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IZOLATION AND CHARACTERIZATION OF DITERPENES FROM PLANT
SPECIES EUPHORBIA PALUSTRIS

Milica Puđa
Center for talented youth Belgrade II, milicapudja@gmail.com
Supervisors: Dr Vele Tešević, associate professor of University of Belgrade Faculty of Chemistry
M. Sc Gordana Krstić, teaching assistant of University of Belgrade Faculty of Chemistry

C 7 (C 6 -133.9 ppm, C 7 –134.8 ppm). It also can be noticed the
1. Introduction presence of oxygenated carbon atoms (C 4 –73.6 ppm, C 13 –
63.2 ppm, C 20 –69.8 ppm) (Picture 2). Based on these data,
the structure of 13–acetoksi–20–izovaleriloksi–12–
Plant species pond spurge (Euphorbia palustris) is very rich
1
source of milky juice-latex. It is known that the latex of deoksiforbol was determined (Picture 3).
these plants contains a large number of organic compounds,
2
such as diterpenes, phenols, and flavonoids. An class of
diterpenes that has been isolated from the plant genus
Euphorbia is a class of tiglianes. It was found that that
tiglianic type of diterpenoids have cytotoxic, analgo-
4
3
antipyretic and sedative effects. They also acts as
5
irritants.
2. Objective

The aim of this work is the isolation of diterpenes from plant
species Euphorbia palustris, and their characterization. Picture 2: H NMR spectrum of fraction 25
1
3. Materials and methods

Lyophilized milk of Euphorbia palustirs was extracted twice
with hexane. The extract was applied to a dry-flash
chromatography column, packed with silica gel. Elution was
carried out by solvent system hexane/ethyl acetate, with the
solvent polarity increase, i.e. percent of ethyl-acetate
increased. Gradient elution was monitored by TLC. Total of
17 fractions from dry–flash column was eluted. Fractions 6,
7 and 8, that were equal on the basis of Rf values, were
13
collected and subsequently separated on column Picture 3: C NMR Picture 4: Structure of 13– acetoxy–20
chromatography. A silica gel was used as stationary phase, spectrum of fraction 25 –isovaleryloxy–12– deoksiforbol
while solvent system hexane/ethyl-acetate was used as a
mobile phase. Elution was isocratic, with a mixture of
hexane/ethyl-acetate (V/V 8/2) and monitored by TLC also. 5. Conclusion
Total of 27 fractions were eluted by the gravity column.
th
Fraction 25 was further analyzed. In this paper a method of a new diterpene ester (13-acetoxy-
20-isovaleryloxy-12-deoxyphorbol) isolation and
4. Results and discussion characterization was described. Characterization of the
isolated compounds was performed by NMR spectroscopy.
In further research the biological activity of the isolated
th
Based on the results of TLC fraction 25 was prepared for compounds will be tested.
13
1
recording NMR spectra. NMR spectrum of H and C were
1
recorded. Based on the appearance of H NMR spectra, it
can be concluded that the compound contains two protons, 6. Literature
2
which are located on the sp -hybridized carbon (H– 1 7.61
ppm, H– 7 5.72). It can also be observed the presence of a 1. Jančić R. (2004):Botanika farmaceutika. Službeni list SCG.
proton which is oxygenated to carbon (H 2 0–4:45 ppm) 2. Eke,T., et al. (2000): Arch. Ophthalmol (AMA). 18, 13–16.
13
(Picture 1). From the C NMR spectrum was observed: keto
group (C 3 –209.1 ppm), ester carbonyl groups (isovaleroxy 3. Quing–Wen S. et al. (2008): Chem. Rev. 108, 4295 – 4327
170.8 ppm, acetoxy 175.2 ppm), as well as two double bonds 4. Qing–Gao M. et al. (1997): J. Phytochemistry. 44, 663–666.
at positions C 1 –C 2 (C 1 -161.4 ppm, C 2 –132.8 ppm), and C 6 - 5. Baloch I. B. et al. (2007): Eur. J. Med. Chem. 43, 274–281.