FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fra`r[ J[ 1999^ 04] 298Ð203

Changes in the essential oil content and its composition

during palmarosa (Cymbopogon martinii (Roxb.) Wats.
var. motia) inflorescence development
V. S. Dubey, G. R. Mallavarapu and R. Luthra
Biochemistry Division, Central Institute of Medicinal & Aromatic Plants, P.O. CIMAP, Lucknow-226015 (U.P.), India

Received 8 November 0888

Revised 06 April 1999
Accepted 06 April 1999

ABSTRACT] The ~owering tops of palmarosa "Cymbopo`on martinii# consist of spikelets and each spikelet is
further composed of racemes and a leaf!like structure called a spathe[ Both racemes and spathe contain essential
oil[ Changes in fresh weight\ dry weight\ chlorophyll and essential oil content and its major constituents\ viz[
geraniol and geranyl acetate\ were examined for both racemes and spathe at various stages of spikelet devel!
opment[ The essential oil content was maximal at the unopened spikelets stage and decreased signi_cantly
thereafter[ At unopened spikelets stage\ the proportion of geranyl acetate "47[5)# in the raceme oil was relatively
greater compared with geraniol "26[1)#\ whereas the spathe oil contained more geraniol "50[8)# compared with
geranyl acetate "22[3)#[ The relative percentage of geranyl acetate in both the oils\ however\ decreased sig!
ni_cantly with development and this is accompanied by a corresponding increase in the percentage of geraniol[
Analysis of the volatile constituents from racemes and spathes "from mature spikelets# and seeds by capillary
GC indicated 17 minor constituents besides the major constituent geraniol[ "E#!Nerolidol was detected for the
_rst time in an essential oil from this species[ The geraniol content predominated in the seed oil\ whereas the
geranyl acetate content was higher in the raceme oil[ Copyright Þ 1999 John Wiley + Sons\ Ltd[

KEY WORDS] Cymbopo`on martinii\ Roxb[ Wats[ var[ motia^ palmarosa^ essential oil^ geraniol^ geranyl acetate

Introduction general\ rich in geranyl acetate\ whereas the leaf oil

Cymbopo`on martini "Family Poaceae#\ commonly
known as palmarosa\ yields an essential oil rich in
geraniol[ Palmarosa oil is of commercial importance\
being extensively used in perfumes\ soaps\ cosmetics\
toiletry and tobacco products[0Ð3 It is used mainly as a
source of high!quality geraniol which is used in high!
grade perfumes and cosmetics[1 The characteristic
odour of palmarosa oil is due to its high content of
total alcohol\ mainly geraniol\ and a small but varying
amount of esters associated with geraniol[4 The trace
constituents present in the oil are responsible for the
characteristic olfactory note of the palmarosa oil[ There
are several reports regarding the chemical composition
of palmarosa oil[5Ð09 The essential oil is present in all
parts of the grass\ viz[ ~owering top "in~orescence#\
leaves and stem\ but the ~owering tops contain the
maximum oil[ The immature in~orescence oil is\ in
 Correspondence to] R[ Luthra\ Biochemistry Division\ Central Institute of
Medicinal + Aromatic Plants\ P[O[ CIMAP\ Lucknow!115904 India[ E!mail]
cimapÝsatyam[net[in
Contract:grant sponsor] CSIR\ New Delhi\ India[
contained more geraniol[00 The maximum yield of oil is
obtained when the plant is at a full ~owering stage\
whereas the geraniol content is found to be maximal
towards the end of blooming[01 Palmarosa seeds have
also been found to contain an appreciable amount of
oil with high geraniol content[02
One of the most important characteristics of essen!
tial oil accumulation is its dependence on the devel!
opmental stage of the organ:plant part concerned[
Essential oil biogenesis has been reported to be associ!
ated with early growth\ and the composition of the
essential oil also varies greatly at di}erent growth
stages[03Ð07 As the oil from immature palmarosa
in~orescences "spikelets# contained more geranyl acet!
ate\ whereas the seed oil was rich in geraniol\ it was
believed that geraniol might be produced from the
hydrolysis of geranyl acetate during the development
process[ We report here the changes in essential oil
content and its major constituents\ geraniol and ger!
anyl acetate\ during palmarosa in~orescence devel!
opment[ The essential oils from the mature
in~orescence parts has also been compared with the
seed oil by capillary GC for their minor constituents[

