Phytochemical analysis of Clinopodium candidissimum (Munby) Kuntze growing in Algeria by an integrated HS-SPME-GC-MS, NMR and HPLC-DAD-MSn approach: valorisation of an endemic natural source of bioactive compounds.

Clinopodium candidissimum (Munby) Kuntze (Lamiaceae) is used in traditional medicine and as a food condiment in Algeria, where it is known as Zaater cheleuh and Nabta elbida. Here, we report the comprehensive characterisation of non-volatile polar constituents extracted from C. candidissimum aerial parts (a mixture of inflorescences, stems and leaves), and their aroma profile. Qualitative 1H-NMR and quali-quantitative HPLC-MSn analyses of fractions obtained with solvents at different polarity revealed the presence of aglyconic and glycosylated flavonoids (3.1%), phenylpropanoids (3.6%), gallic acid derivatives (0.76%), and triterpenoids (0.62%), among the others. On the other hand, HS-SPME-GC-MS allowed to identify 38 volatile constituents, among which the oxygenated monoterpenes pulegone (44.8%), piperitenone (6.6%), isopulegone (5.8%) and neo-menthol (3.8%), and the sesquiterpene hydrocarbons germacrene D (16.2%) and bicyclogermacrene (3.0%) were the most abundant. Overall, results indicate that C. candidissimum represents an endemic natural source of antioxidants and bioactive compounds, and they will be useful for further studies on this species.

In Vitro Antibacterial, Antioxidant, Anticholinesterase, and Antidiabetic Activities and Chemical Composition of Salvia balansae.

CONTEXT: Salvia balansae de Noé (S. balansae) (Lamiaceae) is known to be an important plant used in folk medicine as an herbal remedy in Algeria. OBJECTIVE: The purpose of the present study was to demonstrate the phytochemical composition, antioxidant activities, enzyme inhibitory activities, and antimicrobial activities of S. balansae extracts. MATERIALS AND METHODS: A methanolic extract and a petroleum ether extract from the aerial parts of the plant were assessed for their chemical composition. HPLC-MS and HPLC-DAD assessed the content of phenols, GC-MS the fatty acid composition, and ICP-MS the mineral profiles of the plant. Additionally, we evaluated the bioactivities of S. balansae extracts by the DPPH, ABTS, and CUPRAC assays, including the antioxidant potential against AChE, BChE, α-amylase, and α-glucosidase for enzyme inhibition. The antibacterial and antifungal activities of the methanolic extract were determined by the disc diffusion test against several strains of bacteria and yeasts. RESULTS: Our findings revealed that the aerial parts of S. balansae were rich in phytochemical components and contained large amounts of minerals. Quantitative analysis of phenolic compounds by HPLC-DAD revealed the presence of 12 compounds in three major classes, flavonoids, hydroxycinnamic acid, and phenolic acid derivatives, with 0.61, 0.45, and 0.29 mg/g of extract, respectively. Nine phenolic constituents were quantified by HPLC-MS analysis; catechin (72.5%) was the main compound, followed by myricetin (21.7%). The fatty acid composition of the S. balansae petroleum ether extract by GC-MS analysis was quantified. Seventeen compounds, including palmitic acid, were identified as the major fatty acids. The antioxidant activity of the S. balansae extracts was measured by three different methods: the methanol extract provided better results than the petroleum ether extract, and interesting values were noted for the DPPH, ABTS, and CUPRAC assays of 242.7 ± 7.44, 124.1 ± 9.70, and 222.9 ± 6.05 µg/mL, respectively. The enzyme inhibition activity of the plant could not be determined. The antimicrobial results of the methanolic extract obtained from the disc diffusion method, followed by measurements of MIC, MBC, and MFC against several bacteria and yeasts, indicated that S. balansae exhibited noticeable antimicrobial and antifungal activities. CONCLUSIONS: These results provided new data about the main phenolic compounds and biological activities of extracts of the aerial parts of S. balansae, which might be an alternative source for synthetic bioactive compounds.

