Kniga na apstrakti book of abstrcts

Distribucija i ekologija na humano-toksi~ni makromiceti 
vo Republika Makedonija
Mitko Karadelev
1
, Sami Sulejmani
2,
Biljana Bauer Petrovska
3
1
Institut za Biologija, Prirodno - matemati~ki fakultet, 
Univerzitet „Sv. Kiril i Metodij
“
, Gazi Baba bb, P. Fah 162, 1000 Skopje; 
2
Prirodno - matemati~ki fakultet, Dr`aven Univerzitet vo Tetovo, 1200 Tetovo; 
3
Farmacevtski fakultet, Univerzitet „Sv. Kiril i Metodij
“
, Vodwanska 17, 1000 Skopje, RM
Republika Makedonija e mikolo{ki relativno slabo istra`ena. Vrz baza na dosega{nite istra`u-
vawa vo Makedonija e utvrdeno prisustvo na 1250 vidovi makromiceti od koi 117 vida poka`uvaat poma-
lku ili pove}e toksi~no dejstvo vrz ~ove~kiot organizam. Ovie vidovi pripa|aat na rodovite: Amanita,
Boletus, Clitocybe, Collybia, Coprinus, Cortinarius, Entoloma, Galerina, Gymnopilus, Gyromitra, Hebeloma, Helvella,
Hydnum, Hygrocybe, Hygrophoropsis, Hypholoma, Inocybe, Lactarius, Laetiporus, Leccinum, Lepiota, Lepista, Leptopodia,
Leucoagaricus, Macrolepiota, Mycena, Omphalotus, Otidea, Panaeolus, Panellus, Paxillus, Peziza, Pholiota,Pluteus,
Polyporus, Psathyrella, Psilocybe, Ptichoverpa, Ramaria, Russula, Sarcodon, Scleroderma, Stropharia, Suillus, Tricholoma,
Tricholomopsis i Xerocomus. Ovie gabi se sobrani vo listopadni i ~etinarski {umi, na livadi, na po~vi
bogati so organska materija, na izmet itn. Najgolem del od niv se terikolni, 7 vida se lignikolni, a 3
vida se sobrani na izmet. Kako rezultat na prisustvoto na odredeni halucinogeni materii, tie predi-
zvikuvaat sedum razli~ni sindromi: faloiden, panterinski, giromitrinski, orelaninski, gastrointesti-
nalen, muskarinski i koprinusen. Halucinogenite materii vo ovie gabi se razli~ni falotoksini, ama-
toksini, orelanin, giromitrin, muskarin, koprinmin, kako i razni halucinogeni i psihotropni supstanci. 
Od vidovite koi sodr`at faloidni otrovi (falotoksini i amatoksini) i predizvikuvaat
faloiden sindrom vo Makedonija se prisutni: Amanita phalloides, A.verna, A.virosa, Galerina autumnalis, G.mar-
ginata, G.venenata, Lepiota brunneo-incarnata, L.castanea, L.helveola, L. josserandii i L.ochraceofulva. Sidromot e
so letalitet 20-30% i u~estvuva so 90-95% od micetizmite so smrten ishod. Orelaninski sindrom (letal-
itet 10-15%) predizvikuvaat vidovi od kompleksniot rod Cortinarius (C. orellanus, C. speciosissimus i C. splen-
dens). Od vidovite koi go sodr`at termolabilniot mikotoksin giromitrin kaj nas se prisutni Gyromitra
esculenta, G.gigas i G.infula. Istiot ima letalitet 30%. Muskarinski sindrom (letalitet 8%) predizvikuva-
at gabi od pove}e rodovi. Tuka spa|aat vidovi od rodovite Inocybe (I. patouillardi, I. fastigiata, I. umbrina),
Clitocybe (C. cerussata, C. dealbata, C. candicans, C. rivulosa) i Amanita (A. muscaria). Sli~en, no so mnogu pomal
letalitet (2%) e panterinskiot sindrom koj go predizvikuva Amanita pantherina. Golem broj gabi se javu-
vaat kako gastrointestinalni iritanti. Smrtni slu~ai se mnogu retki, a simptomite obi~no is~eznu-
vaat za nekolku minuti do 4 ~asa. Tuka vleguvaat pretstavnici od slednite rodovi: Agaricus (A. silvicola, A.
