The current study investigates the content of sugars, organic acids, phenolic acids and flavonoids, as well as antioxidant and antimicrobial activity of Balkan-endemic Micromeria frivaldszkyana. Glucose was the most abundant sugar in the plant (2.77%), followed by fructose (1.18%) and galactose (0.82%). Eight organic acids were detected with quinic acid being in the highest content—556.3 mg/100 g DW. From the individual phenolic acids, rosmarinic acid was found in the most significant amounts (2040.1 ± 1.97 mg/100 g) and hesperidin was the major representative of flavonoids with content 131.2 ± 5.6 mg/100 g DW. The antioxidant activity of the plant was studied by six methods: 2,2′-diphenylpicrylhydrazyl (DPPH)—286.4 ± 10.43 mM TE/g, 2,2′azinobis (3)-ethylbenzthiazoline-6-sulfonic acid (ABTS)—358.4 ± 10.4 mM TE/g, ferric reducing antioxidant power (FRAP)—388.0 ± 32.4 mM TE/g, cupric reducing antioxidant capacity (CUPRAC)—905.6 ± 19.2 mM TE/g, Oxygen Radical Absorbance Capacity (ORAC)—3250.5 ± 208.1 µmol TE/g and Hydroxyl Radical Averting Capacity (HORAC)—306.1 ± 23.5 µmol GAE/g. In vitro antimicrobial activity against nine microorganism was evaluated but the extract displayed antimicrobial activity only against Listeria monocytogenes ATCC 19111 with inhibition zone diameter 9 mm and minimal inhibitory concentration (MIC) 10 mg/mL.

Extracts from medicinal plants are widely used in the treatment and prevention of different diseases. Micromeria frivaldszkyana is a Balkan endemic species with reported antioxidant and antimicrobial characteristics; however, its phytochemical composition is not well defined. Here, we examined the metabolome of M. frivaldszkyana by chromatography–mass spectrometry (GC-MS), ultra-performance liquid chromatography–mass spectrometry (UPLC-MS-MS), and inductively coupled plasma mass spectrometry (ICP-MS). Amino acids, organic acids, sugars, and sugar alcohols were the primary metabolites with the highest levels in the plant extract. Detailed analysis of the sugar content identified high levels of sucrose, glucose, mannose, and fructose. Lipids are primary plant metabolites, and the analysis revealed triacylglycerols as the most abundant lipid group. Potassium (K), magnesium (Mg), zinc (Zn), and calcium (Ca) were the elements with the highest content. The results showed linarin, 3-caffeoil-quinic acid, and rosmarinic acid, as well as a number of polyphenols, as the most abundant secondary metabolites. Among the flavonoids and polyphenols with a high presence were eupatorin, kaempferol, and apigenin—compounds widely known for their bioactive properties. Further, the acute toxicity and potential anti-inflammatory activity of the methanolic extract were evaluated in Wistar rats. No toxic effects were registered after a single oral application of the extract in doses of between 200 and 5000 mg/kg bw. A fourteen-day pre-treatment with methanolic extract of M. frivaldszkyana in doses of 250, 400, and 500 mg/kg bw induced anti-inflammatory activity in the 1st, 2nd, and 3rd hours after carrageenan injection in a model of rat paw edema. This effect was also present in the 4th hour only in the group treated with a dose of 500 mg/kg. In conclusion, M. frivaldszkyana extract is particularly rich in linarin, rosmarinic acid, and flavonoids (eupatorin, kaempferol, and apigenin). Its methanolic extract induced no toxicity in male Wistar rats after oral application in doses of up to 5000 mg/kg bw. Additionally, treatment with the methanolic extract for 14 days revealed anti-inflammatory potential in a model of rat paw edema on the 1st, 2nd, and 3rd hours after the carrageenan injection. These results show the anti-inflammatory potential of the plant, which might be considered for further exploration and eventual application as a phytotherapeutic agent.

