Variation in Phenolic Content, Antioxidant Activity and Alpha-amylase and Acetylcholinesterase Inhibitory Capacities of Different Extracts from Tunisian Satureja barceloi (Willk) L.

The antioxidant activity and the anti-α-amylase and anti-acetylcholinesterase capacities of secondary metabolites from different organs (roots, stems, leaves and flowers) of Tunisian Satureja barceloi were determined. The variation in the distribution of phenolic metabolites among roots, stems, leaves and flowers extracts of S. barceloi with various solvent systems (methanol, ethyl acetate, hexane and distilled water) has not been characterized before. Significant variation of phenolic compounds was observed according to organs rather than to extracting solvents. The analyzed organs show a high level of phenolic compounds although the stems contains the highest total polyphenols (132.53±0.48 mg AGE/g Ex), flavonoids (48.99±0.65 mg RE/g Ex) and flavonols (34.93±0.29 mg QE/g Ex) contents. The phenolic fraction was dominated by sagerinic acid, caffeic acid glucoside and epigallocatechin, detected using HPLC-PDA/ESI-MS. The antioxidant activity of all extracts, evaluated by four in vitro tests, was high and varied significantly according to the type of solvent used and the plant organ. The aqueous extracts of leaves exhibited the greatest inhibitory effect on alpha-amylase while the methanolic extract of leaves and stems revealed the most important acetylcholinesterase inhibitory effect. Hence, S. barceloi extracts could be used as a source of various bioactive molecules in pharmaceutical industry.

Deverra triradiata Hochst. ex Boiss. from the Northern Region of Saudi Arabia: Essential Oil Profiling, Plant Extracts and Biological Activities.

Devrra triradiata Hochst. ex Boiss is an occasional plant species in the Northern region of Saudi Arabia. The shrub is favored on sandy desert wadis, gypsaceous substrate, and sandy gravel desert. In folk medicine, the plant is used for many purposes; to relieve stomach pains, against intestinal parasites, and for the regulation of menstruation. The present study describes the chemical composition of the essential oils (EOs) of different plant parts of D. triradiata. In vivo and in vitro biological activities of plant extracts and essential oils were also studied. Phenylpropanoids, elemicin (flowers: 100%), dillapiole (Stems: 82.33%; and seeds: 82.61%), and apiol (roots: 72.16%) were identified as the major compounds. The highest antioxidant activity was recorded for the EOs of roots and stems (IC50 = 0.282 µg/mL and 0.706 µg/mL, respectively). For plant extracts, ethyl acetate showed the highest antioxidant activities (IC50 = 2.47 and 3.18 µg/mL). EOs showed high antifungal activity against yeasts with low azole susceptibilities (i.e., Malassezia spp. and Candida krusei). The MIC values of EOs ranged between 3.4 mg/mL and 56.4 mg/mL. The obtained results also showed phytotoxic potential of plant extracts both on the germination features of Triticum aestivum seeds and the vegetative growth of seedlings.

Allium sativum mitigates oxidative damages induced by Microcystin-LR in heart and liver tissues of mice.

Microcystins (MCs) are hepatotoxic cyanotoxins implicated in several incidents of human and animal toxicity. Microcystin-(Lysine, Arginine) or MC-LR is the most toxic and encountered variant of MCs where oxidative stress plays a key role in its toxicity. This study investigated the oxidative damages induced in the liver and heart of Balb/C mice by an intraperitoneal injected acute dose of MC-LR. Thereafter, the potential protective effect of garlic (Allium sativum) extract supplementation against such damages was assessed through the evaluation of oxidative stress and cytotoxicity markers. Lipid peroxidation (LPO), carbonyl content (CC), glutathione content (GSH), alkaline phosphatase activity (ALP), lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH) activities were measured. Results showed important oxidative damages in hepatic and cardiac cells of mice injected with the toxin. However, these damages have been significantly reduced in mice supplemented with garlic extract. Thus, this study demonstrated for the first time the effective use of garlic as an antioxidant agent against oxidative damages induced by MC-LR. As well, this study supports the use of garlic as a potential remedy against pathologies related to toxic agents.

Ethnopharmacological study of natural products used for traditional cancer therapy in Algeria.

