FLAVOUR A N D FRAGRANCE JOURNAL, VOL.

9,339-342 (1994)

A Comparative Study of the Essential Oils of Five piper

Species from Peninsular Malaysia
Ibrahim bin Jantan, Abdul Rashih Ahmad, Abu Said Ahmad and Nor Azah Mohd Ali
Chemistry Division, Forest Research Institute Malaysia, Kepong, 52109 Kuala Lumpur, Malaysia

The chemical composition of the leaf oils of five Malaysian Piper species ( P . aduncum L., P. lanatum Roxb., P.
pedicellosum Wall., P. penangense C.DC. and P. betle L.) was determined by co-chromatography with authentic
samples on two columns of different polarity, GC-MS and KovAts retention indices. Dillapiole at 64.5% was the
most abundant component in the oil of P. aduncum. Chavibetol constituted 69.0% and 42.7% of the oils of P.
betle and P. lanatum respectively. The major components of the oil of P . pedicellosum were P-phellandrene
(21.9%) and eugenol (17.2%. )E)-Nerolidol (17.5%) and cedrol (14.8%) were found to be the major constituents
of P. penangense.

KEY WORDS Piper aduncum L. Piper lanatum Roxb. Piper pedicellosum Wall. Piper penangense C.DC.
Piper betle L. Piperaceae GC-MS Volatile leaf oil Dillapiole Chavibetol Eugenol

INTRODUCTION ine type, a- yrones, terpenes and phenolic

The essential oils of the economi-
Piper belonging to the family Piperaceae, is a cally more important species such as P. betle and
large genus of shrubs and vines found in all the P. nigrum have been thoroughly investigated. 7-9
moist warm parts of the world. The most popular However, the reports on the other species have
species is Piper nigrum L. which is the source of been brief2-6 and no systematic chemical studies
pepper, a widely used condiment. Some species on their volatile constituents have yet been made.
such as P. clusii DC. ,P. saigonense DC., P. longi- In the course of our work to identify economically
folium Ruiz & Pavon, P. longum L. and P. adun- important essential oils from Peninsular Malaysia
cum L. have been used in several countries on a and towards the development of a taxonomic
small scale as substitutes for pepper. P. methy- guide for species identification, we report on the
sticum Forst. f., and P. medium Jacq. are widely chemical composition of the leaf oils of five Piper
used in the Pacific and tropical America respec- species, viz. P. aduncum L., P. lanatum Roxb.,
tively as ceremonial and social non-intoxicant P. pedicellosum Wall, P. penangense C.DC. and
drinks. The leaves of P. betle L. have been tradi- P. betle L.
tionally used for chewing purposes throughout
the southeast Asian region. In Peninsular Malaysia
some species such as P. caninum Blume, P . chaba
Hunter and P. nigrum L. are used traditionally as
EXPERIMENTAL
ingredients in dart-poison owing to their suffi-
ciently strong irritant properties. The rubefacient
effect of many species which include P . banta- Plant Material
mense Blume, P. porphyrophyllum N.E.Br. and The leaves of the five Piper species were col-
P. sarmentosum Roxb. is so intense that they are lected at the following places: P. penangense, P.
used in folk medicine as counter-irritants in poul- penicellosum and P. lanatum at the Forest Re-
tices for headaches and other pains. Several Piper search Institute of Malaysia, Kepong; P. aduncum
species are taken internally as tonics, indigestion at Selayang, Selangor; P. betle at Masjid Tanah,
mixtures and postnatal medications. Melaka. Voucher specimens were deposited at
Chemical studies on several Piper species have the herbarium of the Forest Research Institute of
resulted in the isolation of alkaloids of the piper- Malaysia.

ccc 0882-5734/94/060339-04 Received 16 May 1994

0 1994 by John Wiley & Sons, Ltd. Accepted 25 July 1994
340 I . bin JANTAN ETAL.