Phytochemicals, Antioxidant and Antimicrobial Potentials and LC-MS Analysis of Centaurea parviflora Desf. Extracts.

Centaurea parviflora (C. parviflora), belonging to the family Asteraceae, is an Algerian medicinal plant used in folk medicine to treat different diseases related to hyperglycemic and inflammatory disorders, as well as in food. The present study aimed to assess the total phenolic content, in vitro antioxidant and antimicrobial activity and phytochemical profile of the extracts of C. parviflora. The extraction of phenolic compounds from aerial parts was conducted using solvents of increasing polarity starting from methanol, resulting in crude extract (CE), to chloroform extract (CHE), ethyl acetate extract (EAE) and butanol extract (BUE). The total phenolic, flavonoid and flavonol contents of the extracts were determined using the Folin-Ciocalteu and AlCl3 methods, respectively. The antioxidant activity was measured with seven methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, galvinoxyl free-radical-scavenging test, 2,2'-Azino-Bis(3-Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS) assay, cupric reducing antioxidant capacity (CUPRAC), reducing power, Fe+2-phenanthroline reduction assay and superoxide-scavenging test. The disc-diffusion method aimed at testing the sensitivity of bacterial strains toward our extracts. A qualitative analysis with thin-layer chromatography of the methanolic extract was performed. Moreover, HPLC-DAD-MS was used to establish the phytochemical profile of the BUE. The BUE was found to contain high amounts of total phenolics (175.27 ± 2.79 µg GAE/mg E), flavonoids (59.89 ± 0.91 µg QE/mg E) and flavonols (47.30 ± 0.51 µg RE/mg E). Using TLC, different components such as flavonoids and polyphenols were noted. The highest radical-scavenging ability was recorded for the BUE against DPPH (IC50 = 59.38 ± 0.72 µg/mL), galvinoxyl (IC50 = 36.25 ± 0.42 µg/mL), ABTS (IC50 = 49.52 ± 1.54 µg/mL) and superoxide (IC50 = 13.61 ± 0.38 µg/mL). The BUE had the best reducing power according to the CUPRAC (A0.5 = 71.80 ± 1.22 µg/mL), phenanthroline test (A0.5 = 20.29 ± 1.16 µg/mL) and FRAP (A0.5 = 119.17 ± 0.29 µg/mL). The LC-MS analysis of BUE allowed us to identify eight compounds including six phenolic acids and two flavonoids: quinic acid, five chlorogenic acid derivatives, rutin and quercetin 3-o-glucoside. This preliminary investigation revealed that the extracts of C. parviflora have a good biopharmaceutical activity. The BUE possesses an interesting potential for pharmaceutical/nutraceutical applications.

Ethnobotanical investigation of Pistacia lentiscus L. grown in El Kala (Algeria), and phytochemical study and antioxidant activity of its essential oil and extracts.

Ethnobotanical investigation was carried out using questionnaires among majority of El Kala (Algeria) inhabitants. The investigation found that lentisk (Pistacia lentiscus L.) is widely used in traditional therapy, especially in gastrointestinal and dermatological problems. Bioactive phytochemicals were determined by LC-MS/MS, and 14, 13, and 12 compounds were identified in leaves, stems and seeds, respectively. Flavonoids and phenolic acids were the most abundant compounds. Moreover, total phenols (306.5 mg GAE/g), flavonoids (95.25 mg RE/g), and condensed tannins (170.75 mg CE/g) contents were determined in leaves. Also, essential oil composition was investigated using GC/MS and 27 aroma compounds were identified. Monoterpene and sesquiterpene hydrocarbons were the most abundant. Besides, antioxidant activity was performed by DPPH, ß-carotene bleaching, and ferrous ions chelating tests; and leaves extracts were more effective (IC50 of 2.75, 3.45, 13.5 µg/ml), than essential oils (IC50 of 10.5, 70.5, 157.25 µg/ml), and standards (trolox, 4.75; BHT, 5.25 µg/ml), respectively.

Food Security beyond Cereals: A Cross-Geographical Comparative Study on Acorn Bread Heritage in the Mediterranean and the Middle East.