xanthodermus), Boletus (B. luridus, B. satanas), Entoloma (E. lividum, E. nidorosum, E. rhodopolium), Hebeloma (H. sinapizans,
H. crustuliniforme), Lactarius (L. chrysorheus, L. glaucescens, L. helvus, L. rufus, L. scrobiculatus, L. torminosus, L. uvidus),
Lepiota (L. clypeolaria, L. cristata, L. naucina), Naematoloma fasciculare, Paxilus involutus, Pholiota (Ph. aurea, Ph.
squarosa), Albatrelus cristatus, Meripilus giganteus, Phaeolus schweinitzii, Laetiporus sulphureus, Ramaria formosa, Russula
emetica, Scleroderma aurantium, Tricholoma (T. pardinum, T. nudum, T. saponaceum, T. sulphureum, T. sejunctum) i Verpa
bohemica. Od vidovite koi predizvikuvaat koprinusen sindrom (se javuva za 5-10 minuti po jadeweto ako
vo prethodnite 24 ~asa e konzumiran alkohol) kaj nas se zastapeni Coprinus atramentarius, Coprinus micaceus
i Cliticybe clavipes. Od halucinogenite gabi (Psilocibin-psilocinski sidrom) registrirani se vidovite:
Psilocybe (P. cyanescens, P.caerulescens), Panaeolus sphinctrinus, Conocybe cyanopus i Gymnopilus aeruginosus. 
Ova, sekako, ne e kone~na lista na toksi~nite gabi vo Makedonija i od pove}eto vidovi postojat samo
mal broj lokaliteti. So ponatamo{nite istra`uvawa na diverzitetot na gabite vo Makedonija bi se dobi-
la pokompletna slika za distribucijata na ovie organizmi i bi se utvrdil to~niot broj na toksi~ni vidovi.
Macedonian pharmaceutical bulletin 53 (1,2) 280-281 (2007)
SOP - 10
281
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

The Effect of Wine Polyphenol Resveratrol on the Isolated 
Human Internal Mammary Artery
A. Novakovic
1
, M. Peric
2
, D. Nezic
2
,  Lj. Gojkovic Bukarica
3
1
Department of Pharmacology, Faculty of Pharmacy, Belgrade, 
2
Institute for Cardiovascular Diseases “Dedinje”, Belgrade, 
3
Department of Clinical Pharmacology, Pharmacology and Toxicology, Faculty of Medicine, Belgrade, Serbia.
Resveratrol is naturally occuring phytoalexin produced by some spermatophytes, such as grapevines, in
response to injury, and is presumed to be beneficial for human health. Resveratrol has been shown to induce vasore-
laxation. The mechanisms by which resveratrol causes vasorelaxation are uncertain.
Aim: The aim of this study was to investigate the mechanism(s) of endothelium-independent resveratrol-
induced vasorelaxation in human internal mammary artery (HIMA) obtained from male patients undergoing coro-
nary artery bypass surgery, and to clarify the contribution of different K+ channel subtypes in resveratrol action in
this blood vessel. 
Methods: HIMA rings without endothelium were precontracted with phenylephrine.
Results: Resveratrol induced a concentration-dependent relaxation of the HIMA. Highly selective blocker
of ATP-sensitive K+ channels, glibenclamide as well as nonselective blockers of Ca-sensitive K+ channels, tetraethy-
lammonium and charybdotoxin did not block resveratrol-induced relaxation of HIMA rings. 4-Aminopyridine, non
selective blocker of voltage-gated K+ (KV) channels, and margatoxin which inhibits KV1.2, KV1.3 and KV1.6 chan-
nels abolished relaxation of HIMA rings induced by resveratrol. 
