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Abstract

In the present study, the chemical composition of the essential oils from aerial parts of Ballota nigra subsp. uncinata (Bég.) Patzak collected in Sicily was evaluated by gas chromatography (GC) and GC-mass spectrometry. The main components of the oil were (E)-phytol (20.0%), α-pinene (9.0%), hexahydrofarnesyl acetone (5.7%), and α-selinene (5.1%). Cluster analysis of the essential oil compositions of all the taxa belonging to B. nigra s.l. group was performed.

Keywords

Lamiaceae Stachydeae essential oil

Ballota L. (Lamiaceae) is a genus belonging to the tribe Stachydeae, subtribe Ballotae. The Plant List, which has been used to validate the scientific names of the species, includes more than 160 scientific plant names of species rank for the genus Ballota. Of these, only 30 are accepted species names. They are native to Macaronesia, Europe, Mediterranean to West Asia, Mauritania, Chad, and South Africa. Ballota species are perennial herbs characterized by flowers held in verticillasters and by an unpleasant aromatic foliage.¹

Lamiaceae taxa have attracted interest by the researcher for the valuable biological properties of their extracts, such as antibacterial, antioxidant, and hypoglycemia.² Furthermore, Lamiaceae essential oils have shown a promising anti-inflammatory potential³ and are widely used in aromatherapy for several minor clinical uses.⁴ In this context, Ballota species have been used in folk medicine as antiulcer, antispasmodic, diuretic, choleretic, antihemorrhoidal, and sedative agents.⁵ The antimicrobial activities^6
-8 and the antioxidant activities⁹ of Ballota species were recently reported as well as the antifungal activities of some flavonoids isolated from 2 species.^10,11 The water extract has been reported to have antinociceptive, antiinflammatory, and hepatoprotective activities.¹² In Europe, the polar extracts of the flowering aerial parts of Ballota are commonly used due to their neurosedative activity.^13,14 More recently, the general antioxidant activity,¹⁵ the in vitro inhibition of LDL (low-density lipoprotein) peroxidation¹⁶ and the antibacterial activity^17,18 of these plants have been published. Phytochemical investigations showed that labdane diterpenoids, flavonoids, and phenylpropanoids are the characteristic features of the genus, and recently the occurrence of non-volatile and volatile metabolites, the ethnopharmacological uses, and the biological properties of all the studied taxa of Ballota have been reviewed.¹⁹

As a continuation of our researches on Ballota species,^20
-22 we decided to investigate the chemical composition of the essential oil B. nigra subsp. uncinata, collected in Sicily.

Ballota nigra subsp. uncinata (Bég.) Patzak (syn. Ballota nigra subsp. ruderalis (Sw.) Briq.) is present in the western part of North Africa, Southern Europe, Turkey, and Mediterranean Middle East.²³ Previous investigations on this taxon reported the occurrence of only 1 labdane diterpenoid, dehydrohispanolone,^24,25 the chemical composition of the essential oil of Turkish accession,²⁶ and its antilisterial activity against L. monocytogenes.²⁷

Results and Discussion

Hydrodistillation of B. nigra subsp. uncinata (B.n.u.) aerial parts gave a pale yellow oil. Overall, 49 compounds have been identified in B.n.u., representing 96.6% of the total components. The components are listed in Table 1 according to their retention indices on an HP 5 MS column and are classified on the basis of their chemical structures into 9 classes.

Table 1

Percent Composition of the Essential Oils of Aerial Parts of Ballota nigra subsp. uncinata Arranged by Class.