Traditional and complementary medicine constitutes an important, and often underestimated, source of healthcare for multiple diseases including cancer. However, little is known about the ethnomedical knowledge and practices in Northern Africa. The main objective of this study is to identify and analyze the variety of natural products used in Algerian ethnopharmacology for cancer therapy. For this purpose, semi-structured interviews with 225 traditional healers, herbalists and practitioners were realized in twelve locations in Algeria throughout field studies performed from June 2015 to July 2019. Interviews covered popular and vernacular names of the natural product, mode of use and administration, dose, period of treatment, toxicity and side effects among other data. The obtained results reveal the use of 113 medicinal plants (belonging to 53 families and 104 genera), 10 animal species and various products and by-products from different origins such as honey, olive oil, thorns, urine, milk, animal fat and the alkaline water of Zamzam. Basing on the frequency of citation (FC), use reports (UR) and use value (UV), the most used natural products for cancer treatment are honey (FC = 181, UR = 194, UV = 0.65), Nigella sativa L. (FC = 131, UR = 152, UV = 0.54), Aristolochia longa L. (FC = 118, UR = 144, UV = 0.51), Berberis vulgaris L. (FC = 111, UR = 142, UV = 0.51), Curcuma longa L. (FC = 107, UR = 121, UV = 0.43), Trigonella foenum-graecum L. (FC = 102; UR = 119, UV = 0.43), Citrus limon (L.) Obseck (FC = 97, UR = 120, UV = 0.43), Artemisia herba-alba Asso (FC = 92, UR = 115, UV = 0.41) and the holy water 'Zamzam' (FC = 110, UR = 110, UV = 0.43) respectively. Mixtures of two or more ingredients were frequently used. The use of Pelophylax saharicus skin' was reported for the first time for the treatment of visible tumors and skin cancer. This is the first study documenting the traditional uses of various natural products for cancer treatment in Algeria. Our findings are relevant to document the traditional uses of numerous natural products and to provide background basis to search for novel compounds for cancer therapy.

Euphorbia honey and garlic: Biological activity and burn wound recovery.

Currently, chronic wounds and microbial resistance to antibiotics have led to search new healing agents. Combinations of natural products are widely practiced in traditional medicine and exhibited synergistic activity with increased efficacy in treating several pathologies. This study assays the antioxidant, synergistic antimicrobial and burn wound healing activities of Euphorbia honey and Allium sativum (garlic). The minimal inhibitory concentration (MIC) of each natural product was determined against microorganisms commonly found in wound infections. The synergistic antimicrobial effect was assessed by mixing different concentrations of honey and garlic extract below their relative MICs. Subsequently, the antioxidant activity, total phenolic (TPC) and flavonoid (TFC) contents of both natural products and a selected mixture of them were evaluated. Efficacy of that mixture was also evaluated as topical application on male and female Wistar rats skin burn wound, compared to Euphorbia honey and two conventional treatments. Results showed that the mixture honey-A. sativum has synergistic antimicrobial effect against all tested strains. Besides, A. sativum presented higher antioxidant activity along with higher TPC and TFC compared to honey and their mixture. However, the mixture showed higher wound healing activity reflected by shorter epithelialization and wound contraction time, as well as, better histological recovery of the treated tissues. Our results also showed that burn wound healing is not affected by gender. Our findings support the idea of combining natural products as an effective therapy.

Polyphenolic Profiling, Quantitative Assessment and Biological Activities of Tunisian Native Mentha rotundifolia (L.) Huds.

Phenolic profiling of ten plant samples of Mentha rotundifolia (L.) Huds. collected from different bioclimatic areas of Tunisia, was for the first time carried out by using a fast ultra-high-performance liquid chromatography (UHPLC)-high resolution tandem mass spectrometry (HRMS/MS) method on a Q Exactive platform equipped with an electrospray ionization (ESI) source. An intraspecific, interpopulation variability was evidenced and a total of 17 polyphenolic metabolites were identified and quantified by using the UHPLC-HRESIMS/MS method, here validated for specificity, linearity, limit of detection, limit of quantitation, accuracy and precision. The quantitative method resulted sensitive at the nM level and reliable for rapid polyphenol quantification in vegetal matrices. The metabolomic study allowed us to identify a new compound, named salvianolic acid W, which was isolated and characterized mainly by NMR and MS analysis. A statistical correlation of the phenolic composition with antioxidant and anti-acetylcholinesterase activities was provided.

