New phytochemical data on Dracocephalum species - Semmelweis ...

<strong>New</strong> <strong>phytochemical</strong> <strong>data</strong> on Dracocephalum species
Theses of the Ph.D. dissertation
András Zoltán Kakasy
Semmelweis University
Doctoral School of Pharmaceutical Sciences
Department of Pharmacognosy
Supervisor:
Prof. Dr. Éva Lemberkovics
Budapest
2006.

Ph.D. Thesis
NEW PHYTOCHEMICAL DATA ON DRACOCEPHALUM SPECIES
by András Zoltán Kakasy
Supervised by Prof. Dr. Éva Lemberkovics
Department of Pharmacognosy, Faculty of Pharmacy, Semmelweis University
SUMMARY
Most recent publications relating to Dracocephalum species have discussed the phytochemistry
and pharmacology of non-volatile and volatile compounds with promising pharmacological effects, such as
rosmarinic and caffeic acids, flavonoids with cytotoxic activity, or diterpenes with trypanocidal activity.
The aim of our work was to contribute new results concerning the <strong>phytochemical</strong> characterization of some
dragonhead species.
We have studied samples of D. moldavica L., D. ruyschiana L., D. argunense Fisch. ex Link, D.
bipinnatum Rupr., D. diversifolium Rupr., D. grandiflorum L., D. peregrinum L., D. renati Emb. and D.
rupestre Hance., all cultivated by ourselves.
In the essential oil of D. renati Emb. by means of GC and GC-MS we detected monoterpenes (limononene,
carvone, neral, geranial and linalyl acetate), methyl chavicol and sesquiterpenes (β-caryophyllene,
caryophyllene oxide and bicyclovetivenol). The essential oil of D. ruyschiana L. contains mainly
pinocamphone and isopinocamphone and smaller amounts of sesquiterpenes (β-caryophyllene,
caryophyllene oxide, β-cubebene, germacrene-D and elemol), β-pinene, myrcene, limonene, p-cymene and
methyl chavicol. The essential oil of D. grandiflorum L. was found to contain mainly sesquiterpenes
(aromadendrene, β-caryophyllene, caryophyllene oxide, β-cubebene and β-bourbonene) and carvacrol,
together with smaller amounts of β-asarone and methyl chavicol.
As compared with species belonging in the Lamiaceae family, in which ursolic and oleanolic acids were
detected in higher amounts (exceeding 3% and 1.5%, respectively), in the dragonhead species we found
only average quantities. Ursolic and oleanolic acids, measured by a GC method (after silylation) were
found in relatively high quantities in D. diversifolium Rupr. (0.81/0.25%) and D. peregrinum L.
(0.72/0.26%) 1 .
The main anthocyanidins of the purified and hydrolysed extract of D. moldavica L. (investigated by HPLC
analysis) proved to be delphinidin and cyaniding; pelargonidin was detected merely in traces. The GC-MS
analysis of the constituents of the extracted matrix (before and after hydrolysis) of D. moldavica L. and D.
ruyschiana L., in the form of their trimethylsilyl (oxime) ether/ester derivatives led to the identification and
quantification of more than 30 newly identified constituents, including sugars, sugar alcohols, flavonoids,
and aliphatic and aromatic carboxylic acids, together with their esters 2 .
Phenoloids, such as rosmarinic acid (26.76 mg/g), tannins (11.2%) and flavonoids (0.5%) accumulate in D.
moldavica L. in the early stages of ontogeny. Higher quantity of caffeic acid (0.5 mg/g) and essential oil
(0.40-0.83% v/m) were measured during flowering.
An infusion prepared from Moldavian dragonhead herb contains large quantity of tannins and up to 20-30%
of the phenolcarboxylic acids and flavonoids present in the herb. Alcohol, used as a solvent to prepare a
tincture, dissolves primarily the essential oils of the herb. The main volatile terpenoid of the infusion is
geraniol, while in the tincture geranyl acetate predominates.
We have established, that during the process of extraction of Moldavian dragonhead herb by supercritical
carbon dioxide, oxygenated monoterpenes are extracted at the beginning of the extraction process; less
volatile components and fatty acids were extracted later 3 .