Copyright Þ 1999 John Wiley + Sons\ Ltd[

209 V[ S[ DUBEY\ G[ R[ MALLAVARAPU AND R[ LUTHRA
Figure 1. Palmarosa inflorescence (flowering top consisting of spikelets) (A) and a spikelet (B) at five consequent
developmental stages (I–V). Each spikelet is composed of racemes (R) and a leaf-like structure, the spathe (S)
Material and Methods
Plant Material
Palmarosa "Cymbopo`on martinii Roxb[ Wats[ var[
motia# plants were raised from seedlings at the exper!
imental farm of Central Institute of Medicinal and Aro!
matic Plants "CIMAP#\ Lucknow\ India[ The ~owering
tops were tagged at the time of emergence[ They consist
of spikelets\ and each spikelet is further comprised of
racemes and a leaf!like structure called a spathe[
Racemes are 0[4Ð0[7 cm long\ occur in pairs "one sub!
sessile and the other pedicelled# and develop into seeds
on maturity[ The ~owering tops take about 21 days to
reach maturity and the samples were taken at 7 day
intervals\ representing _ve developmental stages] I\
unopened spikelets^ II\ partially opened spikelets^ III\
fully opened spikelets^ IV\ partially mature spikelets^
and V\ fully mature spikelets "Figure 0A\ B#[ The spike!
lets were removed from the ~owering heads at each
developmental stage\ and separated into racemes and
spathes for various studies[
Copyright Þ 1999 John Wiley + Sons\ Ltd[
Growth Parameters
For the determination of fresh weight "FW# and dry
weight "DW#\ a known number of the spikelets\ racemes
and spathes were weighed immediately after harvest and
then oven dried at 54>C till constant weight[ Chloro!
phyll was estimated by the method of Arnon[08
Isolation and GC Analysis of Essential Oil
The essential oil was extracted in duplicate from freshly
harvested tissues "spikelets:racemes:spathes# by a
steam distillation technique using a mini!Clevenger
apparatus\19 and collected in small stoppered tubes con!
taining anhydrous sodium sulphate to remove traces of
moisture[ The oil replicates were pooled for gas chro!
matographic analysis[ Oil density was determined to
convert the volume yields into their respective weights\
as described by Guenther[10
Gas chromatographic "GC# analysis of the essential
oil was performed using a Perkin!Elmer "Model 2819
Flavour Fra`r[ J[ 1999^ 04] 298Ð203
 ESSENTIAL OIL OF CYMBOPOGON MARTINII 200

B# apparatus equipped with a ~ame ionization detector

"FID# and a stainless steel column "1 m×2 mm i[d[#
packed with 09) FFAP "free fatty acid phase# on chro!
mosorb WAW "79Ð099 mesh#[ The operating con!
ditions were] column temperature\ isothermal at 054>C^
injector and detector temperature\ 199>C and 149>C\
respectively^ nitrogen and hydrogen ~ow rate adjusted
to 29 ml:min and 17 ml:min\ respectively[ The geraniol
and geranyl acetate peaks were identi_ed by coinjecting
authentic standards and quanti_ed using a Varian Inte!
grator "Model 3399#[
GC analysis for the minor constituents in the oils of
palmarosa seeds\ racemes and spathes were carried out
on PerkinÐElmer Gas Chromatograph\ Model 7499\
equipped with FID\ using BP!0 "SG Engineering Pvt[
Ltd[\ Melbourne\ Australia# capillary column "14
m×9[4 mm i[d[#\ coated with dimethylsiloxane\ as
described in detail previously[02 Nitrogen was used as
carrier gas at 09 psi inlet pressure[ Temperature pro!
gramming was 59>CÐ119>C at 4>C:min[ The split ratio
used was 0 ] 79[
Identi_cation of the components was done by com!
paring the relative retention indices "Kovats# of the
peaks "relative to C!7 to C!12 alkanes# on BP!0 column
with the literature values02 and also by peak enrichment
on coinjection with standards\ wherever possible[