K_i ^a	K_i ^b	Component	B.n.u.^c	Ident. ^d
930	1014	α-Thujene	1.7	1, 2
936	1075	α-Pinene	9.0	1, 2, 3
973	1132	Sabinene	0.6	1, 2
978	1118	β-Pinene	1.1	1, 2, 3
1030	1203	Limonene	0.5	1, 2, 3
		Monterpene hydrocarbons	12.9
1098	1553	Linalool	0.8	1, 2, 3
1143	1532	Camphor	0.4	1, 2, 3
1167	1719	Borneol	0.9	1, 2, 3
1176	1611	Terpinen-4-ol	0.4	1, 2, 3
1187	1706	α-Terpineol	0.6	1, 2, 3
1262	1583	cis-Chrysanthenyl acetate	0.4	1, 2
		Oxygenated monoterpenes	3.5
1363	1492	Cyclosativene	2.1	1, 2
1377	1497	α-Copaene	1.8	1, 2
1385	1535	β-Bourbonene	0.7	1, 2
1404	1666	(Z)-Caryophyllene	4.2	1, 2, 3
1407	1538	α-Gurjunene	0.5	1, 2
1418	1612	(E)-Caryophyllene	2.2	1, 2, 3
1437	1628	Aromadendrene	2.6	1, 2
1463	1661	allo-Aromadendrene	0.6	1, 2
1475	1715	β-Selinene	2.0	1, 2
1477	1726	Germacrene D	3.7	1 ,2
1479	1698	δ-Selinene	0.8	1, 2
1487	1731	α-Selinene	5.1	1, 2
1495	1740	Valencene	2.1	1, 2
1521	1773	7-epi-α-Selinene	1.1	1, 2
1526	1173	δ-Cadinene	1.2	1, 2
1554	1856	Germacrene B	t^e	1, 2
		Sesquiterpene hydrocarbons	30.7
1527	2001	Isocaryophyllene oxide	0.5	1, 2
1581	2008	Caryophyllene oxide	1.1	1, 2, 3
1584	2176	Longiborneol	1.9	1, 2
1586	2099	Globulol	0.8	1, 2
1593	2104	Viridiflorol	2.8	1, 2
1636	2185	γ-Eudesmol	1.5	1, 2
1735	2355	Eremophilone	0.6	1, 2
		Oxygenated sesquiterpenes	9.2
1836	1963	Neophytadiene	0.5	1, 2
2054	2524	Abietatriene	1.0	1, 2
2132	2625	(E)-Phytol	20.0	1, 2
		Diterpenes	21.5
854	1209	(E)−2-Hexenal	0.6	1, 2
1102	1401	Nonanal	0.7	1, 2
1380	1835	β-Damascenone	0.6	1, 2
1845	2131	Hexahydrofarnesyl acetone	5.7	1, 2
1915	2387	(E,E)-Farnesyl acetone	0.6	1, 2
		Carbonylic compounds	8.2
1353	2186	Eugenol	1.5	1, 2, 3
		Phenol	1.5
1094	1095	1-Undecene	5.0	1, 2
2300	2300	Tricosane	0.6	1, 2, 3
2500	2500	Pentacosane	0.7	1, 2, 3
2700	2700	Heptacosane	0.9	1, 2, 3
2900	2900	Nonacosane	0.5	1, 2
		Hydrocarbons	7.7
977	1452	1-Octen-3-ol	0.7	1, 2
1002	1243	2-Pentylfuran	0.7	1, 2
		Others	1.4
		Total	96.6

^aHP-5 MS column.

^bHP Innowax column.

^cB.n.u.: Ballota nigra L. subsp. uncinata (Fiori et Beg.) Patzak.

^d1: Retention index, 2: mass spectrum, 3: co-injection with authentic compound.

^et: Trace, <0.05%.

The main constituents of B.n.u. were found to be (E)-phytol (20.0%), α-pinene (9.0%), hexahydrofarnesyl acetone (5.7%), α-selinene (5.1%), 1-undecene (5.0%), and (Z)-caryophyllene (4.2%). Sesquiterpenes constituted the most abundant fraction of the oil (39.9%), with a prevalence of sesquiterpene hydrocarbons (30.7%) among which α-selinene (5.1%), (Z)-caryophyllene (4.2%), and germacrene D (3.7%) predominated. Among the 7 oxygen-containing sesquiterpenes (9.2%), viridiflorol (2.8%) was the most abundant. With regard to monoterpenes, 6 oxygen-containing monoterpenes accounted for 3.5% of the total oil and monoterpene hydrocarbon represented 12.9% with α-pinene (9.0%) as the main compound. Also, diterpene fraction was noteworthy (21.5%), with (E)-phytol (20.0%) being the main compound of the class and the oil. Hexahydrofarnesyl acetone (5.7%) and 1-undecene (5.0%) were, instead, the principal products among the carbonylic compounds (8.2%) and the hydrocarbons (7.7%), respectively.

