FLAVOUR AND FRAGRANCE JOURNAL, VOL. 8.

11-15 (1993)

The Composition of the Essential Oil from Aristolochia

asclepiadifolia Brandg. Root

h r e n z o Sagrero-Nieves*
Instituro de Ciencias Bhicas, Universidad Veracruzano, Apartado Postal 585, Xalapa 91000, Veracruz, Mexico

George R. Waller
Department of Biochemistry, Oklahoma State Uniuersity, Stillwater, Oklahoma 740784454, U S A

Richard P.Sgaramella
International Flavor and Fragrances, Union Beach, New Jersey 07735, USA

The volatile constituents of the root of Aristolochia asclepiad$olia Brandg. has been investigated by gas chromatogra-
phy-mass spectrometry, by which many compounds were identified during a four-year period. Linalol and its oxides,
borneol and its oxides, guaiol and isomers were the major compounds found, accompanied by a-bisabolol, ledol,
terpineol, limonene and its oxides with vanillin being a minor but important component. The freshly prepared
essential oil has a characteristic pleasant aroma, and smells like many absolutes.

KEY WORDS Aristolochia asclepiadifolia Brandg. Aristolochiaceae Root Essential oil composition

INTRODUCTION which in moderate doses is a gastric stimulant;

however, they were used by the American Indian to
The genus Aristolochia, a member of the Aristolo- check vomiting.6 A. serpentaria has been reported
chiacae, is distributed in tropical and subtropical to be effective in curing cancer when boiled with
xgions of the world. Aristolochia asclepiadifolia rum,' as well as useful for other malfunctions of the
Brandg.' is a perennial endemic plant of Veracruz body.' Crude extracts of the roots of A. indica L. in
State, Mexico. The species is a vine 30-100 m long petroleum ether, chloroform and alcohol showed a
and is locally called 'guaco'. The root is very high contraceptive activity in mature female miceg
aromatic and some people of the region of Veracruz The sesquiterpenes isolated from A. indica showed
use it in alcoholic infusions to combat injuries this high antifertile activity in female mice." This
caused by scorpions and snakes and to treat dysen- work describes the chemical composition of the
tery. essential oil of the root of Aristolochia asclepiadifo-
Previous work on members of the Aristolochia lia, analysed in 1986, 1988, 1989, and 1990.
genus reported the inactivation of some Formosan
snake venoms by aristolochic acid.' This com-
pound was also isolated from A. bracteata Retz. A .
mollisima Hance and A. versicolar S . M. Hwang. It EXPERIMENTAL
is reported to have chemosterilant properties in
insects' and contraceptive activity in female Material
Other such species, A. serpentaria L. (Vir-
ginia snakeroot) and A . reticulata Nutt (Texas Roots were collected from random lianas (vines)
snakeroot), contain a bitter principle in the roots in the final fruiting stage growing near Actopan,
'Author to whom correspondence should be addressed.

0882-5734/93/010011-05SO7.50 Received 20 June 1992

Q 1993 by John Wiley & Sons, Ltd. Accepted 10 August 1992
12 L. SAGRERO-NIEVES,G. R. WALLER AND R. P. SGARAMELLA

