Agro-ecological cultivation, secondary metabolite characteristics and microbiological tests of lemon balm (Melissa officinalis) – the variety Citronella

  • I. Salamon University of Prešov
  • M. V. Kryvtsova Uzhhorod National University
  • K. I. Trush Uzhhorod National University
  • A. I. Fandalyuk Zakarpatian Institute of Agroindustrial Production
  • M. J. Spivak D. K. Zabolotny Institute of Microbiology and Virology of NAS of Ukraine
Keywords: essential oil; chemical composition; antimicrobial activity; aromatic plant.


Lemon balm (Melissa officinalis L.) is a native of the eastern Mediterranean region southward to Israel and Syria and eastward through the Crimea, the Caucasus, and Northern Iran to the Tien Shan and the Pamirs. It appears to have been naturalized in the Central Europe including the Carpathian Ukraine and Eastern Slovakia. The objective of this study was characterization of the chemical composition and antimicrobial activity of essential oils from Melissa officinalis L. variety Citronella in the conditions of cultivation in a lowland region of the Ukrainian Carpathians. Plant material, balm dry leaves and leafy tops of the shoots, were grown in a small-scale cultivation ground at the Zakarpatian Institute of Agroindustrial Production in Velikaja Bakta, Carpathian-Ukraine. The Citronella variety is used for herb production. Essential oil from balm parts (herb and leaves) was isolated by distillation with boiled water. Hydro-distillation lasted for 4 hours, sample weights were 10 g of dry drug matter. A modified Coocking & Middleton distillation apparatus was used. Identification and evaluation of balm essential oil were carried out using the selected analytical method of HPLC-DAD analysis. The final balm extract was analyzed directly on a Dionex UltiMate 3000 system. The analysis of balm essential oil was carried out using a gas chromatograph Varian 3090, connected to a MS Saturn 2100T integrator. The sensitivity of microorganisms to plant extracts was determined by the disk diffusion test (with disk diameter of 6 mm). The quantitative and qualitative characteristics of balm essential oil (0.05–0.12% v/w) and its composition show very high content of α-citrale (geranial) (44%), β-citrale (neral) (29%) and citronellal (9%). Rosmarinic acid can be extracted from leaves and its content equals up to 1.7 ± 0.1%. Composition of the natural substances depends on the balm taxon, origin, soil and climatic growing conditions, and methods of harvest and isolation and effective weed control which are used. Antibacterial effect of the oil showed low activity against all bacterial strains. Our results confirmed higher activity of EOs against Gram-positive bacteria than Gram-negative. The balm essential oil displayed antimitotic effect especially towards Candida glabrata clinical strain. Thus, studies have shown that Melissa officinalis variety of Citronella in the conditions of cultivation in the lowland region of Carpathian-Ukraine according to biochemical characteristics and microbiological properties corresponds to world analogues.


Babulka, P. (2005). La mélisse (Melissa officinalis L.). Phytotherapie, 3(3), 114–117.

Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71–79.

Boonyarikpunchai, W., Sukrong, S., & Towiwat, P. (2014). Antinociceptive and anti-inflammatory effects of rosmarinic acid isolated from Thunbergia laurifolia Lindl. Pharmacology Biochemistry and Behavior, 124(1), 67–73.

Burdock, G. A. (2009). Fenaroli's handbook of flavor ingredients. 6th Edition, CRC Press, Boca Raton.

Cosge, B., Ipek, A., & Gurbuz, B. (2009). GC/MS analysis of herbage essential oil from lemon balms (Melissa officinalis L.) grown in Turkey. Journal of Applied Biological Sciences, 3(2), 149–152.

Couladis, M., & Koutsaviti, A. (2017). Chemical composition of the essential oils of Salvia officinalis, S. fruticosa, Melissa officinalis, and their infusions. Ratarstvo i Povrtarstvo, 54(1), 36–41.

De Sousa, A. C., Gattass, C. R., Alviano, D. S., Alviano, C. S., Blank, A. F., & Alves, P. B. (2004). Melissa officinalisa L. essential oil: Antitumoral and antioxidant activities. Journal of Pharmacy and Pharmacology, 56(5), 677–681.

Fejér, J., & Gruľová, D. (2017). Vplyv exogennych faktorov na tvorbu sekundarnych metabolitov medovky lekarskej (Melissa officinalis L.) [The influence of exogenous factors on the formation of secondary metabolites of lemon balm]. Folia Oecologica, 9(2), 70–79 (in Slovak).

Friedman, T. (2015). The effect of rosmarinic acid on immunological and neurological systems: A basic science and clinical review. Journal of Restorative Medicine, 4(1), 50–59.

Gamaro, G. D., Suyenaga, E., Borsoi, M., Lermen, J., Pereira, P., & Ardenghi, P. (2011). Effect of rosmarinic and caffeic acids on inflammatory and nociception process in rats. International Scholarly Research Notices Pharmacology, 2011, article ID 451682.

Gurčík, Ľ., Dúbravská, R., & Miklovičová, J. (2012). Economics of the cultivation of Salvia officinalis and Melissa officinalis. Agricultural Economics (Zemědělská Ekonomika), 51(8), 348–356.

