# Phytochemistry and Biological Properties ofSalvia verbenaca L.: A Comprehensive Review

Authors: Hanae Naceiri Mrabti; Naoual El Menyiy; Saoulajan Charfi; Mohammed Saber; Saad Bakrim; Reema A. Alyamani; Abdur Rauf; Ahmed M. H. Ali; Emad M. Abdallah; Naserddine El Omari; Abdelhakim Bouyahya; Hamza Assaggaf
Journal: BioMed Research International
 (2022)

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## Abstract

The family Lamiaceae contains several plants used in traditional medicine to fight against different diseases. Salvia verbenaca L. (S. verbenaca) is one of the Lamiaceae species distributed around the Mediterranean regions. This plant exhibits different bioactive properties, including antibacterial, anticancer, antioxidant, antileishmanial, antidiabetic, immunomodulatory, and wound healing. This review was conducted to revise previous studies on S. verbenaca addressing its botanical description, geographical distribution, and phytochemical, pharmacological, and toxicological properties. Moreover, the main pharmacological actions of S. verbenaca major compounds were well investigated. Literature reports have revealed that S. verbenaca possesses a pivotal role in medicinal applications. The findings of this work noted that S. verbenaca was found to be rich in chemical compound classes such as terpenoids, phenolics, fatty acids, sterols, and flavonoids. Numerous studies have found that S. verbenaca essential oils and extracts have a wide range of biological effects. These results support the potential pharmacological properties of S. verbenaca and its traditional uses. This analysis can constitute a scientific basis for further refined studies on its pure secondary metabolites. Therefore, the outcome of the present work may support the perspective of identifying new therapeutical applications with detailed pharmacological mechanisms of S. verbenaca to prevent the development of some diseases such as neurodegenerative disorders. However, toxicological investigations into S. verbenaca are needed to assess any potential toxicity before it can be further used in clinical studies.


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## Body

## 1. Introduction

Since the beginning of time, medicinal plants have been and continue to be the primary source of medicine [1]. Salvia verbenaca L. (S. verbenaca) is a medicinal herb belonging to the family Lamiaceae, which is the most representative genus of Salvia [2, 3]. This plant is endemic to the Mediterranean region, including Morocco, Canaries Islands, Algeria, Tunisia, Libya, Egypt, and Cyprus, and has also spread to Europe and Asia [4]. In traditional medicines, S. verbenaca has been used to fight against numerous diseases; several ancient and current investigations revealed that S. verbenaca presents a chemical diversity in terms of chemical composition according to the chemical characteristics of the extracts from various parts. Indeed, S. verbenaca contains numerous secondary metabolites that belong to a wide variety of phytochemical classes [5]. S. verbenaca terpenoids have been revealed to have a large diversity due to several factors, including genetic, ecological, environmental, edaphic, and diverse plant parts [6]. An antibacterial potential against a wide range of gram-positive and gram-negative bacteria has been documented [7–11]. Consequently, the antibacterial efficacy of extracts and essential oils (EO) from S. verbenaca was remarkable against gram-positive bacteria compared to the gram-negative bacteria. Furthermore, S. verbenaca was found to have an antioxidant effect against free radical damage [12] and significantly reduce the level of intracellular reactive oxygen species (ROS) [13, 14]. According to previous studies, the anticancer properties of S. verbenaca extracts and essential oils have also been reported [15–19]. The antiparasitic properties of S. verbenaca, in particular, antileishmanial effects, have been investigated elsewhere [20].Besides,S. verbenaca was reported to have an inhibitory effect of xanthine oxidase [21] and a healing effect on burns [22]. Furthermore, S. verbenaca revealed immunomodulatory effects [23]. Furthermore, the toxicological tests found that the ethanolic extract of S. verbenaca did not cause any toxic symptoms or death in rats [24].The objective of the current article was to provide a general review ofS. verbenaca such as botanical description, geographical distribution, phytochemistry, and pharmacological properties. Hopefully, this analysis could be a scientific basis for further refined studies on pure compounds from S. verbenaca that may lead to the identification of new therapeutical applications.


## 2. Research Methodology

All data aboutS. verbenaca (botanical description, taxonomy, destruction, phytochemical, and pharmacological properties) were collected using several databases like Web of Science, Google Scholar, Scopus, ScienceDirect, SpringerLink, Wiley Online, PubMed, and SciFinder and were reviewed in order to compile literature on S. verbenaca. The structures of the chemical profiles were identified in S. verbenaca, and the ChemDraw Pro 8.0 software was used to create the illustrations.


### 2.1. Botanical Description

S. verbenaca is a perennial herb that reaches between 10 and 50 cm (in height), hairy at the top, odorous, more or less glandular at the top. It grows in the dry lawns, the slopes, and at the edges of the paths. The slightly branched stems carry bunches of dark blue flowers in spring. Leaves are oblong, 2-3 cm broad, crenelated or incised-lobed, with the upper stalkless (Figure 1). The flowers are quite small, pale blue or whitish, in whorls usually close together, forming a fairly short cluster; the fruiting calyx with almost closed lips, bristling with spread hairs; the corolla is 10–15 mm, twice as long as the calyx, with wide lips, very uneven, the upper one compressed and curved in a false shape, and the style with little or prominent point [25].Figure 1
Salvia verbenaca L.: (a) whole plant; (b) aerial part; (c) flowers.
(a)(b)(c)


### 2.2. Geographic Distribution

S. verbenaca has a very wide geographical distribution around the Mediterranean region, including Morocco, Algeria, Tunisia, Canaries, Egypt, Libya, Turkey, Cyprus, Transcaucasia, and Western and Southern Europe. It is also grown in South West Africa, North America, and Australia [5].


### 2.3. Ethnomedicinal Uses

The ethnobotanical investigations intoS. verbenaca revealed its wide applications in folkloric medicine to treat numerous disorders as listed in Table 1. In Morocco, its application in folk medicinal systems includes the treatment of some digestive disorders such as abdominal colics [26–28]. The most commonly used part of the plant is the aerial part, which is prepared by infusion before being used to treat respiratory problems and genitourinary and skin diseases [27]. Dried leaves are also used for the treatment of wounds, burns, and abscesses [29]. Aerial parts are utilized in decoction or infusion to treat diabetes [30].Table 1
Some medicinal applications ofS. verbenaca.
Part usedPreparationTraditional applicationRef.Aerial partDecoction, infusionDiabetes[30]LeafDecoction, powderAbdominal colics, cold, fever, healing[26]LeafPowderWound treatment[29]LeafDecoction, infusionGenitourinary, skin, digestive, and respiratory problems[27]Aerial partNo informationDigestive problems[28]Leaf, whole plantPowderHealing of burns, wounds, and abscesses[29]NI: no information.