Veracruz, Mexico. A voucher specimen is deposited gram similar to Kovhts indices. The quantitation
at Instituto de Ecologia Herbarium (XAL). was based upon the GC-FID peak integration to
provide area percentage data.
Isolation of Volatile Compounds.
Freshly ground root (100 g) was subjected to
steam distillation for 6 h in an all-glass system
containing no stopcock grease. The distillate was
saturated with sodium chloride (Baker, certified
reagent grade) and extracted with ethyl ether
(ACS-certified reagent grade). The extract was
dried over anhydrous magnesium sulphate, and
filtered, the ether was allowed to evaporate at room
temperature (25°C) and ethereal extracts of the
essential oil were analysed.
Gas Chromatography-Mass SpectrometryData
Analysis Systems (GC-MS-DAS)
Gas chromatography was carried out with a
Hewlett-Packard Model 5880 instrument equipped
with a flame ionization detector and an OV-1
capillary fused-silica column coated with methyl
silicone, 50 m x 0.32 mm (id.). The helium flow
through the column was 1.8 ml/min. The injection
port and detector temperatures were 260°C. The
column temperature was held at 50°C and then
programmed at 2"C/min to 225°C and held there
for 60 min. The essential oil sample (1.5 pl) was
injected in the splitless mode. The capillary gas
chromatograph was interfaced to a Kratos MS-50
mass spectrometer operated in the electron ioniza-
tion mode. Mass spectral scans were taken at
0.33 min/scan at 70 eV. Results were acquired and
analysed with a Kratos DS-55 data system."
The identifications were based on comparisons
of known with unknown spectra, computer com-
parison, visual interpretation of the fragmentation
patterns, and comparison with a proprietary (Inter-
national Flavor and Fragrances) elution order pro-
RESULTS AND DISCUSSIONS
The freshly prepared essential oil had a character-
istically pleasant aroma, smelling like a composite
of many absolutes. The fragrance was evaluated as
dry, sweet and woody, it resembled balsam abso-
lute and when fresh, mate absolute and fig leaf
absolute.
Table 1 lists the many compounds that were
identified completely or tentatively during four
years of study. Table 2 lists the major compounds
found to occur during each year. We have found
compounds reported to have antibacterial roper-
ties: terpene-4-01, a-terpineol and linalol.' P These
compounds were major constituents of A. asclepia-
difolia. a-Terpineol and linalol were also reported
in A. triangularis Chamis~o.'~ Borneol, also a ma-
jor component, was reported in A. zenkeri,14 A.
long^'^ and A. dibilis Sieb et ZUC.'~ Guaiol found
''
in A. asclepiadifolia as a major component is
known to be present in flavourin and is biogeneti-
cally derivable from farnesol. Germacrene-D,
found in 1989 and 1990 has been reported as the
major compound in leaves of A. triangularis and in
minor concentrations in the root and stems of this
specie^.'^ The prescence of the aromadendrane in
A. dibilid6 is important because it is chemically
related to ledol'" which is also found in this work
and in A. indica.lg It was the first aromadendrane
derivative to be identified and its occurrence could
be of significance as a marker for chemotaxonomic
studies for the Aristolochia genus. The content of
vanillin, although quite small, is important from
the aroma and quality of the essential oil viewpoint.
The A. asctepiadfolia root oil is quite variable in

Table 1. Identification of compounds that occur in the essential oil from the roots of Arirfolochio
asclepiad~oliaBrandg. by GC-MS-DA (listed in order of elution)

Relative concentration VA)

No. Compound 1986 1988 1989 1990

1 Ethyl ether (residual solvent) 0.90 0.00 1.51 11.0

2 Acetaldehyde 0.80 0.23 0.22 0.10
3 Ethanol 1.40 2.25 0.51 0.12
4 Acetic acid 0.90 0.01 0.40 0.10
5 Ethyl acetate 0.90 0.01 1.75 0.20
6 Propylene glycol - - 0.15 0.03
 ESSENTIAL OIL OF ARISTOLOCHIA ASCLEPIADIFOLIA ROOT 13

Table 1. (COW.)

Relative concentration (%)

