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Xhmka PACTHTE^BHOTO CBiPBA. 2019. №4. C. 129-134. DOI: 10.14258/j cprm.2019045267
UDC 633.88:576.8
STUDY OF THE COMPOSITION OF EXTRACT AND ANTIMYCOTIC PROPERTIES ANTHEMIS ALTISSIMA L.
© S.J. Mustafayeva1, S. V. Serkerov1", K.F. Bakhshaliyeva2
11nstitute of Botany of Azerbaijan National Academy of Sciences, Badamdar ave., 40, Baku, Az1073 (Azerbaijan), e-mail: s.serkerov@mail.ru 2 Institute of Microbiology of Azerbaijan National Academy of Sciences, Badamdar ave. 40, Baku, Az1073 (Azerbaijan)
The qualitative and quantitative composition of chemical components of ethanol extract of the above-ground part of Anthemis altissima L. (syn. Cota altissima (L.) J.Gay) species (family Asteraceae) collected during flowering phase from the flora of Azerbaijan has been studied.
The chemical composition of ethanol extract was analyzed by chromatography-mass-spectrometry methods. 54 components were identified in the extract. As a result of the study, it was found that the A. altissima contained benzene aromatic compounds (77.21%). Among them Benzene, (1-butylheptyl) - 7.84%, Benzene, (1-propyloctyl) - 7.71%, Benzene, (1-pentyloctyl) - 7.03%, Benzene, (1-pentylheptyl) - 5.69%, Benzene, (1-butyloctyl) - 5.37%, Benzene, (1-propylnonyl) - 5.02% were the main components.
In addition to the above components in the extract of the aerial part other biological active substances, such as alcohols (6.37% - of them prevail Hop-22(29)-en-3. p.-ol - 1.53%, y-Sitosterol - 1.32%), esters of acids (5.3% - of them prevail Hexa-decanoic acid, ethyl ester - 0.88%; Linoleic acid, ethyl ester - 0.63%) and alkanes (1.64% - of them prevail Eicosane - 0.53%; Pentacosane - 0.40%) were also identified. The share of unidentified components was 8.99%.
The article also presents the results of the study of the antifungal properties of A. altissima and its aqueous extract against the culture of pathogenic fungi Fusarium oxysporium and Aspergillus niger. The aqueous extract of A. altissima, in contrast to the plant itself, shows a fungistatic effect on the fungi Fusarium oxysporum and Aspergillus niger.
Keywords: Anthemis altissima; extract; chromatography-mass-spectrometry; biological active compounds; antifungal; cultures of pathogenic fungi Fusarium oxysporium and Aspergillus niger.
Introduction
The genus Anthemis L. of the tribes Anthemideae Cass. of the families Asteraceae Bercht. et J. Presl breed is recognized up to 210 species spread throughout Europe, the Mediterranean, West Asia, and Africa. Anthemis taxa have been commonly used as folk medicine, insecticides and ornamental plant from ancient times up to now. Anthemis is probably, in a phytochemical sense, one of the most repeatedly investigated genera of the Asteraceae family. A lot of work has been done on sesquiterpene lactones, polyacetylenes, flavonoids, essential oils [1-15]. Essential oils from flowers Anthemis is very useful for pharmaceuticals, and food additives, as well as important sources in food, aromatic and cosmetic industries. The species Anthemis has been studied on the chemical composition and diverse biological activities, such as their antioxidant, antifungal, antiplasmodial, antitumor, schistosomicidal, cytotoxic, anthelmintic, phytotoxic, analgesic effects [16-25].
Extracts are used to allay pain and irritation, clean wounds and ulcers and aid prevention, as well as therapy for irradiated skin injuries, treatment of cystitis and dental afflictions. Some species of Anthemis are being used as
~TT~T7 ttz ; T- : 7"T ttt 7 herbal tea to cure anxiety, flatulence stomach disorders
Mustafayeva Sitara Jalal - Leading scientist, candidate of
biological sciences (Ph.D), assistant of professor, Plant and carminative infusions [20].
resources depatoent, e-mail: s.serker°v@mail.ra 13 species of Anthemis are spread in
Serkerov Sirajeddin Veli - Doctor, Professor, Plant resources Azerbaijan [26]. The species are spread mainly in dry department, e-mail: s.serkerov@mail.ru
Bakhshaliyeva Konul Farrukh - Doctor, assistant of cla^ stonY ^ torn slopei5, in sandY placei5, claY stonei5,
professor, Head of laboratory of "Microbiological grasslands, rocks, on stony traps, and sometimes on a Biotechnology", e-mail: s.serkerov@mail.ru
* Corresponding author.
weed. Some species are known as medicines, decorative, dyes and insecticides plants. Antibacterial, antimicrobial and antifungal properties of these species are also known [6, 21].