Conclusions: In conclusion, we have shown that resveratrol potently relaxed HIMA rings with denuded
endothelium. It seems that 4-AP- and margatoxin-sensitive K+ channels located in smooth muscle of HIMA medi-
ated this relaxation.
Macedonian pharmaceutical bulletin 53 (1,2) 282 (2007)
PP - 133
282
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

Secondary metabolite production in Astragalus parnassi Boiss. plant
Elizabeta Markoska
1,3
, Ana Petkovska
2
, Mirko Spasenoski
1
, Marina Stefova
2
, Sonja Gadzovska
1
1
Institute of Biology, Faculty of Natural Sciences and Mathematics, P.O. Box 162, 1000 Skopje, Macedonia.
2
Institute of Chemistry, Faculty of Natural Sciences and Mathematics, P.O. Box 162, 1000 Skopje, Macedonia.
3
AD Jaka 80-Radovis, Ankarska 33, 1000 Skopje, Macedonia.
Astragalus L. (Fabaceae), as the largest genus of vascular plants on earth, contains an estimated number of
3000 annual and perennial species and 245 taxonomic sections. The roots of various Astragalus spp. are used in tra-
ditional medicine for preparation with an antiperspirant, diuretic and tonic actions. They have also been used in the
treatment of diabetes mellitus, nephritis, leukemia and uterine cancer, but also for their hepatoprotective, antioxida-
tive, immunostimulant and antiviral properties. Anti-inflammatory, analgesic, hypotensive, sedative and cardioton-
ic activities are also reported. Antibacterial activity is also ascribed to some Astragalus spp.: A. siculus, A. gummifer,
A. membranaceus and A. melanophrurius exhibit moderate antibacterial effect against Gram-positive and Gram-neg-
ative tested strains.
The present work was done to determine secondary metabolite production in Astragalus parnassi Boiss.
plants. They were collected in Veles: Gradsko on paleogene lapor at the altitude from 170 to 200 m. This study has
been focused on the production of various phenylpropanoids (phenolic compounds and flavonoids) in different plant
organs. The plant material was collected in three seasons (spring, summer and autumn) of 2005-2006 and separated
into roots, root’s neck, stem (basal, middle and apical parts), spines (rachis) and leaves. A qualitative analysis of the
flavonoid aglycones in A. parnassi extracts using high-performance liquid chromatography (HPLC) with UV diode
array detector was performed. Extracts were prepared by acid hydrolysis in acetone and acidified by hydrochloric
acid on a water bath. Aglycones were reextracted in ethyl acetate, evaporated to dryness and the residue dissolved
in methanol was subjected to a HPLC analysis. Secondary metabolite contents (phenolic compounds, flavonols and
flavanols) were quantified in methanolic extracts by spectrophotometric assay. The statistical analyses were per-
formed with the SPSS statistical software program (SPSS version 11.0.1 PC, USA, IL). Means were expressed with
their standard error and compared by one-way ANOVA (GML procedure). All statistical tests were considered sig-
nificant at P
≤0.05. 
A tentative identification of phenolic acids and flavonoid aglycones of flavones, flavonols and isoflavones
was performed according to their UV spectra. A significant amount of the quercetin was found in the leaves collect-
ed in spring, whereas chryseriol was identified in the spines (rachis) from all three seasons. An isoflavone with UV
spectrum identical to the one of biochanin A but with a shorter retention time was identified in the spines collected
in summer and autumn. Phenolic acids were detected in all analysed extracts and, according to their UV spectra, caf-
feic, ferrulic, p-coumarinic and chlorogenic acid and derivatives are present in the analysed plant material. A. par-
nassi leaves exhibit higher yield of secondary metabolite compared with all other plant organs during the vegetation
period. The difference among secondary metabolite contents in whole plant at different development stages was
found to be significant (P<0.01). The highest level of phenylpropanoid contents was detected in the flowering stage
(summer period) in all plant organs. During the vegetative period, positive linear relationships between secondary
metabolite contents in different plant organs were found. In particular, the leaves and the upper parts of stems (api-
cal parts and spines) have comparatively higher levels of phenylpropanoids.
