FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fragr. J. 2001; 16: 212–214

DOI: 10.1002/ffj.985

Chemical composition of the volatiles of Hemidesmus

indicus R. Br.
S. Nagarajan, L. Jagan Mohan Rao and K. N. Gurudutt∗
Plantation Products, Spices and Flavour Technology Department, Central Food Technological Research Institute, Mysore 570 013,
India
Received 21 July 2000
Revised 6 November 2000
Accepted 17 November 2000

ABSTRACT: The fragrant roots of Hemidesmus indicus (Indian sarsaparilla) is used in Indian native medicine
and herbal tea preparations. The volatiles obtained by steam distillation (yield, 0.25%) contained 2-hydroxy-4-
methoxybenzaldehyde (91%) and (
) ledol (4.5%), which are isolable in pure form, as the major constituents.
The GC–MS analysis of the residual oil showed the presence of over 40 minor constituents. Among them,
nerolidol (1.2%), borneol (0.3%), linalyl acetate (0.2%), dihydrocarvyl acetate (0.1%), salicylaldehyde (0.1%),
isocaryophyllene (0.1%), ˛-terpinyl acetate (traces) and 1,8-cineol (traces) are important as aromatic and bio-active
principles. Copyright  2001 John Wiley & Sons, Ltd.
KEY WORDS: Hemidesmus indicus; Asclepiadaceae; volatile oil; GC–MS analysis; 2-hydroxy-4-methoxybenz-
aldehyde; ledol

Introduction (Hemidesmus indicus R.Br./MGM/ LJM/ 8/ 2000) was

Indian sarsaparilla (Hemidesmus indicus R.Br.), which
belongs to the monotypic genus Hemidesmus of the fam-
ily Asclepiadaceae, is a twining slender shrub found in
India. Its root is woody with a diameter of 0.5–1.5 cm,
brownish or purple in colour and of aromatic odour.
The root-bark is easily separable from the inner core,
which is the aromatic part. The dried roots are medic-
inal and well known in Indian pharmacopoeia as the
drug ‘Anantmul’,1 which is a tonic and blood puri-
fier and is used for treating a variety of physiologi-
cal disorders. Syrupy extract made from the roots is
used as a flavouring agent and in the preparation of
‘sherbet’, which is reported to have cooling properties.1
The methanol extract of the root is known to achieve
neutralization of snake venom.2 While 2-hydroxy-4-
methoxybenzaldehyde is reported as the major con-
stituent of its essential oil,3 no information is available
on the minor constituents. Therefore, we undertook a
systematic analysis of the volatile oil and report its com-
position here.
deposited at the herbarium of the University.
Isolation of the Volatiles
The roots were cleaned with water and cut into small
pieces, 2–3 cm long, for distillation by the following
alternative methods:
Clevenger Distillation
The material (275 g) was placed in a round-bottomed
flask containing 1 l distilled water, two drops of ‘anti-
foam’ added, and subjected to Clevenger distillation
for 8 h. The distillate was extracted with diethyl ether
100 ml ð 3, the ethereal layers combined and dried
over anhydrous Na2 SO4 . The extract was concentrated
on a water bath to 10 ml and further evaporated to
dryness by passing argon (yield, 0.11 g).
Likens–Nickerson Method

A 3 l round-bottomed flask containing the material

Experimental
H. indicus root
Fresh roots were collected from the herbal garden of
University of Mysore, Mysore. A voucher specimen
Ł Correspondence to: K. N. Gurudutt, Plantation Products, Spices and
Flavour Technology Department, Central Food Technological Re-
search Institute, Mysore 570 013, India. E-mail: cif@cscftri.ren.nic.in
(250 g) and 1 l water was connected to the one arm
of a Likens–Nickerson apparatus and a 250 ml round-
bottomed flask containing a mixture of diethyl ether
(100 ml) and petroleum ether (50 ml) to the other arm.
Simultaneous distillation and extraction was carried out
for 5 h. After cooling, the organic layer was separated,
dried over anhydrous Na2 SO4 , concentrated in a rotary
evaporator to 10 ml, and further evaporated to dryness
by purging with argon (yield, 0.12 g).