### 2.4. Phytochemistry

Like all medicinal plants belonging to the family Lamiaceae,S. verbenaca contains numerous secondary metabolites with different classes, such as flavonoids, terpenoids, alkaloids, and phenolic acids. Currently, several analytical investigations using different technical tools (GC, GC-SM, GC-MS, GC-FID, HPLC, 1D and 2D NMR, IR, UV, 1H NMR, and 13C NMR) have been applied to identify and isolate bioactive compounds from medicinal plants. Indeed, investigations into the chemical constituents of S. verbenaca revealed the presence of terpenoids, phenolics, fatty acids, flavonoids, and sterols (Table 2). As listed in Table 2, the chemical content of S. verbenaca was investigated in different areas with various medicinal applications by using different analytical tools. The results are different according to numerous factors, such as the study area, plant part used, and adopted methodology.Table 2
Chemical composition of various parts ofS. verbenaca.
Part usedCountryHarvest siteHarvest seasonExtracts/essential oilsChemical compositionAnalysisReferencesAerial parts (dry)TunisiaRass Zebib (subhumid)Flowering periodEO1,8-Cineole (9.7%), p-cymene (8.4%),α-pinene (5.4%), γ-terpinene (3.1%), beta-caryophyllene (5.3%), viridiflorol (7.3%), epi-13-manool (4.7%), thymol (3.7%), limonene (2.8%), camphor (2.7%)GC and GC-MS[6]Aerial parts (dry)TunisiaBir Mroua (subhumid)Flowering periodEOβ-Caryophyllene (15.3%), germacrene D (7.1%), epi-13-manool (6.2%), α-copaene (6.1%), α-humulene (4.3%), α-cadinol (3.9%), viridiflorol (3.4%), p-cymene (3.3%), δ-cadinene (3.1%), p-cymen-8-ol (2.6%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaBeja (higher semiarid)Flowering periodEOβ-Caryophyllene (15.3%), α-humulene (3.0%), viridiflorol (11.6%), 1,8-cineole (3.3%), germacrene D (3.3%), (Z)-β-ocimene (4.0%), T-cadinol (1.9%), p-cymene (2.8%), thymol (2.7%), epi-13-manool (2.5%)GC and GC-MS[6, 31]Aerial parts (dry)TunisiaTunis (higher semiarid)Flowering periodEOViridiflorol (17.7%), 1,8-cineole (8.5%),α-pinene (4.6%), p-cymene (5.2%), β-caryophyllene (5.5%), thymol (4.4%), epi-13-manool (4.0%), α-humulene (2.4%), α-thujone (3.6%), γ-terpinene (2.4%)GC and GC-MS[31, 33, 34]Aerial parts (dry)TunisiaTouiref (moderate semiarid)Flowering periodEOα-Pinene (15.9%), camphor (4.7%), 1,8-cineole (12.8%), viridiflorol (10.0%), (Z)-β-ocimene (5.4%), camphene (2.6%), β-caryophyllene (5.3%), thymol (4.2%), p-cymene (4.2%), α-thujone (3.4%)GC and GC-MS[31, 33, 35]Aerial parts (dry)TunisiaBou Arada (moderate semiarid)Flowering periodEO1,8-Cineole (9.4%), p-cymene (8.7%), viridiflorol (8.3%),α-pinene (4.9%), thymol (2.7%), β-caryophyllene (4.9%), α-humulene (3.5%), γ-terpinene (3.0%), α-thujone (3.0%), epi-13-manool (3.6%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaSers (lower semiarid)Flowering periodEOα-Pinene (14.7%), viridiflorol (10.8%), β-caryophyllene (4.6%), (Z)-β-ocimene (4.5%), epi-13-manool (2.8%), thymol (4.4%), p-cymene (4.1%), camphor (3.5%), α-thujone (2.9%), 1,8-cineole (10.9%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaEnfidha (lower semiarid)Flowering periodEOViridiflorol (10.5%), camphor (2.9%), epi-13-manool (10.5%), 1,8-cineole (8.7%), p-cymene (8.3%),α-terpineol (3.0%), α-pinene (4.5%), thymol (4.2%), γ-terpinene (3.2%), bornyl acetate (3.2%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaChott Meriem (higher arid)Flowering periodEOp-Cymene (14.2),α-pinene (9.6), γ-terpinene (5.1), camphene (3.9), viridiflorol (5.1), limonene (3.4), epi-13-manool (3.2), thymol (2.5), 1,8-cineole (12.8)GC and GC-MS[31–33]Aerial parts (dry)TunisiaHencha (higher arid)Flowering periodEOViridiflorol (10.0%), bicyclogermacrene (2.3%), germacrene D (5.6%), 1,8-cineole (4.9), epi-13-manool (4.7%),α-thujone (3.2%), β-pinene (3.0%), camphor (2.9%), α-humulene (2.5%), β-caryophyllene (7.2%)GC and GC-MS[31–33]Aerial parts (dry)SpainMurciaFlowering stageEOp-Cymene (11.4%), 1,8-cineole (7.7%), viridiflorol (7.0%), camphene (2.7%),β-caryophyllene (4.5%), β-pinene (2.7%), γ-terpinene (4.0%), epi-13-manool (3.9%), camphor (3.7%), α-pinene (8.1%)GC and GC-MS[6]Aerial parts (dry)SpainMurciaEarly fruiting stageEOCaryophyllene oxide (12.4%), bornyl acetate (3.2%), viridiflorol (9.1%),β-caryophyllene (5.6%), p-cymene (5.6%), α-pinene (4.0%), epi-13-manool (2.3%), thymol (2.0%), β-ionone (2.0%), 1,8-cineole (6.3%)GC and GC-MS[6]Aerial parts (dry)SpainMurciaLate fruiting stageEOβ-Caryophyllene (14.2%), α-thujone (8.2%), 8-cineole (4.7%), epi-13-manool (7.1%), bornyl acetate (3.5%), α-humulene (6.7%), 1 α-pinene (4.3%), caryophyllene oxide (3.0%), β-pinene (2.8%), viridiflorol (13.5%)GC and GC-MS[6]Aerial partsAlgeriaBecharApril 2011EOEpi-α-cadinol (11.6%), β-caryophyllene (11.33%), bicyclogermacrene (10.9%), γ-cadinene (7.9%), cis-muurola-4(14),5-diene (7.8%), muurola-3,5-diene (5.2%), spathulenol (3.0%), cis-calamenene (2.0), α-humulene (1.9), 1,10-di-epi-cubenol (20.9%)GC and GC-MS[31]Aerial parts (fresh)JordanShafa-Badran-AmmanFlowering period (April to May 2011)EOLinalool (61.32%),β-elemene (1.50%), (Z)-β-ocimene (4.03%), β-eudesmol (3.66%), spathulenol (3.40%), E-β-ocimene (2.63%), β-caryophyllene (2.98%), α-copaene (2.50%), γ-cadinene (1.55%), bicyclogermacrene (5.94%)GC-MS and GC-FID[32]Aerial parts (dry)JordanShafa-Badran-AmmanFlowering period (April to May 2011)EOLinalool (30.72%), bicyclogermacrene (14.70%),β-caryophyllene (7.42%), germacrene D (25.92%), α-copaene (5.13%), isopentyl isovalerate (0.97%), δ-cadinene (2.05%), (Z)-β-ocimene (1.18%), spathulenol (1.58%), α-gurjunene (1.07%)GC and GC-MS[32]StemJordanMediterraneanFull maturation periodEOZ-β-Ocimene (32.6%), trans-sabinene hydrate acetate (14.5%), α-gurjunene (6.0%), β-bourbonene (1.5%), E-β-ocimene (7.8%), sabinene (2.9%), α-phellandrene (3.1%), germacrene D (1.6%), α-pinene (9.3%), β-pinene (8.1%)GC-MS and GC-FID[32]JordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate acetate (38.1%), E-caryophyllene (9.1%), δ-selinene (5.2%), β-gurjunene (2.5%), sabinene (4.8%), δα-copaene (4.1%), γ-gurjunene (2.9%), cadinene (4.3%), β-selinene (2.2%), germacrene D (13.3%)GC and GC-MS[32]LeavesJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (30.2%), β-bourbonene (7.7%), E-β-ocimene (4.3%), α-pinene (3.0%), α-gurjunene (13.8%), β-selinene (2.8%), δ-cadinene (2.5%), β-pinene (2.4%), myrcene (2.0%), Z-β-ocimene (17.1%)GC and GC-MS[32]LeavesJordanIrano-TuranianFull maturation periodEOδ-Selinene (21.5%), E-caryophyllene (11.4%), terpinolene (4.3%), α-copaene (9.6%), sabinene (9.0%), Z-β-ocimene (4.8%), β-cubebene (4.4%), δ-cadinene (2.7%), cis-β-guaiene (2.0%), germacrene D (19.8%)GC and GC-MS[32]PreflowerJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (56.5%), α-pinene (6.5%), myrcene (1.5%), E-β-ocimene (4.3%), α-gurjunene (3.2%), β-pinene (5.3%), sabinene (1.2%), trans-β-guaiene (1.0%), limonene (0.7%), Z-β-ocimene (13.5%)GC and GC-MS[32]PreflowerJordanIrano-TuranianFull maturation periodEOSabinene (42.7%),α-thujene (7.2%), γ-terpinene (6.1%), E-β-ocimene (1.9%), α-terpinene (3.6%), β-pinene (3.2%), β-phellandrene (6.8%), terpinolene (1.6%), limonene (1.0%), trans-sabinene hydrate (20.4%)GC and GC-MS[32]FlowerJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (58.6%), E-β-ocimene (5.3%), α-pinene (5.2%), sabinene (1.1%), β-pinene (4.9%), α-phellandrene (1.4%), α-gurjunene (1.0%), camphene (0.4%), isobornyl acetate (0.4%), Z-β-ocimene (18.8%)GC and GC-MS[32]JordanIrano-TuranianFull maturation periodEOSabinene (37.5%),Z-β-ocimene (9.9%), α-thujene (4.6%), myrcene (4.2%), β-pinene (3.9%), E-β-ocimene (8.9%), γ-terpinene (3.0%), E-caryophyllene (1.9%), α-terpinene (1.4%), trans-sabinene hydrate (20.0%)GC and GC-MS[32]PetalJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (87.0%), E-β-ocimene (1.5%), germacrene D (1.0%), α-phellandrene (0.5%), β-pinene (0.3%), α-gurjunene (1.7%), n-nonane (0.2%), myrcene (0.2%), β-selinene (0.2%), Z-β-ocimene (7.1%)GC and GC-MS[32]PetalJordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate (18.8%), E-β-ocimene (9.9%), γ-terpinene (2.9%), germacrene D (9.6%), β-copaene (4.1%), α-copaene (3.2%), E-caryophyllene (13.9%), β-selinene (2.9%), γ-gurjunene (2.8%), Z-β-ocimene (9.6%)GC and GC-MS[32]SepalJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (36.6%), β-pinene (14.0%), 8-cineole (3.9%), Z-β-ocimene (4.5%), 1 δ-elemene (2.8%), β-cedrene (8.7%), sabinene (2.7%), camphene (1.9%), β-cubebene (1.9%), α-pinene (18.1%)GC and GC-MS[32]SepalJordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate (58.8%), terpinolene (5.0%), E-β-ocimene (4.1%), p-methyl-acetophenone (3.2%), germacrene D (3.1%), Z-β-ocimene (4.9%), δ-selinene (2.0%), γ-terpinene (1.7%), δ-cadinene (1.3%), E-caryophyllene (5.6%)GC and GC-MS[32]Aerial partsAlgeriaMogheulApril 2011EOGermacrene D (20.5%),β-caryophyllene (3.8%), beta-cubebene (2.7%), δ-cadinene (2.6%), 1,10-di-epi-cubenol (2. 6%), γ-cadinene (2.5%), (E)-β-farnesene (3.5%), bicyclogermacrene (2.2%), α-muurolol (2.1%), α-copaene (10.4%), β-phellandrene (3.8%)GC and GC-MS[36]SeedsSpain——EOCamphor (38.94%), 13-epi-manool (5.61%), 𝛿-elemene (3.93%), beta-eudesmol (3.76%), n-undecane (2.65%), α-terpinyl acetate (4.77%), linalyl acetate (2.53%), neryl acetate (2.40%), α-terpineol (2.03%), caryophyllene oxide (7.28%)GC-MS and GC-FID[33]Aerial partsGreeceCrete IslandBlossoming (April 2004)EOBeta-phellandrene (30.3%), methyl ester of 6-octadecenoic acid (15.0%), camphor (7.0%), (Z)-β-ocimene (6.6%), fenchone (9.4%), isopropyl ester (7.8%), aromadendrene (4.0%), α-humulene (3.7%), (E)-caryophyllene (16.1%)GC and GC-MS[34]Aerial parts (fresh)SicilyPiano BattagliaFull flowering stage (July 2009)EOHexadecanoic acid (23.1%), ethyl hexadecanoate (2.6%), benzaldehyde (7.3%), 9,12,15-octadecatrienal (2.9%), limonene (2.0%), (E)-β-ionone (1.9%), (Z)-9-octadecenoic acid (11.9), phenyl acetaldehyde (1.5%), (E)-caryophyllene (1.2%), β-phellandrene (5.9%)GC and GC-MS[8]Aerial partsAlgeriaDjelfaMarch 2019EOcis-Muurola-3,5-diene (14.6%), unknown (10.5%), bicyclogermacrene (6.8%), bicycloelemene (4.3%), γ-cadinene (4.8%), β-pinene (4.2%), 2,3-dehydro-1,4-cineol (3.7%), α-cubebene (3.0%), α-pinene (2.8%), γ-amorphene (10.5%)GC and GC-MS[35]Leaves and flowers (dried)TurkeyKütahya-Gediz2016-2017EOLinalyl acetate (81.97%),β-myrcene (2.73%), n-pentanal (0.42%), beta-ocimene (0.39%), hexanal (0.34%), α-pinene (0.34%), limonene (1.14%), trans-caryophyllene (0.32%), β-pinene (0.31%), linalool (8.66%)GC and GC-MS[37]Aerial parts (wild)SicilyPiano BattagliaFull flowering stage (July 2009)EOHexadecanoic acid (23.1%), benzaldehyde (7.3%), b-phellandrene (5.9%), limonene (2.0%), 9,12,15-octadecatrienal (2.9%), ethyl hexadecanoate (2.6%), caryophyllene oxide (1.9%), (E)-b-ionone (1.9%), spathulenol (1.7%), (Z)-9-octadecenoic acid (11.1%)GC and GC-MS[19]Aerial parts (cultivated)SicilyPiano BattagliaJuly 2010EOHexadecanoic acid (11.0%), (E)-b-ionone (3.9%), (Z)-9-octadecenoic acid (5.6%), b-phellandrene (4.1%), caryophyllene oxide (2.8%), (E)-caryophyllene (3.8%), methyl hexadecanoate (3.8%), carvacrol (2.4%), spathulenol (2.0%), hexahydrofarnesyl acetone (9.7%)GC and GC-SM[19]FruitsTunisiaSabelet Ben AmmarFull fruit ripening stageEOβ-Caryophyllene (23.1%), camphene (6.5%), α-humulene (5.6%), germacrene D (3.5%), viridiflorol (4.3%), 1-octen-3-ol (3.9%), (E)-β-ocimene (1.5%), 1,8-cineole (3.0%), manool (1.1%), caryophyllene oxide (15.9%)GC and GC-MS[33]StemsTunisiaSabelet Ben AmmarFull fruit ripening stageCamphor (10.9%), terpinolene (6.6%), methyl eugenol (6.1%),α-pinene (5.9%), α-thujone (3.1%), 1,8-cineole (5.8%), caryophyllene oxide (4.5%), aromadendrene (3.6%), epi-13-manool (2.3%), viridiflorol (10.3%)GC and GC-MS[33]LeavesTunisiaSabelet Ben AmmarFull fruit ripening stageepi-13-Manool (13.7%), camphor (3.9%), caryophyllene oxide (3.9%),α-pinene (3.4%), p-cymen-8-ol (3.7%), terpinen-4-ol (3.6%), 1,8-cineole (3.0%), eugenol (2.8%), (E)-β-ocimene (2.6%), manool (11.0%)GC and GC-MS[33]Aerial partsTunisiaSabelet Ben AmmarFull fruiting stage (April 2007)EOViridiflorol (21.6%), methyl eugenol (9.4%),α-terpineol (5.3%), spathulenol (3.7%), β-caryophyllene (7.1%), caryophyllene oxide (2.4%), epi-13-manool (2.2%), germacrene D (1.9%), eugenol (1.8%), camphene (17.6%)GC and GC-MS[33]Aerial partsTunisiaSersFull fruiting stage (April 2007)(Z)-β-ocimene (29.5%), beta-thujone (7.9%), α-pinene (5.5%), tricyclene (5.1%), 18-cineole (1.9%), α-calacorene (2.5%), terpinen-4-ol (2.1%), germacrene D (3.1%), β-caryophyllene (1.8%), β-phellandrene (8.2%)GC and GC-MS[33]Aerial partsTunisiaSomaaTricyclene (18.8%), nonane (10.3%), terpinolene (7.3%), -terpineol (2.2%), bornyl acetate (4.9%), camphor (2.9%),α-terpinyl acetate (3.5%), limonene (2.3%), α β-eudesmol (2.2%), methyl eugenol (7.7%)GC and GC-MS[33]SeedsTunisiaSabelet Ben AmmarFull ripeness (April 2007)EOCamphor (33.83%), caryophyllene oxide (10.11%), octane (4.78%), 13-epi-manool (3.57%), hexanal (2.46%),β-bisabolene (1.84%), α-terpineol (3.24%), tricyclene (5.54%), α-copaene (3.19%), α-thujene (13.36%)GC-MS and GC-FID[33]SeedsTunisiaSabelet Ben AmmarFull ripeness (April 2007)Lipid extractionPalmitic acid (9.25%), stearic acid (2.48%), linolenic acid (45.89%), arachidic acid (0.20%), C18:3n-3/C18:2n-6 (1.67%), SFA (11.93%), USFA (88.07%), oleic acid (14.67%), linoleic acid (27.39%), palmitoleic acid (0.12%)GC and GC-MS[33]SeedsTunisiaSersFull ripeness (April 2007)EOβ-Pinene (48.08%), epi-cubebol (10.74%), β-eudesmol (1.00%), α-bisabolol (2.97%), caryophyllene oxide (2.90%), spathulenol (0.93%), eugenol (0.97%), geraniol (0.95%), borneol (1.97%), germacrene D (2.09%)GC and GC-MS[33]SeedsTunisiaSersFull ripeness (April 2007)Lipid extractionPalmitic acid (9.63%), oleic acid (14.14%), linoleic acid (23.79%), linolenic acid (42.84%), SFA (18.35%), USFA (81.65%), stearic acid (4.22%), arachidic acid (4.50%), C18:3n-3/C18:2n-6 (1.53%), palmitoleic acid (0.89%)GC and GC-MS[33]SeedsTunisiaSomaaFull ripeness (April 2007)EOOctane (27.39%),δ-cadinene (5.77%), p-cymene (1.64%), camphor (3.53%), bicyclogermacrene (1.86%), β-pinene (3.74%), α-terpineol (1.38%), limonene (0.79%) n-nonane (18.01%), epi-cubebol (9.02%)GC and GC-MS[33]SeedsTunisiaSomaaFull ripeness (April 2007)Lipid extractionPalmitic acid (12.11%), stearic acid (3.02%), linoleic acid (25.33%), arachidic acid (1.30%), SFA (16.43%), linolenic acid (41.71%), oleic acid (15.51%), USFA (83.57%), C18:3n-3/C18:2n-6 (1.65%), palmitoleic acid (1.02%)GC and GC-MS[33]Aerial partAlgeriaBordj Bou ArreridjFlowering stage (spring April-May)Crude extract (CrE)Flavonoids (08.40±0.32mg EQ/g E), polyphenols (177.56±2.51mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjChloroform extract (ChE)Flavonoids (14.87±0.81mg EQ/g E), polyphenols (156.81±1.57mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjEthyl acetate extract (EAE)Flavonoids (28.81±0.38mg EQ/g E), polyphenols (661.78±4.00mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjAqueous extract (AqE)Flavonoids (06.74±0.14mg EQ/g E), polyphenols (123.18±4.20mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaLaghouatMay 200480% (v/v) aqueous methanolFlavonoids (3.04±0.01mg RE/g dw), total phenols (7.2±0.04mg GAE/g dw), flavonols (0.85±0.001mg QE/g dw)Spectrophotometrically[12]Aerial partAlgeriaSetif and Batna2016DecoctionTotal phenols (129.02±2.67mg GAE/g DW), total flavonoid contents (18.62±0.06mg QE/g DW), total tannin contents (73.80±2.23mg TAE/g DW)Total carotenoid contents (0.92±0.041mg/g DW)Total chlorophyll A (1.21±0.02mg/g DW)Total chlorophyll B (2.48±0.04mg/g DW)Spectrophotometrically[13]Aerial partAlgeriaSetif and Batna2016Methanol extract (85%)Total phenols (190.16±1.74mg GAE/g DW)Total flavonoid contents (23.50±0.71mg QE/g DW)Total tannin contents (118.88±1.25mg TAE/g DW)Total carotenoid contents (0.58±0.005mg/g DW)Total chlorophyll A (1.67±0.02mg/g DW)Total chlorophyll B (0.63±0.01mg/g DW)Spectrophotometrically[13]Aerial partTurkeyArtvin06th September 2004Methanol extractsRosmarinic acids (29.30±0.24 μg mg_1)Spectrophotometrically[38]Rosmarinic acids (26.12±0.73 μg mg_1)HPLC[6]Aerial partsTunisiaTunis (higher semiarid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoïc acids (229.87±8.60 μg/g), p-coumaric acid (77.65±5.67 μg/g), rosmarinic acid (1688.01±63.42 μg/g), vanillic acid (20.21±0.46 μg/g), caffeic acids (97.29±2.86 μg/g), ferulic acids (40.41±3.32 μg/g)Phenolic diterpenesCarnosic acids (63.52±15.30 μg/g), methyl carnosate contents (633.37±11.66 μg/g), carnosol (25.52±7.27 μg/g)FlavonoidsNaringenins (940.41±22.50 μg/g), cirsiliols (73.16±1.72 μg/g), luteolins (13.84±2.62 μg/g), apigenins (3.01±0.69 μg/g), naringins (57.30±3.55 μg/g), hesperidins (21.74±3.2 μg/g), genkwanins (2.80±0.72 μg/g)HPLC-UV[6]Aerial partsTunisiaBir Mroua (subhumid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoïc acids (382.79±11.98 μg/g), caffeic acid (191.19±27.72 μg/g), rosmarinic acid (2503.96±224.40 μg/g), p-coumaric acid (133.78±1.88 μg/g), ferulic acid (72.89±0.86 μg/g), vanillic acid (14.51±0.46 μg/g)Phenolic diterpenesCarnosic acids (67.95±3.73 μg/g), carnosols (32.09±1.46 μg/g)FlavonoidsNaringenins (1402.07±5.17 μg/g), luteolins (21.14±2.03 μg/g), hesperidins (84.48±4.67 μg/g), apigenins (13.56±0.51 μg/g), cirsiliols (53.18±3.15 μg/g), genkwanins (2.53±0.57 μg/g), naringins (36.79±2.83 μg/g)HPLC-UV[6]Aerial partsTunisiaHencha (higher arid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoic acids (51.18±3.76 μg/g), caffeic acids (50.77±3.04 μg/g), ferulic acids (74.55±16.66 μg/g), p-coumaric acids (22.51±0.84 μg/g), rosmarinic acids (475.74±7.45 μg/g), vanillic acids (9.59±0.91 μg/g)Phenolic diterpenesMethyl carnosate (1159.73±41.68 μg/g), carnosic acids (55.47±1.60 μg/g)FlavonoidsNaringenins (254.82±22.14 μg/g), cirsiliols (57.89±3.81 μg/g), luteolins (51.65±2.42 μg/g), apigenins (23.95±1.00 μg/g), genkwanins (2.65±0.12 μg/g), hesperidins (24.19±1.21 μg/g), naringins (20.26±0.50 μg/g)HPLC[6]Aerial parts (dried)Saudi ArabiaAssir18th February 2001Alcoholic extractVerbenacines and salvinines1D and 2D NMR[39]Roots (dried)AlgeriaBatna—Acetone extract6,7-Dehydroroyleanones, cryptanol, sitosterols, campesterols, 6-hydroxysalvonolones, microstegiols, stigmasterolsIR, UV, 1H 13C NMR, and NMR[40]The terpenoids contained in the essential oils ofS. verbenaca L. mostly consist of α-pinene, β-pinene, sabinene, 1,8-cineole, β-phellandrene, linalool, p-cymene, linalyl acetate, E-β-ocimene, (Z)-β-ocimene, tricyclene, camphor, 1,10-di-epi-cubenol, epi-13-manool, cis-muurola-3,5-diene, δ-selinene, trans-sabinene hydrate acetate, β-caryophyllene, viridiflorol, and germacrene D [31–33] (Table 1, Figure 2).Figure 2
Terpenoid structures identified fromS. verbenaca EO.Belloum et al. [36] evaluated the volatile contents of the essential oil of S. verbenaca aerial parts using GC-MS and GC. In this sense, they recorded the presence of many terpenoids like germacrene D (20.5%), β-phellandrene (3.8%), α-copaene (10.4%), β-caryophyllene (3.8%), epi-α-cadinol (11.6%), and 1,10-di-epi-cubenol (20.9%). These compounds were the major terpenoids identified in S. verbenaca L. as reported elsewhere [31]. Moreover, in Spain, Taârit et al. [34] identified camphor (38.94%), 13-epi-manool (5.61%), and caryophyllene oxide (7.28%), from the essential oils of its seeds. A Greek study on S. verbenaca aerial parts has identified (E)-caryophyllene (16.1%) and β-phellandrene (30.3%) [34]. Moreover, Khemkham et al. [35] revealed cis-muurola-3,5 diene (14.6%) in the dried aerial parts of S. verbenaca as a major compound.Al-Jaber et al. [25] compared the different parts of S. verbenaca volatile compounds collected from two locations in Jordan. Monoterpene hydrocarbons dominated the emission profile of stem, sepal, and leaf samples from the Mediterranean zone (68.0%, 33.7%, and 42.2%, respectively). Oxygenated monoterpenes controlled the production and emission of flowering components, including preflowering buds, fully grown flowers, and petals. Also, Taârit et al. [33] showed that the major compounds in EOs in Salvia aerial parts from the three Algerian regions were the monoterpene hydrocarbons and oxygenated sesquiterpenes. Additionally, the influence of collecting locations and phenophases on the production and chemical composition of S. verbenaca L. essential oils was examined by Farhat et al. [6]. In this study, it was reported that at the floral stage, monoterpene hydrocarbons (31.9%) prevail, whereas oxygenated sesquiterpenes (27.5%) predominate at the early fruiting stage. Sesquiterpene hydrocarbons were the most abundant chemical class at late fruiting (28.2%). Furthermore, Al-Jaber [32] reported that S. verbenaca EO was primarily composed of oxygenated monoterpenes (61.32%), with the monoterpene alcohol linalool serving as the sole monoterpene alcohol, whereas the essential oil obtained from the air-dried plant was primarily composed of sesquiterpene hydrocarbons (62.66%), with germacrene D serving as the major component (25.92%). Chemical heterogeneity of EOs was isolated from three distinct S. verbenaca tissues (leaves, twigs, and stem). In this regard, the EO of S. verbenaca from the fruits contains the highest concentrations of -caryophyllene (23.1%) and caryophyllene oxide (15.9%), while the EO from the stems contains the highest concentrations of camphor and viridiflorol and, and in comparison, the leaf oil contains the highest concentrations of epi-13-manool and manool [33].Regarding phenolic acid compounds, several phenolic compounds were identified in theS. verbenaca methanolic extract, which was the phenolic acid with six compounds: p-hydroxybenzoic acid, vanillic acid, rosmarinic acid, p-coumaric acid, caffeic acid, phenolic diterpenes, and ferulic acid, with three compounds: carnosol, carnosic acid, and methyl carnosate [6] (Table 1, Figure 3). In Turkey, Tepe et al. [38] extracted rosmarinic acid from the dried methanolic extracts of this plant.Figure 3
Phenolic acids and phenolic diterpenes isolated fromS. verbenaca.Moreover, Farhat et al. [6] have identified several flavonoids in methanol extract from aerial parts of Tunisian S. verbenaca L such as luteolin, apigenin, genkwanin, cirsiliol, naringenin, hesperidin, and naringin (Table 1, Figure 4).Figure 4
Flavonoids ofS. verbenaca extracts.Certain fatty acids were found inS. verbenaca (Table 1). Taârit et al. [33] identified approximately eight constituents (oleic acid, linoleic acid, arachidic acid, linolenic acid, palmitic acid, stearic acid, palmitoleic acid, and ethyl palmitate) (Figure 5). Russo et al. [19] isolated several interesting fatty acids from essential oils of S. verbenaca aerial parts, including (Z)-9-octadecenoic acid (oleic acid), hexadecanoic acid (palmitic acid), methyl hexadecanoate (methyl palmitate), and ethyl hexadecanoate (ethyl palmitate).Figure 5
Some fatty acids isolated fromS. verbenaca extracts.Additionally, Kabouche et al. [40] on the roots of S. verbenaca allowed the isolation of other secondary metabolites including five sterols (campesterol, stigmasterol, sitosterol, 6-hydroxysalvonolone, and microstegiol) and two diterpenes (6,7-dehydroroyleanone, cryptanol). Ahmed et al. [39] isolated two new diterpenes, namely, verbenacine and salvinine, from S. verbenaca aerial parts (Table 1, Figure 6).Figure 6
Structure of compounds isolated fromS. verbenaca.


### 2.5. Bioeffective Properties

Different parts ofS. verbenaca exhibit the presence of several bioactive molecules of antibacterial, antileishmanial, antioxidant, and anticancer activities (Figure 7).Figure 7
Pharmacological properties ofS. verbenaca.