Thus, it may be considered that the occurrence of phenylpropanoids in A. parnassi Boiss. plants is of signif-
icant pharmacognostic value especially for further evaluation of their medicinal properties. Moreover, this is the first
report on A. parnassi Boiss. and further studies will be needed to clarify the chemistry of this species.
Macedonian pharmaceutical bulletin 53 (1,2) 283 (2007)
PP - 134
283
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

Chemical composition and antimicrobial activity of the essential oil 
of Trinia glauca
Milica Pavlovic
1
, Marina Milenkovic
2
, Jelena Antic Stankovic
2
, Maria Couladis
3
, 
Olga Tzakou
3,
Nada Kovacevic
1
1
Institute of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, V. Stepe 450, 11000 Belgrade, Serbia 
2
Institute of Microbiology and Immunology, Faculty of Pharmacy, 
University of Belgrade, V. Stepe 450, 11000 Belgrade, Serbia
3
Department of Pharmacognosy and Chemistry of Natural Products, School of Pharmacy, University of Athens,
Panepistimioupoli Zographou 157 71 Athens, Greece
The genus Trinia Hoffm. (Apiaceae) is represented by 9 species in the Flora Europaea, while in the flora of
Serbia 3 species are reported. Trinia glauca (L.) Dumort. is a glabrous, glaucous, biennial or perennial herb, up to
50 cm high, growing on dry grassland, usually on limestone soil.
The aerial parts of T. glauca were collected on the mountain Suva planina in Serbia at an altitude of 1500
m, in June 2005. Semi-crushed air-dried plant material (50 g in 500 ml water) was subjected to hydrodistillation for
3 h using a Clevenger-type apparatus. The essential oil was analyzed by GC and GC-MS. The identification of the
compounds was based on comparison of their Retention indices (RI), their retention times (RT) and mass spectra
with those obtained from authentic samples and/or the NIST/NBS, Wiley libraries and literature. 
Seventy compounds were identified, representing 94.7% of the total oil. The oil was characterized by high
percentage of sequiterpene hydrocarbons (42.2%), followed by oxygenated sesquiterpenes (28.9%). Germacrene D
(14.7%), spathulenol (12.5%) and bicyclogermacrene (5.4%) were found to be the major constituents. 
Antibacterial and antifungal activities of the essential oil were assayed using agar diffusion method against
six bacterial (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis ATCC 122288, Micrococcus luteus
ATCC 10240, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Klebsiella pneumoniae
NCIMB9111 and one strain of yeast Candida albicans ATCC 10259. The essential oil showed antimicrobial activi-
ty against all microorganisms tested but differences in microbial susceptibility were registered. The highest activity
was detected against Micrococcus luteus ATCC 10240 and Staphylococcus epidermidis ATCC 12228, while
Escherichia coli ATCC 25922 and Klebsiella pneumoniae NCIMB 9111 were more resistant (only 4% essential oil
solution was active). 