Copyright  2001 John Wiley & Sons, Ltd.


Steam Distillation
Steam was passed steadily into a suspension of the
material (400 g) in 4 l water for 4 h. The steam distillate
(3 l) was cooled and extracted with dichloromethane
(100 ml ð 3). The combined dichloromethane extracts
were dried over anhydrous Na2 SO4 and evaporated to
dryness (yield, 0.3 g).
Isolation of the Major Constituents
The sticky mass from steam distillation was dissolved in
n-pentane (10 ml) and stored overnight in a refrigerator,
when needle-like crystals were formed. They were sepa-
rated by decanting off the mother liquor. The latter was
reprocessed to obtain two more crops of crystals. The
solids were combined and recrystallized from pentane
(yield, 0.27 g).
The mother liquor on evaporation gave a viscous oil
(28 mg, 9.3%) which was loaded onto a column of silica
gel (100–200 mesh, 5 g) and eluted with hexane–ethyl
acetate (1–5%) mixtures to collect five fractions. The
needle-like crystals (13 mg) that separated out from the
second fraction were filtered and recrystallized from
hexane.
1
H NMR spectra of the crystalline solids were re-
corded on a Varian EM 390 NMR spectrometer
(90 MHz) using CDCl3 as solvent and tetramethylsilane
as the internal standard. Optical rotation was recorded
on Perkin-Elmer 243 polarimeter.
GC–MS Analysis of the Volatiles
The GC–MS analysis of the total volatiles was per-
formed on a Shimadzu GC-17A coupled with a QP 5000
MS by injecting 2 µl of its dichloromethane solution
(100 mg in 2 ml) using a SPB-1 column (Supelco, USA,
30 m ð 0.32 mm, film thickness 0.25 µm) under the fol-
lowing conditions: oven temperature was programmed at
at 50 ° C for 2 min, rising at 2 ° C/min to 250 ° C (5 min);
injection port temperature, 250 ° C; detector tempera-
ture, 250 ° C; carrier gas, helium; flow rate 1 ml/min.
GC–MS analysis of the residue obtained on removal of
the major crystalline constituent was performed on a DB
wax column (J&W Scientific, USA; 30 m ð 0.25 mm,
film thickness 0.25 µm) with the oven temperature pro-
grammed at 60 ° C for 5 min, rising at 2 ° C/min to 220 ° C,
then held for 5 min; injection port temperature, 225 ° C;
detector temperature, 250 ° C.
Retention indices of all the constituents were deter-
mined by the Kovats method,4 using n-alkanes as
standards. The volatile constituents were identified by a
comparison of their Kovats indices with those reported
in literature4,5 and their identification confirmed by com-
puter matching of their mass spectral fragmentation pat-
terns with those of compounds in the NIST–MS Library.
Copyright  2001 John Wiley & Sons, Ltd.
VOLATILE CONSTITUENTS OF HEMIDESMUS INDICUS 213
Results and Discussion
The volatiles from fresh Hemidesmus indicus roots
were isolated by Clevenger distillation, simultaneous
distillation and extraction and steam distillation meth-
ods, with 0.13%, 0.16% and 0.25% yields, respec-
tively. The last method was not only the most effi-
cient but also convenient for large-scale extraction. From
a pentane solution of the distillate, a colourless crys-
talline material was isolated (91%) and characterized
from its m.p. (40–41 ° C; lit.3 41–43 ° C) and 1 H NMR
spectrum6 and confirmed by its mass spectral fragmenta-
tion [m/z (% abundance): 53(28), 63(25), 81(15), 95(30),
108(25), 134(3), 151(100) and 152(20)] as 2-hydroxy-
4-methoxybenzaldehyde. Incidentally, the volatile oil
obtained from the roots of Decalpis hamiltonii, which
also belongs to the family Asclepiadaceae, contains this
compound to an extent of 96%.7
Total ion chromatogram (TIC) of the residual volatile
oil, generated by GC–MS on a polymethylsiloxane cap-
illary column, indicated the presence of at least 40
constituents. The second most abundant constituent,
which eluted closely after nerolidol, did not give a
clear mass spectrum, indicating that it was probably
a mixture. It was then isolated in pure from by SiO2
column chromatography of the residual oil, followed
by crystallization (yield, 4.5%; purity by GC, 95%).
Its mass spectral fragmentation pattern [m/z (% abun-
dance): 43(100), 55(32), 69(30), 81(30), 93(20), 109(28),
122(25), 161(15), 189(10) and 204(8)] and 222(3)] indi-
cated it to be a sesquiterpene alcohol. These data,
together with the physical data [m.p. 104–105 ° C, [˛]D