As known the plants we use for different purposes (medicines, decorative, food, etc.) are also a good media for many bacteria and fungi. Thus, different mushrooms populate on plants that are favorable to them and cause pathogens to the plant. The intensive development of the disease depends on the interaction of the plant and pathogen with one another (plant resistance), but also on the environment. Diseases caused by fungi weaken the plant by infecting separate plant organs, and ultimately, the plant's total destruction. It should be noted that physiological and structural abnormalities in the herb have a great impact on its productivity. Sometimes the plant has a selective feature, thanks to the biological active compounds contained in the plants, and in particular, prevents any fungus from populating itself, which is antimicrobial.
The literature contains a fairly large number of research papers devoted to the study of the antifungal activity of various plant species (Funtumia elastica, Mallotus oppositifolius, Trachystemon orientalis, Smilax excelsa, Rhododendron ponticum, Phytolacca Americana, Prunus laurocerasus, Artemisia herba alba, Cotula cinerea, Aspho-delus tenuifolius, Euphorbia guyoniana and etc.) and extracts, essential oils, saponins, flavonoids, and other biologically active substances obtained from them [27-30].
Considering the beneficial properties of species of the genus Anthemis, we decided to investigate the chemical composition and antifungal properties of Anthemis altissima L. (syn. Cota altissima (L.) J.Gay), common in the territory of Azerbaijan.
In composition A. altissima identified of essential oils, sesquiterpenoids, steroids, aliphatic carbohydrates, aromatic compounds, high fatty acids, aldehydes, ketones, phenolcarbonic acids and others chemical components [6].
Materials and methods
The investigated materials were the above-ground part of species A. altissima. They were gathered in flowering phase in the village Avaxil of Shamakhi region, Pirqulu settlement, Pirsaat river, yard areas and grassland, as well as Shamakhi city. Here A. altissma forms large spots with an abundance mark of 3 among the forbs.
A. altissima is annual plant with a height of 20-80 sm. The leaves are oblong egg-shaped. The basket are one by one. The flowers are white. Seeds are light-brown and compressed. Flowering time of plant from May to July, seeds - June to August.
A. altissima is found everywhere in Azerbaijan except Nakhchivan. From lowland to middle mountain range, it spreads along the road between fields, plantings, gardens, bushes and grass stones. Weed, forest-meadow is a representative of plant species. It is xeromezophyte plant. Geographical type - xerophytes, class - intermediate sea [31].
The harvested plant material of the above-ground part of A.altissima was dried and crushed. The 300 g of plant raw material A. altissima was extracted in 3 days 3 times with 95% ethanol (alcohol) 3 times a day.
The chemical composition of ethanol extract of A. altissima was analyzed by using chromatography-mass-spectrometry methods.
The component composition of the obtained plant extract A. altissima was investigated by gas chromatog-raphy-mass spectrometry (GC/MS) on an Agilent Technologies gas chromatograph 6890N Network CG System, 5975 inert Mass Selective Detector mass spectrometer as a Split/Splitless detector, Injection - Split, Inlet pressure 60.608 kPa, Split/ Splitless detector 100 Low Mass - 40, High Mass - 400, Thresold 150. A 30-meter capillary quartz column "HP-5MS 5% Metil Siloxane" with an internal diameter of 0.25 mm and a slick of a stationary phase of 0.25 ^ was used. Analyzes were carried out in temperature programming mode from 50 °C to 280 °C at a speed of 15 °C/min. Column temperature: initial temperature 50 °C - 2 minutes is stable; raising the temperature 15 °C/min. up to 200 °C - 6 minutes stable; raising the temperature 15 °C/min. up to 280 °C - 10 minutes stable. Vacuum - HiVac - 3.38* 10-5. Extract diluted with methanol : chloroform (1 : 2).Velocity of carrier gas (He) 1 ml/min. Input samples with current division (1 : 50). The analysis time is 45 minutes. To identify the connection using the standard mass spectrometric library NIST (the NIST Mass Spectral Seach Proqramfo the NIST/EPA/NIH/Mass Spectral Library Version 2.0.g.buld May 19 2011) [32].
