Various studies have shown that Hypogymnia physodes are a source of many biologically active compounds, including lichen acids. These lichen-specific compounds are characterized by antioxidant, antiproliferative, and antimicrobial properties, and they can be used in the cosmetic and pharmaceutical industries. The main aim of this study was to optimize the composition of natural deep eutectic solvents based on proline or betaine and lactic acid for the extraction of metabolites from H. physodes. The design of the experimental method and the response surface approach allowed the optimization of the extraction process of specific lichen metabolites. Based on preliminary research, a multivariate model of the experiment was developed. For optimization, the following parameters were employed in the experiment to confirm the model: a proline/lactic acid/water molar ratio of 1:2:2. Such a mixture allowed the efficient extraction of three depsidones (i.e., physodic acid, physodalic acid, 3-hydroyphysodic acid) and one depside (i.e., atranorin). The developed composition of the solvent mixtures ensured good efficiency when extracting the metabolites from the thallus of H. physodes with high antioxidant properties.

The increasing incidence of dermatological diseases prompts the search for new natural methods of treatments, and lichens, with their special symbiotic structure, are a little-known and promising source of biologically active substances. Seven lichen species, Cladonia unicialis (L.) Weber ex F.H. Wigg. (Cladoniaceae), Evernia prunastri (L.) Ach. (Parmeliaceae), Hypogymnia physodes (L.) Nyl. (Parmaliaceae), Parmelia sulcata (Taylor) (Parmeliaceae), Physcia adscendens (Fr.) H. Olivier (Physciaceae), Pseudoevernia furfuracea (L.) Zopf (Parmeliaceae), and Xanthoria parietina (L.) Th. Fr. (Teloschistaceae), were used in our experiment. We identified different metabolites in the acetone extracts of all the lichen species. Based on the high-performance liquid chromatography analysis, the content of lichen substances in the extracts was evaluated. The impact of the individual lichen-specific reference substances, compared to the lichen extracts, on the viability of keratinocytes (HaCaT cell line) and fibroblasts (BJ cell line) and on the activity of selected skin-related enzymes was investigated. Our results revealed that only emodin anthrone at a concentration of 200 mg/L was cytotoxic to keratinocytes and fibroblasts in both cell viability assays. In turn, the C. uncialis extract was only cytotoxic to keratinocytes when used at the same concentration. The other tested treatments showed a positive effect on cell viability and no cytotoxicity or indeterminate cytotoxicity (shown in only one of the tests). Elastase and collagenase activities were inhibited by most of the lichen extracts. In turn, the individual lichen compounds (with the exception of evernic acid) generally had an undesirable stimulatory effect on hyaluronidase and collagenase activity. In addition, almost all the tested compounds and extracts showed anti-inflammatory activity. This suggests that some lichen compounds hold promise as potential ingredients in dermatological and skincare products, but their safety and efficacy require further study. The high cytotoxicity of emodin anthrone highlights its potential use in the treatment of hyperproliferative skin diseases such as psoriasis.

Soybeans are commonly known as a valuable source of biologically active compounds including isoflavones as well as allantoin and alpha-hydroxy acids. Since these compounds exhibit skin therapeutic effects, they are widely used in the cosmetic and pharmaceutical industries. The presented paper shows the optimization of three solvent systems (ethanol, water, and 1,3-propanediol) to increase the extraction efficiency of isoflavones (daidzin, genistin, 6″-O-malonyldaidzin, 6″-O-malonylglycitin, 6″-O-malonylgenistin), allantoin, and alpha-hydroxy acids (citric acid, malic acid) from soybean leaves. A simplex centroid mixture design for three solvents with interior points was applied for the experimental plan creation. Based on the obtained results of metabolite extraction yield in relation to solvent composition, polynomial regression models were developed. All models were significant, with predicted R-squared values between 0.77 and 0.99, while in all cases the model’s lack of fit was not significant. The optimal mixture composition enabling the maximization of extraction efficiency was as follows: 32.9% ethanol, 53.9% water, and 13.3% propanediol (v/v/v). Such a mixture composition provided the extraction of 99%, 91%, 100%, 92%, 99%, 70%, 92%, and 69% of daidzin, genistin, 6″-O-malonyldaidzin, 6″-O-malonylglycitin, 6″-O-malonylgenistin, allantoin, citric acid, and malic acid, respectively. The solvent mixture composition developed provides a good extraction efficiency of the metabolites from soybean leaves and high antioxidant properties.

Lichen-specific natural products exhibit a wide range of biological activities, which makes them potentially useful in the pharmaceutical, cosmetic, and nutritional industries. In the present study, a capillary electrophoresis method was developed and optimized for the separation of seven major metabolites, physodic acid, 3-hydroxyphysodic acid, atranorin, physodalic acid, chloroatranorin, salazinic acid, and protocetraric acid, found in Hypogymnia physodes. The optimization was performed using a design of experiments approach, focusing on four critical parameters: boric acid concentration, deoxycholic acid concentration, methanol content, and buffer pH. The overall separation efficiency was used as the response factor for optimization. The optimal separation conditions were achieved using a buffer composed of 60 mM boric acid, 70 mM deoxycholic acid, and 14% methanol at pH 9.6. The validated method was subsequently applied for the chemophenetic analysis of 28 lichen species belonging to the families Cladoniaceae, Parmeliaceae, Physciaceae, Ramalinaceae, and Teloschistaceae. In addition to the above-mentioned lichen compounds, the lichens examined showed the presence of evernic acid, usnic acid, and physicon. The developed CE method offers a reliable and efficient tool for the characterization of lichen metabolites, with potential applications in both botany and natural product research.

Usnic acid (UA) is one of the most extensively studied specialized metabolites of lichens, attracting considerable interest due to its antimicrobial, anti-inflammatory, and cytotoxic properties. The efficiency of UA extraction from lichens depends on multiple interrelated biological and technological factors. This systematic review aims to synthesize and critically evaluate reported strategies for UA extraction from wild-grown lichen biomass, with particular emphasis on extraction efficiency, practicality, and application potential. This systematic literature review, based on the Scopus database was conducted by including original research articles reporting UA extraction from wild-growing lichens. The analysis covered species selection, sample pre-treatment, solvent type, and extraction methodology. A total of 117 studies were included. Due to the predominantly non-polar nature of UA, higher extraction efficiencies were generally achieved using solvents, including acetone, supercritical CO₂, vegetable oils, and lipophilic green solvent systems. Pre-treatment strategies such as grinding or flaking significantly enhanced extraction performance by improving mass transfer. Alongside conventional methods (maceration, reflux, Soxhlet), non-conventional techniques such as Supercritical Fluid Extraction (SFE), Ultrasound- (UAE), and Microwave-Assisted Extraction (MAE) enabled faster and more selective UA extraction with reduced solvent use. Notably, SFE have been reported as particularly promising in terms of selectivity, process control, and potential suitability for scale-up, with commercially available supercritical CO₂ extracts of Usnea species supporting the feasibility of this approach. This review provides a consolidated and application-oriented overview of UA extraction, highlighting strategies that balance efficiency, selectivity, sustainability, and practical implementation.