5.6° (c,0.6 in CHCl3 ); lit.8 m.p. 103–104 ° C, [˛]D

5.8° (c,1.5 in CHCl3 )] and 1 H NMR spectrum9 led
to its identification as (
) ledol.
Rest of the minor constituents were identified by
a library match of their mass spectra and determi-
nation of their retention indices on the SPB-1 col-
umn, and in some cases, on the DB wax column
(see Table 1). Among them, nerolidol, borneol, cis-
caryophyllene, linalyl acetate, dihydrocarvyl acetate,
salicylaldehyde, isocaryophyllene, caryophyllene, lina-
lyl acetate, dihydrocarvyl acetate, salicylaldehyde, iso-
caryophyllene, ˛-terpenyl acetate and 1,8-cineol are
important as flavour constituents.10 The presence of
dodecanoic and hexadecanoic acids in the volatile oil
was probably due to the vigorous distillation conditions.
Conclusion
For the extraction of aroma volatiles from H. indicus
roots, steam distillation is the most suitable method.
2-Hydroxy-4-methoxybenzaldehyde, which can be iso-
lated in pure crystalline form, accounts for ca. 91% of
the extract. The remaining portion consists of over 40
Flavour Fragr. J. 2001; 16: 212–214
214 K. N. GURUDUTT, L. JAGAN MOHAN RAO AND S. NAGARAJAN

Table 1. Chemical composition of the volatiles from the roots of Hemidesmus indicus
SPB-1 DB WAX
No. Compound KI KI MC (%)
1 Salicylaldehyde 1003 1636 122 0.1
2 1,8-Cineol 1011 154 Traces
3 Camphor 1105 1493 152 0.5
4 Pinocarveol 1109 152 Traces
5 ˇ-pinene oxide 1119 152 Traces
6 Pinocarvone 1124 150 Traces
7 Borneol 1140 154 0.3
8 4-Terpenenol 1152 154 Traces
9 Bornyl acetate 1545 196 Traces
10 Myrtenal 1155 150 Traces
11 Methyl salicylate 1158 1735 152 Traces
12 ˛-Terpineol 1164 154 Traces
13 Verbenone 1167 150 Traces
14 Myrtenol 1170 1767 152 Traces
15 Linalyl acetate 1194 196 0.2
16 Isobornyl acetate 1256 1552 196 Traces
17 2-Hydroxy-4-methoxybenzaldehyde‡ 1296 2145 152 91.0
18 Dihydrocarvyl Acetate 1305 196 0.1
19 ˛-Terpinyl acetate 1325 196 Traces
20 ˇ-Elemene 1370 204 Traces
21 cis-Caryophyllene 1427 1589 204 0.2
22 Isocaryophyllene 1453 204 0.1
23 ˇ-Selinene 1457 204 0.1
24 Nerolidol 1563 222 1.2
25 Ledol‡ 1629 2239 222 4.5
26 Dodecanoic acid 1636 2502 200 0.2
27 Hexadecanoic acid 1957 256 0.2
‡
Isolated by SiO2 column chromatography and characterized by physical and spectral methods.

constituents, among which ledol (4.5%) and nerolidol
(1.2%), the sesquiterpene alcohols, are major. The minor
but olfactorily and biologically significant constituents
are borneol, linalyl acetate, dihydrocarvyl acetate, sali-
cylaldehyde, ˛-terpinyl acetate and 1,8-cineol.
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Copyright  2001 John Wiley & Sons, Ltd. Flavour Fragr. J. 2001; 16: 212–214