Macedonian pharmaceutical bulletin 53 (1,2) 284 (2007)
PP - 135
284
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

Composition and antimicrobial activity 
of Marrubium incanum essential oil
Silvana Petrovic
1
, Milica Pavlovic
1
, Marina Milenkovic
2
, Maria Couladis
3
, Olga Tzakou
3
, 
Zoran Maksimovic
1
, Marjan Niketic
4
1
Institute of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, V. Stepe 450, 11221 Belgrade, Serbia 
2
Institute of Microbiology and Immunology, Faculty of Pharmacy, V. Stepe 450, 11221 Belgrade, Serbia
3
Department of Pharmacognosy and Chemistry of Natural Products, 
School of Pharmacy, Panepistimioupoli Zographou, 15771 Athens, Greece
4
Natural History Museum, Njegoseva 51, 11000 Belgrade, Serbia
Marrubium incanum Desr. (Labiatae) is an oromediterranean species native in Italy (Sardinia, Sicily,
Ape
nnines) and Balkan Peninsula, where it inhabits the hinterland of the Adriatic Sea (Slovenia, Croatia, Montenegro
and Albania). This species prefers dry rocky places on limestone in the zone of coastal mountains. In traditional med-
icine of southern Italian regions, especially in native Albanian (Arbërësh) population, it is considered to be medici-
nally equivalent to Marrubium vulgare and, therefore, one of the most popular medicinal plant species. The aerial
parts of both plant species are used, in form of decoctions, as panacea, appetizer, digestive, diuretic and as means to
combat malaria. However, available literature data about the chemical inventory and pharmacological activity of M.
incanum are still scarce. Therefore, this plant species came into the focus of our scientific interest.
Aerial parts were collected in July 2006 in S.W. Montenegro near Cetinje at Mt. Lovcen (1420 m altitude,
from calcareous rocks), during the period of full flowering. The essential oil was isolated from the air-dried plant
material by hydrodistillation, according to the procedure of the European Pharmacopoeia 4, using n-hexane as a col-
lecting solvent. The isolated essential oil was at the room temperature semisolid, green-yellowish, with pleasant and
aromatic odor. Essential oil yield was 0.05 % (w/w). 
Using GC-FID and GC-MS, forty-six compounds (96.3 % of total oil) was identified. M. incanum essential
oil was characterized by the high amount (94.3 %) of sesquiterpenes (hydrocarbons 84.4 % and oxygenated deriva-
tives 9.9 %), with (E)-caryophyllene (27.0 %), germacrene D (26.2 %) and bicyclogermacrene (11.5 %) being the
most abundant components. Among the other classes of compounds, C13-isoprenoids (1.2 %) were identified. 
The microbial growth inhibitory properties of isolated essential oil were determined using the agar diffusion
and broth microdilution method, in comparison to ampicillin, amikacin and amphotericin,  against Gram(+) bacte-
ria Staphylococcus aureus ATCC 25923, S. epidermidis ATCC 12228, Micrococcus flavus ATCC 10240, Enterococcus
faecalis ATCC 29212, Gram(–) bacteria Escherichia coli ATCC 25922, Klebsiella pneumoniae NCIMB 9111,
Pseudomonas aeruginosa ATCC 27853, and two strains of the yeast Candida albicans (ATCC 10259 and
ATCC24433). Results were expresed as diameters of inhibition zones (mm) for essential oil applied in two concen-
trations (100 and 50 mg/ml), minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations
(MBCs). M. incanum essential oil exhibited antimicrobial activity in the following order: M. flavus, C. albicans
ATCC24433 (MIC=6.25 
µg/ml) > E. coli, K. pneumoniae, C. albicans ATCC10259 (MIC=12.5 µg/ml) > E. faecalis
(MIC=25 
µg/ml) > S. aureus, P. aeruginosa (MIC=50 µg/ml) > S. epidermidis (MIC=100 µg/ml). MBC values were
two time greater than corresponding MIC values.
Macedonian pharmaceutical bulletin 53 (1,2) 285 (2007)
PP - 136
285
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

Cadmium content in Hypericum perforatum L. 
grown in different areas of Serbia
D. Djukic-Cosic
1
, Z. Plamenac Bulat
1
, M. Curcic Jovanovic
1
, A. Stanojevic
2
, 
M. Djekic
2
, I. Djuric
3
, Z. Zoricic
4
, V. Matovic
1
1
Institute of Toxicology, Faculty of Pharmacy, Belgrade, Serbia
2
Pharmaceutical Society “Dr Jovan Tucakov”, Faculty of Pharmacy, Belgrade, Serbia
3
Intstitute of Public Health, Kraljevo, Serbia
4
Institute of Public Health, Podgorica, Montenego
Medicinal plants are consumed worldwide for the treatment of several diseases and are important raw mate-
rials for the pharmaceutical industry. In recent decades the use of medicinal plants in both crude and prepared forms
has greatly increased. Thus, herbal extracts and phytopharmaceuticals derivatives of Hypericum perforatum L. are
now successfully competing for status as a standard antidepressant therapy.