This article aims to contribute to the limited literature on traditional gastronomic knowledge concerning acorn-based bread by ethnographically documenting the ingredients, preparation techniques and consumption practices of baked goods made from acorn seeds and flour that are still used today or at least still present in living memory. A qualitative comparative case method was adopted, and ethnographic data were gathered from 67 people in six selected Mediterranean, Central Asian and Middle Eastern countries. The analysis highlighted distinct trajectories in the development of acorn-based bread, showing some differences in terms of ingredients, preparation techniques and baking methods in the two cultural and geographical macro-regions. By exploring the evolution of the alimentary role of acorn bread in the past century, our findings also support the hypothesis that the product, at least during the last two centuries, has mostly been used as a famine food. By acknowledging the cultural importance of acorn fruits and acorn-based products, this study suggests that the rediscovery of acorn-based products and associated traditional knowledge may foster the sustainable development of rural and marginal regions in the Mediterranean, Middle East and Central Asia. This could help to reinforce the resilience of local communities and thus increase food security. Furthermore, reassessing acorns as a foodstuff may aid in developing innovative products in line with emerging trends in the food sector, which is looking for new non-cereal-based bakery products and other novel culinary applications.

Assessment of artemisinin and antioxidant activities of three wild Artemisia species of Algeria.

Artemisinin, a natural product, has received considerable attention in the last few years as a potent antimalarial drug.This study reports the presence of Artemisinin in three Algerian wild Artemisia species assessed by HPLC method: A. herba-alba (AH), A. campestris subsp. glutinosa (AC), and A. judaica subsp sahariensis (AJ).The HPLC analysis of the hexane extracts, showed a difference in artemisinin content in studied species with a yield of 0.64%, 0.34% and 0.04% for AC, AH and AJ, respectively. Moreover, the level of artemisinin obtained in A. campestris was better than those found in A. sieberi and A. annua. This rate has been reported for the first time.Furthermore, the antiradical activities of methanolic extracts of plants were also tested. There was a remarkable antioxidant capacity found in all Artemisia methanolic extracts analysed.

Medicinal plants used by patients to fight cancer in northwestern Algeria.

Cancer is among the deadliest diseases in the world, especially in developed countries where modern treatments are not available to everyone. In North Africa, and especially in Algeria, few herbal treatments against cancer have been documented despite the richness of flora in these countries. This research aim to documents the medicinal plants used by patient to fight cancer in the northwest of Algeria. Data were collected through ethnobotanical surveys engaging 211 cancer patients in hospitals (departments of medical oncology) in two of the largest provinces in northwest Algeria (Tiaret and Tlemcen). The data were organized into usage reports (UR), while the Informant Consensus Factor (ICF) was calculated to evaluate agreement among informants. The patients investigated were mainly women, of middle age (41-50 years) and illiterate. In total, 53 medicinal plants used against several types of cancer have been identified. These plants are dominated by Lamiaceae, Apiaceae and Rosaceae. The leaves and powder of plants are most often used in traditional preparations often mixed with honey. The plants most often cited in the survey were Aristolochia longa, Aquilaria malaccensis, Ephedra alata subsp. alenda, while the most often treated cancer were breast, cervical, colorectal and stomach. 23 plants are not known as a treatment against cancers in North Africa, while 25 plants already known as treatment for cancer were cited here to treat specific new types of this disease.

Composition and profiling of essential oil, volatile and crude extract constituents of Micromeria inodora growing in western Algeria.

