Essential Oil of Calamintha sylvatica Bromf. and Calamintha vardarensis Silic

Essential Oil of Calamintha sylvatica Bromf. and Calamintha vardarensis Silic

Mimica-Dukic, Neda


The essential oil of three populations of Calamintha sylvatica Bromf. native to the mountain region of southwestern Serbia and one population of Calamintha vardarensis Silic from southern former Yugoslav Republic of Macedonia were studied. The highest oil yields were found in C. vardarensis (1.54%), whereas in C. sylvatica oil yields varied from 0.11% up to 0.94%. In all samples, cis-piperitone oxide was a major compound. The highest percentage was found in the oil of C. vardarensis (65.6%;). In C. sylvatica piperitone oxide varied from 48.9-59.2%. In the examined samples, oil composition appeared to vary more according to geographical rather than genetic factors.

Key Word Index

Calamintha vardarensis, Calamintha sylvatica,Lamiaceae, essential oil composition, cis-piperitone oxide.


Calamintha Miller species are well represented in the flora of the former Yugoslavia. The species Calamintha glandulosa (Reg.) Bentham is widespread in the Mediterranean and Submediteranean areas of Dalmatia and Montenegro, while C. sylvntica Bromf. and C. grandiflom Moench is present in the continental part of Serbia. The newly discovered species C. vardarensis Silic is distributed at the Submediterranean zone of the former Macedonia and East Serbia (1,2). Culamintha species are known for their medicinal uses as a stimulant, diaphoretic, expectorant, antipyretic and sedative (3,4). Some species are reported to possess strong antibacterial and antifungal activities (5-7). The major compounds in the oils o f various Calamintha species belong to the C-3 oxygenated p-menthanes; these are: menthone, isomenthone, pulegone, piperitone, piperitone oxide, piperitenone, piperitenone oxide (8-14). Perez-Alonso et al. (15) reported on Calamintha oil rich in carvone and 1,8cineole. Great variations in oil composition occur both in the Calamintha genus both inter- and infraspecific.

In this paper we report on the oil composition of C. sylvattca in Serbia and C. vardarensis in the former Macedonia. The present study was aimed to contribute to the better knowledge of chemical features of Calamintha species of the Balkan Peninsula.


Calamintha sylvatica Bronii. was collected from three habitats (BeIi Rzav, Mitrovac and the gorge of Derventa) on the Tara mountain in Serbia and C. cardarensis in one habitat (Rodazde) in the former Yugoslav republic of Macedonia. Voucher specimens were deposited in the Herbarium ol The Department of Botany, Faculty of Pharmacy in Belgrade and were determined by the author.

oil isolation: Air-dried leaves and inflorescences of each collection were subjected to hydrodistillation for 3 h using a modified Clevenger-type apparatus and collected in hexane. The obtained oils were dried over anhydrous Na0SO-1, decanted, concentrated under reduced pressure (hexane removed) and weighed. oil samples were stored in a dark glass bottles and kept at 40C until analysis.

oil analysis: The oils were analyzed by GC/MS using a Hewlett Packard 6890/5973 system, with an HP-5 MS capillary column (30 m [chi] 0.25mm, filin thickness 0.25µm). GC oven initial temperature was 6O°C and programmed to 28O°C at the rate of 3°C/min. Helium was the carrier gas and the MS were taken at 70 eV. Library search was carried out using Wiley’s NIST/NBS MS libraries and the best match of their mass spectra with those of the literature (16).

Results and Discussion

oil yields from samples of C. sylvatica varied from 0.11% up to 0.94%. The highest oil content (1.50%) was found in the C. Vardarensis collected in the former Macedonia (Table I). Similar results on the oil content were reported by Pavlovic et al. (17), who found double the oil in C. vardarensis (1.26%) than in C. sylvatica (0.66%). Talcing into consideration the variation in oil content among C. sylvatica samples, it appears that the west side of Tara Mountain’s environmental and climatic conditions are more favorable to the oil synthesis than the east.

The oil composition of examined C. sylvatica and C. vardarensis oil is summarized in Table II. In the oil, 49 volatile constituents were identified. The chemical composition of analyzed samples exhibited only small qualitative differences (menthone, isomenthone and a-terpinyl acetate). The oils of all analyzed Cahimintha samples were comprised mainly of unsaturated oxygenated monoterpenes, among which cispiperitone oxide was dominant, and varied from 48.9% (C. sylvatica, east Tara) up to 65.6% (C. vardarensis). Besides, somewhat higher percentages of limonene (1.8-4.0%), [gamma]-terpinene (1.2-1.8%), cis-sabinene hydrate (4.0-4.4%) and terpincn-4-ol (2.2-4.0%) were recorded. The content of total sesquiterpenes varied from 12-18.2%. The main sesquiterpene in all samples was [beta]-caryophyllene, ranging from 5.0-7.5%. A considerable amount of germacrene D was also found (2.0-7.3%).

Although the oil composition of the examined Calamintha species was similar, differences in the percentage distribution of some compounds were noticeable. Interestingly, the oil composition of C. sylvatica from the east Tara was generally (except for the ratio of major compound) more similar to C. vardarensis than to the other two C. sylvatica collected from the opposite side of Tara Mountain. This similarity was reflected in the higher ratio of limonene (4.0%, both) and isopiperitenone (0.5 and 0.7%, respectively) as well as lower percentages of terpinen-4-ol (2.2% and 2.6%, respectively) and total sesquiterpenes (12%, both) in C. sylvatica (east Tara) and C. vardarensis, compared to both C.sylvatica populations, collected from the West side of Tara mountain. In addition, the oil from C. sylvatica from east Tara differed from the other examined, in regard to the considerable percentage of piperitenone oxide (6.5%). Sevarda et al. (18) reported on the great variation of the oil composition in C. vardarensis in the former Macedonia. The authors collected plants from three different habitats, all at the riverbanks. Depending on localities, dominant compounds were menthone (46%;), pulegone (44%) and piperitone (40%). Piperitone oxide was missing in all reported samples of C. vardarensis. The same authors also reported on the composition of C. sylvatica collected from tlie mountain in west Serbia. As dominant compound they found piperitenone oxide (57%), associated with p-cymene (17%). Recently, in C. stjlvatica, naturally occurring in Croatia. Stanic et al. (19) identified pulegone as the most abundant compound (54.2%). In our previous study, we also found pulegone as the major compound (57.9-71.2%) in two populations of C. sylvatica collected from east Serbia (20).

To the best of our knowledge, this is the first report of C. sylvatica considered as a piperitone oxide chemotype. However, several authors have reported piperitone oxide as a major compound in Calamintha nepeta (L) Savi (21). Obviously, a high overlapping in oil composition, not only among Calamintha species, but also among close genera such are Calamintha, Mentha, Satureja, Microineria etc. are present (17,22,23).

Thus, we assume, that in order to clarify the chemical features of Calamintha species distributed in the Balkan Peninsula, further and more complex study, including detailed cluster analysis should be done.


We thank the Ministry of Sciences, Technology and Development of Republic Serbia for financial support (Grant 1862).


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Neda Mimica-Dukic*

Institute of Chemistry, Faculty of Sciences, University of Novi Sad, 21000 Novi Sad, Trg D. Obradovia 3 Yugoslavia

Maria Couladis and Olga Tzakou

Department of Pharniacognosy, School of Pharmacy, University of Athens, Athens 157 71, Greece

Radiasa Jancic and Violeta Slavkovska

Department of Botany, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe, 450, 11 000 Belgrade, Yugoslavia

* Address for correspondence

Received: March 2002

Revised: July 2002

Accepted: September 2002

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