Table 2 reports the main compounds of the essential oils of the different taxa of B. nigra sl. group studied so far.

Considering the compounds occurring in the oils with an abundance of more than 3% only, the comparison of our data with those reported in the literature (Table 2) allows to point out some interesting considerations.

Table 2

Main Compounds (>3%) of the Essential Oils From the Aerial Parts of Taxa of Ballota nigra sl Group.

Taxa	Origin	Compounds	Ref
B. nigra	1: Mazandaran, Iran	Caryophyllene oxide (7.9), epi-α-muurolol (6.6), δ-cadinene (6.5), α-cadinol (6.3), γ-amorphene (4.3), β-bourbonene (4.1), 6,10,14-trimethyl-2-pentadecanone (4.0), (E)-caryophyllene (4.0), germacrene D (3.8), aromadendrene (3.4), γ-muurolene (3.2), germacrene D-4-ol (3.2), α-bisabolol (3.2), α-amorphene (3.0)	²⁸
	2: Jadovnik Mt., Serbia	β-Caryophyllene (35.4/39.1), germacrene D (27.4/35.7), α-humulene (7.4/10.4), δ-cadinene (3.8/0), (E)-phytol (2.5/3.8)	²⁹
	3: Golestan, Iran	β-Pinene (39.0), α-pinene (34.5), sabinene (7.7), α-phellandrene (4.1)	³⁰
B. nigra L. subsp. anatolica	1: Mazandaran, Iran	Germacrene D (18.1), nerolidol epoxyacetate (15.4), sclareol oxide (12.1), linalyl acetate (11.5), β-caryophyllene (10.5), spathulenol (9.0), linalool (5.2), longipinene epoxide (4.7)	³¹
	2: Muğla, Turkey	Hexadecanoic acid (40.9), β-bisabolene (13.4), hexahydrofarnesyl acetone (7.9), 1-isobutyl-4-isopropyl-2,2-diemethyl succinate (6.6), β-eudesmol (3.5)	²⁶
	3: Western Turkey	1-Hexacosanol (26.7), caryophyllene oxide (9.3), germacrene-D (9.3), α-selinene (8.7), Z-8-octadecen-1-ol acetate (7.1), 2,5-di-tertoctyl-p-benzoquinone (7.3), arachidic acid (6.0), tetracosane (4.5), heneicosane (4.4), heptacosane (4.3), 2-methyl-1-hexadecanol (3.3), octadecane (3.0), butyl phthalate (3.0)	³²
B. nigra subsp. foetida	1: Pisa, Italy	β-Caryophyllene (25.1), germacrene D (24.2), 1-octen-3-ol (7.3), (E)−2-hexenal (6.1), α-humulene (4.3), caryophyllene oxide (4.2)	³³
	2: Urbino, Italy	β-Caryophyllene (20.0), germacrene D (18.0), caryophyllene oxide (15.0), 1-octen-3-ol (6.8), (E)−2-hexenal (6.1), α-humulene (4.5), β-bourbonene (3.2)	³⁴
	3: Urbino, Italy	β-Caryophyllene (22.6), caryophyllene oxide (18.0), germacrene D (16.5), (E)−2-hexenal (6.5), 1-octen-3-ol (5.5)	³⁵
	4: Nis, Serbia	(E)-Phytol (56.9), germacrene D (10.0), β-caryophyllene (4.7), caryophyllene oxide (3.6), (E)-β-ionone (3.4)	³⁶
	5: Brac, Croatia	Germacrene D (23.1), β-caryophyllene (20.3), caryophyllene oxide (6.2), caryophylladienol I (3.3), (E)-2-hexenal (3.1), hexadecanoic acid (3.1), α-humulene (3.0)	³⁷
B. nigra subsp. kurdica	Kurdistan, Iran	Caryophyllene oxide (39.4), β-caryophyllene (24.9), germacrene D (7.6), 1-undecene (4.2), isoaromadendrene epoxide (3.2)	³⁸
B. nigra f. uncinata	Konya, Turkey	Caryophyllene oxide (21.2), hexadecanoic acid (19.9), β-caryophyllene (18.9), germacrene D (4.6), hexahydrofarnesyl acetone (4.4), spathulenol (4.2), caryolphyllenol II (3.8), bicyclogermacrene (3.7)	²⁶