The chemical constituents of the aerial parts of Micromeria inodora (Desf.) Benth. (Lamiaceae) collected in western Algeria, an aromatic shrub traditionally consumed as an herbal remedy, have been characterized. Secondary metabolites were extracted using sequential extractions with solvents at different polarity, and they were determined using an integrated nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) approach. The composition of the essential oil obtained by hydrodistillation was characterized by gas chromatography-mass spectrometry (GC-MS), and the aroma profile was analysed by head-space solid phase micro extraction (HS-SPME) coupled with GC-MS. NMR and LC-MSn analyses revealed the presence of aglyconic and glycosylated flavonoids, phenylpropanoid derivatives and triterpenoid acids, mainly in the methanol, dichloromethane and n-hexane extracts. The analysis of the essential oil allowed to identify 66 components, mainly oxygenated mono- and sesquiterpenes (38.2 % and 32.0 %, respectively) and sesquiterpene hydrocarbons (10.8 %). The aroma profile revealed by HS-SPME-GC-MS was characterized by 45 volatile compounds, among which oxygenated monoterpenes appeared to be the most abundant (65.8 %), followed by sesquiterpene hydrocarbons (16.4 %) and monoterpene hydrocarbons (10.6 %). Overall, these data indicate that M. inodora could be a potential source of antioxidants and bioactive compounds, and they represent a starting point for further research on this plant species.

Total phytochemical analysis of Thymus munbyanus subsp. coloratus from Algeria by HS-SPME-GC-MS, NMR and HPLC-MSn studies.

In this article, we report a comprehensive characterization of volatile and polar constituents extracted from the aerial parts of Thymus munbyanus subsp. coloratus, a shrub that is used as culinary ingredient and as traditional medicine in Algeria, mainly to treat respiratory and gastrointestinal disorders and endocrine dysfunctions. Headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was used to assess volatile constituents, whereas the phytochemical composition of solid residues obtained from extraction with solvents at diffrent polarity was obtained by an integrated Nuclear Magnetic Resonance (NMR) and liquid chromatography coupled with tandem mass spectrometry (LC-MSn) approach. Fourty-five apolar ccompounds were identified, mainly oxygenated monoterpenes (65.8%), sesquiterpene hydrocarbons and nonoterpene hydrocarbons (18.6 and 14.5%, respectively). On the other hand, LC-MSn and NMR analyses revealed the presence of aglyconic and glycosilated flavonoids, phenylpropanoid derivatives and triterpenoid acids related to oleanolic acid, mainly in the methanol, dichloromethane and hexane extracts. Overall, these data indicate that Thymus munbyanus subsp. coloratus could be a potential source of antioxidants and bioactive compounds, and our results represent a starting point for further research on this plant species.

Ethnoveterinary remedies used in the Algerian steppe: Exploring the relationship with traditional human herbal medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Ethnoveterinary medicine is vastly under-documented, especially in North Africa, where livestock care is increasingly medicalised. Despite evidence of the interdependence of ethnoveterinary practices and ethnomedicine for human care, the overlap between these two systems and the possible drivers of similarity are rarely addressed in ethnopharmacological literature. Here, we present the first quantitative comparison of remedies used to treat human and animal health among a pastoral society. AIMS: This study aims to document the plants and other materials used in ethnoveterinary medicine among nomadic herders in the Algerian steppe. We calculate the overlap with remedies used for human health and evaluate some of the possible drivers of similarities between the two interlinked medical systems. METHODS: The field study was conducted in spring 2018 with 201 local knowledge holders in five provinces in the central part of the Algerian steppe. Forty-six camps and ten weekly animal markets were visited. After obtaining prior informed consent, data was collected through structured interviews. Anonymous sociodemographic information was collected along with veterinary use data. Plant specimens were acquired, identified and deposited in the Botanical Laboratory Herbarium of the University of Tiaret. Ethnobotanical information was structured in use reports and therapeutic applications organised into 13 simple categories. Plants used in ethnoveterinary medicine were compared to those used in human health care using a Chi square test, and ethnoveterinary use was predicted using a generalised linear model with use for human care and plant family as predictive variables. Logistic regressions were also used to test if any specific medicinal application predicts shared use in human and veterinary medicine. RESULTS: Sixty-six plant species from 32 botanical families and ten non-vegetable remedies were documented. Plants from the Lamiaceae, Asteraceae, Fabaceae and Apiaceae families were most commonly used. The plants most often cited by interviewees are all harvested from the wild (Echinops spinosissimus Turra, Atriplex halimus L., Artemisia campestris L., Juniperus phoenicea L. and Peganum harmala L.). Leaves are the most commonly used plant part and decoction is the most common preparation method. There are important similarities between remedies used to treat humans and other animals: two thirds of the remedies used in ethnoveterinary medicine are also used in human health care, and these represent half of the human health treatments. Use for human health and plant family both predict the use of a medicinal plant in veterinary medicine, but no correlation is found regarding specific therapeutic applications. CONCLUSIONS: Traditional veterinary knowledge is still key to sheep and goat herders in the Algerian steppe, but a knowledge transmission gap seems to exist between older and younger generations, and ethnoveterinary practices may disappear in the near future. Treatments for human and animal care overlap to a large extent, and a causal relationship possibly exists for, at least, some of them. However, overall a smaller number of remedies are used to treat animals than humans and for less therapeutic applications. This difference in the diversity of therapeutic applications and remedies should be the object of future research.