#### 2.5.1. Antibacterial Activity

The EOs and other organic extracts ofS. verbenaca showed effective antibacterial effects against various gram-negative and gram-positive bacteria [7, 8, 10]. The inhibition zone diameter of S. verbenaca extracts and EOs and/or the minimum inhibitory concentration (MIC) are presented in (Table 3).Table 3
Antibacterial potential ofS. verbenaca extracts and EOs.
Plant sectionExtractsMethodologyTested bacterial strainsAntibacterial resultsRef.Aerial partsEthanolic extractDisc diffusion method dose (20μL)Gram positiveStaphylococcus aureus (ATCC 25923)Micrococcus luteus (NRRL B-4375)Staphylococcus aureus (MU 44)Staphylococcus aureus (MU 38)Staphylococcus epidermidis (MU 30)Bacillus subtilis (ATCC 6633)Streptococcus mutans (CNCTC 8/77)Gram negativeEscherichia coli (ATCC 25922)Pseudomonas aeruginosa (ATCC 27853)Stenotrophomonas maltophilia (MU 99)Chryseomonas luteola (MU65)Pseudomonas fluorescens (MU 87)Stenotrophomonas maltophilia (MU 64)Pseudomonas stutzeri (MU 70)Ф=11mmNo inhibitionФ=10mmФ=9mmФ=9mmФ=9mmNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibition[10]Not specifiedMethanolic extract (80%)Ethanolic extract (80%)Not specifiedNot specifiedThe 80% methanol extract prepared using ultrasound extraction showed the highest antimicrobial activity[41]Aerial partsMethanolic extractMinimum inhibitory concentrationGram positiveMicrococcus sedentarius (L7B5)Staphylococcus xylosus (IP8166)Corynebacterium gr. C (L3C3)Staphylococcus cohnii (L6S3)Corynebacterium gr. D2 (L19C1)Micrococcus luteus (L1C5)Corynebacterium xerosis (IP5216)Staphylococcus epidermidis (L1S2)Staphylococcus intermedius (IP8160)Corynebacterium gr. B (L16C3)Gram negativeAcinetobacter sp. (LH5DC1)MIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC>1000μg/mL[42]Moraxella sp. (LH7SV1)Alcaligenes sp. (LH4TV1)Pseudomonas cepacia (V6108)Pseudomonas aeruginosa (V5791)MIC=700μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLAerial partsMethanol : chloroform (1 : 1,v/v) extractMicrodilution assayGram positiveBacillus cereus (ATCC 11778)Staphylococcus aureus (ATCC 25923)Gram negativeKlebsiella pneumoniae (NTCC 9633)Escherichia coli (ATCC 8739)MIC=2.0mg/mLMIC=3.0mg/mLMIC=2.0mg/mLMIC=8.0mg/mL[17]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)Disc diffusion methodDose (3 mg and 6 mg/disc)Gram positiveStaphylococcus aureus (ATCC 52952)EAEФ3mg/disc=12mmФ6mg/disc=16mmChEФ3mg/disc=11mmФ6mg/disc=10mmCrEФ̲3mgor6mg/disc=11mmAqE: no inhibition[21]Bacillus cereus (ATCC 10876)EAEФ3mg/disc=13mmФ6mg/disc=15mmChEФ3mg/disc=11mmФ6mg/disc=15mmCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionEnterococcus faecalis (ATCC 49452)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrEФ3mg/disc=noinhibitionФ6mg/disc=11mmAqE: no inhibitionListeria monocytogenes (ATCC 15313)EAEФ3mg/disc=10mmФ6mg/disc=14mmChEФ3mg/disc=8.0mmФ6mg/disc=noinhibitionCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionGram negativeEscherichia coli (ATCC 25922)EAEФ3mg/disc=11mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrE and AqENo inhibitionPseudomonas aeruginosa (ATCC 27853)EAEФ3mg/disc=12mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrEФ3mg/disc=noinhibitionФ6mg/disc=9mmAqE: no inhibitionCitrobacter freundii (ATCC 8090)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=11mmФ6mg/disc=noinhibitionAqE: no inhibitionAcinetobacter baumannii (ATCC 19306)EAEФ3mg/disc=10mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=noinhibitionФ6mg/disc=10mmAqE: no inhibitionProteus mirabilis (ATCC 35659)EAEФ3mgor6mg/disc=13mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrE and AqENo inhibitionSalmonella typhi (ATCC 13311)No inhibitionLeavesEthyl acetate extractAgar diffusion testDose (100 μg and 300 μg/disc)Microbroth dilution assayGram positiveBacillus brevis (ATCC 9999)Bacillus subtilis (ATCC 6633)Staphylococcus aureus (ATCC 43300)Gram negativeKlebsiella pneumoniae (ATCC13883)Escherichia coli (ATCC 25922)MIC=50μg/mLMIC=50μg/mLNo inhibitionNo inhibitionNo inhibition[7]Aerial partsEssential oilBroth dilution methodGram positiveBacillus subtilis (ATCC6633)Staphylococcus aureus (ATCC 25923)Staphylococcus epidermidis (ATCC 12228)Streptococcus faecalis (ATTC 29212)MIC=50μg/mLMIC=100μg/mLMIC=50μg/mLMIC=100μg/mL[8]Gram negativeEscherichia coli (ATCC25922)Proteus vulgaris (ATCC13315)Klebsiella pneumoniae (ATCC10031)Pseudomonas aeruginosa (ATCC27853)MIC>100μg/mLMIC>100μg/mLMIC>100μg/mLMIC>100μg/mLФ: diameter of inhibition.In Turkey, Sarac and Ugur [10] investigated the antibacterial potential of the ethanol extract from S. verbenaca aerial parts; they found that the extract showed a weak antibacterial activity, with IZD between 9 and 11 mm against the gram-positive bacteria Staphylococcus epidermidis (MU 30) (Ф=9mm), Bacillus subtilis (ATCC 6633) (Ф=9mm), S. aureus (MU 44) (Ф=10mm), S. aureus (MU 38) (Ф=9mm), and S. aureus (ATCC 25923) (Ф=11mm), and no activity was seen against Streptococcus mutans (CNCTC8/77) and Micrococcus luteus (NRRL B-4375) and nor gram-negative bacteria, P. fluorescens (MU87), Escherichia coli (ATCC25922), Pseudomonas stutzeri (MU70), Pseudomonas aeruginosa (ATCC27853), Stenotrophomonas maltophilia (MU64), Chryseomonas luteola (MU65), and S. maltophilia (MU99). Moreover, the ethanolic extract prepared from 12 S. verbenaca exhibited lower antimicrobial activity than the methanolic extracts, as found by Kostić et al. [9].The investigation of the methanol extract from aerial parts of TunisianS. verbenaca demonstrated that the extract had a high antibacterial potential (MIC=500μg/mL) against six bacteria isolated from the mouths of patients [42]. However, a South African extract of S. verbenaca that was made with methanol and chloroform had strong antibacterial properties against Klebsiella pneumoniae, Bacillus cereus, Escherichia coli, and Staphylococcus aureus [11]. Moreover, Belkhiri et al. [21] compared the antibacterial potential of four fractions from the methanol extract of Algerian S. verbenaca: chloroform extract, crude extract, aqueous extract, and ethyl acetate extract. They have found that the antibacterial efficacy increases with the concentration of the extract. Al-Zereini [7] also found that the ethyl acetate extract prepared from the leaves of S. verbenaca from Jordan had dose-dependent antibacterial properties against Bacillus brevis (ATCC 9999) and Bacillus subtilis (ATCC 6633). On the other hand, the extract had no effect on Klebsiella pneumoniae (ATCC 13883), Staphylococcus aureus (ATCC 43300), and Escherichia coli (ATCC 25922). Canzoneri et al. [8] found that the EO of S. verbenaca aerial parts has potential antibacterial effects, and this activity is much higher against gram-positive bacteria than gram-negatives.


#### 2.5.2. Antioxidant Activity

The antioxidant potential ofS. verbenaca extracts was investigated by several researchers [12, 21, 23, 38, 42–44], and Table 4 summarizes the majority of the investigations that were carried out on different parts of S. verbenaca, collected from different regions.Table 4
Antioxidant activity ofS. verbenaca.
Part usedExtractsMethods usedKey resultsRef.Not specifiedMethanolic extract (80%)Ethanolic extract (80%)DPPH assayβ-Carotene/linoleic acidsThe 80% methanol extract prepared by maceration was highly activeThe 80% of ethanol extract was the most active[41]Aerial parts (stems and leaves)Hydromethanolic extractOxygen consumptionA strong inhibition of oxygen consumption (92%)[44]Conjugated diene formation (CD)A strong inhibition of CD formation of LDL peroxidation (92%)Thiobarbituric acid reactive substance (TBARS) formationA strong inhibition of TBARS formation of linolenic acid oxidation (93%)Not specifiedMethanolic extractDPPH assayβ-Carotene–linoleic acid methodIC50=14.30±1.42μg/mgPercentinhibition=77.03±0.42%[38]Aerial partsMethanolic extractDPPH assayIC50=86.9μg/mL[42]ABTS assayIC50 at 5min=777.3μg/mLTEAC at 5min=0.624IC50 at 20min=499.5μg/mLTEAC at 10min=0.647TEAC at 20min=0.705TEAC at 15min=0.705Aerial partsMethanolic extract from postdistilled plantDPPH methodSers:IC50=24.47±1.87μg/mLTouiref: IC50=25.11±2.97μg/mLBeja: IC50=26.62±0.8μg/mLChott Meriem: IC50=28.28±0.16μg/mLTunis: IC50=30.34±2.28μg/mLRass Zebib: IC50=31.19±2.25μg/mLBou Arada: IC50=33.47±4.13μg/mLBir Mroua: IC50=34.70±2.43μg/mLHencha: IC50=39.85±3.9μg/mLEnfidha: IC50=40.91±0.5μg/mL[6]ABTS methodHencha:TEAC=120.11±6.62μM trolox/mgEnfidha: TEAC=134.45±5.27μM trolox/mgBir Mroua: TEAC=139.26±10.59μM trolox/mgRass Zebib: TEAC=144.02±3.4μM trolox/mgBou Arada: TEAC=154.97±6.79μM trolox/mgTunis: TEAC=190.51±6.71μM trolox/mgChott Meriem: TEAC=196.72±1.61μM trolox/mgSers: TEAC=271.51±4.52μM trolox/mgBeja: TEAC=282.17±6.58μM trolox/mgTouiref: TEAC=287.81±3.65μM trolox/mgFRAPBeja:142.07±1.46mM Fe+2/mgSers: 139.09±11.23mM Fe+2/mgTouiref: 131.86±1.05mM Fe+2/mgChott Meriem: 124.27±0.38mM Fe+2/mgTunis: 122.33±3.7mM Fe+2/mgBou Arada: 120.53±7.53mM Fe+2/mgRass Zebib: 118.02±15.25mM Fe+2/mgBir Mroua: 109.22±5.04mM Fe+2/mgHencha: 104.89±0.37mM Fe+2/mgEnfidha: 101.46±1.97mM Fe+2/mgAerial partsCrude extractDPPH methodIC50=47.50μg/mL[14]Aerial partsMethanolic extractDPPHFRAPIC50=9.79±0.47μg/mLHigh reducing power[43]Not specifiedMethanolic extractDPPH methodIC50=16.92±0.2μM[12]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)DPPH methodEAE:IC50=0.0086mg/mLCrE: IC50=0.0336mg/mLChE: IC50=0.0725mg/mLAqE: IC50=0.0389mg/mL[21]Reducing power methodEAE: EC50: 0.0047 mg/mLCrE: EC50: 0.0453±0.000 mg/mLAqE: EC50: 0.0455 mg/mLMetal chelating methodAqE and CrE reported the highest activityEAE chelation did not exceed 20%Aerial partsMethanolic extracts (85%) (ME)Decoction extract (distilled water) (DE)DPPH methodME: IC50: 24.36±1.13 μg/mLDE: IC50: 27.26±1.05 μg/mL[13]ABTS methodME: IC50: 19.96±1.03 μg/mLDE: IC50: 36.86±1.03 μg/mLAlkaline DMSO superoxide radical scavengingME: IC50: 07.77±1.00 μg/mLDE: IC50: 18.78±1.07 μg/mLβ-Carotene bleachingDE: inhibition:96.12±2.48%ME: inhibition: 82.58±2.39%Reducing power methodDE: EC50: 69.52±3.07 μg/mLME: EC50: 56.64±4.81 μg/mLMetal chelating activity methodME: IC50: 70.39±1.13 μg/mLDE: IC50: 109.70±1.72 μg/mLPhenanthroline methodME: IC50: 27.03±1.54 μg/mLDE: IC50: 40.26±0.59 μg/mLCupric reducing antioxidant capacity (CUPRAC)ME: A0.50: 14.66±2.51 μg/mLDE: A0.50: 33.00±0.30 μg/mLRootMethanolic extractH2DCF-DA methodSignificant reduction in the intracellular reactive oxygen species (ROS) level for both tested values (1 and 10μg/mL)[14]Kostić et al. [41] evaluated the antioxidant potential of different S. verbenaca extracts using the beta-carotene/linoleic acid system and DPPH assay. They found that the methanol extract had the highest activity in the DPPH method, while the ethanolic extract obtained by ultrasound extraction was the most active metabolite of beta-carotene/linoleic acid. The antioxidant activity of hydromethanolic extract prepared from stems and leaves of Moroccan species was carried out by Khlifi et al. [44]. The results showed that the extract had a significant antioxidant effect at 100 μg/mL, with a strong inhibition of oxygen consumption compared to previous studies [38].The antioxidant potential of TunisianS. verbenaca extracts was also studied [42], and the results showed that methanolic extract from aerial parts had lower activity (IC50=86.9μg/mL) compared to the positive control, which was the trolox (IC50=23.12μg/mL) using the DPPH assay. In addition, it was reported that the antioxidant activity over 20 minutes using the ABTS assay increased with time, but was still four times lower than the activity of trolox. Additionally, Farhat et al. [6] studied the efficacy of the collection sites on the antioxidant capacity of methanolic extract prepared from postdistilled aerial parts of Tunisian species. They found that the site had a significant effect on the antioxidant potential by the DPPH, ABTS, and FRAP methods. Likewise, activity was shown to be substantially linked with total phenolic content.The antioxidant activity of some extracts ofS. verbenaca collected from Algeria was mostly studied using the DPPH assay. It was found that the crude extract prepared from aerial parts had good antioxidant activity that increased with increasing the extract concentration [14]. The scavenging activity was 95% at a concentration of 0.1 mg/mL. Also, the methanol extract of S. verbenaca aerial parts revealed a high reducing power in the FRAP test [43] using the DPPH assay. Additionally, it was cited that the methanol extract had a beneficial effect against free-radical damage and exhibited a 5-fold more inhibitory effect than the standard antioxidant trolox (IC50=72.63μM) [12]. They also observed that the radical scavenging activity had no significant correlation with the phenolic content and a low correlation with the flavonoid content. Belkhiri et al. [21] investigated the antioxidant potential of some fractions of the methanol extracts using the DPPH method, metal chelating activity, and reducing power assay, and all extracts showed potent antioxidant activity [13]. The cupric ion reducing capacity (CUPRAC) and Fe3+ reducing capacity (phenanthroline assay) of the extracts were investigated, and the findings exhibited that both extracts had high antioxidant capacity, with methanolic extract exhibiting the highest activity [14].


#### 2.5.3. Anticancer Activity

The different organic essential oils and extracts ofS. verbenaca have been studied for anticancer properties. Numerous laboratory investigations using cell culture have shown that S. verbenaca extracts and essential oils have antiproliferative properties (Table 5) against a variety of cancer cell lines [15–19, 23, 45].Table 5
Anticancer effects ofS. verbenaca.
Plant partTested extractCell linesMajor resultsRef.LeavesEthyl acetateHuman breast adenocarcinomaIC50: 41.3±4.8 μg/mL[7]Aerial partsMethanolHuman colon adenocarcinomaLC50: 60.4 μg/mL[15]Human hepatoblastomaLC50: 68.9 μg/mLHuman breast cancer cellsLC50: 43.1 μg/mLHuman pancreatic carcinomaLC50: 42.2 μg/mLAerial partsHexaneHuman embryonal rhabdomyosarcoma cancerous cell linesIC50: 474.6±1.3 μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsEthyl acetateHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50: 223.6±1.6 μg/mLAerial partsn-ButanolHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsMethanol and chloroformBreast adenocarcinomaIC50: 31.5±13.7 μg/mL[17]Colon adenocarcinomaIC50: 50.0±5.3 μg/mLGlioblastomaIC50 was not calculatedHuman kidney epithelial cell lineIC50: 20.8±2.5 μg/mLLeavesMethanolMonkey kidney cellsCC50: 64 μg/mL[18]Human larynx cancer cellsCC50=64μg/mLRootsMethanolHuman monocytic leukemia cell line70% of apoptosis and 30% of viable cells at a 1000μg/mL concentration[23]Aerial partsEssential oilsHuman melanoma cell lineIC50=8.1±0.6μg/mL[19]The ethyl acetate extract ofS. verbenaca leaves produced after maceration was examined using the MDA cell lines MB-231 (human breast adenocarcinoma, ATCC HTB-26). The findings indicated that all extracts produced cytotoxicity in MDA MB-231 breast cancer cells [7]. However, it was proved that S. verbenaca leaf extracts possessed cytotoxic effect against HEp-2 (human larynx cancer cells) and Vero (monkey kidney cells) [18]. In another investigation, methanolic extracts of S. verbenaca’s aerial component prepared by maceration were evaluated in vitro against four human cancer cell lines, including HCA, HepG2, MCF-7, and HPC. The findings indicate that LC50 levels higher than 75 μg/mL were deemed inactive [15]. Additionally, MTT assays were used to determine the cytotoxic activity of several extracts (methanol, hexane, ethyl acetate, n-butanol, and chloroform extracts) obtained from the aerial portion of S. verbenaca [16]. Methanol and chloroform extracts of S. verbenaca aerial parts were evaluated against colon adenocarcinoma (HT-29), human cancer cell lines (breast adenocarcinoma (MCF-7), human kidney epithelial cell line and glioblastoma (SF-268)) [17]. S. verbenaca exhibited more favorable action against MCF-7, with an IC50 value of 31.50 13.70 μg/mL, but was inactive versus SF-268 and/or HT-29 cell lines [17].A cell viability study was performed to avoid any cytotoxic concentration ofS. verbenaca root extract on THP-1 cells. The MTT assay revealed that the most cytotoxic concentration of the extract was 1000 μg/mL, which caused 70% of cell death and 30% of cell viability [23]. The essential oils of S. verbena were investigated for their ability to suppress the proliferation of human tumor cells using the human M14 melanoma cell line and shown significant efficacy [19]. The antiproliferative effect of S. verbenaca essential oil may be attributed to active sesquiterpenes in combination with other natural chemicals found in the essential oil components. Indeed, carvacrol and thymol exhibited outstanding anticancer properties through a variety of mechanisms [19].


#### 2.5.4. Antiparasitic Activity

Et-Touys et al. [20] investigated the antileishmanial effects of organic extracts (methanol, n-hexane, and dichloromethane extract) from S. verbenaca, and it was reported that the in vitro antileishmanial effect which was evaluated on the culture of three Leishmania species such as Leishmania infantum, Leishmania tropica, and Leishmania major was good (Table 6).Table 6
Antiparasitic activity ofS. verbenaca.
ActivityPart usedExtractParasiteMajor resultsRef.Antileishmanial activityWhole plant partn-HexaneLeishmania majorIC50: 155.4 μg/mL[20]Leishmania tropicaIC50: 148.2 μg/mLLeishmania infantumIC50: 14.1 μg/mLDichloromethaneLeishmania majorIC50: 24.5 μg/mLLeishmania tropicaIC50: 33.7 μg/mLLeishmania infantumIC50: 31.5 μg/mLMethanolLeishmania majorIC50>1000μg/mLLeishmania tropicaIC50: 850.7 μg/mLLeishmania infantumIC50>1000μg/mLAntimalarial activityAerial partsMethanol chloroformPlasmodium falciparum (FCR-3 strain)IC50: 23.9±1.1 μg/mL[11]Belkhiri et al. [21] additionally observed that S. verbenaca has antihemolytic properties. In vitro antihemolytic activity of S. verbenaca extract was determined by inducing oxidative erythrocyte hemolysis. The results indicated that ethyl acetate extract was the most effective in inhibiting hemolysis, followed by crude extract, chloroform extract, and aqueous extract. Additionally, ethyl acetate extract inhibited hemolysis more effectively than vitamin C.