Phytochemical profile and biological activities of Deverra tortuosa (Desf.)DC.: a desert aromatic shrub widespread in Northern Region of Saudi Arabia.

The present study describes the chemical composition of the essential oil of different plant parts of Devrra tortuosa; in vivo and in vitro biological activities of plant extract and essential oils. Apiol was found to be the major component of the oil (between 65.73% and 74.41%). The best antioxidant activities were observed for the oil of flowers (IC50 = 175 µg/ml). The samples of stems and roots exhibit lower antioxidant activity (IC50 = 201 µg/ml and 182 µg/ml, respectively). The values of IC50 showed that the extracts of methanol exhibit the highest antioxidants activities (IC50 = 64.8 102 µg/ml). EOs showed excellent antifungal activity against yeasts with low azole susceptibilities (i.e. Malassezia spp. and Candida krusei). The MIC values of oils varied between 2.85 mg/mL and 27 mg/mL. The obtained results also showed that the plant extracts inhibited the germination and the shoot and root growth of Triticum æstivum seedlings.

Relationship between chemotypic and genetic diversity of natural populations of Artemisia herba-alba Asso growing wild in Tunisia.

A total of 80 individuals collected from eight populations growing wild in different geographic zones were considered to assess the intraspecific variability of essential oil composition, genetic diversity and population structure of Artemisia herba-alba. The essential oil composition varied significantly between populations. Essential oil profiles were classified into four chemotypes (trans-sabinyl acetate, α-thujone/trans-sabinyl acetate, camphor and α-thujone/camphor/ß-thujone). Despite significant correlation between the amount of some essential oil compounds and a set of climatic data, the global chemical divergence among populations was not related to their bioclimatic and geographic appurtenances. A high level of genetic diversity within populations was revealed either with RAPD and ISSR markers (Na = 1.67, PPL = 66.5%, H = 0.26, I = 0.38 and Na = 1.7, PPL = 69.8%, H = 0.26, I = 0.38, respectively). The level of genetic diversity varied across populations and chemotypes. Populations from the α-thujone/trans-sabinyl acetate chemotype exhibited the highest genetic diversity as revealed by the RAPD markers. However, populations from α-thujone/camphor/ß-thujone chemotype showed the important genetic variation determined by ISSR markers. A significant genetic differentiation among populations and among chemotypes was detected. The combined analysis showed a significant correlation (r = 0.484, p = .032) between the chemical and molecular markers. The PCA, performed on percentages of major oil compounds and the frequencies of polymorphic RAPD and ISSR bands, divided populations according to their chemotypic classification. Taking into consideration the current situation of A. herba-alba populations and their endangered habitats, these results are of value in order to ensure the in-situ and ex-situ conservation of this medicinal species.

Essential Oil Variability in Natural Populations of Artemisia campestris (L.) and Artemisia herba-alba (Asso) and Incidence on Antiacetylcholinesterase and Antioxidant Activities.

The intraspecific variability of Artemisia herba-alba and A. campestris essential oils and the evaluation of their antioxidant and antiacetylcholinesterase activities were determined. Artemisia herba-alba essential oil was found rich in camphor (19.61%), α-thujone (19.40%), ß-thujone (9.44%), chrysanthenone (9.26%), and trans-sabinyl acetate (8.43%). The major compounds of A. campestris essential oil were germacrene D (16.38%), ß-pinene (16.33%), and limonene (9.17%). Significant variation in the essential oil composition was observed among populations of each species. The divergence between populations was attributed to the variation of some climatic factors such as altitude, annual rainfall, winter cold stress, summer precipitation, summer drought stress, evapotranspiration, and humidity. Artemisia herba-alba and A. campestris essential oils exhibited promising antioxidant and antiacetylcholinesterase activities. The level of activity varied significantly according to the species and the essential oil. The highest scavenging activity (IC50 = 0.14 mg/ml) and the uppermost capacity to prevent ß-carotene bleaching (IC50 = 0.10 mg/ml) characterized A. campestris from population 6. A. campestris population 3 possessed the uppermost ability to reduce ferric ions (450.7 µmol Fe2+ /g EO). The population 2 of A. campestris showed the strongest antiacetylcholinesterase activity (IC50 = 0.02 mg/ml). The variation of these activities between the essential oils was explained by their composition differences.