Thymus lanceolatus ethanolic extract protects human cells from t-BHP induced oxidative damage.

This study aimed to investigate the ethanolic extract of T. lanceolatus, a species native to north-western Algeria, traditionally used as herbal tea, seasoning and a preservative for meat and poultry. HPLC analysis showed the presence of fourteen bioactive compounds, among which rosmarinic acid is by far the most abundant one (15440.9 mg kg-1). Its biological activity on proliferation, viability and ROS protection was investigated towards K562, CaCo-2 and SH-SY5Y human cancer cell lines. Cell proliferation was inhibited in K562 and SH-SY5Y cells in the first 48 h at 500 µg mL-1 but slowly resumed after 72 h. A weak cytotoxic effect can be observed at 24, 48 and 72 hours: 12.8%, 14.9% and 24.2% reduction in K562 viability, and 11%, 15% and 12.7% in SH-SY5Y. No cytotoxicity was observed in CaCo-2 cells under the same experimental conditions. Even at the lowest concentrations (50 µg mL-1), the extract was efficiently able to protect human cells against t-BHP-induced oxidative damage. For instance, the highest concentration of the extract (100 µg mL-1) decreases ROS generation by about 30% in SH-SY5Y and 70% in CaCo-2 and K562 cells. The study confirms the very low cytotoxicity of the T. lanceolatus ethanolic extract and highlights its nutraceutical properties as an antioxidative and preservative agent and its possible use as an ingredient in functional foods.

Essential oils (EOs), pressurized liquid extracts (PLE) and carbon dioxide supercritical fluid extracts (SFE-CO2) from Algerian Thymus munbyanus as valuable sources of antioxidants to be used on an industrial level.

The aim of this study was to demonstrate the potential of extracts from Algerian Thymus munbyanus as a valuable source of antioxidants for use on an industrial level. To this end, a study was conducted on the composition and antioxidant activities of essential oils (EOs), pressurized liquid extracts (PLE) and supercritical fluid extracts (SFE-CO2) obtained from Thymus munbyanus subsp. coloratus (TMC) and subsp. munbyanus (TMM). EOs and SFE-CO2 extracts were analysed by GC-FID and GC×GC-TOFMS revealing significant differences. A successive extraction of the solid SFE-CO2 residue by PLE extraction with solvents of increasing polarity such as acetone, ethanol and water, was carried out. The extracts were evaluated for total phenolic content by Folin-Ciocalteu assay, while the antioxidant power was assessed by DPPH, FRAP, and ORAC assays. SFE-CO2 extracts were also analysed for their tocopherol content. The antioxidant activity of PLE extracts was found to be higher than that of SFE-CO2 extracts, and this increased with solvent polarity (water > ethanol > acetone). Overall, these results support the use of T. munbyanus as a valuable source of substances to be used on an industrial level as preservative agents.

Ethnobotanical survey of medicinal plants used by nomadic peoples in the Algerian steppe.