Analysis of the Essential Oils
The plant materials were water-distilled for
eight hours and the oily layers obtained were
separated and dried over anhydrous magnesium
sulphate. The yields were averaged over two
experiments and calculated based on dry weight
of the plant material. The oils were pale yellow
to pale green with characteristic spicy-piney
odours.
The oils were analysed on a Shimadzu G C 9A
chromatograph equipped with a FID detector
using fused-silica capillary column SE 30 (25 m x
0.2mm i.d.). The operating parameters were:
nitrogen as carrier gas at 50cm3 min-', injector
temperature 230°C detector temperature 230°C.
The column was programmed initially at 60" for
10 min, then 3°C min-' to 180°C for 1 min. The
oils were also examined using stationary phase
PEG 20M capillary column (25 m x 0.2 mm i.d.)
under the following programme condition; initial
temperature 50°C to 230°C at 3 min-'.
The oils were also analysed by GC-MS with
Hewlett-Packard GCMSD 5890 series 2 mass
spectrometer (70 eV direct inlet) on an SE 30
capillary column (25m x 0.2mm i.d.) initially at
60°C for 10 min, then programmed at 3°C min-l
to 180°C with helium as carrier gas. The consti-
tuents were identified by co-chromatography on
the different columns with authentic samples and
by comparison of their KovAts indices with litera-
ture values and their mass spectral data with those
from the NBS mass spectral database.
KovAts indices were obtained from the gas
chromatogram by logarithmic interpolation be-
tween bracketing alkanes. The homologous series
of n-alkanes were used as standards. lo
RESULTS AND DISCUSSION
Water distillation of the fresh leaves of the Piper
species yielded the following percentage of essen-
tial oils: P. aduncurn (1.30%); P . fanatum
(1.34%); P . pedicelfosum(1.11'70); P. penangense
(0.23%); P. betle (5.10%). Except for P.
penangese, the oil yields from the other species,
especially P. berle, may be considered satisfactory
for commercial exploitation.
The chemical cbmposition of the leaf oils of
Piper species, presented in Table 1, showed that

Table 1. Chemical constituents on the leaf oils of Piper species (column: 25 m SE 30)
Percentage

Piper Piper Piper Piper Piper

Kovhts aduncum lanatum pedicellosum penangense betle L.
Compound index L. Roxb. Wall. C.DC. Identification
a-Pinene 936 tr 0.1 1.9 0.1 - MS,KI,Co
Camphene 948 - 0.1 0.8 - - MS.KI,Co
P-Pinene 974 0.3 - - 0.6 - MS,KI.Co
M yrcene 983 - tr - - - MS,KI.Co
a-Phellandrene 998 - tr 2.9 - - MS.KI.Co
3-Carene 1003 - tr 0.3 - - MS,KI.Co
a-Terpinene 1013 0.2 t 0.5 - - MS,KI,Co
p-Cymene 1015 0.2 1.2 0.8 - - MS.KI,Co
P-Phellandrene 1026 - 0.3 21.9 - - MS,KI,Co
Limonene 1026 - - - 0.3 - MS,KI,Co
(Z)-8-Ocimene 1028 1 .o 0.1 0.8 - - MS.KI,Co
(E)-a-Ocimene 1039 2.3 0.7 1.8 - - MS.KI.Co
y-Terpinene 1057 0.2 0.1 2.5 - - MS, KI .Co
Terpinolene 1081 - - 0.3 - - MS.KI.Co
allo-Ocimene 1083 tr tr - - - MS.KI,Co
Linalol 1085 - 1.1 2.2 3.2 - MS,KI .Co
Camphor 1128 - - 0.4 - - MS,KI,Co
cis-p-Menth-2-en-1-01 1128 - - tr - - MS.KI
tram-Pinocarveol 1129 - - - 0.1 - MS,KI
Terpinen-4-ol 1166 0.2 0.2 4.1 0.1 - MS,KI,Co
M yrtenal 1174 - - - 0.1 - MS.KI.Co
a-Terpineol 1175 - tr 0.3 0.2 - MS + KI .Co
 ESSENTIAL OILS OF PIPER SPECIES 341

