
2>C:min\ 079Ð149>C at 19>C:min\ 149>C "09 min#[
Injector temperature 149>C^ detector temperature
179>C^ injection mode\ split^ split ratio 0 ] 29^ volume
injected\ 9[1 ml of the oil[ Carrier gas was hydrogen\ 44
kPa[
The second was a Carbowax 19M "Ohio Valley\ USA#
bonded fused!silica capillary column "14 m×9[21 mm
i[d[#\ coated with polyethylene glycol "9[14 mm phase
thickness#^ column temperature\ 39>C "7 min# to 079>C
at 2>C:min\ to 129>C at 19>C:min[ Injector temperature
149>C^ detector temperature 149>C^ injection mode\
split^ split ratio 0 ] 29^ volume injected\ 9[1 ml of the oil[
Carrier gas was hydrogen\ 29 kPa[
GC–MS Analysis
GCÐMS analysis were conducted using a Shimadzu QP
4949 equipped with Adams library\8 using two capillary
columns[ The _rst was a SE!41 "Mega\ Legnano\ Italy#
cross!linked fused!silica capillary column "14 m×9[14
mm i[d[#\ coated with 4) phenyl!polymethylsiloxane
"9[14 mm phase thickness#^ column temperature\ 59>C
"7 min# to 079>C at 2>C:min\ to 129>C at 19>C:min[
Injector temperature 149>C^ injection mode\ split^ split
ratio 0[39^ volume injected\ 9[1ml of the oil[ Helium was
used as a carrier\ using 011[1 kPa "40[5 cm:s#^ interface
temperature 149>C^ acquisition mass range 39Ð399^
solvent cut\ 1 min[
The second was a BP 19 "SGE\ Australia# bonded
fused!silica capillary column "14 m×9[14 mm i[d[#\
coated with polyethylene glycol "9[14 mm phase thick!
ness#^ column temperature\ 39>C "7 min# to 079>C at
2>C:min\ to 129>C at 19>C:min[ Injector temperature
Copyright Þ 1999 John Wiley + Sons\ Ltd[
149>C^ injection mode\ split^ split ratio 0[39^ volume
injected\ 9[1 ml of the oil[ Carrier gas was He\ 81[5 kPa
"44[8 cm:s#^ interface temperature 149>C^ acquisition
mass range 39Ð399^ solvent cut\ 1 min[
Individual components were identi_ed by com!
parison of both mass spectrum and their GC retention
data with those of authentic compounds previously ana!
lysed and stored in the data system[ Other identi_cations
were made by comparison of mass spectra with those in
the data system libraries and cited in the literature[8\09
The linear retention index "LRI# of individual com!
ponents were measured on both columns "SE 41 and
Carbowax 19M#[
Chiral Analysis
Enantiomeric ratios of limonene\ linalool\ citronellal
and b!citronellol were obtained by multidimensional
gas chromatography\ using a model under devel!
opment00 set up with two GC ovens\ the _rst equipped
with a column coated with SE!41 and the second with a
chiral column coated with a derivatized b!cyclodextrin\
and a hot interface\ a rotary switching valve and a
system to maintain a constant ~ow during the transfer[
With this system a heart!cut of the relevant fractions
could be made and these fractions transferred from the
non!chiral column to the chiral one in the following
experimental conditions] precolumn\ SE!41 "Mega\
Legnano\ Italy# cross!linked fused!silica capillary col!
umn "29 m×9[21 mm i[d[#\ coated with 4) phenyl!
polymethylsiloxane "9[39Ð9[34 mm phase thickness#^
column temperature^ 34>C "5 min# to 179>C at 1>C:min^
179>C "04 min#[ Two di}erent analytical columns were
used\ the separation of limonene and linalool enanti!
omers was achieved using a fused!silica capillary col!
umn "14 m×9[14 mm i[d[\ 9[14 mm phase thickness#\
coated with 1\2!di!O!ethyl 5!O!t!butyldimethylsilyl b!
cyclodextrin in PS 975 "02) phenylmethyl!poly!
siloxane# "Mega\ Legnano\ Italy# while citronellal and
b!citronellol enantiomers were separated with a fused!
silica capillary column "14 m×9[14 mm i[d[\ 9[14 mm
phase thickness#\ coated 1\2!di!O!acetyl 5!O!t!butyl!
dimethylsilyl b!cyclodextrin in OV 0690 "03) cyano!
propylphenyl!methylpolysiloxane# "Mega\ Legnano\
italy#^ injection temperature\ 149>C^ column tempera!
ture\ 34>C "5 min# to 89>C at 1>C:min\ 89>C "19 min#^
89>C to 079>C at 1>C:min\ 079>C "09 min#^ interface
temperature\ 199>C^ detector FID\ 179>C "for both
chromatographs#[ Volume injected\ 0 ml of an oil
dilution 0 ] 09^ injection mode\ split^ split ratio 0 ] 04[
Carrier gas was Ar\ 89 kPa "precolumn#\ 009 kPa "ana!
lytical column#[ Evaluation of the elution order of the
enantiomers was achieved using monoterpene mixtures
with known enantiomeric ratio[
Flavour Fra`r[ J[ 1999^ 04] 066Ð070
 COMPOSITION AND ANALYSIS OF CYMBOPOGON WINTERIANUS OIL 068
Results and Discussion
Thirty!one components could be identi_ed in the oil[
The basic composition of the oil of C[ winterianus cul!
tivated in Southern Brazil was similar to that reported
for other Brazilian regions[01 Di}erences were observed
due to higher values of citronellyl acetate and geraniol
and the absence of elemicin and a!cadinol[ The volatile
constituents identi_ed in the oils are listed in Table 0[
Figure 0 shows the total ion chromatogram of the oil[
Compositional changes could be explained by extrinsic
conditions associated to the climate and geographical
origin of the cultivation areas[ The enantiomeric ratio
of four components "limonene\ linalool\ citronellal and
b!citronellol# was established by subsequent transfer
during di}erent analysis using two chiral stationary
phases[ Under the experimental conditions\ limonene
and linalool were best resolved on diethyl tert!butylsilyl!
b!cyclodextrin "1\2!DiEtTBS!b!CDX#\ while citronellal
Figure 1. Chromatogram of Cymbopogon winterianus oil in SE-52
Copyright Þ 1999 John Wiley + Sons\ Ltd[
and b!citronellol were resolved on diacetyl tert!butyl!
silyl!b!cyclodextrin "1\2!DiAcTBS!b!CD#[ The heart
cutting and cut!times for the essential oil are listed in
Table 2[ Figures 1 and 2 show the chromatograms
obtained with the chiral analysis[
Enantiomeric ratios of the components analysed
"Table 1# are similar to those reported in the literature[5
The most marked di}erence was observed for linalool\
which showed a "¦#:"−# enantiomeric ratio of
50[2 ] 27[6[ This value was higher than that reported in
the literature[5
Summarizing\ the chiral essential compounds studied
exhibit an enantiomeric composition which is in practice
in correspondence with reported results for this species[
The enantiomeric distribution for limonene in the oil of
C[ winterianus cultivated in Brazil is _rst reported in
this work[
Furthermore\ the results indicate the utility of the
combination of chemical analysis of an oil and chiral
Flavour Fra`r[ J[ 1999^ 04] 066Ð070
079 D[ LORENZO ET AL[