1. INTRODUCTION, AIMS
1
Janicsák, G., Veres, K., Kakasy, A., Máthé, I. (2006) Biochemical Systematics and Ecology, 34: 392-396.
2
Kakasy, A., Füzfai, Zs., Kursinski, L., Molnár-Perl, I., Lemberkovics, É. (2006) Chromatographia 63: S17-S22
3
Kakasy, A., Lemberkovics, É., Simándi, B., Lelik, L., Héthelyi, É., Antal, I., Szőke, É. (2006) Flavour and Fragrance
Journal 21: 598-603.

The genus dragonhead (Dracocephalum, Lamiaceae) contains some 70 species, which to date
have been investigated primarily from the aspect of their essential oils. The Moldavian
dragonhead (D. moldavica L.), the most extensively studied species of the genus, is
naturalized in this area of Europe as a cultivated medicinal plant. The essential oil
composition of the species has been revealed in some detail, in contrast with the knowledge
on other substances in the herb. D. ruyschiana L. and D. austriacum L. are spontaneous in
Hungary, represented in the Hungarian flora by 30-40 specimens. This is a protected and
endangered species throughout Europe.
A survey of the literature on the essential oils of the dragonhead species yields detailed <strong>data</strong>
on some 20 species. Recent publications have discussed the phytochemistry and
pharmacology of the non-volatile and volatile compounds extracted from the dragonhead
species: e.g. rosmarinic and caffeic acids, lipophilic flavonoids with cytotoxic activity (D.
kotschyi Boiss.), monoterpene glycosides and diterpenes, which have trypanocidal activity (D.
komarovi Lipsky), and antinociceptive essential oil components (D. kotschyi Boiss.).
An extensive survey of the literature on the botanical, <strong>phytochemical</strong> and
(ethno)pharmacological aspects of dragonhead species aroused our interest in this genus. Our
aim was to contribute original <strong>phytochemical</strong> research results to the <strong>data</strong> available on this
genus.
The primary aim of our work was to carry out an extended <strong>phytochemical</strong> survey on a large
number of dragonhead species. Besides Moldavian dragonhead (chosen as the model species
of the genus) and the protected and endangered D. ruyschiana L., we planned to cultivated as
many available species as possible. Unfortunately, the cultivation of D. austriacum L. failed
several times. An additional aim was to examine the compositions of some less well studied
species (D. ruyschiana L., D. grandiflorum L. and D. renati Emb.) by GC and GC-MS
methods and to detect and quantify two triterpene carboxylic acids (ursolic and oleanolic
acids) in eight dragonhead species by a GC method after silylation. We also intended to detect
and quantify rosmarinic and caffeic acids in the same species by TLC-densitometry. It seemed
reasonable to examine the anthocyanins (a group of phenoloids with interesting biological
activities) of selected species (D. moldavica L. and D. ruyschiana L.) by means of column
chromatographic fractionation, an electrophoretic survey, TLC, HPLC and GC-MS (after
derivatization). The latter method seemed suitable for identification and quantification of
further biologically active constituents, such as sugars, sugar alcohols, fatty acids, flavonoids,
and aliphatic and aromatic carboxylic acids.

We set out to follow the quantitative fluctuations in some biologically active compound
groups (volatile oils, aromatic carboxylic acids, total polyphenol, tannin and flavonoids) of D.
moldavica L. during ontogeny and to compare the amounts of these substances in samples
collected in the same stage of the ontogeny, but from cultures at different geographic sites.
It is well known that the effect of a phytotherapeutic product is influenced considerably by the
extraction process, and our final aim was therefore to compare the influences of various
extraction methods (steam distillation, traditional extraction with organic solvents and
supercritical extraction) on the compositions of the essential oils obtained and to quantify the
main active constituent groups in infusions and tinctures.
