Natural Deep Eutectic Solvents for the Extraction of Spilanthol from Acmella oleracea (L.) R.K.Jansen.

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.

Larvicidal, Ovicidal, and Repellent Activities of Leucas stachydiformis (Hochst. ex Benth.) Briq Essential Oil against Anopheles arabiensis.

Larvicidal, ovicidal, and repellent activities of the essential oil extracted by hydrodistillation from the leaves of the endemic Ethiopian plant Leucas stachydiformis (Hochst. ex Benth.) Briq were investigated against Anopheles arabiensis, the dominant malaria vector species in Ethiopia with the objective of searching for a plant-based malaria vector control strategy from medicinal plants. The larvicidal effect was tested against the fourth instar An. arabiensis wild larvae whilst freshly laid ova of An. arabiensis were used to determine the ovicidal activity of the essential oil at concentrations ranging from 6.25 to 400 ppm. Concentrations of 41.6-366.7 µg/cm2 were used to evaluate the repellent activity of the essential oil on 3-5 days old adult female An. arabiensis. The oil composition of L. stachydiformis was also analyzed using GC-MS. The study revealed that the oil possesses the highest larvicidal activity at 400 ppm and 200 ppm after 24 h and 48 h of treatment. LC50 values for the fourth larval instar after 24 h and 48 h of treatment were 43.4 ppm and 34.2 ppm, respectively. After 72 h of exposure, the oil displayed 100% ovicidal activity at 400 ppm with an IH50 value of 32.2 ppm. In the repellency test, at concentrations of 366.7, 133.3, and 41.6 µg/cm2, the oil gave a total percentage protection of 67.9 ± 4.2%, 37.2 ± 2.8%, and 32 ± 2.2%, respectively, for 4 h. The highest concentration (366.7 µg/cm2) gave 100% protection up to 90 min. GC-MS analysis of the oil revealed the presence of 24 compounds representing 90.34% of the total oil with caryophyllene oxide, germacrene D, and trans-caryophyllene constituting more than 50% of its components. Results of the present study suggest that the essential oil of L. stachydiformis has the potential to be used for the control of An. arabiensis mosquitoes.

Investigation of Hydrocolloid Plant Polysaccharides as Potential Candidates to Mimic the Functions of MUC5B in Saliva.

The successful substitution of complex physiological fluids, such as human saliva, remains a major challenge in drug development. Although there are a large number of saliva substitutes on the market, their efficacy is often inadequate due to short residence time in the mouth, unpleasant mouthfeel, or insufficient protection of the teeth. Therefore, systems need to be identified that mimic the functions of saliva, in particular the salivary mucin MUC5B and the unique physiological properties of saliva. To this end, plant extracts known to contain hydrocolloid polysaccharides and to have mucus-forming properties were studied to evaluate their suitability as saliva substitutes. The aqueous plant extracts of Calendula officinalis, Fucus sp. thalli, and lichenan from Lichen islandicus were examined for composition using a range of techniques, including GC-MS, NMR, SEC, assessment of pH, osmolality, buffering capacity, viscoelasticity, viscoelastic interactions with human saliva, hydrocolloid network formation, and in vitro cell adhesion. For this purpose, a physiologically adapted adhesive test was developed using human buccal epithelial cells. The results show that lichenan is the most promising candidate to mimic the properties of MUC5B. By adjusting the pH, osmolality, and buffering capacity with K2HPO4, it was shown that lichenan exhibited high cell adhesion, with a maximum detachment force that was comparable to that of unstimulated whole mouth saliva.