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Phytotaxonomic Markers of Propolis: Isolation and Structure Assessment

Sanja Savić

Talent Center Belgrade II, Belgrade savicsanja333@gmail.com
Mentor: Dr Miroslav Novaković, ITHM, Belgrade University

1.Introduction

Propolis is a dense, resinous mixture that honey bees collect
from tree buds and burgeons and process using salivary
glands secretion. Propolis is derived from the Greek words
“pro,” meaning “in defense of” and “polis,” meaning “city.”
Together, the meaning, “in defense of the city,” refers to the
defense of the bee hive. The main use of propolis by bees is
to protect their hive from wind, moisture and outside
invaders. Propolis is also used for covering of the outside
walls of the combs due to its disinfectant properties.
Propolis consists of various constituents, primarily phenols
(58%) and waxes (32%), as well as flavonoids (6%) and
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minerals.
Cinnamic acids and their derivates occur frequently in
several natural products, including propolis. They have Figure 1. А – p-coumaric acid; B – 1,3-di-p-
been reported to have many biological activities, e.g. coumaroyl-2-acetylglycerol
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antioxidant and antimicrobial. Some derivates of cinnamic
acids like glycerol esters can be used as taxonomic markers,
and have a major role in determining the botanical origin of 4.Conclusion
propolis.
2.Methods Cinnamic acids derivatives, apart from being biologically
active natural products, represent important
phytotaxonomic markers. Based on literature data it was
concluded that propolis analyzed in this research originate
The techniques used to isolate compounds from propolis from Populus tremula – aspen, since 1,3-di-p-coumaroyl-2-
are column (CC) and semi-preparative high-performance acetylglycerol shown above (figure 1B) is one of the
liquid chromatography (HPLC), while the process of phytotaxonomic markers for this plant species.
elution was monitored using thin-layer chromatography
(TLC). In the column-chromatography silica gel served as
stationary phase, while the combination of methylene- 5. References
chloride and methanol was used as eluent. The elution
process was gradient, and the fractions obtained were
combined on the basis of TLC, and further separated into [1] FAO/WHO Expert Committee on Food Additives.
pure substances using semi-preparative liquid Meeting Evaluation of certain veterinary drug residues in
chromatography. Separation was performed on a reversed- food (2008). 70th report of the Joint FAO/WHO Expert
phase C18 column, with solvent system 0.1% formic Committee on Food Additives. (WHO technical report
acid/acetonitrile, using gradient program. Pure compounds series; no. 954)
were identified using NMR spectroscopy and MS
spectrometry. [2] R. Kannan, R. Arumugam, T. Thangaradjou, P.
Anantharaman. Phytochemical constituents, antioxidant
properties and p-coumaric acid analysis in some seagrasses,
3. Results and discussion Parangipettai, India Food Research International (2013), 54,
1229-1236.

From the combined fractions obtained after semi- [3] М. Popova, B. Trusheva, R. Khismatullin, N. Gavrilova,
preparative HPLC cinnamic acid (from fractions BP23-30, G. Legotkina, J.Lyapunov, V. Bankova. The Triple
figure 1A), and its glycerol derivate (from fraction BP3-5, Botanical Origin of Russian Propolis from the Perm
figure 1B) were isolated. Their structures were determined Region, Its Phenolic Content and Antimicrobial Activity.
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using C and H NMR spectra, and confirmed by MS Natural Product Comunications (2013), 8 (5), 617 - 620.
spectra.