2. MATERIALS AND METHODS
2.1. Plant material
Phytochemical analyses were carried out on samples of D. moldavica L. and D. ruyschiana
L., cultivated in Baksa, Marosvásárhely/Tîrgu Mureş (Transylvania, Romania) and Vácrátót
in 1999-2002. Samples of D. moldavica L. were gathered in well-determined stages of the
ontogeny. Samples of six further dragonhead species (D. argunense Fisch. ex Link, D.
bipinnatum Rupr., D. diversifolium Rupr., D. grandiflorum L., D. peregrinum L., D. renati
Emb. and D. rupestre Hance.) were obtained from the collection of the Botanical Garden of
the Institute of Ecology and Botany of the Hungarian Academy of Sciences in Vácrátót,
Hungary in 1999. Seeds of D. renati Emb. and D. ruyschiana L. were obtained via seedexchange
contracts (Copenhagen and Gießen).
2.2. Microscopy
Preparations (cross-sections of fresh leaves) were investigated with an Aksziszkop XL
microscope. Different types of covering and glandular trichomes were recorded.
2.3. Extraction methods
An infusion and a tincture (using 80% ethanol) of D. moldavica L. herb were prepared
according to the instructions of the Hungarian Pharmacopoeia (7 th edition). Soxhlet extracts of
the same plant material were made with n-hexane and ethanol as organic solvents. The main
parameters of the supercritical fluid carbon dioxide extraction: pressure 45 MPa, temperature

40 °C, flow rate of the solvent 7 kg/min., pressure in the first separator 8 MPa or 9 MPa, and
pressure in the second separator 2 MPa. We also performed fractionated extraction by
breaking off the extraction process at predetermined time intervals (each 30 min from the
beginning of the extraction process). In this manner, we collected four successive fractions
from the second separator.
Dried and powdered flower samples of D. moldavica L. were extracted with methanolacetone
1:1 acidified with hydrochloric acid (0.05% w/w). The extract obtained was purified
on a Sephadex LH-20 column using a gradient eluent system (water → methanol). Each
solution was acidified with hydrochloric acid (0.05% w/w). The major, intense violet fraction
was freeze-dried.
2.4. Qualitative and quantitative <strong>phytochemical</strong> analyses
Essential oils were obtained by steam distillation, using the apparatus official in the
Hungarian Pharmacopoeia (7 th edition), and quantified both volumetrically and
gravimetrically. The compositions of the essential oils were analysed by GC and GC-
MS. Main parameters: fused silica columns with the following stationary phases:
enantioselective Rt-βDEXm, DB-1701 and MDN-5S, programmed temperature, and
an electron ion trap detector in the MS section. The relative standard deviation
(RSD%) of the relative content for each identified constituent was calculated from the
<strong>data</strong> of three replicate analyses.
Ursolic and oleanolic acids were detected and quantified in purified and silylated
extracts by GC (a fused silica column with HP 5 as stationary phase, isothermal,
internal standard: cholesteryl acetate).
The presence of acylated anthocyanins in the purified and freeze-dried flower extracts
was verified by means of an electrophoretic survey (paper electrophoresis).
The sugar moieties of the anthocyanins (after acid hydrolysis of the freeze-dried
fractions) and the acylating acids (detached from the sugar moieties of the glycoside)
were identified by TLC methods, partly developed by ourselves.
A portion of the freeze-dried fraction was subjected to acid hydrolysis. The solution
containing the aglycones was extracted and passed through a C18 SPE column.
Analyses were performed on a Surveyor system (DAD detector). HPLC conditions: a
Phenomenex Luna (5 µm C18; 250x4.6 mm I.D.) column coupled with a SecurityGard
C18 guard column, isocratic conditions/gradient elution.

GC/MS analyses were carried out directly on the extracts (without Sephadex and SPM
purification) and also after extract hydrolysis (2 M trifluoroacetic acid, 2 h, 4 h).
Constituents were identified and quantified as their trimethylsilyl (oxime) ether/ester
derivatives, from a single run, on the basis of their total (TIC) and selective fragment
ion (SIM) responses. Calculations were related to the dry matter contents of the
extracts.
Rosmarinic and caffeic acids were detected and quantified in purified extracts by
TLC-densitometry.
The total flavonoid content of D. moldavica L. herb was measured by means of the
spectrophotometic method official in the German Pharmacopoeia (10 th edition). The
total polyphenol and tannin contents were quantified by the spectrophotometric
methods official in the Hungarian Pharmacopoeia (7 th edition).