Bioactive compounds from Hypericum humifusum and Hypericum perfoliatum: inhibition potential of polyphenols with acetylcholinesterase and key enzymes linked to type-2 diabetes.

CONTEXT: Natural products are reported to have a wide spectrum of pharmacological properties such as antimicrobial, anti-inflammatory and anti-cholinesterase. The genus Hypericum (Hypericaceae) is a source of a variety of molecules with different biological activities, notably hypericin and various phenolics. OBJECTIVES: The goals of the present work were the determination of total phenolic and flavonoid content, hypericin and hyperforin concentration as well as the evaluation of biological of Hypericum humifusum L. (Hhu) and Hypericum perfoliatum L. (Hper). MATERIALS AND METHODS: The various extracts of aerial parts were powdered, and then extracted with methanol. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) methods against four Gram-positive bacteria, four Gram-negative bacteria and yeast. RESULTS: The results revealed that H. humifusum, bear the highest total phenolic and flavonoid content (48-113 mg GAE/g and 8-41 mg RE/g, respectively) as well as hypericin (60-90 mg/g) and hyperforin (8-30 mg/g) concentration. Both species showed significant antioxidant activity as revealed by DPPH, FRAP, ABTS, and metal chelating assays. H. humifusum exhibited a strong acetylcholinesterase (3.86-4.57 mg GALAEs/g), α-glucosidase (0.73-2.55 mmol ACEs/g) and α-amylase (3-8 mmol ACEs/g) inhibitory activity. The extract of H. humifusum exhibited strong antibacterial activity mainly against Staphylococcus epidermidis, Staphylococus aureus, and Enterococcus faecium (MIC values ranging from 200 to 250 µg/mL). The highest antifungal activity was showed for H. perfoliatum extract (MIC value = 250 µg/mL). CONCLUSION: The data suggest that H. humifusum could be used as valuable new natural agents with functional properties for pharmacology industries.

Asteraceae Artemisia campestris and Artemisia herba-alba Essential Oils Trigger Apoptosis and Cell Cycle Arrest in Leishmania infantum Promastigotes.

We report the chemical composition and anti-Leishmania and antioxidant activity of Artemisia campestris L. and Artemisia herba-alba Asso. essential oils (EOs). Our results showed that these extracts exhibit different antioxidant activities according to the used assay. The radical scavenging effects determined by DPPH assay were of IC50 = 3.3 mg/mL and IC50 = 9.1 mg/mL for Artemisia campestris and Artemisia herba-alba essential oils, respectively. However, antioxidant effects of both essential oils, determined by ferric-reducing antioxidant power (FRAP) assay, were in the same range (2.3 and 2.97 mg eq EDTA/g EO, resp.), while the Artemisia herba-alba essential oil showed highest chelating activity of Fe2+ ions (27.48 mM Fe2+). Interestingly, we showed that both EOs possess dose-dependent activity against Leishmania infantum promastigotes with IC50 values of 68 µg/mL and 44 µg/mL for A. herba-alba and A. campestris, respectively. We reported, for the first time, that antileishmanial activity of both EOs was mediated by cell apoptosis induction and cell cycle arrest at the sub-G0/G1 phase. All our results showed that EOs from A. herba-alba and A. campestris plants are promising candidates as anti-Leishmania medicinal products.

Bioactive compounds contents, antioxidant and antimicrobial activities during ripening of Prunus persica L. varieties from the North West of Tunisia.

Bioactive molecules from fruits of four varieties of Prunus persica at different stages of ripening (green, small orange, red) were studied. For example, contents on polyphenols (20.36mg GAE/g FW) and flavonoids (0.764mg RE/g FW) were high and varied according variety. The antioxidant activity, using four different tests (DPPH radical scavenging activity, reducing power, ß carotene bleaching system and TBARS assay) showed that the variety Chatos exhibited the highest antioxidant activity comparing with others varieties. The antibacterial activity of Prunus persica varieties studied seems to be more sensitive against Staphylococcus aureus and Listeria monocytogenes. The capacity of peach DMSO extracts to inhibit Candida albicans growth was more pronounced, especially, in the presence of Chatos DMSO extract. Enzymes inhibition gives results which correlate with polyphenols, flavonoids and condensed tannins contents, and so, confirm the fascinating bioactivity of this fruit.

