I. M. Sudarma, et al. /Journal of Natural Products, Vol.

8(2015): 27-32

ISSN 0974 – 5211

Journal of Natural Products
Volume 8 (2015)

Research Paper www.JournalofNaturalProducts.Com

Chemical Transformation of Eugenol Isolated from leaves of

Syzygium aromaticum to its new isothiocyanate derivatives
I.M. Sudarma1*, A. Kusnandini1, M.G. Darmayanti1
1
Department of Chemistry, Faculty of Science University of Mataram
Jl. Majapahit 62 Mataram 83125, Indonesia
*Corresponding Author
(Received 14 October 2014; Revised 25 January-03 April 2015; Accepted 15 April 2015)

ABSTRACT
The main aim of this research was to develop new or novel compounds with potential
biological activity from readily accessed natural products, in particular eugenol.
Eugenol (1) has been previously isolated from leaves of Syzygium aromaticum and
transformed to its nitro eugenol in good yield by adding potassium hydrogen sulfate
and ammonium nitrate. The nitro group of eugenol reduced smoothly to amino-
eugenol by treated with zinc and formic acid. Reaction of amino eugenol (3) with
carbon disulfide which mainly analysed by GC-MS produced eugenol (1) (22.46%),
M+. 164; isothiocyanate derivative (4) (4.36%), M+. 221; amino eugenol (3) (32.16%),
M+. 179; thiolisothiocyanate (5) (4.28%), M+. 256; unidentified compound (4.04%);
major isothiocyanate derivative (6) (32.70%), M+. 221.

Keywords: Chemical transformation; Eugenol; New isothiocyanate.

INTRODUCTION
The discovery, development, and application of natural drugs has been done for
thousands of years. As a type of plant extracts, eugenol (1) as a very wide biological
activities and application history. The aim of this research was to develop new or
novel compounds with potential biological activity such as isothiocyanate derivatives
from readily accessed natural products, in particular eugenol. Isothiocyanate is
sulphur-containing phytochemicals and has the chemical group -N=C=S, formed
by substituting the oxygen in the isocyanate group with a sulfur. Isothiocyanates have
been shown to be especially effective in fighting cancers (Cinciripini et al., 1997;
Hecht, 2000). Isothiocyanates can be found in cruciferous or "cabbage family"
vegetables such as broccoli, cauliflower, kale, turnips, collards, Brussels sprouts,
cabbage, kohlrabi, rutabaga, Chinese cabbage, bok choy, horseradish, radish, and
watercress (Drewnowski and Carneros, 2000). The synthesis of isothiocyanates
proceeds through the reaction between a primary amine (e.g. aniline) and carbon
disulfide in aqueous ammonia. This results in precipitation of the ammonium
dithiocarbamate salt, which is then treated with lead nitrate to yield the corresponding
isothiocyanate (Dains et al., 1926). Isothiocyanate derivatives could be also

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 I. M. Sudarma, et al. /Journal of Natural Products, Vol. 8(2015): 27-32

synthesized from readily accessed natural products (e.g. eugenol) which is easily
isolated from Syzygium aromaticum.
Cloves (Syzygium aromaticum) are harvested primarily in Indonesia,
Madagascar, and Zanzibar, India, Pakistan, and Sri Lanka. According to FAO,
Indonesia produced almost 80% of the world's clove output in 2005. Cloves contain
eugenol (4-Allyl-2-methoxyphenol), a main constituent of the essential oil and have
been used for antimicrobial (Eyambe et al., 2011) and anesthetic (Jadhav et al., 2004).
Eugenol is considered as a phenolic compound similar to benzene with three
substituents (hydroxy, methoxy, and allyl) which undergo electrophilic aromatic
substitution reactions through nitration (Sudarma et al., 2014). Nitro-eugenol is of
considerable importance in the production of other fine chemicals such as amino-
eugenol for further chemical synthesis. Nitro compounds can be reduced easily to the
corresponding amino derivatives. Amino eugenol has an amino group (-NH2) which is
also easily reacted with acyl and carbon disulfide. Eugenol has been previously
investigated and reported for its further chemical transformation (Sudarma et al.,
2009a; 2009b; 2013; Sudarma, 2010; Carrasco et al., 2008), but its transformation to
new isothiocyanate derivatives has not been reported yet.