3. RESULTS AND DISCUSSION
3.1. Microscopy
We have described different types of covering trichomes (e.g. D. ruyschiana L.: tooth-like
conical, D. grandiflorum L.: large, multicellular, with striated cuticle) and glandular
trichomes (mainly Lamiaceous with 8 secretory cells) on the leaf surface of 13 dragonhead
species, with special emphasis on distinctive features.
3.2. Essential oil compositions of D. renati Emb., D. grandiflorum L. and
D. ruyschiana L.
In the essential oil of D. renati Emb. (0.5% v/m), we identified 18.3% limonene and
oxygenated monoterpenes (carvone 2.1%, neral 0.53%, geranial 0.65% and linalyl acetate
0.74%), methyl chavicol (0.31%), and sesquiterpenes such as β-caryophyllene (1.07%),
caryophyllene oxide (0.69%) and bicyclovetivenol (0.25%). The main component of the
essential oil is an as yet unidentified compound (M + 150).
A common feature of the species belonging in the subgenera Dracocephalum and Ruyschiana
is their low essential oil content, and this accords well with our measurements (0.08%
essential oil in D. grandiflorum L.). The presence of sesquiterpene hydrocarbons such as

aromadendrene (5.7%), β-caryophyllene (8.7%), caryophyllene oxide (11.7%), β-cubebene
(3.2%) and β-bourbonene (22.1%) is characteristic of the essential oil profile of D.
grandiflorum L. Besides sesquiterpenes, carvacrol was identified in appreciable amount
(13.4%), together with smaller quantities of other phenylpropane compounds such as β-
asarone, methyl chavicol and anethol. These <strong>data</strong> are in reasonable accord with the earlier
classification of D. grandiflorum L., together with D. nutans L. and D. scrobiculatum Regel,
into the chemotype where the essential oil is dominated by sesquiterpenes and monoterpenes.
Nevertheless, our results indicate that the phenylpropane compounds belong more to the
profile of the essential oil of D. grandiflorum L. than monoterpenes. The occurrence of
methyl chavicol, anethol, β-cubebene, carvacrol, aromadendrene, caryophyllene oxide and β-
asarone in this essential oils is reported for the first time by ourselves.
The predominant compounds in the essential oil of D. ruyschiana L. (0.23%) are oxygenated
bicyclic monoterpenes, such as pinocamphone (43.6%) and isopinocamphone (21.5%).
Pinocamphone has been identified as main constituent, together with isopinocamphone in D.
nutans L. and D. nodulosum Rupr., and with trans-pinocamphone in D. fruticulosum Steph. ex
Willd. Besides the main components, we have identified monoterpenes: myrcene (3.12%),
limonene (0.65%), p-cymene (1.5%) and β-pinene (0.89%); sesquiterpenes: β-caryophyllene
(3.8%), caryophyllene oxide (1.6%), β-cubebene (1.6%), germacrene-D (3.6%) and elemol
(4.4%); and a phenylpropane: methyl chavicol (0.62%).
The essential oil compositions of D. ruyschiana L. and D. renati Emb. and the new
constituents of the essential oil of D. grandiflorum L. are reported for the first time in this
paper.
3.3. Ursolic and oleanolic acid contents of some dragonhead species
Both triterpenoid acids were detected in all investigated dragonhead species. As compared
with species belonging in the family Lamiaceae and the subfamily Nepetoideae, in which
ursolic and oleanolic acids were detected in higher amounts (exceeding 3% and 1.5%,
respectively), in the dragonhead species we measured only average quantities. These two
acids, measured by a GC method (after silylation), were found in relatively higher quantities
in D. diversifolium Rupr. (0.81/0.25%), D. peregrinum L. (0.72/0.26%) and D. ruyshiana L.
(0.67/0.24%).
