Phytochemical Investigation of Polyphenols from the Aerial Parts of Tanacetum balsamita Used in Transylvanian Ethnobotany and Parallel Artificial Membrane Permeability Assay.

In this study, based on ethnobotanical data recorded in Transylvania, the polyphenolic compounds and the permeability of the aerial part's extract of Tanacetum balsamita were investigated. Ultrahigh-performance liquid chromatography-tandem mass spectrometry was applied for the analysis of the extracts. Parallel artificial membrane permeability assay (PAMPA) for the gastrointestinal tract and the blood-brain barrier was conducted. In the ethanolic and aqueous extracts of the species traditionally used for wound, furuncle, and liver disorders, 92 polyphenols were characterized (e.g., flavonoid, hydroxycinnamic acid, catechin, dihydroxybenzoyl, lignan derivatives, and a monoterpene) including 54 compounds identified for the first time in the plant. In the PAMPA tests, eight components were shown to be capable of passive diffusion across the studied membranes. These include apigenin and seven methoxylated flavonoid derivatives. Based on these results, methoxylated flavonoids might promote the pharmacological potential of T. balsamita to be applied in the enhancement of novel remedies.

A Comprehensive Study on Lathyrus tuberosus L.: Insights into Phytochemical Composition, Antimicrobial Activity, Antioxidant Capacity, Cytotoxic, and Cell Migration Effects.

In this study, in vitro antioxidant, antimicrobial, cytotoxic, and cell migration effects of phenolic compounds of Lathyrus tuberosus leaves, known in the Transylvanian ethnomedicine, were investigated. Ultra-high-performance liquid chromatography-tandem mass spectrometry was employed for the analysis of the ethanolic and aqueous extracts. The antimicrobial properties were determined using a conventional microdilution technique. Total antioxidant capacity techniques were used using cell-free methods and cell-based investigations. Cytotoxic effects were conducted on 3T3 mouse fibroblasts and HaCaT human keratinocytes using a multiparametric method, assessing intracellular ATP, total nucleic acid, and protein levels. Cell migration was visualized by phase-contrast microscopy, employing conventional culture inserts to make cell-free areas. Together, 93 polyphenolic and monoterpenoid compounds were characterized, including flavonoid glycosides, lignans, hydroxycinnamic acid, and hydroxybenzoic acid derivatives, as well as iridoids and secoiridoids. The ethanolic extract showed high antioxidant capacity and strong antimicrobial activity against Bacillus subtilis (MIC80 value: 354.37 ± 4.58 µg/mL) and Streptococcus pyogenes (MIC80 value: 488.89 ± 4.75 µg/mL). The abundance of phenolic compounds and the results of biological tests indicate the potential for L. tuberosus to serve as reservoirs of bioactive compounds and to be used in the development of novel nutraceuticals.

Evaluation of the Chemical Stability, Membrane Permeability and Antiproliferative Activity of Cyclic Diarylheptanoids from European Hornbeam (Carpinus betulus L.).

Four cyclic diarylheptanoids-carpinontriols A (1) and B (2), giffonin X (3) and 3,12,17-trihydroxytricyclo [12.3.1.12,6]nonadeca-1(18),2(19),3,5,14,16-hexaene-8,11-dione (4)-were isolated from Carpinus betulus (Betulaceae). Chemical stability of the isolated diarylheptanoids was evaluated as a function of storage temperature (-15, 5, 22 °C) and time (12 and 23 weeks). The effect of the solvent and the pH (1.2, 6.8, 7.4) on the stability of these diarylheptanoids was also investigated. Compounds 2 and 4 showed good stability both in aqueous and methanolic solutions at all investigated temperatures. Only 2 was stable at all three studied biorelevant pH values. Degradation products of 1 and 3 were formed by the elimination of a water molecule from the parent compounds, as confirmed by ultrahigh-performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-HR-MS). The permeability of the compounds across biological membranes was evaluated by the parallel artificial membrane permeability assay (PAMPA). Compound 3 possesses a logPe value of -5.92 ± 0.04 in the blood-brain barrier-specific PAMPA-BBB study, indicating that it may be able to cross the blood-brain barrier via passive diffusion. The in vitro antiproliferative activity of the compounds was investigated against five human cancer cell lines, confirming that 1 inhibits cell proliferation in A2058 human metastatic melanoma cells.

Characterization of alkaloid profile of Hyoscyamus reticulatus L. and Atropa belladonna subsp. caucasica (Kreyer) Avet by LC-MS and NMR.