#### 2.5.5. Insecticidal Activity

In most cases, the application of synthetic pesticides is the primary approach for controlling insect pests, which produces excellent effects in a short period of time. Meanwhile, their irrational usage has resulted in global issues such as pollution, nontarget toxicity, biodiversity loss, and the development of pest resistance [46–48]. This need arose from a desire to provide alternatives to synthetic insecticides, which can have negative environmental consequences [49–52]. The insecticidal capabilities of S. verbenaca extracts and essential oils have been documented to have potential impact against several pests in previous studies [53]. Insecticidal action has been shown in several experiments using some Salvia species [54].Essential oils fromSalvia species revealed 100% repellency activity against adults of Aedes albopictus [55]. The oil of S. verbenaca drastically shortened the lifespan of cowpea weevil and prevented females from laying eggs [56]. Several crude extracts and essential oils from Salvia species were tested for pesticide activity against the test pest larvae [57–59]. Insectistatic and insecticidal properties of chloroform extracts from the aerial portions of four Salvia species were examined [60]. S. verbenaca extracts are very effective against Culex quinquefasciatus mosquitos [61]. Caryophyllene oxide was the major component in the essential oil of S. verbenaca with 7.28 [62]. The insecticidal activity and fumigant toxicity of caryophyllene oxide were tested against two insect pests, and it was shown to be effective [63].


### 2.6. Other Biological Effects

Different extracts fromS. verbenaca have also exhibited other biological activities such as antihemolytic, immunomodulatory, and enzyme inhibitory effects (Table 7).Table 7
Other biological activities ofS. verbenaca.
ActivityPart usedExtractsExperimentMajor resultsRef.AntihemolyticAerial partEthyl acetate2,2-Azobis (2-amidinopropane) dihydrochloride induces erythrocyte oxidative hemolysis (AAPH)HT50: 165 min[21]CrudeHT50: 125.1 minChloroformHT50: 111.5 minAqueousHT50: 111.5 minXanthine oxidase inhibitionAerial partChloroformColorimetric approach based on uric acid generation at 295 nm in the presence of 100 mM xanthine in phosphate bufferIC50: 0.0088±0.0 mg/mLEthyl acetateIC50: 0.0165±0.001 mg/mLCrudeIC50: 0.0520±0.003 mg/mLAqueousIC50: 0.9800±0.004 mg/mLPorcine liver carboxylesterase inhibitionAerial partAqueous methanolEnzyme inhibition by spectrophotometric assayCE (carboxylesterase) inactivation with apI=5.1 and a Ki value of 38 Mm[12]Healing of burnsLeavesHexaneSecond-degree burn injury induced by a hot metal cylinder in ratsAccelerated healing process with 44.34%[16]Ethyl acetateAccelerated healing process with 47.55%n-ButanolAccelerated healing process with 49.16%AnticholinesterasesAerial partMethanolCholinesterase inhibitionInhibition effect of AChE at 100μg/mL[13]Anti-α-amylaseα-Amylase inhibitionIC50: 01.3±0.08 μg/mLAnti-α-glucosidaseα-Glucosidase inhibitionIC50=150.5±1.4μg/mLImmunomodulatoryAerial partsMethanolPhagocytic activity used carbon clearance rate testSignificantly increased phagocytic index (0.095±0.012) at a dose of 200 mg/kgIncreased corrected phagocytic index α (0.095±1.71)[14]


#### 2.6.1. Xanthine Oxidase Inhibitory Effect

Xanthine oxidase, abbreviated as XO, is an oxidoreductase that catalyzes the conversion of hypoxanthine to xanthine and xanthine to uric acid. Xanthine oxidase is generally present in the liver and in an inactive form in the blood in humans. A blood test for XO may identify liver impairment because xanthine oxidase is released into the blood in situations of severe liver injury [21].


#### 2.6.2. Burn Recovery Activities

Guaouguaou et al. [24] evaluated the impact of three S. verbenaca extracts on the healing of burns in rats using hexane, ethyl acetate, and n-butanol. The results indicated that various Salvia verbenaca plant extracts were more effective than silver sulfadiazine (SSD) and that it is the most widely used topical treatment for injury, with healed areas of 29.17% (base), 44.34% (hexane), 47.55% (ethyl acetate), 49.16% (n-butanol), and 41.09% SSD.


#### 2.6.3. In Vitro Antidiabetic Activity

Several earlier studies have shownS. verbenaca’s antidiabetic activity in vitro [13]. Additional studies are shown in Section 2.8 and Figure 8.Figure 8
Antidiabetic mechanism insights ofS. verbenaca main compounds.


#### 2.6.4. Immunomodulatory Effects

Previous studies investigated the immunomodulatory effects ofS. verbenaca aerial parts [14]. The carbon clearance rate test was used to determine the immunostimulant potential of this plant on phagocytic activity. The phagocytic index was much higher in rats who were given S. verbenaca at a dose of 200 mg/kg than in rats who were not given the herb.


### 2.7. Toxicological Investigations ofS. verbenaca

The toxicological investigations ofS. verbenaca have not been well studied. However, some studies carried out recently have confirmed the safety of these plant extracts (Table 8). Indeed, a report by Guaouguaou et al. [64] focused on the acute and subchronic effects of S. verbenaca toxicity in mice and rats through oral and topical administration. The findings of the acute toxicity of the fractions derived from S. verbenaca (n-butanol, hexane, and ethyl acetate) demonstrated that the LD50 of this plant after oral administration at 2000 mg kg-1 is not deadly [64]. In order to complete the toxicity profile of this plant, more research should be done to find out how toxic it is over a long period of time.Table 8
Toxicological studies ofS. verbenaca.
ActivitiesPart usedExtractExperimental approachMajor resultsRef.Acute oral toxicityAerial partsn-Butanol, hexane, ethyl acetateOrally delivered at a dose of 2000 mg/kg in a volume of 0.25 mL per 20 g of body weight to mice and examined for 14 daysLD50>2000mg/kg body weight[64]Acute dermal toxicityAerial partsHexane, ethyl acetate,n-butanolFor 14 days, daily topical application ofS. verbenaca extracts at a dose of 2000 mg/kg body weightThere are no adverse effects, behavioral problems, or fatalities[64]Subchronic dermal toxicityAerial partsHexane, ethyl acetate,n-butanolFor 28 days, daily topical application ofS. verbenaca extracts at a dose of 2000 mg/kg body weightThere are no harmful symptoms or changes in the amount of water or food consumedThere is no lethalityThere is no change in the parameters of fasting blood circulationThere were no morphological alterations in the main vital organs investigated[64]


### 2.8. Pharmacological Properties ofS. verbenaca Main Volatile Compounds

Several studies examined the major volatile chemicals found inS. verbenaca, including carvacrol, thymol, and linalool. Studies showed that carvacrol has hypoglycemic properties through intrinsic mechanisms such as blood glucose and insulin level lowering [65]. Additionally, carvacrol resulted in a drop in glucose levels. Additionally, these substances were shown to enhance the activity of glucokinase and glucose-6-phosphate dehydrogenase in the liver [66]. Carvacrol inhibits the enzymes α-amylase and alpha-glucosidase in vitro [67] and beta-galactosidase in vitro [68]. Thymol was also able to normalize blood sugar, plasma insulin, HbA1c, and the insulin resistance index in patients with hyperglycemia [69]. The levels of expression of genes involved in the production of insulin have been studied and reported in STZ-induced diabetic mice [70, 71], and a rise in Mafa and Pdx1 gene expression has been reported. Limonene is another major constituent of S. verbenaca that has been shown to improve glucose homeostasis. Indeed, this substance boosts hepatic glycogen and plasma glucose levels [72] (Figure 8).The antidiabetic effect has been also revealed by linalool (another main compound ofS. verbenaca) [73, 74]. Indeed, linalool lowered blood glucose, hemoglobin A1c, fructosamine, interleukin-6, and tumor necrosis factor-α (TNF-α), while it increased insulin levels [74].The major phytochemical compounds ofS. verbenaca exhibited remarkable antibacterial effects [75–77]. Rhayour et al. [70] investigated the impact of thymol on gram-positive and gram-negative microorganisms, including Bacillus subtilis and Escherichia coli [75–78]. Antibacterial activity is demonstrated by modifying cell shape, damaging cell walls and membranes, and limiting the development of some types of bacteria, including P. aeruginosa [79]. In addition, limonene was found to be antibacterial because it targeted microorganisms’ cytoplasmic membranes, weakened membrane integrity, blocked respiratory enzymes, and lost the proton motive force (Figure 9).Figure 9
Suggested antibacterial mechanisms ofS. verbenaca compounds.The anticancer properties of the major components inS. verbenaca (carvacrol, limonene, and thymol) have also been reported recently [80–82]. Thymol has been shown to have anticancer properties via a variety of mechanisms, including inducing severe DNA damage, including the production of reactive oxygen species (ROS) and subsequent increase in oxidative stress and/or mitochondrial dysfunction, or via the nuclear factor of activated T cell (NFAT-2) route [81]. Additionally, carvacrol increased apoptosis in cells, perhaps via activating mitochondrial apoptotic and signaling pathways [83].


## 2.1. Botanical Description

S. verbenaca is a perennial herb that reaches between 10 and 50 cm (in height), hairy at the top, odorous, more or less glandular at the top. It grows in the dry lawns, the slopes, and at the edges of the paths. The slightly branched stems carry bunches of dark blue flowers in spring. Leaves are oblong, 2-3 cm broad, crenelated or incised-lobed, with the upper stalkless (Figure 1). The flowers are quite small, pale blue or whitish, in whorls usually close together, forming a fairly short cluster; the fruiting calyx with almost closed lips, bristling with spread hairs; the corolla is 10–15 mm, twice as long as the calyx, with wide lips, very uneven, the upper one compressed and curved in a false shape, and the style with little or prominent point [25].Figure 1
Salvia verbenaca L.: (a) whole plant; (b) aerial part; (c) flowers.
(a)(b)(c)


## 2.2. Geographic Distribution

S. verbenaca has a very wide geographical distribution around the Mediterranean region, including Morocco, Algeria, Tunisia, Canaries, Egypt, Libya, Turkey, Cyprus, Transcaucasia, and Western and Southern Europe. It is also grown in South West Africa, North America, and Australia [5].


## 2.3. Ethnomedicinal Uses

The ethnobotanical investigations intoS. verbenaca revealed its wide applications in folkloric medicine to treat numerous disorders as listed in Table 1. In Morocco, its application in folk medicinal systems includes the treatment of some digestive disorders such as abdominal colics [26–28]. The most commonly used part of the plant is the aerial part, which is prepared by infusion before being used to treat respiratory problems and genitourinary and skin diseases [27]. Dried leaves are also used for the treatment of wounds, burns, and abscesses [29]. Aerial parts are utilized in decoction or infusion to treat diabetes [30].Table 1
Some medicinal applications ofS. verbenaca.
Part usedPreparationTraditional applicationRef.Aerial partDecoction, infusionDiabetes[30]LeafDecoction, powderAbdominal colics, cold, fever, healing[26]LeafPowderWound treatment[29]LeafDecoction, infusionGenitourinary, skin, digestive, and respiratory problems[27]Aerial partNo informationDigestive problems[28]Leaf, whole plantPowderHealing of burns, wounds, and abscesses[29]NI: no information.