No. Compound 1986 1988 1989 1990

7 cis-Hex-3en-1-01 - 0.10 0.04

8 Acetaldehyde diethyl acetal 0.02 0.14 -
9 Hexanal 0.01 - -
10 Hexan-2-one 0.01 - -
11 Hexa-2.4dienal - - -
12 1.2-Dimethyl-1,3dioxolane4rnethanol 0.08 - -
13 cis-Hept-2-enal - 0.37 -
14 2-Ethoxyethyl acetate 0.05 - -
15 a-Pinene - 0.07 0.07
16 Camphene 0.12 0.74 0.06
17 a-Myrcene (+ isomers) - 0.15 0.70
18 Hexanoic acid 0.10 0.07 0.04
19 Sabinene - 0.26 0.05
20 Limonene 0.03 3.74 0.40
21 2-Methylheptan-1-01 - - -
22 1-Pinene - 0.26 0.30
23 8-Myrcene - 0.15 -
24 a-Fenchene - 0.18 0.04
25 Benzyl alcohol 0.2 1 - -
26 pCymene 0.13 1.84 -
27 2,3,3-Trimethyl-2-norbornanol 0.43 0.09 -
28 pCresoI 0.17 - -
29 1,8-Cineoie - 0.05 0.21
30 Acetophenone - 0.07 0.03
31 y-Terpinene - 0.09 0.04
32 trans-Linalol oxide (furan form) 0.23 0.81 0.53
33 cis-Linalol oxide (furan form) 0.12 0.85 0.51
34 Linaloi 0.21 20.84 14.00
35 Phenethyl alcohol 0.25 0.10 0.03
36 cis-Sabinene hydrate - 0.07 0.03
37 Camphor 0.14 0.34 0.03
38 1.3-Dimethoxybenzene 0.11 - -
39 Linalol oxide (dehydrated) 0.04 - -
40 Borneo1 2.04 1.60 0.05
41 cis-Hex-knyl acetate - 0.15 0.11
42 CIsopropylcyclohex-2-en-I-one 0.12 0.50 0.03
43 Neomenthyl acetate 0.37 - -
44 cis-Hex-3cnyl butyrate 0.02 - -
45 CTerpineol 0.20 1.15 0.3 1
46 Hex-3cnyl butyrate - - 0.31
47 a-Terpineol 1.04 0.28 0.18
48 Limonene oxide 0.4 1 0.10 0.04
49 Fenchene 0.41 0.70 -
50 4-Phenylbutan-2-one 0.79 4.80 2.20
51 Anisyl acetate 0.24 - -
52 Carveol isomer - 0.16 0.05
53 cis-8-Ocimene - 0.20 0.33
54 Carveol - 0.18 0.3 1
55 Bornyl formate 0.21 0.10 0.03
56 Carvone 0.15 0.15 0.03
57 Hex-3cnyl 2-methylvalerate 0.19 - -
58 trans-Dec-2-enal 2.48 - -
59 2,6-Dimethylocta-2,7diene-l,6diol 1.79 - -
60 Undeca-2,4-dienal 0.05 - -
61 CPhenyl-2-butanol acetate - - 0.20
62 Geraniol - 0.12 0.07
63 Phellandral - 0.08 0.12
64 1-Vinylhexyl acetate - 0.07 -
14 L. SAGRERO-NIEVES,G. R. WALLER AND R. P.SGARAMELLA

Table 1. (Cont.)

Relative concentration (%)