The antifungal activity of the A. altissima and aqueous extracts obtained from them was studied by conventional methods against the culture of microscopic pathogenic fungi Fusarium oxysporum and Aspergillus niger, stored in the museum of the Institute of Microbiology ANAS [33].
Antifungal activity was measured in two steps as follows: development of fungus in solid media and development of fungus in aqueous extract of different concentrations from A. altissima.
Development of fungi in solid media. The material obtained from dried A. altissima plants was ground into small pieces of 0.5 to 1 cm in size and wetted with 55-60% water. Therefore, fungi can develop in a solid medium. The pH level was relatively constant in the range of 6.5 to 7. The substrate, widely washed with culture, was placed in Petri dishes and sterilized at 1 atm pressure for 45 minutes. After sterilization, the fungal biomass was also inoculated into Petri dishes and incubated at 25-27 °C. The number of colonies was performed on each of the dishes 3, 5 and 7 days after the inoculation.
The development of fungi in different extracts. For study antifungal effects of extracts from aboveground parts species A. altissima was taken various coagulation extracts (5, 10, 15 q) and it has been heated. The extraction pour into glass bottles. Then, extraction was cooled, filtered and poured into glass bottles. It was sterilized at 0.5 atm pressure for 30 minutes. The test was conducted at pH levels of 6, 5 and 7. Fungi strains were inoculated into extract inside a test tube. Then they were incubated at 25-27 °C for seven days. Czapek's media was used for comparing results.
Experimental part
As mentioned above, 300 grams of plant raw material A. altissima was extracted with ethanol and total of 26 grams of extractable ingredients were obtained, which means 8.7% of the production. It was identified 54 components from the ethanol extract of the A. altissima by using chromato-mass-spectroscopy method. The results are given in the table.
As can be seen from the table, the extract contains aromatic compounds, alcohols, acids, alkanes, lactone, coumarine, triterpenoids and terpenes. Preference is given to benzene-cyclic aromatic compounds - 77.21%. Among them are Benzene, (1-butylheptyl) - 7.84%, Benzene, (1-propyloctyl) - 7.71%, Benzene, (1-pentyloctyl) - 7.03%, Benzene, (1-pentylheptyl) - 5.69%, Benzene, (1-butyloctyl) - 5.37%, Benzene, (1-propylnonyl) - 5.02% the main components.
in addition to the above components in the extract of the aerial part other biological active substances, such as alcohols (6.37% - of them prevail Hop-22(29)-en-3 p-ol - 1.53%, y-Sitosterol - 1.32%), esters of acids (5.3% -of them prevail Hexadecanoic acid, ethyl ester 0.88%; Linoleic acid, ethyl ester 0.63%) and alkanes (1.64% - of them prevail Eicosane - 0.53%; Pentacosane - 0.40%) have been identified. The share of unidentified components was 8.99%.
As we noted A. altissima species antifungal activity was measured in two steps as follows: development of fungus in solid media and development of fungus in aqueous extract of different concentrations from A. altissima.
Figure 1 shows the results of the growth of phytopathogen fungi Fusarium oxysporum and Aspergillus niger on A. altissima.
As shown in figure 1 in solid food environment consisting of A. altissima, the size of fungal colonies of Fusarium oxysporium on 3-rd day were 1.0 sm, 5-th day 2.6, and 7-th day 5.6 sm, the colonies of Aspergillus niger fungi, respectively, were 4.2; 6.9; 8.0 sm.
The control options size of fungi colonies was 8.0 sm.
Thus, the results show that aboveground parts A. altissima can not be used as an antifungal vehicle.
When analyzing literature sources, we encountered similar studies on the antifungal activity of water extract of A. altissima. Aqueous extract A. altissima growing in the Kermanshah acts antifungal against Phytophthora drechsleri and prevented the growth [28]. Out of 54 identified components of the extract, 20 components make up the aromatic compounds. besides, among the components found in the extract there are no compounds that are contained in large quantities, and therefore it is not possible to connect the antifungal properties with any predominant components. According to our assumptions, the antifungal activity of the extract is probably due to the synergistic active components of the aromatic compounds.
Figure 2 presents the results of the effect of different concentrations of water extract of A. altissima on the development of the fungi Fusarium oxysporum and Aspergillus niger.