Almost all the B. nigra species contain the sesquiterpenes caryophyllene and/or caryophyllene oxide among the main compounds with the exception of 2 cases: B. nigra 3 collected in Iran³⁰ and B. nigra subsp. anatolica 2 collected in Turkey.²⁶

The B. nigra subsp. uncinata, we examined, contains on the contrary to the previously investigated B. nigra subsp. uncinata, also a relevant amount (9%) of monoterpene α-pinene exclusively contained in the B. nigra 3 collected in Iran.³⁰ A cluster statistical analysis (Figure 1) of these data, based on a comparison of the species according to the relative amounts of compounds, shows a certain resemblance of our sample with B. nigra subsp. foetida 4 collected in Serbia.³⁶ In fact, both species, apart from caryophyllene and germacrene D, are the only ones containing the diterpene phytol as the most abundant compound.

Figure 1

The dendrogram of the clusters obtained by statistical analysis.

Experimental

Plant Material

Aerial parts (flowers, stems, and leaves) of B. nigra subsp. uncinata (Bég.) Patzak (B.n.u.) were collected at Lipari (38°29′51.652″N, 14°57′1.519″E), Aeolian Islands, Sicily (Italy), at the beginning of June 2018. The authentication was carried out by Mr Emanuele Schimmenti, Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Italy. A voucher specimen (PAL MB-2018/86) was deposited in Department STEBICEF, University of Palermo.

Isolation of the Essential Oil

The air-dried samples (200 g) were ground in a Waring blender and then subjected to a single hydrodistillation for 3 hours using n-hexane as the solvent, according to the standard procedure previously described.³⁹ The extracts were dried over anhydrous sodium sulfate and then stored in sealed vials, at −20°C, ready for the gas chromatography (GC) and GC-mass spectrometry (MS) analyses. The samples yielded 0.016% (B.n.u.) of yellow oils (w/w).

Gas Chromatography–Mass Spectrometry

Analytical GC was carried out on a Perkin-Elmer Sigma 115 GC fitted with an HP-5 MS capillary column (30 m × 0.25 mm), 0.25 µm film thickness. Column temperature was initially kept at 40°C for 5 minutes, then gradually increased to 250°C at 2°C/min rate, held for 15 minutes and finally raised to 270°C at 10°C/min. Diluted samples (1/100 v/v, in n-pentane) of 1 µL were injected at 250°C, manually, and in the splitless mode. Flame ionization detection (FID) was performed at 280°C. GC-MS analysis was performed on an Agilent 6850 Ser. II apparatus, fitted with a fused silica DB-5 capillary column (30 m × 0.25 mm), 0.33 µm film thickness, coupled to an Agilent Mass Selective Detector MSD 5973; ionization voltage 70 eV; electron multiplier energy 2000 V. GC conditions were as given; transfer line temperature, 295°C. Analysis was also run by using a fused silica HP Innowax polyethyleneglycol capillary column (50 m × 0.20 mm, 0.20 µm film thickness). In both cases, helium was used as carrier gas (1 mL/min). Identification of compounds was carried out using NIST 11, Wiley 9, FFNSC 2, and Adams databases.⁴⁰ These identifications were confirmed by linear retention indices with those available in literature by the SciFinder database. Some of the compounds were also confirmed by comparison of mass spectra and retention times with standard compounds available in the laboratory. The retention indices were determined in relation to a homologous series of n-alkanes (C8–C30) injected under the same operating conditions. Component relative concentrations were calculated based on GC peak areas without using correction factors.