The ferns Asplenium ceterach L., Asplenium scolopendrium L. and Asplenium trichomanes L. have wide application in traditional medicine worldwide. However, the scientific research on their anticancer and antibacterial properties is insufficient. The present article aims to provide more information on this topic. Extracts derived from the aerial parts of A. ceterach, A. scolopendrium and A. trichomanes were examined using a panel of in vitro assays with different bacterial and mammalian cells. The cytotoxicity and anticancer activity of the samples were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trypan blue assays with three human (A549, FL, HeLa) and three murine (3T3, TIB-71, LS48) cell lines. Inhibitory effects on the growth of Gram-positive (Bacillus cereus) and Gram-negative (Pseudomonas aeruginosa) bacteria were determined by the agar diffusion assay. Apoptosis-inducing properties of the extracts were analyzed by flow cytometry. Superoxide dismutase (SOD) activity in extract-treated cells was investigated by ELISA. The obtained results demonstrate selective anticancer activity of all three Asplenium species. The extract from A. ceterach displayed the strongest inhibitory properties against human cervical cancer cells and bacterial cells. It induced a lower level of cytotoxicity against mouse cell lines, indicating a species-specific effect. The extract from A. trichomanes demonstrated better anticancer and antibacterial properties than the sample from A. scolopendrium. Further experiments linked the mechanism of action of A. ceterach extract with oxidative stress-inducing potential and strong proapoptotic potential against the cervical cancer cell line HeLa. A. trichomanes and A. scolopendrium extracts appeared to be potent inducers of necrotic cell death.

Background: Ferns are underestimated as medicinal plants and their use in traditional medicine is limited despite their diversity. Most of the research on their healing properties and phytochemical composition related to their biological activity has been focused on the secondary metabolites synthesized by ferns. Objective: In this study, we aimed to make an in-depth chemical characterization of three ferns widely spread in Bulgaria - Asplenium ceterach L., Asplenium scolopendrium L. and Asplenium trichomanes L. Methods: Micro elemental analysis was carried out using ICP-MS. Standard laboratory methods were used to determine the content of proteins, fats and fatty acids. Chromatographic methods were applied to quantify some secondary metabolites. Two antioxidant methods and two antimicrobial tests were used to evaluate the biological properties of the ferns tested. Results: Micro elemental analysis showed that these ferns could be useful source of zinc and iron. Protein content slightly varied among the species (13.6% - 18.4% of the dried plant weight). Negligible was the variation in oil content (3.1% - 4.0%, dry weight base); oil composition was dominated by saturated fatty acids. Some principal classes secondary metabolites, individual phenolic acids and flavonoids were quantified. The extract obtained from Asplenium ceterach L. demonstrated the highest antioxidant capacity. Fern extracts showed moderate antimicrobial activity against six of the 14 microorganisms tested. Conclusion: The results obtained may reveal new areas of application for ferns and are a solid basis for comparison with the same species growing in other geographical and climatic conditions, which may affect their healing properties.

Marrubium species have been used since ancient times as food additives and curative treatments. Their phytochemical composition and various pharmacological activities were the focus of a number of scientific investigations but no comprehensive metabolome profiling to identify the numerous primary and secondary metabolites has been performed so far. This study aimed to generate a comprehensive picture of the total metabolite content of two Marrubium species—M. peregrinum and M. friwaldskyanum—to provide detailed information about the main primary and secondary metabolites. In addition, the elemental composition was also evaluated. For this purpose, non-targeted metabolomic analyses were conducted using GC-MS, UPLC-MS/MS and ICP-MS approaches. Nearly 500 compounds and 12 elements were detected and described. The results showed a strong presence of phenolic acids, flavonoids and their glucosides, which are generally of great interest due to their various pharmacological activities. Furthermore, tissue-specific analyses for M. friwaldskyanum stem, leaves and flowers were carried out in order to outline the sources of potentially important bioactive molecules. The results generated from this study depict the Marrubium metabolome and reveal its dual scientific importance—from one side, providing information about the metabolites that is fundamental and vital for the survival of these species, and from the other side, defining the large diversity of secondary substances that are a potential source of phytotherapeutic agents.

The present study is focused on the chemical and lipid composition of seed oil of the European ornamental and invasive wood plant Ailanthus altissima (Simaroubaceae). Total lipids, proteins, carbohydrates, ash, and moisture in the seeds were determined. A high yield of glyceride oil (30.7%) was found, as well as a high content of fibers (29.6%) and proteins (18.7%). Physicochemical properties of the oil define it as semi-dry (129.4 g I₂/100 g Iodine value) with oxidative stability, refractive index, saponification value, and relative density similar to widely used oils with nutritional value and health benefits. The composition of the seed oil was determined chromatographically. Unsaturated fatty acids (95.3%) predominated in the seed oil, of which linoleic acid (48.6%) and oleic acid (44.8%) were the major ones. The main lipid-soluble bioactive components were β-sitosterol (72.6%), γ-tocopherol (74.6%), phosphatidylinositol (29.5%), and phosphatidic acids (25.7%). The proven in vitro DNA-protective ability of seed oil is reported for the first time. The seed oil exhibited a weak antiproliferative effect on HT-29 and PC3 tumor cell lines and showed no cytotoxicity on the BALB/c 3T3 cell line. In brief, the present study reveals that A. altissima seed oil can be used as a healthy food.