Figure 2. Contribution of raceme (Ž) and spathe (R) to

Results and Discussion
Ontogenic Changes in Essential Oil Content and
Composition
Racemes and spathes were separated from the spikelets
at each developmental stage and analysed for essential
oil content and composition[ During palmarosa in~or!
escence development\ the fresh weight "FW# and dry
weight "DW# of the racemes increased until the partially
opened "II# and fully opened spikelets "III# stages\
respectively "Figure 1A\ B#[ The chlorophyll content of
the racemes\ which is an indicator of photosynthetic
capacity of the tissue\ increased until stage II "Figure
2A#[ A signi_cant decrease in the DW of the racemes
occurred after stage III\ suggesting that the in~orescence
had entered into the maturation phase[
The FW and DW of the spathe decreased signi_cantly
until stage II\ and this caused a decrease in the FW
and DW of the spikelets "Figure 1A\ B#[ However\ the
contribution of the spathe to the FW and DW of the
spikelet after stage II was small compared to that of the
racemes[ The chlorophyll content of the spathe
decreased until stage III and then remained more or less
constant "Figure 2A#[ The appreciable decrease in FW\
DW and chlorophyll content of the spathe until stage
II suggests that the spathe entered into the maturation
phase immediately after spikelet opening[ The essential
the spikelet (ž) fresh weight (A) and dry weight (B)
during inflorescence development
oil content of both racemes and spathe decreased with
in~orescence development "Figure 2B#[ The decrease in
the essential oil content was signi_cant until stage III[
The relative percentage of geranyl acetate in the
raceme oil decreased signi_cantly until stage III\
whereas that in the spathe decreased from stage II to
stage III "Figure 3A\ B#[ The decrease in the percentage
of geranyl acetate in both raceme and spathe oils is
accompanied by a corresponding increase in the per!
centage of geraniol in their oil[ Thus\ the ger!
aniol ] geranyl acetate ratio increased with in~orescence
development "Figure 4#\ suggesting the role of an ester!
ase in producing geraniol[ We have recently identi_ed
and isolated an esterase enzyme which is involved in the
production of geraniol by hydrolysing geranyl acetate
during palmarosa in~orescence development[11
Volatile Constituents from Palmarosa Seed,
Raceme and Spathe Oils
Twenty!three components were identi_ed in the pal!
marosa seed oil by capillary GC "Table 0#[ Geraniol
"73[05)# was the major constituent and the other con!
stituents "×0[9)# in the oil were linalool "1[99)#\ b!