## 2.4. Phytochemistry

Like all medicinal plants belonging to the family Lamiaceae,S. verbenaca contains numerous secondary metabolites with different classes, such as flavonoids, terpenoids, alkaloids, and phenolic acids. Currently, several analytical investigations using different technical tools (GC, GC-SM, GC-MS, GC-FID, HPLC, 1D and 2D NMR, IR, UV, 1H NMR, and 13C NMR) have been applied to identify and isolate bioactive compounds from medicinal plants. Indeed, investigations into the chemical constituents of S. verbenaca revealed the presence of terpenoids, phenolics, fatty acids, flavonoids, and sterols (Table 2). As listed in Table 2, the chemical content of S. verbenaca was investigated in different areas with various medicinal applications by using different analytical tools. The results are different according to numerous factors, such as the study area, plant part used, and adopted methodology.Table 2
Chemical composition of various parts ofS. verbenaca.
Part usedCountryHarvest siteHarvest seasonExtracts/essential oilsChemical compositionAnalysisReferencesAerial parts (dry)TunisiaRass Zebib (subhumid)Flowering periodEO1,8-Cineole (9.7%), p-cymene (8.4%),α-pinene (5.4%), γ-terpinene (3.1%), beta-caryophyllene (5.3%), viridiflorol (7.3%), epi-13-manool (4.7%), thymol (3.7%), limonene (2.8%), camphor (2.7%)GC and GC-MS[6]Aerial parts (dry)TunisiaBir Mroua (subhumid)Flowering periodEOβ-Caryophyllene (15.3%), germacrene D (7.1%), epi-13-manool (6.2%), α-copaene (6.1%), α-humulene (4.3%), α-cadinol (3.9%), viridiflorol (3.4%), p-cymene (3.3%), δ-cadinene (3.1%), p-cymen-8-ol (2.6%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaBeja (higher semiarid)Flowering periodEOβ-Caryophyllene (15.3%), α-humulene (3.0%), viridiflorol (11.6%), 1,8-cineole (3.3%), germacrene D (3.3%), (Z)-β-ocimene (4.0%), T-cadinol (1.9%), p-cymene (2.8%), thymol (2.7%), epi-13-manool (2.5%)GC and GC-MS[6, 31]Aerial parts (dry)TunisiaTunis (higher semiarid)Flowering periodEOViridiflorol (17.7%), 1,8-cineole (8.5%),α-pinene (4.6%), p-cymene (5.2%), β-caryophyllene (5.5%), thymol (4.4%), epi-13-manool (4.0%), α-humulene (2.4%), α-thujone (3.6%), γ-terpinene (2.4%)GC and GC-MS[31, 33, 34]Aerial parts (dry)TunisiaTouiref (moderate semiarid)Flowering periodEOα-Pinene (15.9%), camphor (4.7%), 1,8-cineole (12.8%), viridiflorol (10.0%), (Z)-β-ocimene (5.4%), camphene (2.6%), β-caryophyllene (5.3%), thymol (4.2%), p-cymene (4.2%), α-thujone (3.4%)GC and GC-MS[31, 33, 35]Aerial parts (dry)TunisiaBou Arada (moderate semiarid)Flowering periodEO1,8-Cineole (9.4%), p-cymene (8.7%), viridiflorol (8.3%),α-pinene (4.9%), thymol (2.7%), β-caryophyllene (4.9%), α-humulene (3.5%), γ-terpinene (3.0%), α-thujone (3.0%), epi-13-manool (3.6%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaSers (lower semiarid)Flowering periodEOα-Pinene (14.7%), viridiflorol (10.8%), β-caryophyllene (4.6%), (Z)-β-ocimene (4.5%), epi-13-manool (2.8%), thymol (4.4%), p-cymene (4.1%), camphor (3.5%), α-thujone (2.9%), 1,8-cineole (10.9%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaEnfidha (lower semiarid)Flowering periodEOViridiflorol (10.5%), camphor (2.9%), epi-13-manool (10.5%), 1,8-cineole (8.7%), p-cymene (8.3%),α-terpineol (3.0%), α-pinene (4.5%), thymol (4.2%), γ-terpinene (3.2%), bornyl acetate (3.2%)GC and GC-MS[31–33]Aerial parts (dry)TunisiaChott Meriem (higher arid)Flowering periodEOp-Cymene (14.2),α-pinene (9.6), γ-terpinene (5.1), camphene (3.9), viridiflorol (5.1), limonene (3.4), epi-13-manool (3.2), thymol (2.5), 1,8-cineole (12.8)GC and GC-MS[31–33]Aerial parts (dry)TunisiaHencha (higher arid)Flowering periodEOViridiflorol (10.0%), bicyclogermacrene (2.3%), germacrene D (5.6%), 1,8-cineole (4.9), epi-13-manool (4.7%),α-thujone (3.2%), β-pinene (3.0%), camphor (2.9%), α-humulene (2.5%), β-caryophyllene (7.2%)GC and GC-MS[31–33]Aerial parts (dry)SpainMurciaFlowering stageEOp-Cymene (11.4%), 1,8-cineole (7.7%), viridiflorol (7.0%), camphene (2.7%),β-caryophyllene (4.5%), β-pinene (2.7%), γ-terpinene (4.0%), epi-13-manool (3.9%), camphor (3.7%), α-pinene (8.1%)GC and GC-MS[6]Aerial parts (dry)SpainMurciaEarly fruiting stageEOCaryophyllene oxide (12.4%), bornyl acetate (3.2%), viridiflorol (9.1%),β-caryophyllene (5.6%), p-cymene (5.6%), α-pinene (4.0%), epi-13-manool (2.3%), thymol (2.0%), β-ionone (2.0%), 1,8-cineole (6.3%)GC and GC-MS[6]Aerial parts (dry)SpainMurciaLate fruiting stageEOβ-Caryophyllene (14.2%), α-thujone (8.2%), 8-cineole (4.7%), epi-13-manool (7.1%), bornyl acetate (3.5%), α-humulene (6.7%), 1 α-pinene (4.3%), caryophyllene oxide (3.0%), β-pinene (2.8%), viridiflorol (13.5%)GC and GC-MS[6]Aerial partsAlgeriaBecharApril 2011EOEpi-α-cadinol (11.6%), β-caryophyllene (11.33%), bicyclogermacrene (10.9%), γ-cadinene (7.9%), cis-muurola-4(14),5-diene (7.8%), muurola-3,5-diene (5.2%), spathulenol (3.0%), cis-calamenene (2.0), α-humulene (1.9), 1,10-di-epi-cubenol (20.9%)GC and GC-MS[31]Aerial parts (fresh)JordanShafa-Badran-AmmanFlowering period (April to May 2011)EOLinalool (61.32%),β-elemene (1.50%), (Z)-β-ocimene (4.03%), β-eudesmol (3.66%), spathulenol (3.40%), E-β-ocimene (2.63%), β-caryophyllene (2.98%), α-copaene (2.50%), γ-cadinene (1.55%), bicyclogermacrene (5.94%)GC-MS and GC-FID[32]Aerial parts (dry)JordanShafa-Badran-AmmanFlowering period (April to May 2011)EOLinalool (30.72%), bicyclogermacrene (14.70%),β-caryophyllene (7.42%), germacrene D (25.92%), α-copaene (5.13%), isopentyl isovalerate (0.97%), δ-cadinene (2.05%), (Z)-β-ocimene (1.18%), spathulenol (1.58%), α-gurjunene (1.07%)GC and GC-MS[32]StemJordanMediterraneanFull maturation periodEOZ-β-Ocimene (32.6%), trans-sabinene hydrate acetate (14.5%), α-gurjunene (6.0%), β-bourbonene (1.5%), E-β-ocimene (7.8%), sabinene (2.9%), α-phellandrene (3.1%), germacrene D (1.6%), α-pinene (9.3%), β-pinene (8.1%)GC-MS and GC-FID[32]JordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate acetate (38.1%), E-caryophyllene (9.1%), δ-selinene (5.2%), β-gurjunene (2.5%), sabinene (4.8%), δα-copaene (4.1%), γ-gurjunene (2.9%), cadinene (4.3%), β-selinene (2.2%), germacrene D (13.3%)GC and GC-MS[32]LeavesJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (30.2%), β-bourbonene (7.7%), E-β-ocimene (4.3%), α-pinene (3.0%), α-gurjunene (13.8%), β-selinene (2.8%), δ-cadinene (2.5%), β-pinene (2.4%), myrcene (2.0%), Z-β-ocimene (17.1%)GC and GC-MS[32]LeavesJordanIrano-TuranianFull maturation periodEOδ-Selinene (21.5%), E-caryophyllene (11.4%), terpinolene (4.3%), α-copaene (9.6%), sabinene (9.0%), Z-β-ocimene (4.8%), β-cubebene (4.4%), δ-cadinene (2.7%), cis-β-guaiene (2.0%), germacrene D (19.8%)GC and GC-MS[32]PreflowerJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (56.5%), α-pinene (6.5%), myrcene (1.5%), E-β-ocimene (4.3%), α-gurjunene (3.2%), β-pinene (5.3%), sabinene (1.2%), trans-β-guaiene (1.0%), limonene (0.7%), Z-β-ocimene (13.5%)GC and GC-MS[32]PreflowerJordanIrano-TuranianFull maturation periodEOSabinene (42.7%),α-thujene (7.2%), γ-terpinene (6.1%), E-β-ocimene (1.9%), α-terpinene (3.6%), β-pinene (3.2%), β-phellandrene (6.8%), terpinolene (1.6%), limonene (1.0%), trans-sabinene hydrate (20.4%)GC and GC-MS[32]FlowerJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (58.6%), E-β-ocimene (5.3%), α-pinene (5.2%), sabinene (1.1%), β-pinene (4.9%), α-phellandrene (1.4%), α-gurjunene (1.0%), camphene (0.4%), isobornyl acetate (0.4%), Z-β-ocimene (18.8%)GC and GC-MS[32]JordanIrano-TuranianFull maturation periodEOSabinene (37.5%),Z-β-ocimene (9.9%), α-thujene (4.6%), myrcene (4.2%), β-pinene (3.9%), E-β-ocimene (8.9%), γ-terpinene (3.0%), E-caryophyllene (1.9%), α-terpinene (1.4%), trans-sabinene hydrate (20.0%)GC and GC-MS[32]PetalJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (87.0%), E-β-ocimene (1.5%), germacrene D (1.0%), α-phellandrene (0.5%), β-pinene (0.3%), α-gurjunene (1.7%), n-nonane (0.2%), myrcene (0.2%), β-selinene (0.2%), Z-β-ocimene (7.1%)GC and GC-MS[32]PetalJordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate (18.8%), E-β-ocimene (9.9%), γ-terpinene (2.9%), germacrene D (9.6%), β-copaene (4.1%), α-copaene (3.2%), E-caryophyllene (13.9%), β-selinene (2.9%), γ-gurjunene (2.8%), Z-β-ocimene (9.6%)GC and GC-MS[32]SepalJordanMediterraneanFull maturation periodEOtrans-Sabinene hydrate acetate (36.6%), β-pinene (14.0%), 8-cineole (3.9%), Z-β-ocimene (4.5%), 1 δ-elemene (2.8%), β-cedrene (8.7%), sabinene (2.7%), camphene (1.9%), β-cubebene (1.9%), α-pinene (18.1%)GC and GC-MS[32]SepalJordanIrano-TuranianFull maturation periodEOtrans-Sabinene hydrate (58.8%), terpinolene (5.0%), E-β-ocimene (4.1%), p-methyl-acetophenone (3.2%), germacrene D (3.1%), Z-β-ocimene (4.9%), δ-selinene (2.0%), γ-terpinene (1.7%), δ-cadinene (1.3%), E-caryophyllene (5.6%)GC and GC-MS[32]Aerial partsAlgeriaMogheulApril 2011EOGermacrene D (20.5%),β-caryophyllene (3.8%), beta-cubebene (2.7%), δ-cadinene (2.6%), 1,10-di-epi-cubenol (2. 6%), γ-cadinene (2.5%), (E)-β-farnesene (3.5%), bicyclogermacrene (2.2%), α-muurolol (2.1%), α-copaene (10.4%), β-phellandrene (3.8%)GC and GC-MS[36]SeedsSpain——EOCamphor (38.94%), 13-epi-manool (5.61%), 𝛿-elemene (3.93%), beta-eudesmol (3.76%), n-undecane (2.65%), α-terpinyl acetate (4.77%), linalyl acetate (2.53%), neryl acetate (2.40%), α-terpineol (2.03%), caryophyllene oxide (7.28%)GC-MS and GC-FID[33]Aerial partsGreeceCrete IslandBlossoming (April 2004)EOBeta-phellandrene (30.3%), methyl ester of 6-octadecenoic acid (15.0%), camphor (7.0%), (Z)-β-ocimene (6.6%), fenchone (9.4%), isopropyl ester (7.8%), aromadendrene (4.0%), α-humulene (3.7%), (E)-caryophyllene (16.1%)GC and GC-MS[34]Aerial parts (fresh)SicilyPiano BattagliaFull flowering stage (July 2009)EOHexadecanoic acid (23.1%), ethyl hexadecanoate (2.6%), benzaldehyde (7.3%), 9,12,15-octadecatrienal (2.9%), limonene (2.0%), (E)-β-ionone (1.9%), (Z)-9-octadecenoic acid (11.9), phenyl acetaldehyde (1.5%), (E)-caryophyllene (1.2%), β-phellandrene (5.9%)GC and GC-MS[8]Aerial partsAlgeriaDjelfaMarch 2019EOcis-Muurola-3,5-diene (14.6%), unknown (10.5%), bicyclogermacrene (6.8%), bicycloelemene (4.3%), γ-cadinene (4.8%), β-pinene (4.2%), 2,3-dehydro-1,4-cineol (3.7%), α-cubebene (3.0%), α-pinene (2.8%), γ-amorphene (10.5%)GC and GC-MS[35]Leaves and flowers (dried)TurkeyKütahya-Gediz2016-2017EOLinalyl acetate (81.97%),β-myrcene (2.73%), n-pentanal (0.42%), beta-ocimene (0.39%), hexanal (0.34%), α-pinene (0.34%), limonene (1.14%), trans-caryophyllene (0.32%), β-pinene (0.31%), linalool (8.66%)GC and GC-MS[37]Aerial parts (wild)SicilyPiano BattagliaFull flowering stage (July 2009)EOHexadecanoic acid (23.1%), benzaldehyde (7.3%), b-phellandrene (5.9%), limonene (2.0%), 9,12,15-octadecatrienal (2.9%), ethyl hexadecanoate (2.6%), caryophyllene oxide (1.9%), (E)-b-ionone (1.9%), spathulenol (1.7%), (Z)-9-octadecenoic acid (11.1%)GC and GC-MS[19]Aerial parts (cultivated)SicilyPiano BattagliaJuly 2010EOHexadecanoic acid (11.0%), (E)-b-ionone (3.9%), (Z)-9-octadecenoic acid (5.6%), b-phellandrene (4.1%), caryophyllene oxide (2.8%), (E)-caryophyllene (3.8%), methyl hexadecanoate (3.8%), carvacrol (2.4%), spathulenol (2.0%), hexahydrofarnesyl acetone (9.7%)GC and GC-SM[19]FruitsTunisiaSabelet Ben AmmarFull fruit ripening stageEOβ-Caryophyllene (23.1%), camphene (6.5%), α-humulene (5.6%), germacrene D (3.5%), viridiflorol (4.3%), 1-octen-3-ol (3.9%), (E)-β-ocimene (1.5%), 1,8-cineole (3.0%), manool (1.1%), caryophyllene oxide (15.9%)GC and GC-MS[33]StemsTunisiaSabelet Ben AmmarFull fruit ripening stageCamphor (10.9%), terpinolene (6.6%), methyl eugenol (6.1%),α-pinene (5.9%), α-thujone (3.1%), 1,8-cineole (5.8%), caryophyllene oxide (4.5%), aromadendrene (3.6%), epi-13-manool (2.3%), viridiflorol (10.3%)GC and GC-MS[33]LeavesTunisiaSabelet Ben AmmarFull fruit ripening stageepi-13-Manool (13.7%), camphor (3.9%), caryophyllene oxide (3.9%),α-pinene (3.4%), p-cymen-8-ol (3.7%), terpinen-4-ol (3.6%), 1,8-cineole (3.0%), eugenol (2.8%), (E)-β-ocimene (2.6%), manool (11.0%)GC and GC-MS[33]Aerial partsTunisiaSabelet Ben AmmarFull fruiting stage (April 2007)EOViridiflorol (21.6%), methyl eugenol (9.4%),α-terpineol (5.3%), spathulenol (3.7%), β-caryophyllene (7.1%), caryophyllene oxide (2.4%), epi-13-manool (2.2%), germacrene D (1.9%), eugenol (1.8%), camphene (17.6%)GC and GC-MS[33]Aerial partsTunisiaSersFull fruiting stage (April 2007)(Z)-β-ocimene (29.5%), beta-thujone (7.9%), α-pinene (5.5%), tricyclene (5.1%), 18-cineole (1.9%), α-calacorene (2.5%), terpinen-4-ol (2.1%), germacrene D (3.1%), β-caryophyllene (1.8%), β-phellandrene (8.2%)GC and GC-MS[33]Aerial partsTunisiaSomaaTricyclene (18.8%), nonane (10.3%), terpinolene (7.3%), -terpineol (2.2%), bornyl acetate (4.9%), camphor (2.9%),α-terpinyl acetate (3.5%), limonene (2.3%), α β-eudesmol (2.2%), methyl eugenol (7.7%)GC and GC-MS[33]SeedsTunisiaSabelet Ben AmmarFull ripeness (April 2007)EOCamphor (33.83%), caryophyllene oxide (10.11%), octane (4.78%), 13-epi-manool (3.57%), hexanal (2.46%),β-bisabolene (1.84%), α-terpineol (3.24%), tricyclene (5.54%), α-copaene (3.19%), α-thujene (13.36%)GC-MS and GC-FID[33]SeedsTunisiaSabelet Ben AmmarFull ripeness (April 2007)Lipid extractionPalmitic acid (9.25%), stearic acid (2.48%), linolenic acid (45.89%), arachidic acid (0.20%), C18:3n-3/C18:2n-6 (1.67%), SFA (11.93%), USFA (88.07%), oleic acid (14.67%), linoleic acid (27.39%), palmitoleic acid (0.12%)GC and GC-MS[33]SeedsTunisiaSersFull ripeness (April 2007)EOβ-Pinene (48.08%), epi-cubebol (10.74%), β-eudesmol (1.00%), α-bisabolol (2.97%), caryophyllene oxide (2.90%), spathulenol (0.93%), eugenol (0.97%), geraniol (0.95%), borneol (1.97%), germacrene D (2.09%)GC and GC-MS[33]SeedsTunisiaSersFull ripeness (April 2007)Lipid extractionPalmitic acid (9.63%), oleic acid (14.14%), linoleic acid (23.79%), linolenic acid (42.84%), SFA (18.35%), USFA (81.65%), stearic acid (4.22%), arachidic acid (4.50%), C18:3n-3/C18:2n-6 (1.53%), palmitoleic acid (0.89%)GC and GC-MS[33]SeedsTunisiaSomaaFull ripeness (April 2007)EOOctane (27.39%),δ-cadinene (5.77%), p-cymene (1.64%), camphor (3.53%), bicyclogermacrene (1.86%), β-pinene (3.74%), α-terpineol (1.38%), limonene (0.79%) n-nonane (18.01%), epi-cubebol (9.02%)GC and GC-MS[33]SeedsTunisiaSomaaFull ripeness (April 2007)Lipid extractionPalmitic acid (12.11%), stearic acid (3.02%), linoleic acid (25.33%), arachidic acid (1.30%), SFA (16.43%), linolenic acid (41.71%), oleic acid (15.51%), USFA (83.57%), C18:3n-3/C18:2n-6 (1.65%), palmitoleic acid (1.02%)GC and GC-MS[33]Aerial partAlgeriaBordj Bou ArreridjFlowering stage (spring April-May)Crude extract (CrE)Flavonoids (08.40±0.32mg EQ/g E), polyphenols (177.56±2.51mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjChloroform extract (ChE)Flavonoids (14.87±0.81mg EQ/g E), polyphenols (156.81±1.57mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjEthyl acetate extract (EAE)Flavonoids (28.81±0.38mg EQ/g E), polyphenols (661.78±4.00mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaBordj Bou ArreridjAqueous extract (AqE)Flavonoids (06.74±0.14mg EQ/g E), polyphenols (123.18±4.20mg EGA/g E)Spectrophotometrically[21]Aerial partAlgeriaLaghouatMay 200480% (v/v) aqueous methanolFlavonoids (3.04±0.01mg RE/g dw), total phenols (7.2±0.04mg GAE/g dw), flavonols (0.85±0.001mg QE/g dw)Spectrophotometrically[12]Aerial partAlgeriaSetif and Batna2016DecoctionTotal phenols (129.02±2.67mg GAE/g DW), total flavonoid contents (18.62±0.06mg QE/g DW), total tannin contents (73.80±2.23mg TAE/g DW)Total carotenoid contents (0.92±0.041mg/g DW)Total chlorophyll A (1.21±0.02mg/g DW)Total chlorophyll B (2.48±0.04mg/g DW)Spectrophotometrically[13]Aerial partAlgeriaSetif and Batna2016Methanol extract (85%)Total phenols (190.16±1.74mg GAE/g DW)Total flavonoid contents (23.50±0.71mg QE/g DW)Total tannin contents (118.88±1.25mg TAE/g DW)Total carotenoid contents (0.58±0.005mg/g DW)Total chlorophyll A (1.67±0.02mg/g DW)Total chlorophyll B (0.63±0.01mg/g DW)Spectrophotometrically[13]Aerial partTurkeyArtvin06th September 2004Methanol extractsRosmarinic acids (29.30±0.24 μg mg_1)Spectrophotometrically[38]Rosmarinic acids (26.12±0.73 μg mg_1)HPLC[6]Aerial partsTunisiaTunis (higher semiarid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoïc acids (229.87±8.60 μg/g), p-coumaric acid (77.65±5.67 μg/g), rosmarinic acid (1688.01±63.42 μg/g), vanillic acid (20.21±0.46 μg/g), caffeic acids (97.29±2.86 μg/g), ferulic acids (40.41±3.32 μg/g)Phenolic diterpenesCarnosic acids (63.52±15.30 μg/g), methyl carnosate contents (633.37±11.66 μg/g), carnosol (25.52±7.27 μg/g)FlavonoidsNaringenins (940.41±22.50 μg/g), cirsiliols (73.16±1.72 μg/g), luteolins (13.84±2.62 μg/g), apigenins (3.01±0.69 μg/g), naringins (57.30±3.55 μg/g), hesperidins (21.74±3.2 μg/g), genkwanins (2.80±0.72 μg/g)HPLC-UV[6]Aerial partsTunisiaBir Mroua (subhumid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoïc acids (382.79±11.98 μg/g), caffeic acid (191.19±27.72 μg/g), rosmarinic acid (2503.96±224.40 μg/g), p-coumaric acid (133.78±1.88 μg/g), ferulic acid (72.89±0.86 μg/g), vanillic acid (14.51±0.46 μg/g)Phenolic diterpenesCarnosic acids (67.95±3.73 μg/g), carnosols (32.09±1.46 μg/g)FlavonoidsNaringenins (1402.07±5.17 μg/g), luteolins (21.14±2.03 μg/g), hesperidins (84.48±4.67 μg/g), apigenins (13.56±0.51 μg/g), cirsiliols (53.18±3.15 μg/g), genkwanins (2.53±0.57 μg/g), naringins (36.79±2.83 μg/g)HPLC-UV[6]Aerial partsTunisiaHencha (higher arid)Flowering stage (March and April 2008)Methanolic extractsPhenolic acidsp-Hydroxybenzoic acids (51.18±3.76 μg/g), caffeic acids (50.77±3.04 μg/g), ferulic acids (74.55±16.66 μg/g), p-coumaric acids (22.51±0.84 μg/g), rosmarinic acids (475.74±7.45 μg/g), vanillic acids (9.59±0.91 μg/g)Phenolic diterpenesMethyl carnosate (1159.73±41.68 μg/g), carnosic acids (55.47±1.60 μg/g)FlavonoidsNaringenins (254.82±22.14 μg/g), cirsiliols (57.89±3.81 μg/g), luteolins (51.65±2.42 μg/g), apigenins (23.95±1.00 μg/g), genkwanins (2.65±0.12 μg/g), hesperidins (24.19±1.21 μg/g), naringins (20.26±0.50 μg/g)HPLC[6]Aerial parts (dried)Saudi ArabiaAssir18th February 2001Alcoholic extractVerbenacines and salvinines1D and 2D NMR[39]Roots (dried)AlgeriaBatna—Acetone extract6,7-Dehydroroyleanones, cryptanol, sitosterols, campesterols, 6-hydroxysalvonolones, microstegiols, stigmasterolsIR, UV, 1H 13C NMR, and NMR[40]The terpenoids contained in the essential oils ofS. verbenaca L. mostly consist of α-pinene, β-pinene, sabinene, 1,8-cineole, β-phellandrene, linalool, p-cymene, linalyl acetate, E-β-ocimene, (Z)-β-ocimene, tricyclene, camphor, 1,10-di-epi-cubenol, epi-13-manool, cis-muurola-3,5-diene, δ-selinene, trans-sabinene hydrate acetate, β-caryophyllene, viridiflorol, and germacrene D [31–33] (Table 1, Figure 2).Figure 2
Terpenoid structures identified fromS. verbenaca EO.Belloum et al. [36] evaluated the volatile contents of the essential oil of S. verbenaca aerial parts using GC-MS and GC. In this sense, they recorded the presence of many terpenoids like germacrene D (20.5%), β-phellandrene (3.8%), α-copaene (10.4%), β-caryophyllene (3.8%), epi-α-cadinol (11.6%), and 1,10-di-epi-cubenol (20.9%). These compounds were the major terpenoids identified in S. verbenaca L. as reported elsewhere [31]. Moreover, in Spain, Taârit et al. [34] identified camphor (38.94%), 13-epi-manool (5.61%), and caryophyllene oxide (7.28%), from the essential oils of its seeds. A Greek study on S. verbenaca aerial parts has identified (E)-caryophyllene (16.1%) and β-phellandrene (30.3%) [34]. Moreover, Khemkham et al. [35] revealed cis-muurola-3,5 diene (14.6%) in the dried aerial parts of S. verbenaca as a major compound.Al-Jaber et al. [25] compared the different parts of S. verbenaca volatile compounds collected from two locations in Jordan. Monoterpene hydrocarbons dominated the emission profile of stem, sepal, and leaf samples from the Mediterranean zone (68.0%, 33.7%, and 42.2%, respectively). Oxygenated monoterpenes controlled the production and emission of flowering components, including preflowering buds, fully grown flowers, and petals. Also, Taârit et al. [33] showed that the major compounds in EOs in Salvia aerial parts from the three Algerian regions were the monoterpene hydrocarbons and oxygenated sesquiterpenes. Additionally, the influence of collecting locations and phenophases on the production and chemical composition of S. verbenaca L. essential oils was examined by Farhat et al. [6]. In this study, it was reported that at the floral stage, monoterpene hydrocarbons (31.9%) prevail, whereas oxygenated sesquiterpenes (27.5%) predominate at the early fruiting stage. Sesquiterpene hydrocarbons were the most abundant chemical class at late fruiting (28.2%). Furthermore, Al-Jaber [32] reported that S. verbenaca EO was primarily composed of oxygenated monoterpenes (61.32%), with the monoterpene alcohol linalool serving as the sole monoterpene alcohol, whereas the essential oil obtained from the air-dried plant was primarily composed of sesquiterpene hydrocarbons (62.66%), with germacrene D serving as the major component (25.92%). Chemical heterogeneity of EOs was isolated from three distinct S. verbenaca tissues (leaves, twigs, and stem). In this regard, the EO of S. verbenaca from the fruits contains the highest concentrations of -caryophyllene (23.1%) and caryophyllene oxide (15.9%), while the EO from the stems contains the highest concentrations of camphor and viridiflorol and, and in comparison, the leaf oil contains the highest concentrations of epi-13-manool and manool [33].Regarding phenolic acid compounds, several phenolic compounds were identified in theS. verbenaca methanolic extract, which was the phenolic acid with six compounds: p-hydroxybenzoic acid, vanillic acid, rosmarinic acid, p-coumaric acid, caffeic acid, phenolic diterpenes, and ferulic acid, with three compounds: carnosol, carnosic acid, and methyl carnosate [6] (Table 1, Figure 3). In Turkey, Tepe et al. [38] extracted rosmarinic acid from the dried methanolic extracts of this plant.Figure 3
Phenolic acids and phenolic diterpenes isolated fromS. verbenaca.Moreover, Farhat et al. [6] have identified several flavonoids in methanol extract from aerial parts of Tunisian S. verbenaca L such as luteolin, apigenin, genkwanin, cirsiliol, naringenin, hesperidin, and naringin (Table 1, Figure 4).Figure 4
Flavonoids ofS. verbenaca extracts.Certain fatty acids were found inS. verbenaca (Table 1). Taârit et al. [33] identified approximately eight constituents (oleic acid, linoleic acid, arachidic acid, linolenic acid, palmitic acid, stearic acid, palmitoleic acid, and ethyl palmitate) (Figure 5). Russo et al. [19] isolated several interesting fatty acids from essential oils of S. verbenaca aerial parts, including (Z)-9-octadecenoic acid (oleic acid), hexadecanoic acid (palmitic acid), methyl hexadecanoate (methyl palmitate), and ethyl hexadecanoate (ethyl palmitate).Figure 5
Some fatty acids isolated fromS. verbenaca extracts.Additionally, Kabouche et al. [40] on the roots of S. verbenaca allowed the isolation of other secondary metabolites including five sterols (campesterol, stigmasterol, sitosterol, 6-hydroxysalvonolone, and microstegiol) and two diterpenes (6,7-dehydroroyleanone, cryptanol). Ahmed et al. [39] isolated two new diterpenes, namely, verbenacine and salvinine, from S. verbenaca aerial parts (Table 1, Figure 6).Figure 6
Structure of compounds isolated fromS. verbenaca.