ETHNOPHARMACOLOGICAL RELEVANCE: This study is the first ethnobotanical survey focusing on the herbal medicines traditionally used by the nomadic community of the Algerian steppe, identifying new medicinal plants and uses from one of the most characteristic indigenous populations in Algeria. Moreover, the study contributes to the understanding of transmission of medicinal plant knowledge in the Mediterranean basin. AIM OF THE STUDY: This work aims to document the phytotherapeutical knowledge and practice of the nomadic community of the Algerian steppe, and compare it with neighbouring sedentary populations and Mediterranean historical texts. Through this, the study strives to evaluate processes of transmission of knowledge among this population, for whom written sources have been largely unavailable. METHODS: Ethnobotanical surveys were carried out during two years (2015-2017). In total, 73 informants from nomadic populations were interviewed in several steppe regions including areas in the administrative departments of Tiaret, Saida, Naama, Djelfa and M'sila. Structured interviews about medicinal plant knowledge were combined with guided tours with the informants. Prior informed consent was always obtained. The surveys allowed for the collection of sociodemographic data and traditional knowledge about medicinal plants and their uses. Informant Consensus Factor (FIC) was calculated to evaluate agreement among informants. Results were compared to existing literature to evaluate similarities between this nomadic medicinal flora, that of neighbouring communities and historical texts and identify new plant citations and uses. RESULTS: Among Algerian nomadic communities, herbal remedies are used mostly by women and elders, who are often illiterate. We identified 97 taxa of medicinal plants belonging to 42 botanical families, importantly Lamiaceae, Asteraceae and Apiaceae, like in neighbouring communities. The most common plant parts and method of preparation are also shared with neighbouring populations. New uses are described for 25 known medicinal taxa, and nine species with undocumented medicinal uses in recent literature have been reported. However, some of these have been reported in Mediterranean materia medica. In total, 60% of the medicinal plant diversity used by Algerian nomads are well-known plants of the Mediterranean ethnopharmacological heritage. CONCLUSION: This study highlights the importance of traditional medicine for Algerian nomad communities. This indigenous population has specific knowledge about plants from their steppe environment, but also shares a pool of knowledge with sedentary Algerian populations and Mediterranean people as a whole. Our research shows that a common North African and Mediterranean ethnobotanical heritage exists.

NMR, HS-SPME-GC/MS, and HPLC/MSn Analyses of Phytoconstituents and Aroma Profile of Rosmarinus eriocalyx.

In this work, a comprehensive study on the chemical constituents of the aerial parts of Rosmarinus eriocalyx (Lamiaceae), an aromatic shrub traditionally consumed as a food and herbal remedy in Algeria, is presented. The aroma profile was analysed by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC/MS), whereas the crude extract constituents were analyzed by 1 H-NMR and by high performance liquid chromatography coupled with mass spectrometry (HPLC/MSn ). Thirty-nine volatile compounds, most of them being monoterpenes, have been identified, with camphor, camphene, and α-pinene as the most abundant constituents. 1 H-NMR analysis revealed the presence of phenolic compounds and betulinic acid while HPLC/MSn allowed the identification of glycosilated and aglyconic flavonoids as well as phenylpropanoid derivatives. Some of these constituents, namely as betulinic acid, rosmanol, and cirsimaritin were reported for the first time in R. eriocalyx.

Essential Oil of Thymus munbyanus subsp. coloratus from Algeria: Chemotypification and in vitro Biological Activities.

Thymus munbyanus subsp. coloratus (Lamiaceae) is a small shrub endemic to Algeria and Morocco where is found in lawns, rockeries and mountainous regions. From a phytochemical point of view this taxon has never been characterized. In this work we have analysed the chemical compositions of the essential oils obtained from inflorescences and vegetative parts by GC/MS. A new chemotype, i.e. borneol-chemotype, was characterized for the first time in the species. Furthermore, we assessed the biological activities of essential oils, namely the antioxidant, antimicrobial and cytotoxicity on tumor cells that were evaluated by the DPPH, ABTS, and FRAP, disc diffusion, and MTT methods, respectively. Biological assays highlighted a moderate inhibitory effect on Staphylococcus aureus, Escherichia coli and Candida albicans (inhibition zone diameter in the range 9 - 10 mm), and noteworthy cytotoxicity on A375 human melanoma cells (IC50 of 46.95 µg/ml).