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 298Ð203
201 V[ S[ DUBEY\ G[ R[ MALLAVARAPU AND R[ LUTHRA
Figure 3. Changes in chlorophyll (A) and essential oil (B)
content of the raceme (Ž), spathe (R) and spikelets (ž)
during inflorescence development
caryophyllene "0[03)#\ "E\Z#!farnesol "6[1)# and ger!
anyl hexanoate "0[64)#[ Geranyl acetate\ which is the
major constituent "besides Geraniol# in the herb oil\
was present only in a minor amount "9[36)# in the
palmarosa seed oil[ The composition of the seed oil was
essentially the same as described earlier[02
The composition of raceme and spathe oils is being
reported for the _rst time "Table 0#[ The major con!
stituents in the raceme oil from mature palmarosa
in~orescence were geraniol "61[12)#\ geranyl acetate
"07[11)#\ linalool "2[03)# and "E\Z#!farnesol "1[72)#
and other constituents were present in minor amounts
"³0[9)#\ whereas in the spathe oil the major con!
stituents were geraniol "62[3)#\ geranyl acetate
"09[43)#\ linalool "0[97)#\ "E\Z#!farnesol "7[45)# and
geranyl hexanoate "1[30)#\ and other constituents were
present in minor amounts "³0[9)#[ b!bisabolene\
which is normally present in trace amounts in palmarosa
oil\02 was found to be present in 9[04) in the spathe
oil[ Furthermore\ isolation of spathe oil has led to the
identi_cation of "E#!nerolidol for the _rst time from
palmarosa oil[
Several investigations on the composition of pal!
Copyright Þ 1999 John Wiley + Sons\ Ltd[
Figure 4. Relative percentage of geraniol (ž) and geranyl
acetate (Ž) in the raceme (A) and spathe (B) oil during
inflorescence development
marosa oil\ obtained from the ~owering herbs\1\5Ð09
seeds02 and seedlings\12 resulted in the identi_cation of
more than 099 compounds\ which include mono!
terpenes\ sesquiterpenes\ oxygenated sesquiterpenes\
phenyl propanoids and pyrazines[ Comparison of vol!
atile constituents from seed\ raceme and spathe oils
from mature palmarosa in~orescences indicated that
terpinene!3!ol\ b!elemene\ b!selinene\ d!cadinene and
geranyl heptanoate were exclusively present in the seed
oil[ Citronellol and nerol\ the minor components in the
raceme and spathe oil\ were present in trace amounts in
the seed oil[ a!Pinene\ b!bisabolene\ and "E#!nerolidol
were present only in the spathe oil[ The proportion of
geraniol in the seed oil "73[05)# was considerably
higher as compared to that of spathe "62[39)# and
raceme "61[12)# oils[ The proportion of geranyl acetate
was higher in the raceme "07[11)# oil than the spathe
"09[43)# oil[ The seed oil\ however\ contained very little
"9[36)# geranyl acetate "Table 0#[ The composition of
spathe oil is similar to that reported for herb oil in
palmarosa[02
In conclusion\ the results indicate that the geraniol ]
Flavour Fra`r[ J[ 1999^ 04] 298Ð203
 ESSENTIAL OIL OF CYMBOPOGON MARTINII 202

tained mainly geraniol[ Thus\ overmaturing the in~or!

escence will lead to increased production of geraniol\
which is an indicator of oil quality in palmarosa[

Acknowled`ements * The authors are grateful to Dr Sushil Kumar\

Director\ CIMAP\ for his interest and encouragement[ The _nancial
support from CSIR\ New Delhi\ to V[S[D[ as Senior Research Fel!
lowship is duly acknowledged[

Figure 5. Changes in geraniol : geranyl acetate ratio in
the essential oil from raceme (ž) and spathe (Ž) during
inflorescence development
geranyl acetate ratio in the raceme and spathe oil
increased as the in~orescence matured\ and much less
geranyl acetate was present in the seed oil\ which con!
References
0[ Virmani OP\ Srivastava GN\ Singh DV[ Palmarosa and Its Cul!
tivation in India[ Farm Bull[ No[ 6\ Central Institute of Medicinal
and Aromatic Plants] Lucknow\ India\ 0876[
1[ Husain A[ Essential Oil Bearin` Plants and their Cultivation[ Cen!
tral Institute of Medicinal and Aromatic Plants] Lucknow\ India\
0883^ 47[
2[ Boelens MH[ Perfum[ Flavor[ 0883^ 08] 18[
3[ Sharma JR\ Lal RK\ Mishra HO[ Pafai J[ 0886^ 08] 26[
4[ Nigam MC\ Srivastava HK\ Siddiqui MS[ Pafai J[ 0876^ 8] 02[
5[ Saxena DB\ Maheshwari ML[ Ind[ Perfum[ 0879^ 13] 004[
6[ Maheshwari ML\ Mohan J[ Pafai J[ 0874^ 6] 10[
7[ Patil KB\ Jayappa V[ Ind[ Perfum[ 0875^ 29] 175[
8[ Gaydou EM\ Randriamiharisoa RP[ Phytochemistry 0876^ 15]
072[
09[ Siddiqui N\ Garg SC[ J[ Essent[ Oil Res[ 0889^ 1] 82[
00[ Luthra R[\ Dubey VS\ Sangwan RS[ 53th Annual Meetin` of
the Society of Biolo`ical Chemists "SBC!I#[ Society of Biological
Chemists\ Lucknow\ India\ 0884^ 016[
01[ Akhila A\ Tyagi BR\ Naqvi AA[ Ind[ Perfum[ 0873^ 17] 015[
02[ Mallavarapu GR\ Rao BRR\ Kaul PN\ Ramesh S\ Bhattacharya
AK[ Flavour Fra`r[ J[ 0887^ 02] 056[

