Isoquinoline alkaloids such as sanguinarine and chelerythrine have demonstrated antimicrobial activity against a range of microorganisms; however, their effects on Leptospira spp. remain poorly characterized. In this study, we evaluated the in vitro susceptibility of multiple Leptospira strains to sanguinarine chloride, chelerythrine chloride, and a combined sanguinarine–chelerythrine fraction. Bacterial responses were assessed using flow cytometry, metabolic activity assays, and optical density measurements. The tested com-pounds exhibited limited and strain-dependent effects. The most pronounced response was observed in Leptospira interrogans serovar Australis (strain Balico), where chelerythrine chloride consistently reduced viability and metabolic activity under the conditions tested. In contrast, other strains showed minimal or inconsistent responses. The combined fraction did not demonstrate enhanced activity compared to individual compounds. These findings indicate that isoquinoline alkaloids exert variable and generally modest effects on Leptospira spp., highlighting substantial inter-strain heterogeneity in susceptibility. While the observed activity is limited, this work provides a preliminary assessment of these compounds in Leptospira and underscores the need for further studies to clarify their mechanisms of action and potential relevance.

Acorns represent a traditional seasonal feed resource for pigs in Mediterranean production systems; however, comparative information on the nutritional composition and safety of acorns from Central European oak species remains scarce. The present study comprehensively characterized acorns of Quercus robur, Q. petraea, and Q. rubra to assess their potential relevance to pig nutrition based on their chemical composition. Proximate composition, mineral content, fatty acid and amino acid profiles, phenolic compounds, tannins, triterpenoid acids, and heavy metal concentrations were determined using analytical methods. Differences among the analyzed acorn samples were observed in nutritional composition and phytochemical profile. Acorns from all three species were characterized by a high proportion of unsaturated fatty acids, particularly oleic acid, a favourable amino acid profile, and the presence of phenolic compounds, tannins, and triterpenoid acids. Heavy metal concentrations remained below the maximum levels established by European legislation for feed materials, indicating that the analyzed acorns showed no heavy-metal-related exceedances of the applicable European limits for feed materials. The results indicate that acorns from Central European oak species represent a locally available feed resource of potential relevance to pig nutrition. This comprehensive chemical characterization provides a scientific basis for future nutritional studies and further evaluation of acorns as sustainable feed materials.

Background: Pain is a major clinical and socioeconomic problem worldwide. The available therapies are not always effective and are often associated with the multiple adverse effects that reduce their clinical application. Natural compounds are an important group of pharmaceuticals that may be used in pain management. We aimed to investigate the analgesic activity of the sanguinarine–chelerythrine from Coptis chinensis. Methods: The analgesic and anti-inflammatory activity of the sanguinarine–chelerythrine fraction of C. chinensis extract (SC 5 and 10 mg/kg), sanguinarine (SAN 1 and 2 mg/kg) and chelerythrine (CHEL 4 and 8 mg/kg) was assessed in tail flick and formalin tests. A microscopic and macroscopic examination of stomach mucosae was performed. TNFα and MMP-9 levels were measured with ELISA kits. Results: Morphine (MORF), CHEL and SC prolongated the tail withdrawal latency, with comparable analgesic activity between MORF and CHEL 8 mg/kg. MORF, CHEL 8 mg/kg, and SAN 2 mg/kg ameliorated the pain reaction in the neurogenic phase of the formalin test. In the inflammatory phase of the formalin test, all tested substances exerted analgesic activity. SAN, CHEL and SC additionally reduced TNFα and MMP-9 secretion. Conclusions: Our results confirmed analgesic effects of CHEL and SC with CHEL analgesic activity comparable to MORF. All investigated substances exerted significant anti-inflammatory activity without concomitant gastrotoxicity.