Table 1. continued
Percentage

Piper Piper Piper Piper

Kovhts aduncum lanarum pedicellosum penangense Piper
Compound index L. Roxb. Wall C.DC. betle L. Identification
Methyl chavicol 1179 - - - - MS.KI
Myrtenol 1182 - - - 0.1 MS,KI,Co
Decanal 1187 - 0.4 0.3 - MS,KI.Co
p-Menthane-1.3-diol 1205 - - - 0.2 MS, KI ,Co
Citronellol 1216 0.3 - - - MS.KI,Co
Chavicol 1230 - - - - MS.KI
Bornyl acetate 1272 - - - 0.4 MS.KI.Co
Undecan-2-one 1275 - - - 1.1 MS,KI
Piperitone 1228 0.2 - - - MS,KI
6-Elemene 1325 0.3 0.2 - 7.4 MS,KI
Eugenol 1334 - - 17.2 2.7 MS,KI ,Co
Chavibetol 1344 - 42.7 - - MS,KI,Co
a-Cubebene 1350 0.3 - - 2.7 MS,KI.Co
a-Copaene 1365 0.1 1.2 1.3 3.5 MS,KI,Co
Methyl eugenol 1372 - 1 .o - - MS.KI
p-Elemene 1376 0.3 0.2 - - MS,KI
p-Patchoulene 1378 0.1 - - - MS,KI,Co
p-Bourbonene 1380 0.7 - 0.3 - MS,KI
Eremophilene 1388 - - 0.8 - MS,KI
a-Gurjunene 1896 - - 2.4 - MS.KI,Co
p-Cubebene 1387 0.1 - - - MS,KI,Co
y-Elemene 1390 0.9 5.4 - 2.3 MS,KI
p-Caryophyllenc 1405 5.1 6.8 7.5 4.4 MS.KI,Co
a-Bergamotene 1435 - - - - MS, KI ,Co
Aromadendrcne 1444 0.2 - - - MS,KI.Co
c-Muurolene 1445 - 0.1 1.2 1.2 MS,KI
a-Humulene 1453 1.6 1.1 1.5 0.9 MS.KI,Co
a-Guaicne 1455 - - - 0.4 MS,KI,Co
allo-Aromadendrene 1463 - 0.5 1.7 - MS.KI.Co
a-Cadinene 1469 - 6.6 2.9 - MS,KI,Co
y-Muurolene 1473 - 1.2 - - MS,KI
6-Selinene 1482 5.2 - 2.9 4.2 MS,KI
a-Muurolene 1483 0.2 6.2 1.2 2.8 MS,KI
Viridiflorene 1485 2.0 - - - MS.KI
Eugenyl acetate 1486 - 5.9 - - MS,KI
a-Elemene 1490 - - 3.9 - MS,KI
a-Selinene 1494 0.2 - - - MS.KI
y-Cadinene 1509 0.8 1.9 2.7 - MS.KI,Co
Calamenene 1510 - 1.4 0.3 0.7 MS.KI,Co
6-Cadinene 1516 1.o 0.3 0.3 0.6 MS,KI,Co
(E)-Nerolidol 1550 0.9 0.4 0.7 17.5 MS.KI.Co
Palustrol 1556 - 0.2 0.7 8.9 MS,KI
p-Guaiene 1560 - - - 0.9 MS,KI ,Co
Spathulenol 1569 - 0.3 1.4 1.1 MS.KI
4-Allyl-1.2-diacetoxybenzene 1582 - 2.1 - - MS,KI
Led01 1584 - - 0.6 - MS,KI
Dillapiole 1603 64.5 - - - MS.KI
Cedrol 1609 - - - 14.8 MS,KI.Co
Laevojuvenol 1620 - - 1.8 - MS.KI
p-Eudesmol 1639 - - - 8.1 MS.KI
Apiole 1641 0.3 - - - MS.KI
Guaiazulene 1742 - - 0.4 0.8 MS.KI
Phytol 2084 1.1 0.4 0.6 1.1 MS.KI,Co
Yields were calculated based on the concentration obtained on column SE 30;the Kovhts index of compounds on the PEG
20M column was also determined: MS = mass fragmentation; KI = retention index; Co = co-chromatography with authen-
tic sample.
342 1. bin JANTAN E T A L .
the qualitative and quantitative distribution of the
oil constituents among different species may be of
chemotaxonomic importance.
The most abundant component in the leaf oil
of P. aduncum was dillapiole, which at 64.5% is
greater than the 58% reported in the oil of similar
species from Fiji. The oil yield reported by us for
this species (1.30%) is also substantially higher
than the 0.46% reported for the Fijian sample.6