No. Compound 1986 1988 1989 1990

65 Cuminyl alcohol 0.02 0.35 0.05 0.03

66 Bornyl acetate 1.30 0.85 0.13 0.07
67 Carvacrol 0.70 0.48 0.21 0.04
68 Dodeca-2,edienal - 0.32 - -
69 3,7-Dimethylocta-l,5diene-3,7-diol - 0.49 - -
70 Perillyl acetate 1.70 1.28 0.07 0.30
71 a-Terpinyl acetate (isomer) - - - 0.11
72 a-Terpinyl acetate 0.80 0.39 0.17 0.14
73 irons-8-Terpinylacetate 0.90 0.55 0.12 0.05
74 Vanillin 200 2.10 - -
75 a-Cubebene - 0.59 0.15 0.15
76 a-Copaene 1.00 2.00 0.70 0.52
77 8-Cubebene isomer - - 0.12 0.13
78 a-Santalene - - 0.04 0.04
79 8-Elemene(isomer) - - - 0.04
80 8-Elemene 0.80 0.29 0.05 0.03
81 8-Cary ophyllene - - 1.15 1.10
82 B-Cubebene - - 0.29 0.30
83 Bergamotene - - 0.30 0.20
84 a-Caryophyllene - - 0.50 0.51
85 fi-Selinene 0.70 0.92 1.71 1.10
86 Germacrene-D - - 0.88 1.10
87 Valencene 1.00 0.15 - -
88 Calamenene - 0.12 0.58 0.24
89 Vetiverol 0.80 0.90 1.60 0.21
90 Sesquiterpene hydrocarbons - - 0.69 0.65
(unidentified)
91 a-Murolene - - 0.55 0.50
92 4-Bisabolene - - 2.24 1.88
93 Sesquiterpene hydrocarbons - - 1.68 0.4
(unidentified)
94 1,5-Di-tert-butyl-3,3dimethyl- 1 .00 0.42 - -
bicyclo[3.l.0]hexan-2-one
95 p-Elemol(+ isomers) - - 1.10 1 .50
96 y-Cadinene - 0.76 - 0.60
97 Patchouli alcohol (tentative) - 200 - -
98 Spathulenol - 1.22 - 0.82
99 a-Bisabolol 0.90 1.54 6.20 6.60
100 Pogostol - 0.73 2.27 3.05
101 Guaiol 3.40 9.70 10.3 13.00
102 Guaiol isomer - - 0.32 250
103 Cadin4en-1-01 - - 0.39 0.28
104 y-Cadinol - - 2.74 0.18
105 Guaiol isomer - - 0.39 0.20
106 8-Eudesmol - - 1.24 2.00
107 2,6-Di-rert-butyl-4 methylphenol 9.00 8.90 - -
108 Led01 8.30 10.1 1.12 0.50
109 Bubnesol - - 1.67 2.80
110 Seaquitcrpene alcohols - - 4.65 2.60
(unidentified)
111 Scsquiterpenehydrocarbons - - 1.68 4.90
(unidentified)
112 Sesquiterpenoidsand sesquiterpsne 29.1 29.7 - -
hydrocarbons (unidentified in
1986 and 1988 oil)
~

Total ncovery (%) 96.69 92.32 94.31 84.46

 ESSENTIAL OIL O F ARISTOLOCHIA ASCLEPIADIFOLIA ROOT 15

Table 2. Major compounds and types identified in essential oil of A. asclepiadifolia

Brandg.

Relative concentration (%)

Compound 1986 1988 1989 1990

Linalol and its oxides 10.22 0.60 2240 15.04

Borneol, esters and isomers 17.00 3.95 290 0.15
CPhenylbutan-Zone - 0.79 4.80 2.20
Guaiol and isomers 3.40 9.70 11.01 15.70
Led01 8.30 10.1 1.12 0.50
Vanillin 2.00 2.10 - -
Perillyl acetate 1.70 1.28 0.07 0.30
Terpineol (4 and a-plus other isomers, 4.60 2.20 2.50 0.97
their acetates and hydrocarbons)
Limonene and its oxide 1.40 0.44 4.02 0.58
Caryophyllene isomers (a + 8) - - 1.65 1.61
B-Selinene 0.70 - 0.88 4.10
Germacrene-D - - 0.88 4.10
Vetiverol 0.80 0.90 1.60 0.21
a-Bisabolol 0.90 1.54 6.20 6.60
8-Bisabolene - - 2.24 1.80
Pogostol - 0.73 2.27 3.05
Bu bnesol - - 1.67 280

composition on a year by year basis. We are unable
to pinpoint the reason, for this variation; it could
be due to climatic, nutritional genetic, environmen-
tal, age or other factors.
Acknowledgements-We thank 0.C. Dermer, G. V. Odell and E.
D. Mitchell, Jr, for their critical review of this manuscript. ms
Journal Article No. 5611 of the Oklahoma Agricultural
Experiment Station, Oklahoma State University, Stillwater,
Oklahoma, 74078. USA.
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