3.4. Identification and quantification of rosmarinic and caffeic acids

Rosmarinic acid was detected in all investigated dragonhead species. In comparison with
Melissa officinalis L., official in the Hungarian Pharmacopoeia (8 th edition), with a total
hydroxycinnamic acid derivative content expressed in terms of rosmarinic acid of not less
than 40 mg/g, only D. moldavica L. (28.45 mg/g), D. renati Emb. (21.28 mg/g) and D.
argunense Fisch. ex Link (18.45 mg/g) contain rosmarinic acid in concentrations of the same
order of magnitude. With the exception of D. bibinnatum Rupr. and D. argunense Fisch. ex
Link., in each of the investigated dragonhead species caffeic acid was detected in measurable
amount, but in much lower quantity than rosmarinic acid (e.g. D. grandiflorum L. 5.13 mg/g
rosmarinic acid and 1.14 mg/g caffeic acid).
3.5. Anthocyanins
The main anthocyanidins identified in the purified and hydrolysed D. moldavica L. flower
extract by HPLC (spectral analysis, authentic standards) proved to be delphinidin and
cyanidin, while pelargonidin was detected only in traces.
From the electrophoretic mobility (2-2.3 cm) of the extracted pigments, it can be concluded
that the anthocyanins of D. moldavica L. (flower and stem) and D. ruyschina L. (flower) are
aclyted on the sugar moiety. By means of TLC methods, we detected glucose and rhamnose
as the sugar moieties of anthocyanins, and p-coumaric acid and malic acid as the acylating
acids of the sugar moieties.
3.6. Further non-volatile constituents in D. moldavica L. (flower and stem) and D.
ruyschiana L. (flower)
GC-MS analysis of the constituents of the extracted matrices (before and after hydrolysis) of
D. moldavica L. and D. ruyschiana L., in the form of their trimethylsilyl (oxime) ether/ester
derivatives resulted in the identification and quantification of more than 30 constituents,
quantitated partly via their TIC and partly via their SIM responses.
Identification/quantification proved to relate to a total of 35-69% (expressed as the dry matter
content of the extracts).
Identified as main constituents were aliphatic (succinic, malic, tartaric and citric acids) and
aromatic carboxylic acids (chlorogenic, caffeic, ferulic, 3,4-dihydroxyphenyllactic,
rosmarinic, hydroxybenzoic, trimethoxybenzoic and vanillic acids), together with their
corresponding esters (ferulic acid-3,4-dihydroxyphenyllactic acid ester), saccharides
(arabinose, rhamnose, glucose, galactose, disaccharides and tetrasaccharides), sugar alcohols

and acids (levulinic acid, arabitol, galacturonic acid and inositol), fatty acids (palmitic, stearic
and margarinic acids), apigenin and luteolin flavon aglycones and tocopherol, phosphoric acid
and quinic acid. As result of hydrolysis, the amounts of some constituents (succinic acid,
citric acid, trimethoxybenzoic acid, arabinose, glucose and galacturonic acid) increased, due
to the cleavage of the glycosidic linkages of saccharides and the decomposition of esters.
The apigenin content of the extract of D. ruychiana L. is only the one-tenth of to the amount
measured in the flower extract of the D. moldavica L. sample. Even in the hydrolysed sample
(0.035%), it proved to be one-twentieth of the corresponding value measured in the
hydrolysed extract of D. moldavica L. (0.76%). At the end of the hydrolysis, luteolin and
apigenin could not be detected, presumably because of their decomposition. Unfortunately,
the presence of the anthocyanidins could not be confirmed by GC-MS, very probably since
they are present in concentrations below the detection limits of our measurements: < 0.01%,
expressed as a percentage of the dry matter content of the extract.
The accuracy of the process (including extraction, derivatization and GC-MS quantitation),
characterized by the RSD% values of the constituents, proved to be < 5 RSD% in the case of
the major constituents (present in > 1%) and 5-10 RSD% for the minor constituents (< 1%).
3.7. Quantitative fluctuation of the main phenoloids and volatile terpenoids of
Dracocephalum moldavica L. during ontogeny
Phenoloids such as rosmarinic acid (26.76 mg/g), tannins (11.2%) and flavonoids (0.5%)
accumulate in D. moldavica L. in the early stages of ontogeny. Higher quantities of caffeic
acid (0.5 mg/g) and essential oil (0.40-0.83% v/m) were measured during flowering. The herb
can be considered a valuable tannin source when collected before flowering. Caffeic acid
reaches its maximum level during full flowering (0.5 mg/g). Independently of the placement
of cultures (with noteworthy climatic differences), the highest essential oil content was
measured in samples collected during flowering. On comparing the essential oil contents of
samples gathered in the same phase of the ontogeny, but from cultures at different geographic
sites, we concluded that higher the elevation, the better is the essential oil production of
culture.