The alkaloid profile of Hyoscyamus reticulatus L. and Atropa belladonna subsp. caucasica (Kreyer) Avet have not been characterized yet. UHPLC-PDA-Q-TOF-MS/MS and LC-DAD-QqQ-MS/MS methods were used herein to characterize the metabolite profiles of these plants. Flash chromatography in combination with preparative- and semi preparative HPLC were utilized for the isolation of the compounds of interest. The structure of the isolated compounds was proposed based on their MS/MS fragmentation and NMR characteristics. As a total of 19 tropane derivatives, two tyramine derivatives (N-cis- and N-trans- feruloyl tyramine), a lignanamide (grossamide), an alkylamide (pellitorine) were identified in the root and herb extracts of H. reticulatus. Moreover, rutin and caffeoylquinic acids were found in the leaves and fruits, while kaempferol-3-O-glucoside-7-O-rhamnoside and quercetin-3-O-glucoside-rhamnoside-rhamnoside were exclusively characteristic for the leaves of H. reticulatus. The root and herb extracts of A. caucasica contained 16 tropane alkaloid derivatives along with methyl tropate and two tyramine derivatives.

Membrane Permeability and Aqueous Stability Study of Linear and Cyclic Diarylheptanoids from Corylus maxima.

Seven diarylheptanoids were isolated from Corylus maxima by flash chromatography and semipreparative high-performance liquid chromatography (HPLC) and identified by Orbitrap® mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy as linear diarylheptanoids: hirsutanonol-5-O-ß-D-glucopyranoside (1), platyphyllonol-5-O-ß-D-xylopyranoside (4), platyphyllenone (5); and cyclic derivatives: alnusonol-11-O-ß-D-glucopyranoside (6), alnusone (7), giffonin F (8), carpinontriol B (9). Cyclic diarylheptanoids are reported in C. maxima for the first time. The aqueous stability of the isolated compounds and other characteristic constituents of C. maxima, oregonin (2), hirsutenone (3), quercitrin (10) and myricitrin (11) was evaluated at pH 1.2, 6.8 and 7.4. The passive diffusion of the constituents across biological membranes was investigated by parallel artificial membrane permeability assay for the gastrointestinal tract (PAMPA-GI) and the blood-brain barrier (PAMPA-BBB) methods. The cyclic diarylheptanoid aglycones and quercitrin were stable at all investigated pH values, while a pH-dependent degradation of the other compounds was observed. A validated ultrahigh-performance liquid chromatography-diode-array detection (UHPLC-DAD) method was utilized for the determination of compound concentrations. The structures of the degradation products were characterized by UHPLC-Orbitrap® MS. Platyphyllenone and alnusone possessed log Pe values greater than -5.0 and -6.0 in the PAMPA-GI and PAMPA-BBB studies, respectively, indicating their ability to cross the membranes via passive diffusion. However, only alnusone can be considered to have both good aqueous stability and satisfactory membrane penetration ability.

Isolation and quantification of diarylheptanoids from European hornbeam (Carpinus betulus L.) and HPLC-ESI-MS/MS characterization of its antioxidative phenolics.

Detailed polyphenol profiling of European hornbeam (Carpinus betulus L.) bark, leaf, male and female catkin extracts was performed by high-performance liquid chromatography-diode array detection coupled to tandem mass spectrometry (HPLC-DAD-MS/MS). A total of 194 compounds were characterized and tentatively identified. Gallo- and ellagitannins dominated in the methanol extracts, while flavonol glycosides and methoxylated flavones prevailed in the ethyl acetate samples. In the quest for diarylheptanoids, twelve compounds were isolated by the combination of subsequent reversed-phase flash chromatographic and high-performance liquid chromatographic methods. The structural elucidation of the isolated components was performed by ultrahigh-performance liquid chromatography-Orbitrap mass spectrometry (UHPLC-Orbitrap-MS) as well as 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. Six known cyclic diarylheptanoids, together with a new compound were described in Carpinus betulus for the first time. The occurrence of a linear diarylheptanoid and a lignan has also been unprecedented in the genus Carpinus. Moreover, three known flavonol glycosides were isolated. Based on the identification of characteristic fragment ions, a new mass spectrometric fragmentation pathway for meta,meta-cyclophane-type diarylheptanoids was proposed. Quantities of the four major cyclic diarylheptanoids in European hornbeam were determined by a validated UHPLC-DAD method for the first time. The antioxidant properties of the extracts and the isolated compounds were assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Contribution of the individual constituents to the total radical scavenging activity of the samples was evaluated by an off-line DPPH-HPLC-DAD method. This allowed the identification of gallo- and ellagitannin derivatives as the constituents being primarily responsible for the antioxidant capacity of the extracts.