## 2.5. Bioeffective Properties

Different parts ofS. verbenaca exhibit the presence of several bioactive molecules of antibacterial, antileishmanial, antioxidant, and anticancer activities (Figure 7).Figure 7
Pharmacological properties ofS. verbenaca.


### 2.5.1. Antibacterial Activity

The EOs and other organic extracts ofS. verbenaca showed effective antibacterial effects against various gram-negative and gram-positive bacteria [7, 8, 10]. The inhibition zone diameter of S. verbenaca extracts and EOs and/or the minimum inhibitory concentration (MIC) are presented in (Table 3).Table 3
Antibacterial potential ofS. verbenaca extracts and EOs.
Plant sectionExtractsMethodologyTested bacterial strainsAntibacterial resultsRef.Aerial partsEthanolic extractDisc diffusion method dose (20μL)Gram positiveStaphylococcus aureus (ATCC 25923)Micrococcus luteus (NRRL B-4375)Staphylococcus aureus (MU 44)Staphylococcus aureus (MU 38)Staphylococcus epidermidis (MU 30)Bacillus subtilis (ATCC 6633)Streptococcus mutans (CNCTC 8/77)Gram negativeEscherichia coli (ATCC 25922)Pseudomonas aeruginosa (ATCC 27853)Stenotrophomonas maltophilia (MU 99)Chryseomonas luteola (MU65)Pseudomonas fluorescens (MU 87)Stenotrophomonas maltophilia (MU 64)Pseudomonas stutzeri (MU 70)Ф=11mmNo inhibitionФ=10mmФ=9mmФ=9mmФ=9mmNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibition[10]Not specifiedMethanolic extract (80%)Ethanolic extract (80%)Not specifiedNot specifiedThe 80% methanol extract prepared using ultrasound extraction showed the highest antimicrobial activity[41]Aerial partsMethanolic extractMinimum inhibitory concentrationGram positiveMicrococcus sedentarius (L7B5)Staphylococcus xylosus (IP8166)Corynebacterium gr. C (L3C3)Staphylococcus cohnii (L6S3)Corynebacterium gr. D2 (L19C1)Micrococcus luteus (L1C5)Corynebacterium xerosis (IP5216)Staphylococcus epidermidis (L1S2)Staphylococcus intermedius (IP8160)Corynebacterium gr. B (L16C3)Gram negativeAcinetobacter sp. (LH5DC1)MIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC>1000μg/mL[42]Moraxella sp. (LH7SV1)Alcaligenes sp. (LH4TV1)Pseudomonas cepacia (V6108)Pseudomonas aeruginosa (V5791)MIC=700μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLAerial partsMethanol : chloroform (1 : 1,v/v) extractMicrodilution assayGram positiveBacillus cereus (ATCC 11778)Staphylococcus aureus (ATCC 25923)Gram negativeKlebsiella pneumoniae (NTCC 9633)Escherichia coli (ATCC 8739)MIC=2.0mg/mLMIC=3.0mg/mLMIC=2.0mg/mLMIC=8.0mg/mL[17]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)Disc diffusion methodDose (3 mg and 6 mg/disc)Gram positiveStaphylococcus aureus (ATCC 52952)EAEФ3mg/disc=12mmФ6mg/disc=16mmChEФ3mg/disc=11mmФ6mg/disc=10mmCrEФ̲3mgor6mg/disc=11mmAqE: no inhibition[21]Bacillus cereus (ATCC 10876)EAEФ3mg/disc=13mmФ6mg/disc=15mmChEФ3mg/disc=11mmФ6mg/disc=15mmCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionEnterococcus faecalis (ATCC 49452)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrEФ3mg/disc=noinhibitionФ6mg/disc=11mmAqE: no inhibitionListeria monocytogenes (ATCC 15313)EAEФ3mg/disc=10mmФ6mg/disc=14mmChEФ3mg/disc=8.0mmФ6mg/disc=noinhibitionCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionGram negativeEscherichia coli (ATCC 25922)EAEФ3mg/disc=11mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrE and AqENo inhibitionPseudomonas aeruginosa (ATCC 27853)EAEФ3mg/disc=12mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrEФ3mg/disc=noinhibitionФ6mg/disc=9mmAqE: no inhibitionCitrobacter freundii (ATCC 8090)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=11mmФ6mg/disc=noinhibitionAqE: no inhibitionAcinetobacter baumannii (ATCC 19306)EAEФ3mg/disc=10mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=noinhibitionФ6mg/disc=10mmAqE: no inhibitionProteus mirabilis (ATCC 35659)EAEФ3mgor6mg/disc=13mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrE and AqENo inhibitionSalmonella typhi (ATCC 13311)No inhibitionLeavesEthyl acetate extractAgar diffusion testDose (100 μg and 300 μg/disc)Microbroth dilution assayGram positiveBacillus brevis (ATCC 9999)Bacillus subtilis (ATCC 6633)Staphylococcus aureus (ATCC 43300)Gram negativeKlebsiella pneumoniae (ATCC13883)Escherichia coli (ATCC 25922)MIC=50μg/mLMIC=50μg/mLNo inhibitionNo inhibitionNo inhibition[7]Aerial partsEssential oilBroth dilution methodGram positiveBacillus subtilis (ATCC6633)Staphylococcus aureus (ATCC 25923)Staphylococcus epidermidis (ATCC 12228)Streptococcus faecalis (ATTC 29212)MIC=50μg/mLMIC=100μg/mLMIC=50μg/mLMIC=100μg/mL[8]Gram negativeEscherichia coli (ATCC25922)Proteus vulgaris (ATCC13315)Klebsiella pneumoniae (ATCC10031)Pseudomonas aeruginosa (ATCC27853)MIC>100μg/mLMIC>100μg/mLMIC>100μg/mLMIC>100μg/mLФ: diameter of inhibition.In Turkey, Sarac and Ugur [10] investigated the antibacterial potential of the ethanol extract from S. verbenaca aerial parts; they found that the extract showed a weak antibacterial activity, with IZD between 9 and 11 mm against the gram-positive bacteria Staphylococcus epidermidis (MU 30) (Ф=9mm), Bacillus subtilis (ATCC 6633) (Ф=9mm), S. aureus (MU 44) (Ф=10mm), S. aureus (MU 38) (Ф=9mm), and S. aureus (ATCC 25923) (Ф=11mm), and no activity was seen against Streptococcus mutans (CNCTC8/77) and Micrococcus luteus (NRRL B-4375) and nor gram-negative bacteria, P. fluorescens (MU87), Escherichia coli (ATCC25922), Pseudomonas stutzeri (MU70), Pseudomonas aeruginosa (ATCC27853), Stenotrophomonas maltophilia (MU64), Chryseomonas luteola (MU65), and S. maltophilia (MU99). Moreover, the ethanolic extract prepared from 12 S. verbenaca exhibited lower antimicrobial activity than the methanolic extracts, as found by Kostić et al. [9].The investigation of the methanol extract from aerial parts of TunisianS. verbenaca demonstrated that the extract had a high antibacterial potential (MIC=500μg/mL) against six bacteria isolated from the mouths of patients [42]. However, a South African extract of S. verbenaca that was made with methanol and chloroform had strong antibacterial properties against Klebsiella pneumoniae, Bacillus cereus, Escherichia coli, and Staphylococcus aureus [11]. Moreover, Belkhiri et al. [21] compared the antibacterial potential of four fractions from the methanol extract of Algerian S. verbenaca: chloroform extract, crude extract, aqueous extract, and ethyl acetate extract. They have found that the antibacterial efficacy increases with the concentration of the extract. Al-Zereini [7] also found that the ethyl acetate extract prepared from the leaves of S. verbenaca from Jordan had dose-dependent antibacterial properties against Bacillus brevis (ATCC 9999) and Bacillus subtilis (ATCC 6633). On the other hand, the extract had no effect on Klebsiella pneumoniae (ATCC 13883), Staphylococcus aureus (ATCC 43300), and Escherichia coli (ATCC 25922). Canzoneri et al. [8] found that the EO of S. verbenaca aerial parts has potential antibacterial effects, and this activity is much higher against gram-positive bacteria than gram-negatives.


### 2.5.2. Antioxidant Activity

The antioxidant potential ofS. verbenaca extracts was investigated by several researchers [12, 21, 23, 38, 42–44], and Table 4 summarizes the majority of the investigations that were carried out on different parts of S. verbenaca, collected from different regions.Table 4
Antioxidant activity ofS. verbenaca.
Part usedExtractsMethods usedKey resultsRef.Not specifiedMethanolic extract (80%)Ethanolic extract (80%)DPPH assayβ-Carotene/linoleic acidsThe 80% methanol extract prepared by maceration was highly activeThe 80% of ethanol extract was the most active[41]Aerial parts (stems and leaves)Hydromethanolic extractOxygen consumptionA strong inhibition of oxygen consumption (92%)[44]Conjugated diene formation (CD)A strong inhibition of CD formation of LDL peroxidation (92%)Thiobarbituric acid reactive substance (TBARS) formationA strong inhibition of TBARS formation of linolenic acid oxidation (93%)Not specifiedMethanolic extractDPPH assayβ-Carotene–linoleic acid methodIC50=14.30±1.42μg/mgPercentinhibition=77.03±0.42%[38]Aerial partsMethanolic extractDPPH assayIC50=86.9μg/mL[42]ABTS assayIC50 at 5min=777.3μg/mLTEAC at 5min=0.624IC50 at 20min=499.5μg/mLTEAC at 10min=0.647TEAC at 20min=0.705TEAC at 15min=0.705Aerial partsMethanolic extract from postdistilled plantDPPH methodSers:IC50=24.47±1.87μg/mLTouiref: IC50=25.11±2.97μg/mLBeja: IC50=26.62±0.8μg/mLChott Meriem: IC50=28.28±0.16μg/mLTunis: IC50=30.34±2.28μg/mLRass Zebib: IC50=31.19±2.25μg/mLBou Arada: IC50=33.47±4.13μg/mLBir Mroua: IC50=34.70±2.43μg/mLHencha: IC50=39.85±3.9μg/mLEnfidha: IC50=40.91±0.5μg/mL[6]ABTS methodHencha:TEAC=120.11±6.62μM trolox/mgEnfidha: TEAC=134.45±5.27μM trolox/mgBir Mroua: TEAC=139.26±10.59μM trolox/mgRass Zebib: TEAC=144.02±3.4μM trolox/mgBou Arada: TEAC=154.97±6.79μM trolox/mgTunis: TEAC=190.51±6.71μM trolox/mgChott Meriem: TEAC=196.72±1.61μM trolox/mgSers: TEAC=271.51±4.52μM trolox/mgBeja: TEAC=282.17±6.58μM trolox/mgTouiref: TEAC=287.81±3.65μM trolox/mgFRAPBeja:142.07±1.46mM Fe+2/mgSers: 139.09±11.23mM Fe+2/mgTouiref: 131.86±1.05mM Fe+2/mgChott Meriem: 124.27±0.38mM Fe+2/mgTunis: 122.33±3.7mM Fe+2/mgBou Arada: 120.53±7.53mM Fe+2/mgRass Zebib: 118.02±15.25mM Fe+2/mgBir Mroua: 109.22±5.04mM Fe+2/mgHencha: 104.89±0.37mM Fe+2/mgEnfidha: 101.46±1.97mM Fe+2/mgAerial partsCrude extractDPPH methodIC50=47.50μg/mL[14]Aerial partsMethanolic extractDPPHFRAPIC50=9.79±0.47μg/mLHigh reducing power[43]Not specifiedMethanolic extractDPPH methodIC50=16.92±0.2μM[12]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)DPPH methodEAE:IC50=0.0086mg/mLCrE: IC50=0.0336mg/mLChE: IC50=0.0725mg/mLAqE: IC50=0.0389mg/mL[21]Reducing power methodEAE: EC50: 0.0047 mg/mLCrE: EC50: 0.0453±0.000 mg/mLAqE: EC50: 0.0455 mg/mLMetal chelating methodAqE and CrE reported the highest activityEAE chelation did not exceed 20%Aerial partsMethanolic extracts (85%) (ME)Decoction extract (distilled water) (DE)DPPH methodME: IC50: 24.36±1.13 μg/mLDE: IC50: 27.26±1.05 μg/mL[13]ABTS methodME: IC50: 19.96±1.03 μg/mLDE: IC50: 36.86±1.03 μg/mLAlkaline DMSO superoxide radical scavengingME: IC50: 07.77±1.00 μg/mLDE: IC50: 18.78±1.07 μg/mLβ-Carotene bleachingDE: inhibition:96.12±2.48%ME: inhibition: 82.58±2.39%Reducing power methodDE: EC50: 69.52±3.07 μg/mLME: EC50: 56.64±4.81 μg/mLMetal chelating activity methodME: IC50: 70.39±1.13 μg/mLDE: IC50: 109.70±1.72 μg/mLPhenanthroline methodME: IC50: 27.03±1.54 μg/mLDE: IC50: 40.26±0.59 μg/mLCupric reducing antioxidant capacity (CUPRAC)ME: A0.50: 14.66±2.51 μg/mLDE: A0.50: 33.00±0.30 μg/mLRootMethanolic extractH2DCF-DA methodSignificant reduction in the intracellular reactive oxygen species (ROS) level for both tested values (1 and 10μg/mL)[14]Kostić et al. [41] evaluated the antioxidant potential of different S. verbenaca extracts using the beta-carotene/linoleic acid system and DPPH assay. They found that the methanol extract had the highest activity in the DPPH method, while the ethanolic extract obtained by ultrasound extraction was the most active metabolite of beta-carotene/linoleic acid. The antioxidant activity of hydromethanolic extract prepared from stems and leaves of Moroccan species was carried out by Khlifi et al. [44]. The results showed that the extract had a significant antioxidant effect at 100 μg/mL, with a strong inhibition of oxygen consumption compared to previous studies [38].The antioxidant potential of TunisianS. verbenaca extracts was also studied [42], and the results showed that methanolic extract from aerial parts had lower activity (IC50=86.9μg/mL) compared to the positive control, which was the trolox (IC50=23.12μg/mL) using the DPPH assay. In addition, it was reported that the antioxidant activity over 20 minutes using the ABTS assay increased with time, but was still four times lower than the activity of trolox. Additionally, Farhat et al. [6] studied the efficacy of the collection sites on the antioxidant capacity of methanolic extract prepared from postdistilled aerial parts of Tunisian species. They found that the site had a significant effect on the antioxidant potential by the DPPH, ABTS, and FRAP methods. Likewise, activity was shown to be substantially linked with total phenolic content.The antioxidant activity of some extracts ofS. verbenaca collected from Algeria was mostly studied using the DPPH assay. It was found that the crude extract prepared from aerial parts had good antioxidant activity that increased with increasing the extract concentration [14]. The scavenging activity was 95% at a concentration of 0.1 mg/mL. Also, the methanol extract of S. verbenaca aerial parts revealed a high reducing power in the FRAP test [43] using the DPPH assay. Additionally, it was cited that the methanol extract had a beneficial effect against free-radical damage and exhibited a 5-fold more inhibitory effect than the standard antioxidant trolox (IC50=72.63μM) [12]. They also observed that the radical scavenging activity had no significant correlation with the phenolic content and a low correlation with the flavonoid content. Belkhiri et al. [21] investigated the antioxidant potential of some fractions of the methanol extracts using the DPPH method, metal chelating activity, and reducing power assay, and all extracts showed potent antioxidant activity [13]. The cupric ion reducing capacity (CUPRAC) and Fe3+ reducing capacity (phenanthroline assay) of the extracts were investigated, and the findings exhibited that both extracts had high antioxidant capacity, with methanolic extract exhibiting the highest activity [14].