Breast cancer is the second most commonly diagnosed cancer worldwide, with the highest mortality of all cancers. In most cases, breast cancer is estrogen-dependent, which means cancer cells express estrogen receptors. Phytoestrogens, due to their structural similarities with estrogen, are capable of binding to estrogen receptors and are therefore also referred to as selective estrogen receptor modulators. They can be either estrogen agonists or antagonists and influence the risk of breast cancer. Two prominent representatives of phytoestrogens are isoflavones, which are found in soy products, and lignans, which are found in flaxseed, vegetables, and certain nuts. Phytoestrogens may be used in breast cancer prevention. They can reduce the risk of breast cancer incidence and its recurrence, yet mostly in postmenopausal women, and the reasons for this have not yet been explained. Plausible mechanisms of cancer risk-lowering properties of phytoestrogens include antiestrogenic action, influencing estrogen metabolism, sex hormone concentrations, and menstrual cycle length. These actions are not observed in every consumer. Phytoestrogens are found in plants in glycoside form (bound with sugar molecules), and at least part of them must be metabolized to their biologically active metabolites (equol for isoflavones and enterolactone for lignans) by intestinal microbiota. Bacterial strains capable of phytoestrogen metabolism have been identified. This article thoroughly describes phytoestrogens' metabolism and their associations with breast cancer risk and outcomes.

Buds of poplar trees (Populus species) are often covered with sticky, usually polyphenol-rich, exudates. Moreover, accessible data showed that some Populus bud extracts may be excellent antibacterial agents, especially against Gram-positive bacteria. Due to the fragmentary nature of the data found, we conducted a systematic screening study. The antimicrobial activity of two extract types (semi-polar—ethanolic and polar—ethanolic-water (50/50; V/V)) from 27 bud samples of different poplar taxons were compared. Antimicrobial assays were performed against Gram-positive (five strains) and Gram-negative (six strains) bacteria as well as fungi (three strains) and covered the determination of minimal inhibitory, bactericidal, and fungicidal concentrations. The composition of extracts was later investigated by ultra-high-performance liquid chromatography coupled with ultraviolet detection (UHPLC-DAD) and with electrospray-quadrupole-time-of-flight tandem mass spectrometry (UHPLC-ESI-qTOF-MS). As a result, most of the extracts exhibited good (MIC ≤ 62.5 µg/mL) or moderate (62.5 < MIC ≤ 500 µg/mL) activity against Gram-positives and Helicobacter pylori, as well as fungi. The most active were ethanolic extracts from P. trichocarpa, P. trichocarpa clone ‘Robusta’, and P. tacamahaca × P. trichocarpa. The strongest activity was observed for P. tacamahaca × P. trichocarpa. Antibacterial activity was supposedly connected with the abundant presence of flavonoids (pinobanksin, pinobanksin 3-acetate, chrysin, pinocembrin, galangin, isosakuranetin dihydrochalcone, pinocembrin dihydrochalcone, and 2′,6′-dihydroxy-4′-methoxydihydrochalcone), hydroxycinnamic acids monoesters (p-methoxycinnamic acid cinnamyl ester, caffeic acid phenethylate and different isomers of prenyl esters), and some minor components (balsacones).

The leaves of Ilex paraguariensis (known as Yerba mate), used as a popular beverage, are a very well-recognized plant material with various biological activities, including analeptic (because of caffeine), anti-obesity (phenolics, saponins), antimicrobial, and antiviral (phenolics, saponins). Here, the chemical compositions of the leaves of two European Ilex species (× meserveae and aquifolium) with three varieties each were investigated. The terpenoid, saponin, and polyphenolic fractions were submitted for LC-MS or GC-MS analysis against a standard Mate leaf. In addition, the aroma profiles of all the species were analysed using HS-SPME-Arrow prior to GC-MS analysis. All fractions were subjected to antiviral and cytotoxic assays. We found 86 compounds in all accessions, with limonene, linalool, and p-cymene being predominant. There were minor similarities between the volatile compositions of the European and South American species. We found ursolic and oleanolic acid to be the main compounds in the terpenoid fraction. Mono-caffeoylquinic acids and di-caffeoylquinic acids were the main constituents of the polar fractions. About 180 compounds from the saponin group were tentatively identified, of which 9 and 3 were selected as distinctive markers for I. meserveae and I. aquifolium, respectively. Based on chemical screening, I. aquifolium Silver Queen was chosen as the source of terpenoid and saponin fractions and polyphenol extracts. The most substantial inhibition of cancer cell growth was observed with saponin in the case of the MCF7 (human breast cancer) cell line, while for LoVo and L929 cell lines (human colorectal cancer and reference mouse fibroblasts), it was slightly weaker. These results should be analysed further as a promising chemoprevention of colorectal and gastrointestinal cancers. Saponin and polyphenolic extracts exhibited similar activities against HSV-1 and HAdV-5, with 4-log reduction in virus titres. This study focuses our attention on a field of potential antiviral formulations derived from European holly.