Table 1. Percentage composition of the essential oil of Cymbopogon win-

terianus Jowitt
Compound ")# L[R[I[
SE!41 CW
0 Myrcene 9[0 876 0019
1 Limonene 2[9 0915 0042
2 b!Phellandrene 9[0 0044
3 Terpinolene 9[0 0122
4 Linalool 9[5 0099 0402
5 Isopulegol 9[0 0031 0424
6 Citronellal 25[0 0047 0321
7 3!Terpineol ³9[0 0066
8 a!Terpineol ³9[0 0078
09 Decanal 9[0 0193
00 b!Citronellol 8[8 0120 0639
01 Nerol 9[2 0120 0656
02 Neral 9[3 0128 0548
03 Geraniol 08[8 0148 0712
04 Geranial 9[5 0169 0573
05 Citronellyl acetate 2[4 0243 0509
06 Geranyl acetate 2[7 0277 0699
07 b!Bourbonene 9[1 0274
08 b!Elemene 0[5 0414
19 "E#!Caryophyllene 9[0 0303
10 a!Humulene 9[0 0335 0599
11 Germacrene D 1[5 0366 0531
12 a!Murolene 9[3 0499
13 g!Cadinene 9[3 0409
14 d!Cadinene 0[8 0410 0574
15 Elemol 4[7 0438 1941
16 Germacrene D!3!ol 0[6 0462 1997
17 09!epi!g!Eudesmol 9[1 0506
18 epi!a!Muurolol 0[9 0539 1199
29 a!Eudesmol 0[5 0542
20 Farnesol 9[1 0610 0833

Identi_ed components 85[2

Grouped components
Monoterpene hydrocarbons 2[1
Oxygen!containing monoterpenes 64[0
Sesquiterpene hydrocarbons 6[3
Oxygen!containing sesquiterpenes 09[5
Others 9[0
 These percentages were obtained on SE!41 except for those of b!phellandrene\ terpinolene\ nerol and b!
elemene which were obtained on Carbowax 19M[
 The components are reported according to their elution order on SE!41[

Table 2. Enantiomeric ratios for limonene, linalool, citronellal and b-citronellol

in Cymbopogon winterianus Jowitt
Limonene Linalool Citronellal b!citronellol

3R!"¦# 3S!"−# 2S!"¦# 2R!"−# 2R!"¦# 2S!"−# 2R!"¦# 2S!"−#

02[9 76[9 50[2 27[6 80[1 7[7 73[6 04[2

Table 3. Heart-cutting times on the essential oil

Cut!times "min#
1\2!DiEtTBS!b!CD 1\2!DiAcTBS!b!CD
Limonene 02[79Ð03[94
Linalool 07[39Ð07[69 07[39Ð07[69
Citronellal 11[09Ð11[24
b!citronellol 16[39Ð16[54

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 066Ð070
 COMPOSITION AND ANALYSIS OF CYMBOPOGON WINTERIANUS OIL 070
Figure 2. Chiral gas chromatogram of limonene and linalool of Cymbopogon winterianus oil on DiEtBuSilb CDX (PS
086) column
Figure 3. Chiral gas chromatogram of linalool, citronellal
and b-citronellol of Cymbopogon winterianus oil on
DiAcTBuSilb CDX (OV 1701) column
analysis of selected optically active ingredients as a
powerful tool to describe an essential oil[
Acknowled`ements * The research was supported by Ministero per
gli A}ari Esteri Italiano\ Direzione Generale per la Cooperazione e
lo Sviluppo[ under Instituto Italo Latino Americano\ and by the
International Foundation for Science "Grant No[ F:1085!1#[ The
Brazilian authors are thankful to Conselho Nacional de Desen!
volvimento Cient(_co e Tecnologico "CNPq\ PDST:DTI programme#
for research support[
Copyright Þ 1999 John Wiley + Sons\ Ltd[
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Flavour Fra`r[ J[ 1999^ 04] 066Ð070