Essential oil composition and antibacterial activity of Origanum vulgare subsp. glandulosum Desf. at different phenological stages.

Variation in the quantity and quality of the essential oil (EO) of wild population of Origanum vulgare at different phenological stages, including vegetative, late vegetative, and flowering set, is reported. The oils of air-dried samples were obtained by hydrodistillation. The yield of oils (w/w%) at different stages were in the order of late vegetative (2.0%), early vegetative (1.7%), and flowering (0.6%) set. The oils were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). In total, 36, 33, and 16 components were identified and quantified in vegetative, late vegetative, and flowering set, representing 94.47%, 95.91%, and 99.62% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follows: carvacrol (61.08-83.37%), p-cymene (3.02-9.87%), and γ-terpinene (4.13-6.34%). Antibacterial activity of the oils was tested against three Gram-positive and two Gram-negative bacteria by the disc diffusion method and determining their diameter of inhibition and the minimum inhibitory concentration (MIC) values. The inhibition zones and MIC values for bacterial strains, which were sensitive to the EO of O. vulgare subsp. glandulosum, were in the range of 9-36 mm and 125-600 µg/mL, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis was the most sensitive and resistant strain, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems.

Myrtus communis L. infusions: the effect of infusion time on phytochemical composition, antioxidant, and antimicrobial activities.

In traditional medicine, myrtle (Myrtus communis L.) is frequently consumed as an infusion and decoction. In this study, we investigate the phenolic and volatile compositions and antioxidant and antibacterial activities of leaf infusions prepared during 3 different times. The total phenolics contents (146.74 to 179.55 mg GAE/g DM) varied significantly between infusions. Eleven phenolic compounds were identified by reversed-phase high-performance liquid chromatography. Phenolic acids (7.64 to 14.28 µmol/g DM) and flavonol glycosides (7.05 to 12.11 µmol/g DM) were the major phenolic fractions of infusions. Significant quantitative variation in 6 phenolic components was observed between infusions. Sixteen volatile components were identified by gas chromatography (GC) and GC mass spectrometry analyses. The main constituents were 1,8-cineole (42.58% to 51.39%), α-terpineol (9.45% to 9.72%), methyl eugenol (6.69% to 7.11%), and linalool (5.91% to 6.06%). Quantitative variations of the volatile components of the analyzed oils in relation to the infusion time were observed. The antioxidant properties of infusions, assayed through DPPH (2,2- diphenyl-1-picrylhydrazyl) method, ß-carotene bleaching test, chelating effect on ferrous ions, and ferric reducing power method, were considerable and varied according to the infusion time. Myrtle infusions exhibited a substantial antimicrobial activity against 6 tested bacteria.

Myrtus communis berry color morphs: a comparative analysis of essential oils, fatty acids, phenolic compounds, and antioxidant activities.

Extracts of mature dark blue and white berries from two Tunisian Myrtus communis morphs growing at the same site were assessed for their essential-oil and fatty-acid compositions, phenolic contents, and antioxidant activities. The GC and GC/MS analyses of the essential oils allowed the identification of 33 constituents. The oils from the dark blue fruits showed high percentages of α-pinene (11.1%), linalool (11.6%), α-terpineol (15.7%), methyl eugenol (6.2%), and geraniol (3.7%). Myrtenyl acetate (20.3%) was found to be the major compound in the oils from white berries. GC Analysis of the pericarp and seed fatty acids showed that the polyunsaturated fatty acids constituted the major fraction (54.3-78.1%). The highest percentages of linoleic acid (78.0%) and oleic acid (20.0%) were observed in the seeds and the pericarps of the white fruits, respectively. The total phenol, flavonoid, and flavonol contents and the concentration of the eight anthocyanins, identified by HPLC analysis, were significantly higher in the dark blue fruits. All extracts showed a substantial antioxidant activity, assessed by the free radical-scavenging activity and the ferric reducing power, with the dark blue fruit extracts being more effective.