### 2.5.3. Anticancer Activity

The different organic essential oils and extracts ofS. verbenaca have been studied for anticancer properties. Numerous laboratory investigations using cell culture have shown that S. verbenaca extracts and essential oils have antiproliferative properties (Table 5) against a variety of cancer cell lines [15–19, 23, 45].Table 5
Anticancer effects ofS. verbenaca.
Plant partTested extractCell linesMajor resultsRef.LeavesEthyl acetateHuman breast adenocarcinomaIC50: 41.3±4.8 μg/mL[7]Aerial partsMethanolHuman colon adenocarcinomaLC50: 60.4 μg/mL[15]Human hepatoblastomaLC50: 68.9 μg/mLHuman breast cancer cellsLC50: 43.1 μg/mLHuman pancreatic carcinomaLC50: 42.2 μg/mLAerial partsHexaneHuman embryonal rhabdomyosarcoma cancerous cell linesIC50: 474.6±1.3 μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsEthyl acetateHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50: 223.6±1.6 μg/mLAerial partsn-ButanolHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsMethanol and chloroformBreast adenocarcinomaIC50: 31.5±13.7 μg/mL[17]Colon adenocarcinomaIC50: 50.0±5.3 μg/mLGlioblastomaIC50 was not calculatedHuman kidney epithelial cell lineIC50: 20.8±2.5 μg/mLLeavesMethanolMonkey kidney cellsCC50: 64 μg/mL[18]Human larynx cancer cellsCC50=64μg/mLRootsMethanolHuman monocytic leukemia cell line70% of apoptosis and 30% of viable cells at a 1000μg/mL concentration[23]Aerial partsEssential oilsHuman melanoma cell lineIC50=8.1±0.6μg/mL[19]The ethyl acetate extract ofS. verbenaca leaves produced after maceration was examined using the MDA cell lines MB-231 (human breast adenocarcinoma, ATCC HTB-26). The findings indicated that all extracts produced cytotoxicity in MDA MB-231 breast cancer cells [7]. However, it was proved that S. verbenaca leaf extracts possessed cytotoxic effect against HEp-2 (human larynx cancer cells) and Vero (monkey kidney cells) [18]. In another investigation, methanolic extracts of S. verbenaca’s aerial component prepared by maceration were evaluated in vitro against four human cancer cell lines, including HCA, HepG2, MCF-7, and HPC. The findings indicate that LC50 levels higher than 75 μg/mL were deemed inactive [15]. Additionally, MTT assays were used to determine the cytotoxic activity of several extracts (methanol, hexane, ethyl acetate, n-butanol, and chloroform extracts) obtained from the aerial portion of S. verbenaca [16]. Methanol and chloroform extracts of S. verbenaca aerial parts were evaluated against colon adenocarcinoma (HT-29), human cancer cell lines (breast adenocarcinoma (MCF-7), human kidney epithelial cell line and glioblastoma (SF-268)) [17]. S. verbenaca exhibited more favorable action against MCF-7, with an IC50 value of 31.50 13.70 μg/mL, but was inactive versus SF-268 and/or HT-29 cell lines [17].A cell viability study was performed to avoid any cytotoxic concentration ofS. verbenaca root extract on THP-1 cells. The MTT assay revealed that the most cytotoxic concentration of the extract was 1000 μg/mL, which caused 70% of cell death and 30% of cell viability [23]. The essential oils of S. verbena were investigated for their ability to suppress the proliferation of human tumor cells using the human M14 melanoma cell line and shown significant efficacy [19]. The antiproliferative effect of S. verbenaca essential oil may be attributed to active sesquiterpenes in combination with other natural chemicals found in the essential oil components. Indeed, carvacrol and thymol exhibited outstanding anticancer properties through a variety of mechanisms [19].


### 2.5.4. Antiparasitic Activity

Et-Touys et al. [20] investigated the antileishmanial effects of organic extracts (methanol, n-hexane, and dichloromethane extract) from S. verbenaca, and it was reported that the in vitro antileishmanial effect which was evaluated on the culture of three Leishmania species such as Leishmania infantum, Leishmania tropica, and Leishmania major was good (Table 6).Table 6
Antiparasitic activity ofS. verbenaca.
ActivityPart usedExtractParasiteMajor resultsRef.Antileishmanial activityWhole plant partn-HexaneLeishmania majorIC50: 155.4 μg/mL[20]Leishmania tropicaIC50: 148.2 μg/mLLeishmania infantumIC50: 14.1 μg/mLDichloromethaneLeishmania majorIC50: 24.5 μg/mLLeishmania tropicaIC50: 33.7 μg/mLLeishmania infantumIC50: 31.5 μg/mLMethanolLeishmania majorIC50>1000μg/mLLeishmania tropicaIC50: 850.7 μg/mLLeishmania infantumIC50>1000μg/mLAntimalarial activityAerial partsMethanol chloroformPlasmodium falciparum (FCR-3 strain)IC50: 23.9±1.1 μg/mL[11]Belkhiri et al. [21] additionally observed that S. verbenaca has antihemolytic properties. In vitro antihemolytic activity of S. verbenaca extract was determined by inducing oxidative erythrocyte hemolysis. The results indicated that ethyl acetate extract was the most effective in inhibiting hemolysis, followed by crude extract, chloroform extract, and aqueous extract. Additionally, ethyl acetate extract inhibited hemolysis more effectively than vitamin C.


### 2.5.5. Insecticidal Activity

In most cases, the application of synthetic pesticides is the primary approach for controlling insect pests, which produces excellent effects in a short period of time. Meanwhile, their irrational usage has resulted in global issues such as pollution, nontarget toxicity, biodiversity loss, and the development of pest resistance [46–48]. This need arose from a desire to provide alternatives to synthetic insecticides, which can have negative environmental consequences [49–52]. The insecticidal capabilities of S. verbenaca extracts and essential oils have been documented to have potential impact against several pests in previous studies [53]. Insecticidal action has been shown in several experiments using some Salvia species [54].Essential oils fromSalvia species revealed 100% repellency activity against adults of Aedes albopictus [55]. The oil of S. verbenaca drastically shortened the lifespan of cowpea weevil and prevented females from laying eggs [56]. Several crude extracts and essential oils from Salvia species were tested for pesticide activity against the test pest larvae [57–59]. Insectistatic and insecticidal properties of chloroform extracts from the aerial portions of four Salvia species were examined [60]. S. verbenaca extracts are very effective against Culex quinquefasciatus mosquitos [61]. Caryophyllene oxide was the major component in the essential oil of S. verbenaca with 7.28 [62]. The insecticidal activity and fumigant toxicity of caryophyllene oxide were tested against two insect pests, and it was shown to be effective [63].


## 2.5.1. Antibacterial Activity

The EOs and other organic extracts ofS. verbenaca showed effective antibacterial effects against various gram-negative and gram-positive bacteria [7, 8, 10]. The inhibition zone diameter of S. verbenaca extracts and EOs and/or the minimum inhibitory concentration (MIC) are presented in (Table 3).Table 3
Antibacterial potential ofS. verbenaca extracts and EOs.
Plant sectionExtractsMethodologyTested bacterial strainsAntibacterial resultsRef.Aerial partsEthanolic extractDisc diffusion method dose (20μL)Gram positiveStaphylococcus aureus (ATCC 25923)Micrococcus luteus (NRRL B-4375)Staphylococcus aureus (MU 44)Staphylococcus aureus (MU 38)Staphylococcus epidermidis (MU 30)Bacillus subtilis (ATCC 6633)Streptococcus mutans (CNCTC 8/77)Gram negativeEscherichia coli (ATCC 25922)Pseudomonas aeruginosa (ATCC 27853)Stenotrophomonas maltophilia (MU 99)Chryseomonas luteola (MU65)Pseudomonas fluorescens (MU 87)Stenotrophomonas maltophilia (MU 64)Pseudomonas stutzeri (MU 70)Ф=11mmNo inhibitionФ=10mmФ=9mmФ=9mmФ=9mmNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibitionNo inhibition[10]Not specifiedMethanolic extract (80%)Ethanolic extract (80%)Not specifiedNot specifiedThe 80% methanol extract prepared using ultrasound extraction showed the highest antimicrobial activity[41]Aerial partsMethanolic extractMinimum inhibitory concentrationGram positiveMicrococcus sedentarius (L7B5)Staphylococcus xylosus (IP8166)Corynebacterium gr. C (L3C3)Staphylococcus cohnii (L6S3)Corynebacterium gr. D2 (L19C1)Micrococcus luteus (L1C5)Corynebacterium xerosis (IP5216)Staphylococcus epidermidis (L1S2)Staphylococcus intermedius (IP8160)Corynebacterium gr. B (L16C3)Gram negativeAcinetobacter sp. (LH5DC1)MIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC=500μg/mLMIC=500μg/mLMIC>1000μg/mLMIC>1000μg/mL[42]Moraxella sp. (LH7SV1)Alcaligenes sp. (LH4TV1)Pseudomonas cepacia (V6108)Pseudomonas aeruginosa (V5791)MIC=700μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLMIC>1000μg/mLAerial partsMethanol : chloroform (1 : 1,v/v) extractMicrodilution assayGram positiveBacillus cereus (ATCC 11778)Staphylococcus aureus (ATCC 25923)Gram negativeKlebsiella pneumoniae (NTCC 9633)Escherichia coli (ATCC 8739)MIC=2.0mg/mLMIC=3.0mg/mLMIC=2.0mg/mLMIC=8.0mg/mL[17]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)Disc diffusion methodDose (3 mg and 6 mg/disc)Gram positiveStaphylococcus aureus (ATCC 52952)EAEФ3mg/disc=12mmФ6mg/disc=16mmChEФ3mg/disc=11mmФ6mg/disc=10mmCrEФ̲3mgor6mg/disc=11mmAqE: no inhibition[21]Bacillus cereus (ATCC 10876)EAEФ3mg/disc=13mmФ6mg/disc=15mmChEФ3mg/disc=11mmФ6mg/disc=15mmCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionEnterococcus faecalis (ATCC 49452)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrEФ3mg/disc=noinhibitionФ6mg/disc=11mmAqE: no inhibitionListeria monocytogenes (ATCC 15313)EAEФ3mg/disc=10mmФ6mg/disc=14mmChEФ3mg/disc=8.0mmФ6mg/disc=noinhibitionCrEФ3mg/disc=noinhibitionФ6mg/disc=12mmAqE: no inhibitionGram negativeEscherichia coli (ATCC 25922)EAEФ3mg/disc=11mmФ6mg/disc=14mmChEФ3mg/disc=09mmФ6mg/disc=12mmCrE and AqENo inhibitionPseudomonas aeruginosa (ATCC 27853)EAEФ3mg/disc=12mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrEФ3mg/disc=noinhibitionФ6mg/disc=9mmAqE: no inhibitionCitrobacter freundii (ATCC 8090)EAEФ3mg/disc=12mmФ6mg/disc=14mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=11mmФ6mg/disc=noinhibitionAqE: no inhibitionAcinetobacter baumannii (ATCC 19306)EAEФ3mg/disc=10mmФ6mg/disc=15mmChEФ3mg/disc=noinhibitionФ6mg/disc=14mmCrEФ3mg/disc=noinhibitionФ6mg/disc=10mmAqE: no inhibitionProteus mirabilis (ATCC 35659)EAEФ3mgor6mg/disc=13mmChEФ3mg/disc=noinhibitionФ6mg/disc=13mmCrE and AqENo inhibitionSalmonella typhi (ATCC 13311)No inhibitionLeavesEthyl acetate extractAgar diffusion testDose (100 μg and 300 μg/disc)Microbroth dilution assayGram positiveBacillus brevis (ATCC 9999)Bacillus subtilis (ATCC 6633)Staphylococcus aureus (ATCC 43300)Gram negativeKlebsiella pneumoniae (ATCC13883)Escherichia coli (ATCC 25922)MIC=50μg/mLMIC=50μg/mLNo inhibitionNo inhibitionNo inhibition[7]Aerial partsEssential oilBroth dilution methodGram positiveBacillus subtilis (ATCC6633)Staphylococcus aureus (ATCC 25923)Staphylococcus epidermidis (ATCC 12228)Streptococcus faecalis (ATTC 29212)MIC=50μg/mLMIC=100μg/mLMIC=50μg/mLMIC=100μg/mL[8]Gram negativeEscherichia coli (ATCC25922)Proteus vulgaris (ATCC13315)Klebsiella pneumoniae (ATCC10031)Pseudomonas aeruginosa (ATCC27853)MIC>100μg/mLMIC>100μg/mLMIC>100μg/mLMIC>100μg/mLФ: diameter of inhibition.In Turkey, Sarac and Ugur [10] investigated the antibacterial potential of the ethanol extract from S. verbenaca aerial parts; they found that the extract showed a weak antibacterial activity, with IZD between 9 and 11 mm against the gram-positive bacteria Staphylococcus epidermidis (MU 30) (Ф=9mm), Bacillus subtilis (ATCC 6633) (Ф=9mm), S. aureus (MU 44) (Ф=10mm), S. aureus (MU 38) (Ф=9mm), and S. aureus (ATCC 25923) (Ф=11mm), and no activity was seen against Streptococcus mutans (CNCTC8/77) and Micrococcus luteus (NRRL B-4375) and nor gram-negative bacteria, P. fluorescens (MU87), Escherichia coli (ATCC25922), Pseudomonas stutzeri (MU70), Pseudomonas aeruginosa (ATCC27853), Stenotrophomonas maltophilia (MU64), Chryseomonas luteola (MU65), and S. maltophilia (MU99). Moreover, the ethanolic extract prepared from 12 S. verbenaca exhibited lower antimicrobial activity than the methanolic extracts, as found by Kostić et al. [9].The investigation of the methanol extract from aerial parts of TunisianS. verbenaca demonstrated that the extract had a high antibacterial potential (MIC=500μg/mL) against six bacteria isolated from the mouths of patients [42]. However, a South African extract of S. verbenaca that was made with methanol and chloroform had strong antibacterial properties against Klebsiella pneumoniae, Bacillus cereus, Escherichia coli, and Staphylococcus aureus [11]. Moreover, Belkhiri et al. [21] compared the antibacterial potential of four fractions from the methanol extract of Algerian S. verbenaca: chloroform extract, crude extract, aqueous extract, and ethyl acetate extract. They have found that the antibacterial efficacy increases with the concentration of the extract. Al-Zereini [7] also found that the ethyl acetate extract prepared from the leaves of S. verbenaca from Jordan had dose-dependent antibacterial properties against Bacillus brevis (ATCC 9999) and Bacillus subtilis (ATCC 6633). On the other hand, the extract had no effect on Klebsiella pneumoniae (ATCC 13883), Staphylococcus aureus (ATCC 43300), and Escherichia coli (ATCC 25922). Canzoneri et al. [8] found that the EO of S. verbenaca aerial parts has potential antibacterial effects, and this activity is much higher against gram-positive bacteria than gram-negatives.


## 2.5.2. Antioxidant Activity

The antioxidant potential ofS. verbenaca extracts was investigated by several researchers [12, 21, 23, 38, 42–44], and Table 4 summarizes the majority of the investigations that were carried out on different parts of S. verbenaca, collected from different regions.Table 4
Antioxidant activity ofS. verbenaca.
Part usedExtractsMethods usedKey resultsRef.Not specifiedMethanolic extract (80%)Ethanolic extract (80%)DPPH assayβ-Carotene/linoleic acidsThe 80% methanol extract prepared by maceration was highly activeThe 80% of ethanol extract was the most active[41]Aerial parts (stems and leaves)Hydromethanolic extractOxygen consumptionA strong inhibition of oxygen consumption (92%)[44]Conjugated diene formation (CD)A strong inhibition of CD formation of LDL peroxidation (92%)Thiobarbituric acid reactive substance (TBARS) formationA strong inhibition of TBARS formation of linolenic acid oxidation (93%)Not specifiedMethanolic extractDPPH assayβ-Carotene–linoleic acid methodIC50=14.30±1.42μg/mgPercentinhibition=77.03±0.42%[38]Aerial partsMethanolic extractDPPH assayIC50=86.9μg/mL[42]ABTS assayIC50 at 5min=777.3μg/mLTEAC at 5min=0.624IC50 at 20min=499.5μg/mLTEAC at 10min=0.647TEAC at 20min=0.705TEAC at 15min=0.705Aerial partsMethanolic extract from postdistilled plantDPPH methodSers:IC50=24.47±1.87μg/mLTouiref: IC50=25.11±2.97μg/mLBeja: IC50=26.62±0.8μg/mLChott Meriem: IC50=28.28±0.16μg/mLTunis: IC50=30.34±2.28μg/mLRass Zebib: IC50=31.19±2.25μg/mLBou Arada: IC50=33.47±4.13μg/mLBir Mroua: IC50=34.70±2.43μg/mLHencha: IC50=39.85±3.9μg/mLEnfidha: IC50=40.91±0.5μg/mL[6]ABTS methodHencha:TEAC=120.11±6.62μM trolox/mgEnfidha: TEAC=134.45±5.27μM trolox/mgBir Mroua: TEAC=139.26±10.59μM trolox/mgRass Zebib: TEAC=144.02±3.4μM trolox/mgBou Arada: TEAC=154.97±6.79μM trolox/mgTunis: TEAC=190.51±6.71μM trolox/mgChott Meriem: TEAC=196.72±1.61μM trolox/mgSers: TEAC=271.51±4.52μM trolox/mgBeja: TEAC=282.17±6.58μM trolox/mgTouiref: TEAC=287.81±3.65μM trolox/mgFRAPBeja:142.07±1.46mM Fe+2/mgSers: 139.09±11.23mM Fe+2/mgTouiref: 131.86±1.05mM Fe+2/mgChott Meriem: 124.27±0.38mM Fe+2/mgTunis: 122.33±3.7mM Fe+2/mgBou Arada: 120.53±7.53mM Fe+2/mgRass Zebib: 118.02±15.25mM Fe+2/mgBir Mroua: 109.22±5.04mM Fe+2/mgHencha: 104.89±0.37mM Fe+2/mgEnfidha: 101.46±1.97mM Fe+2/mgAerial partsCrude extractDPPH methodIC50=47.50μg/mL[14]Aerial partsMethanolic extractDPPHFRAPIC50=9.79±0.47μg/mLHigh reducing power[43]Not specifiedMethanolic extractDPPH methodIC50=16.92±0.2μM[12]Aerial partsMethanolic extract subfractions:Crude extract (CrE)Chloroform extract (ChE)Ethyl acetate extract (EAE)Aqueous extract (AqE)DPPH methodEAE:IC50=0.0086mg/mLCrE: IC50=0.0336mg/mLChE: IC50=0.0725mg/mLAqE: IC50=0.0389mg/mL[21]Reducing power methodEAE: EC50: 0.0047 mg/mLCrE: EC50: 0.0453±0.000 mg/mLAqE: EC50: 0.0455 mg/mLMetal chelating methodAqE and CrE reported the highest activityEAE chelation did not exceed 20%Aerial partsMethanolic extracts (85%) (ME)Decoction extract (distilled water) (DE)DPPH methodME: IC50: 24.36±1.13 μg/mLDE: IC50: 27.26±1.05 μg/mL[13]ABTS methodME: IC50: 19.96±1.03 μg/mLDE: IC50: 36.86±1.03 μg/mLAlkaline DMSO superoxide radical scavengingME: IC50: 07.77±1.00 μg/mLDE: IC50: 18.78±1.07 μg/mLβ-Carotene bleachingDE: inhibition:96.12±2.48%ME: inhibition: 82.58±2.39%Reducing power methodDE: EC50: 69.52±3.07 μg/mLME: EC50: 56.64±4.81 μg/mLMetal chelating activity methodME: IC50: 70.39±1.13 μg/mLDE: IC50: 109.70±1.72 μg/mLPhenanthroline methodME: IC50: 27.03±1.54 μg/mLDE: IC50: 40.26±0.59 μg/mLCupric reducing antioxidant capacity (CUPRAC)ME: A0.50: 14.66±2.51 μg/mLDE: A0.50: 33.00±0.30 μg/mLRootMethanolic extractH2DCF-DA methodSignificant reduction in the intracellular reactive oxygen species (ROS) level for both tested values (1 and 10μg/mL)[14]Kostić et al. [41] evaluated the antioxidant potential of different S. verbenaca extracts using the beta-carotene/linoleic acid system and DPPH assay. They found that the methanol extract had the highest activity in the DPPH method, while the ethanolic extract obtained by ultrasound extraction was the most active metabolite of beta-carotene/linoleic acid. The antioxidant activity of hydromethanolic extract prepared from stems and leaves of Moroccan species was carried out by Khlifi et al. [44]. The results showed that the extract had a significant antioxidant effect at 100 μg/mL, with a strong inhibition of oxygen consumption compared to previous studies [38].The antioxidant potential of TunisianS. verbenaca extracts was also studied [42], and the results showed that methanolic extract from aerial parts had lower activity (IC50=86.9μg/mL) compared to the positive control, which was the trolox (IC50=23.12μg/mL) using the DPPH assay. In addition, it was reported that the antioxidant activity over 20 minutes using the ABTS assay increased with time, but was still four times lower than the activity of trolox. Additionally, Farhat et al. [6] studied the efficacy of the collection sites on the antioxidant capacity of methanolic extract prepared from postdistilled aerial parts of Tunisian species. They found that the site had a significant effect on the antioxidant potential by the DPPH, ABTS, and FRAP methods. Likewise, activity was shown to be substantially linked with total phenolic content.The antioxidant activity of some extracts ofS. verbenaca collected from Algeria was mostly studied using the DPPH assay. It was found that the crude extract prepared from aerial parts had good antioxidant activity that increased with increasing the extract concentration [14]. The scavenging activity was 95% at a concentration of 0.1 mg/mL. Also, the methanol extract of S. verbenaca aerial parts revealed a high reducing power in the FRAP test [43] using the DPPH assay. Additionally, it was cited that the methanol extract had a beneficial effect against free-radical damage and exhibited a 5-fold more inhibitory effect than the standard antioxidant trolox (IC50=72.63μM) [12]. They also observed that the radical scavenging activity had no significant correlation with the phenolic content and a low correlation with the flavonoid content. Belkhiri et al. [21] investigated the antioxidant potential of some fractions of the methanol extracts using the DPPH method, metal chelating activity, and reducing power assay, and all extracts showed potent antioxidant activity [13]. The cupric ion reducing capacity (CUPRAC) and Fe3+ reducing capacity (phenanthroline assay) of the extracts were investigated, and the findings exhibited that both extracts had high antioxidant capacity, with methanolic extract exhibiting the highest activity [14].