The European Food Safety Authority recommends C. incanus as a natural source of antioxidants. Its activity is essentially determined by polyphenols, although specific compounds are not finally indicated. The available plant material comes from different subspecies and locations, which can lead to differences in chemical composition and potency. For this reason, we conducted a detailed analysis of the polyphenol content and antioxidant activity of 52 different C. incanus teas from Turkey, Albania, Greece, and unspecified regions. We focused special attention on ellagitannins, which have not been properly determined so far. Besides oxidative stress, hyperglycemia is an essential component of cardiometabolic diseases. Therefore, in subsequent experiments, we evaluated the ability of C. incanus extracts and individual polyphenols to inhibit α-glucosidase. Using statistical methods, we analyzed how differences in chemical composition affect activity. The results showed that C. incanus is a rich source of ellagitannins (2.5–19%), which dominate among polyphenols (5.5–23%). Turkish-origin products had higher ellagitannin content and a greater antioxidant effect (FRAP, ABTS) than Albanian and Greek products. In contrast, the flavonoid and phenolic acid contents and DPPH values were at similar levels in all products. An in-depth analysis of their composition indicated that all groups of polyphenols are involved in the antioxidant effect, but a significant contribution can be attributed to ellagitannins and flavonoids. C. incanus extracts showed a high capacity to inhibit α-glucosidase activity (IC50 125–145 μg/mL). Ellagitannins were the most effective inhibitors (IC50 0.7–1.1 μM), with a potency exceeding acarbose (3.3 mM). In conclusion, C. incanus, due to the presence of ellagitannins and flavonoids, exhibits powerful antioxidant and α-glucosidase inhibitory effects.

Inflammatory diseases are a common therapeutic problem and nonsteroidal anti-inflammatory drugs are not deprived of side effects, of which ulcerogenic activity is one of the most frequent. The aim of the study was to evaluate the anti-inflammatory activity of the sanguinarine-chelerythrine (SC) fraction of Coptis chinensis and its influence on the integrity of gastric mucosa. The study was conducted on sixty male rats randomly divided into six experimental groups: two control groups (a negative control group CON and a positive control group CAR); three groups receiving an investigational fraction of C. chinensis (1, 5, 10 mg/kg i.g.) named SC1, SC5, and SC10, respectively; and a group receiving indomethacin (IND) (10 mg/kg i.g.) as a reference drug. In all animals, the carrageenan-induced paw oedema was measured; PGE2 release, TNFα production, and MMP-9 concentration in inflamed tissue were determined. Additionally, the macroscopic and microscopic damage of gastric mucosa was evaluated. Administration of SC dose-dependently inhibited the second phase of carrageenan rat paw oedema and PGE2 release, decreased the production of TNFα, and reduced the concentration of MMP-9, and the efficacy of the highest dose was comparable to the effect of IND. Contrary to IND, no gastrotoxic activity of SC was detected. The investigated sanguinarine-chelerythrine fraction of C. chinensis seems to be a promising candidate for further research on new anti-inflammatory and analgesic drugs characterized with a safer gastric profile compared to existing NSAIDs.

Strawberry leaves are considered a valuable waste material; so far, mainly due to their antioxidant properties. Since the annual production of this crop is high, our study aimed to thoroughly examine the chemical composition and antidiabetes-related bioactivity of Fragaria × ananassa leaf of its popular and productive cultivar Senga Sengana. Leaves from three different seasons, collected after fruiting, were extensively analyzed (UHPLC-qTOF-MS/MS, HPLC-DAD). Some individual components were isolated and quantified, including specific flavonol diglycosides (e.g., 3-O-[β-xylosyl(1‴→2″)]-β-glucuronosides). The separated quercetin glycosides were tested in an antiglycation assay, and their methylglyoxal uptake capacity was measured. In addition, the biodegradable polyester precursor 2-pyrone-4,6-dicarboxylic acid (PDC) was confirmed at relatively high levels, providing further opportunity for strawberry leaf utilization. We want to bring to the attention of the food, pharmaceutical, and cosmetic industries the Senga Sengana strawberry leaf as a new botanical raw material. It is rich in PDC, ellagitannins, and flavonols—potent glycation inhibitors.