MATERIALS AND METHODS

Materials: The material used included: dry fall leaves of clove (Syzygium
aromaticum) were collected in july 2012, in Gangga village (northern of Lombok),
dichloromethane, hexane, methanol, tetrahydrofuran, sodium hydroxide pellet,
acetonitrile, ammonium nitrite, potassium hydrogen sulfate, zinc powder, formic acid,
silica gel, sodium carbonate anhydrous, acetyl chloride, carbon disulfide, analytical
thin layer chromatography.
Instrumentation: GC-MS were recorded on GC-MS QP-5050A, BC-17A and MS
5050A Merk Shimadzu. The original 1H nmr, 13C nmr, and DEPT spectra are directly
reproduced throughout. They were generally recorded in CDCl3 on a Bruker
spectrometer at 400MHz.
Procedure: Dried leaves of clove was chopped and grounded to fine particles (125g)
and percolated with dichloromethane (250ml) and kept for 24 hours. Then the liquid
extract was filtered and evaporated to afford yellowish oil (12.5g). This oil was
analyzed by GC-MS to confirm the presence of eugenol. Eugenol was obtained from
the leaves clove oil, according to standard procedure and identified by GC-MS and
NMR analyses. M+. 164, cal for C10H12O2 Major fragments: 49 (M+. –CH3), 131, 121,
103, 91, 77 (C6H6, base peak). 1H NMR (400.1 MHz, CDCl3): δ 3.35 (2H, d, J 6.6 Hz,
H1'); 3.93 (3H, s, OCH3); 5.13 (2H, m, H3'); 5.50 (1H, s, OH); 5.91 (1H, m, H2'); 6.5
- 6.96 (3H, aromatic protons).
Nitration of eugenol using NH4NO3/KHSO4: A round bottomed flask (50ml) with
magnetic stirrer was charged 1.00g eugenol (6.10mmol) and acetonitrile (20ml) then
stirred for 5 min. Potassium hydrogen sulfate (0.64g) and ammonium nitrate (1.4g)
were added and stirred at room temperature for 30 min then refluxed for 5 hour.
Worked up was based on Baghernejad et al., 2009, to afford yellowish to redish oil
(1.1g, 86.28%, pure by tlc analysis). Compound (2) (oil): M+. 209, cal for C10H11NO4
Major fragments : 195 (M+. –CH2), 178, 163, 147, 131, 119, 103, 91 (base peak). IR
(film) nmax/cm-1: 3232 (O-H), 3084 (C=CH-Ar), 3014 (CH=CH2), 2936, 2829, 1634
(C=C), 1547 (NO2), 1399, 1327, 1260 (C-O), 1127 (C-O), 1066, 999, 912, 764. 1H
NMR (400.1 MHz, CDCl3): δ 3.35 (2H, d, J 6.6 Hz, H1'); 3.93 (3H, s, OCH3); 5.13
(2H, m, H3'); 5.91 (1H, m, H2'); 6.96 (1H, s, H3); 7.50 (1H, d, J 0.9 Hz, H5); 10.67

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 I. M. Sudarma, et al. /Journal of Natural Products, Vol. 8(2015): 27-32

(1H, s, OH). 13C NMR (100.6 MHz, CDCl3): δ 39.4 (C1’); 56.7 (OCH3); 115.1 (C5);
117.1 (C3’); 118.6 (C3); 131.2 (C4); 133.6 (C6); 135.9 (C2’); 144.9 (C1); 149.8 (C2).
Preparation of amino-eugenol (3): Nitro-eugenol (620mg, 2.96mmol) was dissolved
in methanol (10ml) and the solution stirred with zinc powder (1g) and formic acid
(2.5ml) for 10 minute. Worked to afford the desired amino eugenol compound (85%
yield) (Gowda et al., 2001). GC-MS: M+. 179, cal for C10H13NO2 Major fragments:
164, 152, 136, 118, 106, 91 (base peak). 1H NMR (400.1 MHz, CDCl3): δ 3.21 (2H, d,
CH2), 3.48 (1H, s, NH); 3.68 (1H, s, NH), 3.85 (3H, s, OCH3); 5.05 (2H,m,CH2), 5.25
(1H,s,OH); 5.91 (1H, m, H2'); 6.18 (1H,s, ArH), 6.25 (1H,s,ArH). FT-IR cm-1: 3389
(O-H), 3306 (NH2), 2943 (C=CH-Ar), 2848 (CH=CH2), 1608 (C=C), 1131 (C-O)
Carbon disulfide reaction of amino eugenol: Amino eugenol (250mg, 1.40 mmol)
was dissolved in a minimum of tetrahydrofuran and stirred for 10 min, then carbon
disulfide (0.2 ml) was added. The solution was stirred overnight. The precipitate was
filtered and washed by methanol to afford amorphous solid (245mg). GC-MS analysis
gave eugenol (1) (22.46%), isocyanate derivative (4) (4.36%) M+. 221; amino eugenol
(3) (32.16%); thiolisothiocyanate (5) (4.28%) M+. 256; unidentified compound (4.04);
major isothiocyanate derivative (6) (32,70%). GCMS: M+. 221, cal for C11H11NO2S
Major fragments: 206 (M+-CH3), 179, 164, 147, 131, 118, 91, and 77.