Thus, as shown in Figure 2, the aqueous extract of A. altissima in different concentrations (5, 10, 15 g) had a fungistatic effect on both fungi (Fusarium oxysporium and Aspergillus niger), inhibition was observed to one degree or another. In contrast to the control (5.12-5.52 g/l), biomass ranged from 0.09 to 0.58 g/l.
The results obtained from the aqueous extracts of A. altissima can be used in the preparation of new antifungal
drugs.
The content of etanol extract of the above-ground part of Anthemis altissima
No. Retention Name of the compound Molecular for- Peak (%)
time mula
1 7.169 2-Hydroxy-gamma-butyrolactone C4H6O3 1.44
2 8.216 Thymine C5H6N2O2 0.49
3 8.492 Butanoic acid, 3-amino- C4H9NO2 0.60
4 9.039 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- C6H8O4 0.67
5 9.516 Catechol C6H6O2 0.24
6 9.775 Benzofuran, 2,3-dihydro- C8H8O 0.17
7 11.557 Naphthalene, 2-methyl- C11H10 0.45
8 11.851 2,1,3-Benzothiadiazole C6H4N2S 1.16
9 12.098 Silane, dimethyl(4-acetylphenoxy)isobutoxy- C14H22O3Si 1.42
10 12.692 Benzene, (1-butylhexyl)- C16H26 1.29
11 12.780 Benzene, (1-propylheptyl)- C16H26 0.97
12 12.951 Benzene, (1-ethyloctyl)- C16H26 1.24
13 13.133 Sorbitol C6H14O6 0.82
14 13.263 Benzene, (1-methylnonyl)- C16H26 2.00
15 13.374 Benzene, (1-pentylheptyl)- C18H30 0.25
16 13.521 Benzene, (1-butylheptyl)- C17H28 7.84
17 13.621 Benzene, (1-propyloctyl)- C17H28 7.71
18 13.839 Benzene, (1-ethylnonyl)- C17H28 3.60
19 14.221 Benzene, (1-methyldecyl)- C17H28 3.87
20 14.304 Bicyclo[6.3.0]undeca-1(8),9-diene, 11,11-dimethyl- C13H20 0.24
21 14.439 Benzene, (1-pentylheptyl)- C18H30 5.69
22 14.492 Benzene, (1-butyloctyl)- C18H30 5.37
23 14.639 Benzene, (1-propylnonyl)- C18H30 5.02
24 14.915 Benzene, (1-ethyldecyl)- C18H30 4.23
25 15.421 Benzene, (1-methylundecyl)- C18H30 4.74
26 15.645 Benzene, (1-pentyloctyl)- C19H32 7.03
27 15.739 Benzene, (1-butylnonyl)- C19H32 4.79
28 15.939 Benzene, (1-propyldecyl)- C19H32 3.85
29 16.315 Benzene, (1-ethylundecyl)- C19H32 3.55
30 16.939 Benzene, (1-methyldodecyl)- C19H32 3.45
31 17.692 Scopoletin C10H8O4 0.38
32 18.151 Hexadecanoic acid, ethyl ester C18H36O2 0.88
33 19.992 Phytol C20H40O 0.72
34 20.786 Linoleic acid ethyl ester C20H36O2 0.63
35 20.874 Ethyl 9,12,15 -octadecatrienoate C20H34O2 0.62
36 22.827 3-Oxatricyclo [4.1.1.0 (2,4)] octane, 2,7,7-trimethyl- C10H16O 0.25
37 22.903 Heptadecane C17H36 0.39
38 23.550 3-Cyclohexene-1-methanol, 6-methyl C8H14O 1.08
39 24.162 Benzylcarbamate C8H9NO2 0.84
40 24.421 Phenol, 2,2'-methylenebis[6-( 1, 1 -dimethylethyl)-4-methyl- C23H32O2 0.35
41 24.556 2,3,4-Trimethylpyrrole C7H11N 0.85
42 25.709 Pentacosane C25H52 0.40
43 25.815 Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester C19H38O4 0.59
44 26.079 Phthalic acid, di(2-propylpentyl) ester C24H38O4 0.51
45 28.103 9,12-Octadecadienoic acid (Z,Z)-2,3-dihydroxypropyl ester C21H38O4 0.36
46 28.209 Linolenic acid, 2-hydroxy1- (hydroxymethyl) ethyl ester C21H36O4 0.46
47 28.285 Heneicosane C21H44 0.32
48 29.432 Decanedioic acid, bis(2-ethylhexyl) ester C26H50O4 0.30
49 30.661 Eicosane C20H42 0.53
50 35.544 Stigmasterol C29H48O 0.57
51 36.726 y-Sitosterol C29H50O 1.32
52 37.561 P-Amyrin C30H50O 0.62
53 40.743 Taraxasterol C30H50O 0.82
54 41.049 Hop-22(29)-en-3 P-ol C30H50O 1.53
HH
3day 5day 7day Kontrol
Fig. 1. Development processes for Fusarium oxysporum and Aspergillus niger colonies in solid media prepared with A. Altissima by abscissa -developments of colonies fungus, sm; by horizontals -days
___I :
5gr lOgr 15gr kontrol
Fig. 2. Effects of extracts from A. altissima on Fusarium oxysporum and Aspergillus niger fungal colonies by abscissa - dry biomass, q/l; by horizontals - aqueous extact
Conclusion
1. The above-ground part of species A. altissima were gathered in flowering phase from the village Avaxil of Shamakhi region, Pirqulu settlement, Pirsaat river, yard areas and grassland, as well as Shamakhi city.