Of the other 31 components identified in the oil,
only (E)-a-ocimene (2.3%), P-caryophyllene
(5.1%), a-humulene (1.6%), p-selinene (5.2%),
viridiflorene (2.0%) and phytol (1.1%) were pre-
sent at greater than 1 % concentration.
Chavibetol was the major component in the
leaf oils of P. betle and P. lanatum, accounting for
69.0% and 42.7% of the oils respectively.
Eugenyl acetate at 8.3% in the former and 5.9%
in the latter was another compound of chemo-
taxonomic interest present in both oils. Other
compounds that were present in appreciable
amounts in P. betle were chavicol (6.0%) and y-
muurolene (5.2%). No monoterpene was de-
tected in the oil, and sesquiterpenes, represented
by eight compounds, constituted only 11.9% of
the oil. Sesquiterpenes were present at more than
34% concentration in the oil of P. lanatum and
the major representatives were P-caryophyllene
(6.8% ), a-cadinene (6.6%),a-muurolene (6.2%)
and p-elemene (5.4%), arranged in decreasing
order of concentration. There were 15 mono-
terpenoids identified but they accounted for only
3.9% of the oil.
The oil of P. pedicellosum was characterized by
its richness in monoterpenes (41.5%) with the
major ones being p-phellandrene (21.9%) and
terpinen-4-01 (4.1%). Eugenol (17.2%) repre-
sents the major non-terpenoid constituent of the
oil. Many of the sesquiterpenes were present in
minor amounts, although those present in amounts
greater than 2% were p-caryophyllene (7.5%), a-
elemene (3.9%), a-cadinene (2.9%), a-selinene
(2.9%), y-cadinene (2.7%) and p-elemene
(2.3%). In contrast, sesquiterpenoids were pre-
sent at more than 85% concentration in the oil of
P. penangense where (E)-nerolidol (17.5%) and
cedrol (14.8%) were the major components.
Other major representatives of sesquiterpenoids
were palustrol (8.0%), P-eudesmol (8.1%),
8-elemene (7.4%), a-bergamotene (4.4%) and
p-selinene (4.2%), arranged in decreasing order
of concentration. Monoterpenes constituted only
5.5% of the oil with linalol (3.2%) as the major
representative. Also, eugenol (2.7%) represents
the major non-terpenoid constituent of the oil.
The discernibly different chemical composition
of the leaf oils of the five Piper species may sug-
gest the use of the essential oils as a taxonomic
marker for species characterization. That is
assuming that ontogenetic variations or the exist-
ence of intraspecificchemicaldifferences (chemical
races) do not interfere with species differentiation.
Acknowledgements -The authors are grateful to the Forest
Research Institute Malaysia and Overseas Development
Agency (FRIM-ODA) Programme for providing the neces-
sary funds for the project. They also wish to thank Mat Asri
Ngah Sanah and Saw Leng Guan of the FRIM’s herbarium
for their help in identifying plant materials.
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