RESULTS AND DISCUSSION

Herein, we report our results on Chemical Transformation of Eugenol Isolated from
leaves of Syzygium aromaticum to its new isothiocyanate derivatives (Scheme-1). In
the synthetic approach based on amino-eugenol (3) as a main material for the
synthesis of isothiocyanate derivatives, required nitro-eugenol (2) as a precursor. This
was prepared by isolation of eugenol (1) from clove oil then converted to nitro-
eugenol (2) using KHSO4/NH4NO3.
H O
Is o la t io n
C lo v e o il
M eO
E u g e n o l (1 )

N itra ta tio n

N O 2
H O

S M eO
N itro -e u g e n o l (2 )
N
H O R e d u c t io n
S
N H2
M eO
HN SH H O
(4 ) (4 .2 8 % ) CS 2
H O
S + M eO
H A m in o -e u g e n o l (3 )
N M eO
(5 ) (4 .3 6 % )
O

M eO
(6 ) (3 2 .7 0 % )

Scheme-1: Synthesis of new isothiocyanate derivatives from eugenol (1).

The structure of the isolated eugenol (1) and nitro-eugenol (2) were confirmed by
their mass spectrum and 1H NMR. The GC-MS analysis of (1) showed the molecular
ion at m/z 164, consistent with the molecular formula C10H12O2. The 1H NMR.

Copyright © 2015, Journal of Natural Products, INDIA, Dr. Sudhanshu Tiwari, All rights reserved 29
 I. M. Sudarma, et al. /Journal of Natural Products, Vol. 8(2015): 27-32

spectrum of (1) confirmed 12 protons, with the methoxy singlet at 3.93 ppm; three
aromatic proton of benzene ring at 6.5 - 6.96 ppm; one phenolic proton at 5.50 ppm;
and the remaining five allyl proton at 3.35, 5.13, and 5.91 ppm. The GC-MS analysis
of (2) showed molecular ion at m/z 209, calculated for C10H11NO4 The FT-IR of (2)
gave a characteristic –OH phenol dan nitro groups stretching bands at frequency 3232
(O-H), and 1547 (NO2). The 1H NMR. spectrum of (2) based on Carrasco et al., 2008,
gave signal –OH shifted downfield at 10.67 ppm due to the formation of hydrogen
bonding between hydrogen from –OH with oxygen from NO2.
Further stage in the synthesis of the amino-eugenol (3) was to reduce the nitro-
eugenol (2) to the corresponding amine. Typical reducing agents such as tin/HCl,
Zn/CaCl2, Zn/formic acid have been used for the reduction of nitro aromatic (Godwa,
et al., 2001). It has been reported that nitro aromatic compound was reduced smootly
with zinc powder and formic acid in methanol to afford amino-group. The structure of
amino-eugenol (3) was confirmed spectroscopically. The GC-MS analysis observed
molecular ion at m/z 179 consistent with the molecular formula C10H13NO2 .
Comparison of the 1H NMR spectra of (2) and (3) revealed that the chemical shifts of
the protons were very similar except the amino group of (3) showed a two protons
singlet at 3,48 and 3,68 ppm. The remaining protons were assigned to the phenolic as
a signal at 5.25 ppm, a methoxy at 3.85 ppm, the protons of methylene which attached
on benzene ring at 3.21 ppm, and a methylene terminal at 5.05 ppm. An olefinic
proton gave signal at 5.91 ppm and two aromatic protons gave signal at 6.18 and 6.25
ppm. In the FT-IR spectrum of (3) a characteristically sharp bands for the aromatic
primary amine group was present at slightly higher frequencies (shorter wavelength,
3306 cm-1 compare to aliphatic primary amine (Silverstein et al., 1981).
Carbon disulfide treated with amino-eugenol (3) for overnight reaction and
analysed by GC-MS produced isothiocyanate derivatives (4), (5), (6), unreacted
amino-eugenol (3), and eugenol (1) (Figure-1).

Figure-1: GC-MS analysis the reaction of amino eugenol and CS2.

The GC-MS analysis showed peak 6 as a major isothiocyanate derivative which has
molecular ion at m/z 221 corresponding to the molecular formula C11H11NO2S.
Mechanistically, the formation of the compound (6) can be explained as follows
(Scheme-2): nucleophilic attack of the amino group from amino eugenol (3) on the
carbon of carbon disulfide to produced intermediated (3b), followed by hydroksi
addition to give (3c). Elimination of hydrogen sulfide would then produced (6).

Copyright © 2015, Journal of Natural Products, INDIA, Dr. Sudhanshu Tiwari, All rights reserved 30
 I. M. Sudarma, et al. /Journal of Natural Products, Vol. 8(2015): 27-32

S H

S S

.. S
NH2 NH
HO HO

MeO MeO
(3) (3b)

SH
S H S H
H N
N O
O - H2S

MeO
MeO (3c)
(6)
Scheme-2: The formation of the compound (6)

The reaction of amino eugenol (3) with CS2 gave side products eugenol (1)
and unreacted amino eugenol (3). These products occur was due to elimination of
amino group by acid (H2S) which was generated during the formation of compound
(6). This acid (H2S) could be react with amino group through nucleophilic substitution
reaction.

Acknowledgements: Authors thank Directorate General Higher Education (DIKTI), Indonesian

Education and Culture Department through “Penelitian Kompetensi ” for financial support.

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Copyright © 2015, Journal of Natural Products, INDIA, Dr. Sudhanshu Tiwari, All rights reserved 32