2. After ethanol extracting of 300 g of A. altissima plant, was obtained 26 g of extractable ingredients, which means 8.7% of output.
3. 54 components from the ethanol extract of the A. altissima were identified by the chromato-mass-spectros-copy method.
4. The extract of A. altissima contains aromatic compounds, alcohols, esters of acids, alkanes, lactone, cou-marine and terpenes. Preference is given to benzene-cyclic aromatic compounds - 77.21%. Among them Benzene, (1-butylheptyl) - 7.84%, Benzene, (1-propyloctyl) - 7.71%, Benzene, (1-pentyloctyl) - 7.03% are the main components. Of the identified components can be also noted alcohols (6.37%), esters of acids (5.3%), and alkanes (1.64%). The share of unidentified components was 8.99%.
5. Unlike the control variant, water extracts of A. altissima have a rather high fungistatic effect on the pathogenic fungi Fusarium oxysporium and Aspergillus niger, which allows them to be used for the preparation of new antifungal drugs.
References
1. Al-Humaidi J. Oriental J. Chem., 2015, vol. 31, no. 4, pp. 2265-2270.
2. Konstantinopoulou M., Karioti A., Skaltsas S., Skaltsa H. J. Nat. Prod., 2003, vol. 66, pp. 699-702. DOI: 10.1021/np020472m.
3. Masterova I., Grancai D., Grancaiova Z., Pour M., Ubik K. Pharmazie, 2005, vol. 60 pp. 956-957.
4. Pavlovic M., Kovacevic N., Tzakou O., Couladis M. Chem. Nat. Compd., 2007, vol. 43, pp. 512-514. DOI: 10.1007/s10600-007-0179-x
5. Vuckovic I., Vujusic L., Milosavljevic S. Serb. Chem. Soc., 2005, vol. 71, pp. 127-133.
6. Rastitel'nyye resursy Rossii: dikorastushchiye rasteniya, ikh komponentnyy sostav i biologicheskaya aktivnost'. [Plant resources of the Russia: Wild flowering plants, their component composition and biological activity]. St. Petersburg, Moscow, 2012, vol. 5, 317 p. (in Russ.).
7. Vujisic L., Vuckovic I., Vajs V., Tesevic V., Janackovic P., Milosavljevic S. Chemistry of Natural Compounds, 2011, vol. 47, no. 3, p. 459.
8. Staneva J.D., Trendafilova-Savkova A., Todorova M.N., Evstatieva L.N., Vitkova A. Z. Naturforsch. C., 2004, vol. 59, no. 3-4, pp. 161-165.
9. Saroglou V., Dorizas N., Kypriotakis Z., Skaltsa H.D. J. Chromatogr. A, 2006, vol. 1104, no. 1-2, pp. 313-322, DOI: 10.1016/j.chroma.2005.11.087.
10. Rezaee M.B., Jaimand K., Assareh M.H. Iranian Journal of Pharmaceutical Research, 2004, vol. 3, suppl. 2, p. 89.
11. Rezaee M.B., Jaimand K., Assareh M.H. Journal of Essential Oil Research, 2006, vol. 18, no. 2, pp. 152-153.
12. Rezaee M.B., Jaimand K. Journal of Essential Oil Research, 2008, vol. 20, no. 2, pp. 172-173.
13. Rezaee M.B., Jaimand K. Faslnamah-I Giyahan-I Daruyi., 2007, vol. 6, no. 22, pp. 99-105; Chem. Abstr., 2009, vol. 150, no. 393461.