3.8. Selectivity of extraction methods
Boiling water, used as a solvent to prepare the infusion of the Moldavian dragonhead herb,
dissolves nearly the whole quantity of tannins, and up to 20-30% of the phenolcarboxylic
acids, flavonoids and essential oil contained by the herb. Ethanol (80%), used as a solvent to
prepare a tincture, dissolves 68% of the essential oil, 27.6% of the total flavonoids, 46.5% of
the tannins, 3.3% of the rosmarinic acid and 2.3% of the caffeic acid contained by the herb.
The main volatile terpenoid in the infusion is geraniol, while that in the tincture is geranyl
acetate.
From a comparison of the yields and selectivities of different extraction methods
(hydrodistillation, extraction with organic solvents, and SFE) referred to the amounts of
extracted volatile terpenoids, the conclusion can be drawn that the selectivity is inversely
proportional to the yield. During the process of extraction of Moldavian dragonhead herb by
supercritical carbon dioxide, oxygenated monoterpenes are extracted at the beginning of the
extraction process; less volatile components and fatty acids are extracted later.

4. ACKNOWLEDGEMENTS
I am indebted to Prof. Éva Szőke, Director of the Department of Pharmacognosy at
Semmelweis University, who provided the facilities for me to prepare my Ph.D. work.
I wish to express my gratitude to Prof. Éva Lemberkovics, supervisor of my Ph.D. work, for
her magnanimous guidance and for her continuous professional and human encouragement.
I would like to thank Prof. Ágnes Kéry, Dr. László Kursinszki, Dr. Gabriella Marczal,
Éva Héthelyi, Kálmán Ditrói, Dr. Andrea Balázs and Dr. Mária Then for their
professional guidance, and the staff of the Department of Pharmacognosy at Semmelweis
University – especially Boross-Szabados – for their support.
I am pleased to take this opportunity to thank members of other universities and departments:
Prof. Imre Máthé, Dr. Gábor Janicsák, Dr. Vilmos Miklóssy-Vári,
Dr. Miklós Galántai † , Prof. Ibolya Molnár-Perl, Zsófia Füzfai, Dr. Béla Simándi, Dr.
György Stampf, Dr. Zoltán Kisgyörgy † , Prof. Árpád Gyéresi and Prof. Károly Csedő,
who all contributed so much to the present work.
I am grateful to my parents, to my sister and her family, and to my friends for their unbroken
encouragement.
This work was supported financially by a scholarship of the Hungarian Ministry of
Education, by the Hungarian Academy of Sciences (OTKA T.30034; F. 029249) and
GVOP tender 3.1.1.-2004-05-0397/3.0.

5. REFERENCES
5.1. Publications in the topics of the Ph.D. work
Articles
Kakasy, A., Lemberkovics, É., Kursinszki, L., Janicsák, G., Máthé, I., Szőke, É. (2000)
Adatok a Dracocephalum moldavica L. fitokémiai értékeléséhez. Orvostudományi értesítő
73: 410-417.
Kakasy, A., Lemberkovics, É., Kursinszki, L., Janicsák, G., Szőke, É. (2002) Data to the
<strong>phytochemical</strong> evaluation of Moldavian Dragonhead (Dracocephalum moldavica L.,
Lamiaceae). Herba Polonica 48: 112-119.
Lemberkovics, É., Kéry, Á., Simándi, B., Kakasy, A., Balázs, A., Héthelyi, É., Szőke É.
(2004) Extrakciós módszerek hatása az illóolajok összetételére. Acta Pharmaceutica
Hungarica 74: 166-170.
Héthelyi, É., Szarka, Sz., Kakasy, A., Galambosi, B., Szőke, É., Lemberkovics, É. (2005)
Dracocephalum speciesek illóolajának kémiai karaktere. Olaj, szappan, kozmetika 54: 75-81.