Essential oils composition in two Rosmarinus officinalis L. varieties and incidence for antimicrobial and antioxidant activities.

The essential oil composition of Rosmarinus officinalis var. typicus and var. troglodytorum endemic to Tunisia, and growing wild in different bioclimates, was determined by GC and GC-MS. Oils were assessed for their antimicrobial and antioxidant activity. A variation of the chemical composition attributed to varieties rather than to bioclimates was revealed. 1.8-Cineole (47.2-27.5%) and camphor (12.9-27.9%) were identified as the main constituents of var. typicus and var. troglodytorum, respectively. The principal component analysis performed on oil constituents for all the populations allowed the distinction of two distinct population groups in accordance to the varietal subdivision. Based on the determination of the diameter of inhibition and the determination of the minimum inhibitory concentration, a low to moderate antimicrobial activity according to oils was revealed against eight bacteria tested. However, oils from var. troglodytorum showed higher bactericidal effect than those from var. typicus. The oils' antioxidant activity, determined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and ß-carotene bleaching test, was relatively high. The highest activity was found in oils from var. troglodytorum and in one population of var. typicus from the upper semi-arid bioclimate.

Variation of the chemical composition of essential oils in Tunisian populations of Thymus algeriensis Boiss. et Reut. (Lamiaceae) and implication for conservation.

The variation of the essential-oil composition among 14 Tunisian natural populations of Thymus algeriensis Boiss. et Reut. (=Thymus hirtus Willd. ssp. algeriensis Boiss. et Reut.) was assessed by GC (RI) and GC/MS. The populations were collected from different geographical regions belonging to the sub-humid, upper semi-arid, mean semi-arid, lower semi-arid, and upper-arid bioclimates. A total of 47 constituents, representing 81.0 to 96.5% of the total oil, were identified. The main volatiles at the species level were 1,8-cineole (17.7%), alpha-pinene (15.5%), and camphor (8.2%). A high variation among populations for the majority of the compounds was shown. Camphor (0.2-14.0%), linalool (0.2-22.4%), borneol (<0.01-24.3%), caryophyllene oxide (<0.01-18.8%), thymol (<0.01-54.9%), gamma-terpinene (0.4-6.5%), alpha-copaene (0.4-7.6%), linalyl acetate (<0.01-6.4%), and methyl eugenol (<0.01-6.9%) were the main constituents differentiating the populations. The chemical differentiation among populations, assessed by principal component analysis (PCA) and a UPGMA (unweighted pair-group method with averaging) cluster analysis performed on all populations and compounds, was high. Six chemotypes according to the main compounds have been distinguished, i.e., caryophyllene oxide/1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene/camphor, borneol/1,8-cineole/alpha-pinene, linalool, and thymol chemotypes. The spatial chemotype distribution was linked to the geographic distance among populations rather than to bioclimates, indicating that local selective environmental factors act on the chemotype diversity. The high chemical variation among populations according to their geographical and bioclimatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. The in situ and ex situ conservation strategies should concern all populations representing the different chemotypes.

Essential oil variation among natural populations of Lavandula multifida L. (Lamiaceae).

Volatiles from twelve wild Tunisian populations of Lavandula multifida L. growing in different bioclimatic zones were assessed by GC (RI) and GC/MS. Thirty-six constituents, representing 83.48% of the total oil were identified. The major components at the species level were carvacrol (31.81%), beta-bisabolene (14.89%), and acrylic acid dodecyl ester (11.43%). These volatiles, together with alpha-pinene, were also the main compounds discriminating the populations. According to these dominant compounds, one chemotype was revealed, a carvacrol/beta-bisabolene/acrylic acid dodecyl ester chemotype. However, a significant variation among the populations was observed for the majority of the constituents. A high chemical-population structure, estimated both by principal component analysis (PCA) and unweighted pair group method with averaging (UPGMA) cluster analysis based on Euclidean distances, was observed. Both methods allowed separation of the populations in three groups defined rather by minor than by major compounds. The population groups were not strictly concordant with their bioclimatic or geographic location. Conservation strategies should concern all populations, because of their low size and their high level of destruction. Populations exhibiting particular compounds other than the major ones should be protected first.