## 2.5.3. Anticancer Activity

The different organic essential oils and extracts ofS. verbenaca have been studied for anticancer properties. Numerous laboratory investigations using cell culture have shown that S. verbenaca extracts and essential oils have antiproliferative properties (Table 5) against a variety of cancer cell lines [15–19, 23, 45].Table 5
Anticancer effects ofS. verbenaca.
Plant partTested extractCell linesMajor resultsRef.LeavesEthyl acetateHuman breast adenocarcinomaIC50: 41.3±4.8 μg/mL[7]Aerial partsMethanolHuman colon adenocarcinomaLC50: 60.4 μg/mL[15]Human hepatoblastomaLC50: 68.9 μg/mLHuman breast cancer cellsLC50: 43.1 μg/mLHuman pancreatic carcinomaLC50: 42.2 μg/mLAerial partsHexaneHuman embryonal rhabdomyosarcoma cancerous cell linesIC50: 474.6±1.3 μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsEthyl acetateHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50: 223.6±1.6 μg/mLAerial partsn-ButanolHuman embryonal rhabdomyosarcoma cancerous cell linesIC50>500μg/mL[16]Vero (monkey kidney cancerous cell lines)IC50>500μg/mLAerial partsMethanol and chloroformBreast adenocarcinomaIC50: 31.5±13.7 μg/mL[17]Colon adenocarcinomaIC50: 50.0±5.3 μg/mLGlioblastomaIC50 was not calculatedHuman kidney epithelial cell lineIC50: 20.8±2.5 μg/mLLeavesMethanolMonkey kidney cellsCC50: 64 μg/mL[18]Human larynx cancer cellsCC50=64μg/mLRootsMethanolHuman monocytic leukemia cell line70% of apoptosis and 30% of viable cells at a 1000μg/mL concentration[23]Aerial partsEssential oilsHuman melanoma cell lineIC50=8.1±0.6μg/mL[19]The ethyl acetate extract ofS. verbenaca leaves produced after maceration was examined using the MDA cell lines MB-231 (human breast adenocarcinoma, ATCC HTB-26). The findings indicated that all extracts produced cytotoxicity in MDA MB-231 breast cancer cells [7]. However, it was proved that S. verbenaca leaf extracts possessed cytotoxic effect against HEp-2 (human larynx cancer cells) and Vero (monkey kidney cells) [18]. In another investigation, methanolic extracts of S. verbenaca’s aerial component prepared by maceration were evaluated in vitro against four human cancer cell lines, including HCA, HepG2, MCF-7, and HPC. The findings indicate that LC50 levels higher than 75 μg/mL were deemed inactive [15]. Additionally, MTT assays were used to determine the cytotoxic activity of several extracts (methanol, hexane, ethyl acetate, n-butanol, and chloroform extracts) obtained from the aerial portion of S. verbenaca [16]. Methanol and chloroform extracts of S. verbenaca aerial parts were evaluated against colon adenocarcinoma (HT-29), human cancer cell lines (breast adenocarcinoma (MCF-7), human kidney epithelial cell line and glioblastoma (SF-268)) [17]. S. verbenaca exhibited more favorable action against MCF-7, with an IC50 value of 31.50 13.70 μg/mL, but was inactive versus SF-268 and/or HT-29 cell lines [17].A cell viability study was performed to avoid any cytotoxic concentration ofS. verbenaca root extract on THP-1 cells. The MTT assay revealed that the most cytotoxic concentration of the extract was 1000 μg/mL, which caused 70% of cell death and 30% of cell viability [23]. The essential oils of S. verbena were investigated for their ability to suppress the proliferation of human tumor cells using the human M14 melanoma cell line and shown significant efficacy [19]. The antiproliferative effect of S. verbenaca essential oil may be attributed to active sesquiterpenes in combination with other natural chemicals found in the essential oil components. Indeed, carvacrol and thymol exhibited outstanding anticancer properties through a variety of mechanisms [19].


## 2.5.4. Antiparasitic Activity

Et-Touys et al. [20] investigated the antileishmanial effects of organic extracts (methanol, n-hexane, and dichloromethane extract) from S. verbenaca, and it was reported that the in vitro antileishmanial effect which was evaluated on the culture of three Leishmania species such as Leishmania infantum, Leishmania tropica, and Leishmania major was good (Table 6).Table 6
Antiparasitic activity ofS. verbenaca.
ActivityPart usedExtractParasiteMajor resultsRef.Antileishmanial activityWhole plant partn-HexaneLeishmania majorIC50: 155.4 μg/mL[20]Leishmania tropicaIC50: 148.2 μg/mLLeishmania infantumIC50: 14.1 μg/mLDichloromethaneLeishmania majorIC50: 24.5 μg/mLLeishmania tropicaIC50: 33.7 μg/mLLeishmania infantumIC50: 31.5 μg/mLMethanolLeishmania majorIC50>1000μg/mLLeishmania tropicaIC50: 850.7 μg/mLLeishmania infantumIC50>1000μg/mLAntimalarial activityAerial partsMethanol chloroformPlasmodium falciparum (FCR-3 strain)IC50: 23.9±1.1 μg/mL[11]Belkhiri et al. [21] additionally observed that S. verbenaca has antihemolytic properties. In vitro antihemolytic activity of S. verbenaca extract was determined by inducing oxidative erythrocyte hemolysis. The results indicated that ethyl acetate extract was the most effective in inhibiting hemolysis, followed by crude extract, chloroform extract, and aqueous extract. Additionally, ethyl acetate extract inhibited hemolysis more effectively than vitamin C.


## 2.5.5. Insecticidal Activity

In most cases, the application of synthetic pesticides is the primary approach for controlling insect pests, which produces excellent effects in a short period of time. Meanwhile, their irrational usage has resulted in global issues such as pollution, nontarget toxicity, biodiversity loss, and the development of pest resistance [46–48]. This need arose from a desire to provide alternatives to synthetic insecticides, which can have negative environmental consequences [49–52]. The insecticidal capabilities of S. verbenaca extracts and essential oils have been documented to have potential impact against several pests in previous studies [53]. Insecticidal action has been shown in several experiments using some Salvia species [54].Essential oils fromSalvia species revealed 100% repellency activity against adults of Aedes albopictus [55]. The oil of S. verbenaca drastically shortened the lifespan of cowpea weevil and prevented females from laying eggs [56]. Several crude extracts and essential oils from Salvia species were tested for pesticide activity against the test pest larvae [57–59]. Insectistatic and insecticidal properties of chloroform extracts from the aerial portions of four Salvia species were examined [60]. S. verbenaca extracts are very effective against Culex quinquefasciatus mosquitos [61]. Caryophyllene oxide was the major component in the essential oil of S. verbenaca with 7.28 [62]. The insecticidal activity and fumigant toxicity of caryophyllene oxide were tested against two insect pests, and it was shown to be effective [63].


## 2.6. Other Biological Effects

Different extracts fromS. verbenaca have also exhibited other biological activities such as antihemolytic, immunomodulatory, and enzyme inhibitory effects (Table 7).Table 7
Other biological activities ofS. verbenaca.
ActivityPart usedExtractsExperimentMajor resultsRef.AntihemolyticAerial partEthyl acetate2,2-Azobis (2-amidinopropane) dihydrochloride induces erythrocyte oxidative hemolysis (AAPH)HT50: 165 min[21]CrudeHT50: 125.1 minChloroformHT50: 111.5 minAqueousHT50: 111.5 minXanthine oxidase inhibitionAerial partChloroformColorimetric approach based on uric acid generation at 295 nm in the presence of 100 mM xanthine in phosphate bufferIC50: 0.0088±0.0 mg/mLEthyl acetateIC50: 0.0165±0.001 mg/mLCrudeIC50: 0.0520±0.003 mg/mLAqueousIC50: 0.9800±0.004 mg/mLPorcine liver carboxylesterase inhibitionAerial partAqueous methanolEnzyme inhibition by spectrophotometric assayCE (carboxylesterase) inactivation with apI=5.1 and a Ki value of 38 Mm[12]Healing of burnsLeavesHexaneSecond-degree burn injury induced by a hot metal cylinder in ratsAccelerated healing process with 44.34%[16]Ethyl acetateAccelerated healing process with 47.55%n-ButanolAccelerated healing process with 49.16%AnticholinesterasesAerial partMethanolCholinesterase inhibitionInhibition effect of AChE at 100μg/mL[13]Anti-α-amylaseα-Amylase inhibitionIC50: 01.3±0.08 μg/mLAnti-α-glucosidaseα-Glucosidase inhibitionIC50=150.5±1.4μg/mLImmunomodulatoryAerial partsMethanolPhagocytic activity used carbon clearance rate testSignificantly increased phagocytic index (0.095±0.012) at a dose of 200 mg/kgIncreased corrected phagocytic index α (0.095±1.71)[14]


### 2.6.1. Xanthine Oxidase Inhibitory Effect

Xanthine oxidase, abbreviated as XO, is an oxidoreductase that catalyzes the conversion of hypoxanthine to xanthine and xanthine to uric acid. Xanthine oxidase is generally present in the liver and in an inactive form in the blood in humans. A blood test for XO may identify liver impairment because xanthine oxidase is released into the blood in situations of severe liver injury [21].


### 2.6.2. Burn Recovery Activities

Guaouguaou et al. [24] evaluated the impact of three S. verbenaca extracts on the healing of burns in rats using hexane, ethyl acetate, and n-butanol. The results indicated that various Salvia verbenaca plant extracts were more effective than silver sulfadiazine (SSD) and that it is the most widely used topical treatment for injury, with healed areas of 29.17% (base), 44.34% (hexane), 47.55% (ethyl acetate), 49.16% (n-butanol), and 41.09% SSD.


### 2.6.3. In Vitro Antidiabetic Activity

Several earlier studies have shownS. verbenaca’s antidiabetic activity in vitro [13]. Additional studies are shown in Section 2.8 and Figure 8.Figure 8
Antidiabetic mechanism insights ofS. verbenaca main compounds.


### 2.6.4. Immunomodulatory Effects

Previous studies investigated the immunomodulatory effects ofS. verbenaca aerial parts [14]. The carbon clearance rate test was used to determine the immunostimulant potential of this plant on phagocytic activity. The phagocytic index was much higher in rats who were given S. verbenaca at a dose of 200 mg/kg than in rats who were not given the herb.


## 2.6.1. Xanthine Oxidase Inhibitory Effect

Xanthine oxidase, abbreviated as XO, is an oxidoreductase that catalyzes the conversion of hypoxanthine to xanthine and xanthine to uric acid. Xanthine oxidase is generally present in the liver and in an inactive form in the blood in humans. A blood test for XO may identify liver impairment because xanthine oxidase is released into the blood in situations of severe liver injury [21].


## 2.6.2. Burn Recovery Activities

Guaouguaou et al. [24] evaluated the impact of three S. verbenaca extracts on the healing of burns in rats using hexane, ethyl acetate, and n-butanol. The results indicated that various Salvia verbenaca plant extracts were more effective than silver sulfadiazine (SSD) and that it is the most widely used topical treatment for injury, with healed areas of 29.17% (base), 44.34% (hexane), 47.55% (ethyl acetate), 49.16% (n-butanol), and 41.09% SSD.


## 2.6.3. In Vitro Antidiabetic Activity

Several earlier studies have shownS. verbenaca’s antidiabetic activity in vitro [13]. Additional studies are shown in Section 2.8 and Figure 8.Figure 8
Antidiabetic mechanism insights ofS. verbenaca main compounds.


## 2.6.4. Immunomodulatory Effects

Previous studies investigated the immunomodulatory effects ofS. verbenaca aerial parts [14]. The carbon clearance rate test was used to determine the immunostimulant potential of this plant on phagocytic activity. The phagocytic index was much higher in rats who were given S. verbenaca at a dose of 200 mg/kg than in rats who were not given the herb.


## 2.7. Toxicological Investigations ofS. verbenaca

The toxicological investigations ofS. verbenaca have not been well studied. However, some studies carried out recently have confirmed the safety of these plant extracts (Table 8). Indeed, a report by Guaouguaou et al. [64] focused on the acute and subchronic effects of S. verbenaca toxicity in mice and rats through oral and topical administration. The findings of the acute toxicity of the fractions derived from S. verbenaca (n-butanol, hexane, and ethyl acetate) demonstrated that the LD50 of this plant after oral administration at 2000 mg kg-1 is not deadly [64]. In order to complete the toxicity profile of this plant, more research should be done to find out how toxic it is over a long period of time.Table 8
Toxicological studies ofS. verbenaca.
ActivitiesPart usedExtractExperimental approachMajor resultsRef.Acute oral toxicityAerial partsn-Butanol, hexane, ethyl acetateOrally delivered at a dose of 2000 mg/kg in a volume of 0.25 mL per 20 g of body weight to mice and examined for 14 daysLD50>2000mg/kg body weight[64]Acute dermal toxicityAerial partsHexane, ethyl acetate,n-butanolFor 14 days, daily topical application ofS. verbenaca extracts at a dose of 2000 mg/kg body weightThere are no adverse effects, behavioral problems, or fatalities[64]Subchronic dermal toxicityAerial partsHexane, ethyl acetate,n-butanolFor 28 days, daily topical application ofS. verbenaca extracts at a dose of 2000 mg/kg body weightThere are no harmful symptoms or changes in the amount of water or food consumedThere is no lethalityThere is no change in the parameters of fasting blood circulationThere were no morphological alterations in the main vital organs investigated[64]


## 2.8. Pharmacological Properties ofS. verbenaca Main Volatile Compounds

Several studies examined the major volatile chemicals found inS. verbenaca, including carvacrol, thymol, and linalool. Studies showed that carvacrol has hypoglycemic properties through intrinsic mechanisms such as blood glucose and insulin level lowering [65]. Additionally, carvacrol resulted in a drop in glucose levels. Additionally, these substances were shown to enhance the activity of glucokinase and glucose-6-phosphate dehydrogenase in the liver [66]. Carvacrol inhibits the enzymes α-amylase and alpha-glucosidase in vitro [67] and beta-galactosidase in vitro [68]. Thymol was also able to normalize blood sugar, plasma insulin, HbA1c, and the insulin resistance index in patients with hyperglycemia [69]. The levels of expression of genes involved in the production of insulin have been studied and reported in STZ-induced diabetic mice [70, 71], and a rise in Mafa and Pdx1 gene expression has been reported. Limonene is another major constituent of S. verbenaca that has been shown to improve glucose homeostasis. Indeed, this substance boosts hepatic glycogen and plasma glucose levels [72] (Figure 8).The antidiabetic effect has been also revealed by linalool (another main compound ofS. verbenaca) [73, 74]. Indeed, linalool lowered blood glucose, hemoglobin A1c, fructosamine, interleukin-6, and tumor necrosis factor-α (TNF-α), while it increased insulin levels [74].The major phytochemical compounds ofS. verbenaca exhibited remarkable antibacterial effects [75–77]. Rhayour et al. [70] investigated the impact of thymol on gram-positive and gram-negative microorganisms, including Bacillus subtilis and Escherichia coli [75–78]. Antibacterial activity is demonstrated by modifying cell shape, damaging cell walls and membranes, and limiting the development of some types of bacteria, including P. aeruginosa [79]. In addition, limonene was found to be antibacterial because it targeted microorganisms’ cytoplasmic membranes, weakened membrane integrity, blocked respiratory enzymes, and lost the proton motive force (Figure 9).Figure 9
Suggested antibacterial mechanisms ofS. verbenaca compounds.The anticancer properties of the major components inS. verbenaca (carvacrol, limonene, and thymol) have also been reported recently [80–82]. Thymol has been shown to have anticancer properties via a variety of mechanisms, including inducing severe DNA damage, including the production of reactive oxygen species (ROS) and subsequent increase in oxidative stress and/or mitochondrial dysfunction, or via the nuclear factor of activated T cell (NFAT-2) route [81]. Additionally, carvacrol increased apoptosis in cells, perhaps via activating mitochondrial apoptotic and signaling pathways [83].


## 3. Conclusions and Perspectives

S. verbenaca, a medicinal plant used in traditional medicine to cure a variety of ailments, was found to be abundant in bioactive chemicals such as flavonoids, terpenoids, and phenolic acids. Numerous pharmacological studies have demonstrated that S. verbenaca extracts and essential oils have extraordinarily beneficial effects on a variety of diseases, including those caused by microbes and those caused by dysregulation of homeostasis. Indeed, this plant demonstrated antibacterial, antidiabetic, anticancer, and immunomodulatory properties via a variety of mechanisms. However, further research should be conducted to find other pharmacodynamic targets. Additionally, pharmacokinetic studies should be conducted to ascertain the absorption, metabolism, and elimination of S. verbenaca bioactive components. Additionally, toxicological studies should be conducted to validate the safety of S. verbenaca extracts at various doses and delivery methods.



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