Kakasy, A., Lemberkovics, É., Simándi, B., Lelik, L., Héthelyi, É., Antal, I., Szőke, É.
(2006) Comparative study of traditional essential oil and supercritical fluid extracts of
Moldavian Dragonhead (Dracocephalum moldavica L.). Flavour and Fragrance Journal 21:
598-603. (If: 0,648)
Kakasy, A., Füzfai, Zs., Kursinski, L., Molnár-Perl, I., Lemberkovics, É. (2006) Analysis of
non-volatile constituents in Dracocephalum species by HPLC and GC/MS. Chromatographia
63: S17-S22 (If: 1,145)
Janicsák, G., Veres, K., Kakasy, A., Máthé, I. (2006) Study of the oleanolic and ursolic acid
contents of some species of the Lamiaceae. Biochemical Systematics and Ecology 34: 392-
396. (If: 0,704)
Oral presentations and posters
Kakasy, A., Lemberkovics, É., Kursinszki, L., Janicsák, G., Máthé, I., Szőke, É. (2000) A
Dracocephalum moldavica L. fenoloidjainak értékmérése. Semmelweis Egyetem Doktori
Iskola, II. Ph. D. Tudományos Napok, Budapest. Abstract: Book of Abstracts p. 61.
Kakasy, A., Lemberkovics, É., Janicsák, G., Kursinszki, L., Máthé, I., Szőke, É. (2000)
Adatok a Dracocephalum moldavica L. fitokémiai értékeléséhez. Erdélyi Múzeum Egyesület
Orvostudományi és Gyógyszerészeti Szakosztály X. Tudományos Ülésszaka,
Székelyudvarhely, Romania. Abstract: Book of Abstracts p. 26.
Kakasy, A., Kursinszki, L., Bihátsi-Karsai, É., Lelik, L., Janicsák, G., Máthé, I., Szőke, É.,
Lemberkovics, É. (2000) Phytochemical evaluation of Dracocephalum moldavica L. by
chromatographic methods. 2 nd International Symposium on Chromatography of natural
products, Lublin – Kazimierz Dolny, Poland. Abstract: Book of Abstracts p. 26

Kakasy, A., Janicsák, G., Kursinszki, L., Szőke, É., Lemberkovics, É. (2000) Phytochemical
analysis of Dracocephalum moldavica L. „Plante medicinale – realizări şi perspective”, Piatra
Neamţ, Romania. Abstract: in Acta Phytoterapica Romanica 6: 62-63.
Kakasy, A., Janicsák, G., Kursinszki, L., Szőke, É., Lemberkovics, É. (2000) Quantitative
variation of phenoloids and volatile terpenoids of Dracocephalum moldavica L. during
ontogeny. Lippay János & Vas Károly Scientific Symposium, Budapest. Abstract: Book of
Abstracts p. 204-205
Lemberkovics, É., Kéry, Á., Simándi, B., Kristó, TSz., Kakasy, A., Szőke, É. (2001) Effect
of extraction methods on the composition of essential oils. World Conference on Medicinal
and Aromatic Plants, Budapest. Abstract: Book of Abstracts p. 38
Lemberkovics, É., Kéry, Á., Simándi, B., Kakasy, A., Szőke, É. (2002) Influence of
extraction methods on the composition of essential oils. 33 rd International Symposium on
Essential Oils, Lisbon, Portugal. Abstract: Book of Abstracts p. 116
Lemberkovics, É., Kéry, Á., Simándi, B., Kakasy, A., Szőke, É. (2002) Quality of traditional
and supercritical extracts from essential oil plants. Medicinal plant research and utilization.
10 th National Conference on Medicinal Plants/8 th National Conference on Drug Research/5 th
National Conference on Phytotherapy. Kecskemét, Hungary. Abstract: Book of Abstracts p.
102, 203
Kakasy, A., Simándi, B., Lemberkovics, É., Szőke, É. (2003) A moldvai sárkányfű
(Dracocephalum moldavica L.) herbájából különböző extrakciós módszerekkel kivont
illóolajok összetételének vizsgálata. Semmelweis Egyetem Doktori Iskola, V. Ph.D.