Table 1. Voltatile constituents from palmarosa seed, raceme and spathe

No[ Compound Retention index Relative percentage in the oil

Seed Raceme Spathe

0[ a!Pinene 822 Ð Ð 9[92

1[ Myrcene 873 9[92 9[98 Ð
2[ Limonene¦0\7!cineole 0913 Ð 9[93 Ð
3[ "E#!b!ocimene 0939 9[93 9[06 Ð
4[ "Z#!linalool oxide 0952 9[94 9[92 Ð
5[ Linalool 0977 1[99 2[03 0[97
6[ Citronellol 0023 9[96 9[92 Ð
7[ Terpinene!3!ol 0052 9[02 Ð Ð
8[ Methyl chavicol 0074 Ð 9[09 !
09[ Citronellol¦nerol 0100 tr 9[11 9[18
00[ Geraniol 0149 73[05 61[12 62[39
01[ Geranyl formate 0172 9[92 9[92 Ð
02[ Geranyl acetate 0246 9[36 07[11 09[43
03: b!elemene 0278 9[94 Ð Ð
04[ b!caryophyllene 0311 0[03 9[33 9[48
05[ a!humulene 0344 9[97 9[00 Ð
06[ b!selinene 0371 9[92 Ð Ð
07[ Geranyl isobutyrate 0381 9[95 Ð 9[97
08[ b!bisabolene 0494 Ð Ð 9[04
19[ d!cadinene 0405 9[93 Ð Ð
10[ Geranyl butyrate 0424 9[05 9[04 9[13
11[ "E#!nerolidol 0437 Ð Ð 9[10
12[ Caryophyllene oxide 0467 9[72 9[05 9[28
13[ Geranyl isovalerate 0473 9[97 9[95 Ð
14[ Geranyl valerate 0529 9[01 9[94 9[97
15[ "Z\Z#!farnesol 0541 9[98 9[94 9[05
16[ "E\Z#!farnesol 0699 6[19 1[72 7[45
17[ Geranyl hexanoate 0615 0[64 9[56 1[30
18[ Geranyl heptanoate 0729 9[94 Ð Ð

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 298Ð203
203 V[ S[ DUBEY\ G[ R[ MALLAVARAPU AND R[ LUTHRA
03[ Singh N\ Luthra R\ Sangwan RS\ Thakur RS[ Curr[ Res[ Med[
Arom[ Plants "CROMAP# 0878^ 00] 063[
04[ Singh N\ Luthra R\ Sangwan RS[ Planta Med[ 0878^ 44] 143[
05[ Singh N\ Luthra R\ Sangwan RS[ Plant Physiol[ Biochem[ 0889^
17] 692[
06[ Luthra R\ Singh N\ Sharma S[ J[ Essent Oil Res[ 0880^ 2] 238[
07[ Luthra R\ Bahal JR\ Tyagi BR[ Curr[ Res[ Med[ Arom[ Plants
"CROMAP# 0884^ 06] 10[
08[ Arnon DI[ Plant Physiol[ 0838^ 13] 0[
Copyright Þ 1999 John Wiley + Sons\ Ltd[
19[ Clevenger JF[ J[ Am[ Pharm[ Assoc[ 0817^ 06] 235[
10[ Guenther E[ The Essential Oil\ Vol[ 0[ Van Nostrand] New York\
0837^ 125[
11[ Dubey VS\ Luthra R[ National Conference on Plant Bio!
technolo`y] Towards Strate`ic A`riculture and Dru` Development\
Lucknow\ India\ 0888^ 001[
12[ Kaul PN\ Bhattacharya AK\ Rao BRR[ Ind[ Perfum[ Essential
Oil Association of India\ 0887^ 31] 095[
Flavour Fra`r[ J[ 1999^ 04] 298Ð203