Statistical Analysis

It was carried out using the cluster method by Primer 6⁴¹ using a matrix composed of the amount (%) of the 50 compounds occurring in 14 subspecies of B. nigra with abundance >3%.

Footnotes

Acknowledgments

The GC–MS spectra were performed at the “C.S.I.A.S.” of the University “Federico II” of Napoli.

Declaration of Conflicting Interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: the assistance of the staff is gratefully appreciated. This work was supported by grant from MIURITALY PRIN 2017 (Project N. 2017A95NCJ).

ORCID iD

Sergio Rosselli

References

World Checklist of Selected Plant Families (WCSP), Kew Science. Accessed 13 01 2020. http://apps.kew.org/wcsp/qsearch.do

Iauk

Acquaviva

Mastrojeni

et al. Antibacterial, antioxidant and hypoglycaemic effects of Thymus capitatus (L.) Hoffmanns. et link leaves’ fractions. J Enzyme Inhib Med Chem. 2015;30(3):360-365.doi:10.3109/14756366.2014.930453

http://www.ncbi.nlm.nih.gov/pubmed/25032744

Zuzarte

Alves-Silva

JM.

Alves

Cavaleiro

Salgueiro

Cruz

. New insights on the anti-inflammatory potential and safety profile of Thymus carnosus and Thymus camphoratus essential oils and their main compounds. J Ethnopharmacol. 2018;225:10-17.doi:10.1016/j.jep.2018.06.025

http://www.ncbi.nlm.nih.gov/pubmed/29933014

Bagetta

Cosentino

Sakurada

. Aromatherapy: basic mechanisms and evidence-based clinical use. 2016. Boca Raton: CRC Press; 2016.

Citoğlu

Tanker

Sever

Englert

Anton

Altanlar

. Antibacterial activities of diterpenoids isolated from Ballota saxatilis subsp. saxatilis . Planta Med. 1998;64(5):484-485.doi:10.1055/s-2006-957494

http://www.ncbi.nlm.nih.gov/pubmed/9690356

Çitoğlu

GS.

Yilmaz

BS.

Altanlar

. Antimicrobial activity of Ballota species growing in. Turkey. J Fac Pharm Ankara. 2003;32(2):93-97.

Dulger

. Antimicrobial activity of the leaves of Ballota acetabulosa on microorganisms isolated from urinary tract infection. Turk J Pharm Sci. 2012;9(3):257-262.

Askun

Tekwu

EM.

Satil

Modanlioglu

Aydeniz

. Preliminary antimycobacterial study on selected Turkish plants (Lamiaceae) against Mycobacterium tuberculosis and search for some phenolic constituents. BMC Complement Altern Med. 2013;13:365.doi:10.1186/1472-6882-13-365

http://www.ncbi.nlm.nih.gov/pubmed/24359458

Citoğlu

GS.

Çoban

Sever

İşcan

. Antioxidant properties of Ballota species growing in Turkey. J Ethnopharmacol. 2004;92(2-3):275-280.doi:10.1016/j.jep.2004.03.012

http://www.ncbi.nlm.nih.gov/pubmed/15138012

10.

Çitoğlu

GS.

Sever

Antus

Baitz-Gács

Altanlar

. Antifungal Flavonoids from Ballota glandulosissima . Pharm Biol. 2003;41(7):483-486.doi:10.1080/13880200308951339

11.

Çitoğlu

GS.

Sever

Antus

Baitz-Gács

Altanlar

. Antifungal Diterpenoids and Flavonoids from Ballota inaequidens . Pharm Biol. 2005;42(8):659-663.doi:10.1080/13880200490902626

12.

Özbek

Citoğlu

GS.

Dülger

Uğraş

Sever

. Hepatoprotective and anti-inflammatory activities of Ballota glandulosissima . J Ethnopharmacol. 2004;95(2-3):143-149.doi:10.1016/j.jep.2004.06.030

http://www.ncbi.nlm.nih.gov/pubmed/15507327

13.

Racz-Kotilla

Racz

Jozsa

. Activity of some species belonging to Labiatiae on the central nervous systems of mice. Herb Hung. 1980;19:49-53.