14. Pavlovic M., Kovacevic N., Tzakou O., Couladis M. Flavour and Fragrance Journal, 2006, vol. 21, no. 2, pp. 297299, DOI: 10.1002/ffj.1592
15. Hanganu D., Pintea A., Marculescu A., Toma C., Mirel S. Farmacia, 2008, vol. 56, no. 3, pp. 344-351.
16. Amjad L., Madani M., Rezvani Z. World Acad. Sci. Eng. Technol., 2012, vol. 61, pp. 901-902.
17. Burim R.V., Canalle R., Lopes J.L.C., Takahashi C.S. Genet. Mol. Biol., 1999, vol. 22, pp. 401-406, DOI: 10.1590/S1415-47571999000300020.
18. Hajdu Z., Zupko I., Rethy B., Forgo P., Hohmann J. PlantaMed., 2010, vol. 76, pp. 94-96.
19. Papaioannoua P., Lazaria D., Kariotib A., Soulelesa C., Heilmannc J., Hadjipavlou-Litinad D., Skaltsab H. Z. Naturforsch., 2007, vol. 62, pp. 326-330.
20. Vakili E., Amjad L., Karbasizade V. Int. J. Agric. Crop Sci., 2013, vol. 5, pp.1464-1467.
21. Mustafayeva S.J., Bakhshaliyeva K.F. Materialy V mezhdunarodnoy nauchnoy konferentsii «Printsipy i sposoby sokhraneniya bioraznoobraziya». [Materials V Int. Scientific. conf., "Principles and retain of Biodiversity Conservation"], Yoshkar-Ola, 2013, pp. 280-284. (in Russ.).
22. Vuckovic I., Vujisic L., Klaas C.A., Merfort I., Milosavljevic S. Nat. Prod. Res., 2011, vol. 25, no. 8, pp. 800-805.
23. Samadi N., Manayi A., Vazirian M., Samadi M., Zeinalzadeh Z., Saghari Z., Abadian N., Mozaffarian V.O-A., Khanavi M. Natural Product Research: Formerly Natural Product Letters, 2012, vol. 26, no. 20, pp. 1931-1934.
24. Quarenghi M.V., Tereschuk M.L., Baigori M.D., Abdala L.R. Fitoterapia, 2000, vol. 71, no. 6, pp. 710-712.
25. Akgul C., Saglikoglu G. indian J. Biochem. Biophys., 2005, vol. 41, no. 6, pp. 395-397.
26. Mustafayeva S.C. News ANAS (Biology and Medical Sciences), 2013, vol. 68, no. 1, pp. 17-23.
27. Adekunle1 A.A, Ikumapayi A.M. West Indian Med. J., 2006, vol. 55, no. 4, pp. 219-223, DOI: 10.1590/S0043-31442006000400003.
28. Abdolmaleki M., Bahraminejad S., Abbasi S. J. Med. Plant., 2011, vol. 10, no. 38, pp. 148-155.
29. Satish S., Raghavendra M.P., Raveesha K.A. Journal Archives of Phytopathology and Plant Protection, 2009, vol. 42, no. 7, pp. 618-625, DOI: 10.1080/03235400701287578.
30. Nasrine S., Sultan A., Mohammed S., Valeria T., Iman B., Naima G., Samia B. Biomed. Res. Int., 2017, vol. 2017, article 7526291, DOI: 10.1155/2017/7526291.
31. FloraAzerbaydzhana. [Flora of Azerbaijan]. Baku, 1961, vol. VIII, pp. 248-265. (in Russ.).
32. Adams R.P. Identification of Essential oil Components by Gas chromatography /Mass Spectroscopy, Alluned Publishing Corp., Carol Stream, IL, USA, 2007, 803 p.
33. Metody eksperimental'noymikologii. [Methods of experimental mycology], ed. V.I. Bilai, Kiev, 1982, 500 p. (in Russ.).
Received March 11, 2019 Revised May 20, 2019 Accepted May 27, 2019
For citing: Mustafayeva S.J., Serkerov S.V., Bakhshaliyeva K.F. Khimiya Rastitel'nogo Syr'ya, 2019, no. 4, pp. 129134. (in Russ.). DOI: 10.14258/jcprm.2019045267.
