Antibiotic resistance is a growing problem and may become the next major global health crisis if no timely actions are taken. Mycobacterial infections are widespread and, due to antibiotic resistance, also hard to treat and a major cause of mortality. Natural compounds have the potential to increase antibiotic effectiveness due to their resistance modulatory and antimicrobial effects. In this study, Peucedanum ostruthium extracts, fractions, and isolated compounds were investigated regarding their antimicrobial and resistance-modulatory effects as well as efflux pump inhibition in Mycobacterium smegmatis. P. ostruthium extracts were found to have anti-mycobacterial potential and resistance modulating effects on ethidium bromide activity. The major antibacterial effect was attributed to ostruthin, and we found that the more lipophilic the substrate, the greater the antimicrobial effect. Imperatorin caused potent modulatory effects by interfering with the action of the major LfrA efflux pump in M. smegmatis. The plant P. ostruthuim has a complex effect on M. smegmatis, including antibacterial, efflux pump inhibition, resistance modulation, and membrane permeabilization, and its major constituents, ostruthin and imperatorin, have a distinct role in these effects. This makes P. ostruthium and its coumarins promising therapeutics to consider in the fight against drug-resistant mycobacteria.

Reynoutria japonica Houtt. is a critical invasive alien plant in Europe and North America with a drastic impact on native flora. However, R. japonica has medicinal potential, especially as a source of stilbenes. In order to explore the potential of simple extractions of R. japonica, we conducted qualitative and quantitative analyses of fresh R. japonica rhizome infusion, decoction, and macerates with ethanol by UHPLC-DAD-ESI-MSⁿ and UHPLC-DAD, with a focus on major constituent groups of stilbenes and anthranoids. Since R. japonica rhizome extracts showed antimicrobial potential in the past, we also evaluated the antimycobacterial effect of raw R. japonica extracts for the first time against Mycobacterium smegmatis. Of thirty-four characterized substances, six were stilbenes and twelve anthranoids. The main constituents, four trans-stilbenes and eight anthranoids, were quantified in a validated UHPLC-DAD method. The 38% ethanol macerate showed high stilbene (155.078 mg/100 g fluid extract) and low anthranoid content (5.420 mg/100 g fluid extract), while decoction showed the highest anthranoids. Antimycobacterial testing gave good results for all macerates (MIC 256 µg/mL) and trans-resveratrol (64 µg/mL). Extraction and enrichment of stilbenes from fresh plant material by simple extraction methods with food-grade solvents might encourage consideration of wild harvest of rhizomes over classic means of eradication of R. japonica.

In course of our studies of the aerial parts of Artemisia abrotanum the major methoxyflavonol could be isolated. However, by NMR structural analysis it became obvious that the substitution pattern in ring B differs from reports for casticin (2). The position of methoxyl and hydroxyl groups are interchanged, i.e., the major flavone is actually chrysosplenetin (1). Three structures in A. abrotanum and A. frigida had to be revised. Use of pyridine-d₅ instead of DMSO‑d₆ made the resolution of the B-ring ¹H and ¹³C NMR signals possible and enabled correct structural assignment by 2D NMR experiments.

The heartwood of the Swiss Stone Pine, Pinus cembra L., has been scarcely investigated for secondary metabolites for a long period of time. Considering age and relative simplicity of heartwood investigations dating back to the 1940s to 1960s, we conducted the first investigation of P. cembra heartwood by HPLC, using UHPLC-DAD-ESI-MSⁿ and HPLC-DAD techniques in combination with isolation and NMR spectroscopy, with focus on stilbenes, bibenzyls and flavonoids. Analytical problems in the HPLC analysis of Pinus stilbenes and flavonoids on reversed stationary phases were also challenged, by comparing HPLC on pentafluorophenyl (PFP) and C18 stationary phases. Seven flavonoids (1, 2, 3, 7, 8, 11, 12), four stilbenes (4, 6, 10, 13), two bibenzyls (5, 9), three fatty acids (14, 16, 17) and one diterpenic acid (15) were detected in an ethanolic extract of Pinus cembra heartwood. HPLC comparison of reversed stationary phases in HPLC showed that the antifungal, antibacterial and chemosensitizing dihydropinosylvin monomethyl ether (9) and pinosylvin monomethyl ether (10) can be separated on PFP, but not on C18 material, when eluting with a screening gradient of 20–100% acetonitrile. Flavonoid separation showed additional benefits of combining analyses on different stationary phases, as flavonoids 7 and 8 could only be separated on one of two C18 stationary phases. Earlier phytochemical results for heartwood investigations were shown to be mostly correct, yet expandable. Substances 5 to 12 were found in alignment with these references, proving remarkable phytochemical analyses at the time. Evidence for the described presence of pinobanksin could not be found. Substances 1 to 4 and 13 have to our knowledge not yet been described for P. cembra.

Introduction: Peels are an abundant but still underutilized waste product in the Citrus fruit industry. They contain coumarins with antiadipogenic potential that could be promising targets in new valorization strategies for Citrus peels.Objectives: In this study, these coumarins, that is, citropten, bergamottin, and 5‐geranyloxy‐7‐methoxycoumarin (5G7MC), were investigated in Citrus limon peels of different commercial varieties by HPLC‐DAD after extraction with ethanol and choline chloride‐based natural deep eutectic solvents (NADES) as alternative extraction agents in green natural product extraction.Results: Target coumarins were almost exclusively found in the flavedo (exocarp) peel layer and in whole peel samples, but not in the albedo (mesocarp) layer. Flavedo samples from varieties “Verdelli” (ES) and “Lisbon” (ZA) showed particularly high concentrations in ethanolic extracts. When applying NADES extraction, best results for citropten were achieved with malonic acid, 1,2‐propanediol, and citric acid as hydrogen bond donors (HBDs), with higher yields than the reference solvent ethanol. For bergamottin and 5G7MC, promising results were obtained for 1,2‐propanediol as HBD, though lower yields than with ethanol. The possible chlorination of coumarin epoxides byakangelicol and oxypeucedanin was recognized for acidic NADES in LC–MS analysis.Conclusion: Overall, C. limon peels proved to be a relevant starting material for valorization through coumarin extraction with ethanol and NADES.

With a growing focus on green chemistry, the extraction of natural products with natural deep eutectic solvents (NADES), which are eutectic mixtures of hydrogen bond donors and acceptors, has become an ever-expanding field of research. However, the use of NADES for the extraction of spilanthol from Acmella oleracea (L.) R.K.Jansen has not yet been investigated. Therefore, in this study, 20 choline chloride-based NADES, and for comparison, ethanol, were used as green extraction agents for spilanthol from Acmella oleracea flower heads. The effects of time, water addition, and temperature on NADES extractions were investigated and analysed by HPLC-DAD quantification. Additionally, UHPLC-DAD-ESI-MSⁿ results for dichloromethane extracts, as well as the isolation of spilanthol and other main constituents as reference compounds, are reported. The best green extraction results were achieved by choline chloride (ChCl) with 1,2-propanediol (P, 1:2 molar ratio, +20% water) at 244.58 µg/mL, comparable to yields with ethanol (245.93 µg/mL). Methylurea (MeU, 1:2, +20% water) also showed promising results as a hydrogen bond donor in combination with choline chloride (208.12 µg/mL). In further experiments with NADES ChCl/P (1:2) and ChCl/MeU (1:2), extraction time had the least effect on spilanthol extraction with NADES, while yield decreased with water addition over 20% and increased with extraction temperature up to 80 °C. NADES are promising extraction agents for the extraction of spilanthol, and these findings could lead to applicable extracts for medicinal purposes, due to their non-toxic constituents.