The World Health Organization (WHO) has established standards for the quality control of medicinal plants
including the classification, botanical identification, determination of active principles and identification of contam-
inants. The WHO recommends qualitative and quantitative assays of heavy metals in phytotherapeutics, especially
in raw materials because the presence of toxic metals in medicinal plants may be detrimental to health.
The aim of this work was to determine cadmium (Cd) concentration in the herb Hypericum perforatum L.
grown in different areas of Serbia: fields of mountains Rtanj and Ozren. Herbs were dried at room temperature and
homogenized. After mineralization with digestion mixture (conc. HNO
3
and conc. HClO
4
in ratio 4:1) cadmium con-
tent was determined by AAS (apparatus GBC932AA).
The obtained results show that Cd concentrations in Hypericum herbs varied from 0.32 to 1.24 mg Cd/kg
dried plant materials. These differences in Cd concentrations are attributed to the mineral composition of the soil
from selected locality. Anyhow, in all investigated samples of herbs Hypericum perforatum L. Cd levels were high-
er than ones proposed by WHO (0.3 mg Cd/kg dried plant materials).
Further investigations are needed in order to find out the correlation between cadmium pollution and con-
tent of pharmacologically active substances of Hypericum perforatum L.
Macedonian pharmaceutical bulletin 53 (1,2) 286 (2007)
PP - 137
286
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION

Importance of researches on aflatoxins presence in herbal drugs 
Dzenita Softic, Sasa Pilipovic, Amar Elezovic
Institute for Quality Control of Medicines FBiH, Titova 9, Sarajevo, Bosnia and Herzegovina
Herbal drugs can be contaminated with various kinds of microorganisms of soil, air and animal origin. It is
of special importance the presence of toxicogenic mould, which number is being significantly increased due to the
inadequate drying, transport and storing. Majority of the present mould is being removed in the process of thermal
processing-preparation of teas. However, even after this processing a portion of those microorganisms as well as
products of its metabolism still remain present in herbal drugs and preparations. 
Micotoxins are widely known as contaminators of cereals and nuts especially those originating from warm
and humid areas. However, earlier researches show that those micotoxins may be present in herbal drugs samples.
Particular problem represents xerophylic moulds, that can develop by itself or in medium with low free water con-
tents (a
w
0,80). Most of those kinds of moulds belong to the genus: Aspergillus and Penicillium, the most frequent
contaminators of herbal drugs. Due to the high toxic level it is of highly significance the presence of the following:
aflotoxins, secondary products of the mould metabolism genus Aspergillus, especially species flavus and parasiticus.
In developed countries of the world a significant attention is being paid to the following up of the aflotoxin
presence and its quantity in the food in order to protect the human health. At the moment in Bosnia and Herzegovina
Rulebook from 1983 is still valid. Over viewing the available data on conducted researches on aflatoxin presence in
herbal drugs at the territory of the Region and making of this paper offered us the opportunity to give our contribu-
tion to rising of awareness on significance of those researches. Also, we would like to point out that harmonization
of domestic rules with rules of European Union is of outmost importance and necessary.  
Macedonian pharmaceutical bulletin 53 (1,2) 287 (2007)
PP - 138
287
^ETVRTI KONGRES NA FARMACIJATA NA MAKEDONIJA SO ME\UNARODNO U^ESTVO
FOURTH CONGRESS OF PHARMACY OF MACEDONIA WITH INTERNATIONAL PARTICIPATION