14.

Vural

Ezer

Erol

Samin

. Anxioloytic and antidepressant activities of some Ballota species. Journal of Faculty of Pharmacy of Gazi University. 1996;13(1):29-32.

15.

Couladis

Tzakou

Verykokidou

Harvala

. Screening of some Greek aromatic plants for antioxidant activity. Phytother Res. 2003;17(2):194-195.doi:10.1002/ptr.1261

http://www.ncbi.nlm.nih.gov/pubmed/12601688

16.

Seidel

Verholle

Malard

et al. Phenylpropanoids from Ballota nigra L. inhibit in vitro LDL peroxidation. Phytother Res. 2000;14(2):93-98.doi:10.1002/(SICI)1099-1573(200003)14:2<93::AID-PTR558>3.0.CO;2-X

http://www.ncbi.nlm.nih.gov/pubmed/10685104

17.

Didry

Seidel

Dubreuil

Tillequin

Bailleul

. Isolation and antibacterial activity of phenylpropanoid derivatives from Ballota nigra . J Ethnopharmacol. 1999;67(2):197-202.doi:10.1016/S0378-8741(99)00019-7

http://www.ncbi.nlm.nih.gov/pubmed/10619384

18.

Dulger

Kilcik

. Antibacterial activity of Ballota acetabulosa against methicillin-resistant Staphylococcus aureus . Asian J Chem. 2011;23(1):416-418.

19.

Rosselli

Fontana

Bruno

. A review of the phytochemistry, traditional uses, and biological activities of the Genus Ballota and Otostegia . Planta Med. 2019;85(11-12):869-910.doi:10.1055/a-0953-6165

http://www.ncbi.nlm.nih.gov/pubmed/31216581

20.

Savona

Bruno

Piozzi

Barbagallo

. Diterpenes from Ballota species. Phytochemistry. 1982;21(8):2132-2133.doi:10.1016/0031-9422(82)83067-7

21.

Bruno

Savona

Pascual

Rodríguez

. Preleosibirin, a prefuranic labdane diterpene from ballota nigra subsp. foetida . Phytochemistry. 1986;25(2):538-539.doi:10.1016/S0031-9422(00)85521-1

22.

Bruno

Bondí

ML.

Piozzi

Arnold

NA.

Simmonds

MSJ

. Occurrence of 18-hydroxyballonigrine in Ballota saxatilis ssp. saxatilis from Lebanon. Biochem Syst Ecol. 2001;29(4):429-431.doi:10.1016/S0305-1978(00)00068-5

http://www.ncbi.nlm.nih.gov/pubmed/11182492

23.

Euro+Med PlantBase - the information resource for Euro-Mediterranean plant diversity. Accessed 13 01 2020. https://ww2.bgbm.org/EuroPlusMed/

24.

Yilmaz

BS.

Çitoğlu

. High performance liquid chromatographic analysis of some diterpenoids of the Ballota species. FABAD J Pharm Sci. 2003;28(1):13-17.

25.

Çitoğlu

GS.

Yilmaz

BS.

Tarikahya

Tipirdamaz

. Chemotaxonomy of Ballota species. Chem Nat Compd. 2005;41(3):299-302.doi:10.1007/s10600-005-0134-7

26.

Kaya

Kürkçüoğlu

Dinç

Doğu

. Compositions of the essential oils of Ballota nigra subsp. uncinata and subsp. anatolica from Turkey. Indian J Pharm Educ. 2017;51(3s):s185-s189.doi:10.5530/ijper.51.3s.9

27.

Sever Yilmaz

Altanlar

Saltan Çitoğlu

. Antilisterial activity of Ballota species growing in Turkey. J Fac Pharm Ankara. 2005;34(3):155-164.

28.

Morteza-Semnani

Saeedi

Akbarzadeh

. The essential oil composition of Ballota nigra . Chem Nat Compd. 2007;43(6):722-723.doi:10.1007/s10600-007-0245-4

29.