Tudományos Napok, Abstract: Book of Abstracts p. 58-59
Kakasy, A., Lemberkovics, É., Marczal, G., Simándi, B., Szőke, É. (2003) A moldvai
sárkányfű (Dracocephalum moldavica L.) illóolajának és szuperkritikus extraktumainak
összehasonlító vizsgálata. Congressus Pharmaceuticus Hungaricus XII., Budapest.
Abstract: in Gyógyszerészet, (special issue 2003): 74.
Kakasy, A,, Lemberkovics, É,, Balázs, A., Kursinszki, L., Janicsák, G., Szőke, É. (2003)
Extraction and <strong>phytochemical</strong> investigation of the essential oil and phenoloids of Moldavian
Dragonhead (Dracocephalum moldavica L.). 45 th Annual Meeting of the Hungarian Society
of Gastroenterology, Balatonaliga, Hungary.
Abstract: in <strong>New</strong>sletter of the Hungarian Society of Gastroenterology 12: 97.
Kakasy, A., Marczal, G., Héthelyi, É., Lemberkovics, É. (2004) Dracocephalum fajok
mikromorfológiai és fitokémiai jellemzése. A gyógynövénykutatás aktuális kérdései 2004. Az
MGyT Gyógynövény Szakosztályának, a Magyar Biológiai Társaság Botanikai
Szakosztályának és az MTA Növénykémiai-Kemotaxonómiai Munkabizottságának közös
előadóülése, Budapest.
Lemberkovics, É., Kakasy, A., Böszörményi, A., Balázs, A., Héthelyi, É., Simándi, B., Kéry,
Á., Szők, É. (2005) Influence of technological and biological conditions on essential oil
composition. 36 th International Symposium on Essential Oils, Budapest. Abstract: Programme
and Book of Abstracts p. 7

Kakasy, A., Füzfai, Zs., Kursinszki, L., Molnár-Perl, I., Lemberkovics, É. (2005) Analysis of
non-volatile constituents in Dracocephalum moldavica L., D. ruyschiana L. and D.
ruyschiana L. by GC-MS and HPLC methods. 6 th Balaton Symposium on High-Performance
Separation Methods, Siófok. Abstract: Book of Abstracts p. 30
Kakasy, A., Janicsák, G., Veres, K., Lemberkovics, É. (2005) Triterpene and
phenolcarboxylic acids in Dracocephalum species. 1 st BBBB Conference on Pharmaceutical
Sciences, Siófok. Abstract: Book of Abstracts p.
Janicsák, G., Veres, K., Kakasy, A., Máthé, I. (2005) Az oleánol- és urzolsav mennyiségi
viszonyai a Lamiaceae családban. Gyógynövények napjaink gyógyszerészetében – tradíciók
és újdonságok. 11 th Hungarian Conference on Medicinal Plants, Dobogókő, Hungary.
Abstract: Programme and Book of Abstracts p. 52.
5.2. Publications in related topics
Article
Lemberkovics, É., Kéry, Á., Simándi, B., Kristó, TSz., Kakasy, A., Szőke, É. (2001)
Evaluation of supercritical plant extracts on volatile and non volatile biologically active
lipophil components. International Journal of Horticultural Science 7(2): 78-83.
Article published in proceeding book
Lemberkovics, É., Kéry, Á., Simándi, B., Kakasy, A., Szőke, É. (2003) Effect of extraction
methods on the composition of essential oils. In: Proceedings of the International Conference
on Medicinal and Aromatic Plants Part II. (Acta Horticulturae 597) Eds.: É. Szőke, I. Máthé,
G. Blunden, Á. Kéry. p. 49-56.
Oral presentation
Lemberkovics, É., Kéry, Á., Simándi, B., Kakasy, A., Czinner, E., Szőke, É. (2003)
Comparative study on supercritical extracts of aromatic plants. The 3 rd Word Congress on
Medicinal and Aromatic Plants for Human Welfare, Chiang-Mai, Thailand. Abstract: Book of
Abstracts p. 363