Helmy, S. (2006). Antioxidative properties and antimicrobial activity of different aromatic extracts of Melissa officinalis L. Arab Universities Journal of Agricultural Sciences, 14(1), 299–316.

Hendawy, S. F., & Khalid, K. A. (2005). Response of sage (Salvia officinalis L.) plants to zinc application under different salinity levels. Journal of Applied Sciences Research, 1(2), 147–155.

Ieri, F., Cecchi, L., Vignolini, P., Belcaro, M. F., & Romani, A. (2017). HPLC/DAD, GC/MS and GC/GC/TOF analysis of lemon balm (Melissa officinalis L.) samples as standardized raw material for food and nutraceutical uses. Advances Horticulturae Sciences, 31(2), 141–147.

Khalid, K. A., & Ahmed, A. M. A. (2011). Effect of harvest time on the lemom balm essential oils. Acta Horticulturae, 925, 237–242.

Kryvtsova, M. V., Kohuch, T. T., Salamon, I., & Spivak, M. J. (2018). Antimicrobial activity of some essential oils on Candida genus clinical isolates. Mikrobiolohichnyi Zhurnal, 80(4), 3–12.

Kryvtsova, M. V., Kostenko, Y. Y., & Salamon, I. (2018). Compositions of essential oils with antimicrobial properties against isolates from oral cavities of patients with inflammatory diseases of the periodontium. Regulatory Mechanisms in Biosystems, 9(4), 491–494.

Lust, J. (2014). The most complete catalog of the herbs ever published. The herb book. Dover Publication, Mineola, New York.

Macura, R., Michalczyk, M., & Banaś, J. (2011). Effect of essential oils of coriander (Coriandrum sativum L.) and lemon balm (Melissa officinalis L.) on quality of stored ground veal. Zywnosc Nauka Technologia Jakosc(Poland), 18(4), 127–137.

Murarikova, A., Kaffkova, K., Raab, S., & Neugebauerova, J. (2015) Evaluation of content of phenolics in Salvia species cultivated in South Moravian Region. Acta Facultatis Pharmaceuticae Universitatis Comenianae, 13(9), 18–22.

Nurzyńska-Wierdak, R., Bogucka-Kocka, A., & Szymczak, G. (2014). Volatile constituents of Melissa officinalis leaves determined by plant age. Natural Product Communication, 9(5), 703–706.

Ondrejovič, M., Kraic, F., Benkovičová, H., & Šilhár, S. (2012). Optimisation of antioxidant extraction from lemon balm (Melissa officinalis). Czech Journal of Food Sciences, 30(4), 385–393.

Patora, J., Majda, T., Gora, J., & Klimek, B. (2003). Variability in the content and composition of essential oil from lemon balm (Melissa officinalis L.) cultivated in Poland. Acta Poloniae Pharmaceutica – Drug Research, 60(5), 395–340.

Queiroz, R. M. de, Takiya, C. M., Guimarães, L. P. T. P., Rocha, G. da G., Alviano, D. S., Blank, A. F., & Gattass, C. R. (2014). Apoptosis-inducing effects of Melissa officinalis L. essential oil in glioblastoma multiforme cells. Cancer Investigation, 32(6), 226–235.

Rocha, J., Eduardo-Figueira, M., Barateiro, A., Fernandes, A., Brites, D., Bronze, R., & Fernandes, E. (2015). Anti-inflammatory effect of rosmarinic acid and an extract of Rosmarinus officinalis in rat models of local and systemic inflammation. Basic and Clinical Pharmacology and Toxicology, 116(5), 398–413.

Rostami, H., Kazemi, M., & Shafiei, S. (2012). Antibacterial activity of Lavandula officinalis and Melissa officinalis against some human pathogenic bacteria. Asian Journal of Biochemistry, 7(3), 133–142.

Safaeian, L., Sajjadi, S., Javanmard, S., Montazeri, H., & Samani, F. (2016). Protective effect of Melissa officinalis extract against H2O2-induced oxidative stress in human vascular endothelial cells. Research in Pharmaceutical Sciences, 11(5), 383.

Salamon, I., Kryvtsova, M., Bucko, D., & Tarawneh, A. (2018). Chemical characterization and antimicrobial activity of some essential oils after their industrial large-scale distillation. The Journal of Microbiology, Biotechnology and Food Sciences, 8(3), 965–969.

Salamon, I., Kryvtsova, M., Bucko, D., & Tatawneh, A. (2018). Natural components and antimicrobial activity of some essential oils after their industrial large-scale distillation. “International congress on natural products from plants and co-products to medicaments and Bio-agriculture”. Hammamet, Tunisia, 8–10 Novembwer, 2018.

Zazharskyi, V. V., Davydenko, P. О., Kulishenko, O. М., Borovik, I. V., & Brygadyrenko, V. V. (2019). Antimicrobial activity of ethanol extracts of 50 plant species. Biosystems Diversity, 27(2), in print.

How to Cite
Salamon, I., Kryvtsova, M. V., Trush, K. I., Fandalyuk, A. I., & Spivak, M. J. (2019). Agro-ecological cultivation, secondary metabolite characteristics and microbiological tests of lemon balm (Melissa officinalis) – the variety Citronella . Regulatory Mechanisms in Biosystems, 10(2), 264-268.