Vukovic

Sukdolak

Solujic

Niciforovic

. Antimicrobial activity of the essential oil obtained from roots and chemical composition of the volatile constituents from the roots, stems, and leaves of Ballota nigra from Serbia. J Med Food. 2009;12(2):435-441.doi:10.1089/jmf.2008.0164

http://www.ncbi.nlm.nih.gov/pubmed/19459749

30.

Jamzad

Rustaiyan

Jamzad

Masoudi

. Essential oil composition of Salvia indica L., Thymus caucasicus Wind. Ex Ronniger subsp. grossheimii (Ronniger) Jalas. and Ballota nigra L. three Labiatae species from Iran. J Ess Oil Bearing Plants. 2011;14(1):76-83.doi:10.1080/0972060X.2011.10643903

31.

Kazemizadeh

Amini

Nazari

Habibi

. Volatile constituents of Ballota nigra subsp. anatolica from Iran. Chem Nat Compd. 2009;45(5):737-738.doi:10.1007/s10600-009-9415-x

32.

Ertaş

Boğa

Yeşil

. Phytochemical profile and ABTS cation radical scavenging, cupric reducing antioxidant capacity and anticholinesterase activities of endemic Ballota nigra L. subsp. anatolica P. H. Davis from Turkey. J Coast Life Med. 2014;2(7):555-559.

33.

Bader

Caponi

Cioni

PL.

Flamini

Morelli

. Composition of the essential oil of Ballota undulata, B. nigra ssp. foetida and B. saxatilis . Flavour Fragr J. 2003;18(6):502-504.doi:10.1002/ffj.1257

34.

Fraternale

Bucchini

Giamperi

Ricci

. Essential oil composition and antimicrobial activity of Ballota nigra L. ssp. foetida . Nat Prod Commun. 2009;4(4):585-588.doi:10.1177/1934578X0900400429

http://www.ncbi.nlm.nih.gov/pubmed/19476011

35.

Fraternale

Ricci

. Essential oil composition and antifungal activity of aerial parts of Ballota nigra ssp foetida collected at flowering and fruiting times. Nat Prod Commun. 2014;9(7):1015-1018.doi:10.1177/1934578X1400900733

http://www.ncbi.nlm.nih.gov/pubmed/25230517

36.

Đorđević

AS.

Jovanović

OP.

Zlatković

BK.

Stojanović

. Chemical composition of Ballota macedonica Vandas and Ballota nigra L. ssp. foetida (Vis.) Hayek essential oils – the chemotaxonomic approach. Chem Biodiv. 2016;13(6):782-788.

37.

Rigano

Marrelli

Formisano

et al. Phytochemical profile of three Ballota species essential oils and evaluation of the effects on human cancer cells. Nat Prod Res. 2017;31(4):436-444.doi:10.1080/14786419.2016.1185722

http://www.ncbi.nlm.nih.gov/pubmed/27189840

38.

Majdi

Dastan

Maroofi

. Chemical composition and antimicrobial activity of essential oils of Ballota nigra subsp. kurdica from Iran. Jundishapur J Nat Pharm Prod. 2017;12(3):e36314.

39.

De Martino

Bruno

Formisano

et al. Chemical composition and antimicrobial activity of the essential oils from two species of Thymus growing wild in Southern Italy. Molecules. 2009;14(11):4614-4624.doi:10.3390/molecules14114614

http://www.ncbi.nlm.nih.gov/pubmed/19924089

40.

Adams

. Identification of Essential Oil Components by Gas Chromatography/ Mass Spectroscopy. 4th ed. Allured Publishing. Carol Stream USA; 2007.

41.

Clarke

KR.

Gorley

; 2006. PRIMER (6.0). Plymouth UK: PRIMER-E.

GC and GC-–MS Analysis of Volatile Compounds From Ballota nigra subsp. uncinata Collected in Aeolian Islands,Sicily (Southern Italy)

Abstract

Keywords

Results and Discussion

Experimental

Plant Material

Isolation of the Essential Oil

Gas Chromatography–Mass Spectrometry

Statistical Analysis

Footnotes

Acknowledgments

Declaration of Conflicting Interests

Funding

ORCID iD

References