Poplar bark and leaves can be an attractive source of salicylates and other biologically active compounds used in medicine. However, the biochemical variability of poplar material requires a standardization prior to processing. The official analytical protocols used in the pharmaceutical industry rely on the extraction of active compounds, which makes their determination long and costly. An analysis of plant materials in their native state can be performed using vibrational spectroscopy. This paper presents for the first time a comparison of diffuse reflectance in the near- and mid-infrared regions, attenuated total reflection, and Raman spectroscopy used for the simultaneous determination of salicylates and flavonoids in poplar bark and leaves. Based on 185 spectra of various poplar species and hybrid powdered samples, partial least squares regression models, characterized by the relative standard errors of prediction in the 4.5-9.9% range for both calibration and validation sets, were developed. These models allow for fast and precise quantification of the studied active compounds in poplar bark and leaves without any chemical sample treatment.

Ilex paraguariensis, the holly tree, is a plant with recognized biological properties, whose aqueous infusions are known as “Yerba mate”, that regulate lipid metabolism, reduce obesity, and improve brain stimulation. In the present study, the effect of standardized saponin and terpenoid fractions of a European taxon, Ilex aquifolium, on blood biochemical parameters in a rat model of metabolic disorder, (fa/fa) Zucker, are presented. The profiles of the volatile fractions of two species and six European varieties of Ilex were investigated. After selecting the best variety, the saponin and terpenoid fractions were isolated and standardized, and animals were fed 10 mg kg−1 b.w. for 8 weeks. A statistically significant decrease in liver adiposity was observed, confirmed by histology and quantitative identification (gas chromatography–mass spectrometry analyses of hepatic lipids. RT-qPCR analysis of gene expression in the aorta revealed that the administration of the terpenoid fraction downregulated LOX-1, suggesting a reduction in atherosclerotic stimuli. In addition, a statistically significant reduction (p < 0.05) in PPARγ for the saponin fraction was observed in the liver. The expression of the ACAT-1 gene in the liver, responsible for the formation of cholesterol esters, increased significantly in the group receiving the terpenoid fraction compared to the control, which was also confirmed by the analysis of individual blood biochemical parameters. The opposite effect was observed for saponins. Taking the above into account, it is shown for the first time that Ilex aquifolium can be a source of compounds that positively influence lipid metabolism.

Plants from the Ilex genus are known for properties such as antimicrobial and anti-inflammatory activity, can act as antiobesity agents and thus can be helpful in medicine. Some holly species, such as Ilex paraguariensis (widely known in the form of popular beverage: yerba mate), have been investigated, while others have been partially researched or remain unknown. Therefore, we performed qualitative and quantitative phytochemical analyses and screened antimicrobial properties of lesser-studied species (I. aquifolium L., I. aquifolium ‘Argentea Marginata’ and I. × meserveae ‘Blue Angel’). I. paraguariensis was used as a standard species for comparison purposes. Investigations were performed on water extracts due to their expected activity and composition. Antimicrobial research included evaluating minimal inhibitory, bactericidal (Staphylococcus aureus and Escherichia coli) and fungicidal concentration (Candida albicans, Alternaria alternata, Fusarium oxysporum, and Aspergillus niger) of extracts. The influence of the extracts on the production, eradication, and viability of bacterial biofilms was also analysed. It was established that Ilex paraguariensis possesses the richest profile of hydroxycinnamic acids derivatives in terms of component concentration and diversity. Ilex spp., especially I. × meserveae, contain a slightly higher amount of flavonoids and more different flavonoid derivatives than I. paraguariensis. However, the strongest antibacterial activity was shown by I. aquifolium L. and its cultivar ‘Argentea Marginata’ in terms of minimal inhibitory, bactericidal and fungicidal concentration, and biofilm assays. Extracts from both species significantly reduced the biofilm viability of S. aureus as well, which may be of use in the production of multicomponent lavaseptics, antiseptics, diuretics (supporting urinary tract infection therapy) and, due to their action on fungi, additives to growth media for specific fungi. The significant content of saponins enables Ilex extracts to be used as natural emulsifiers, for example, in cosmetics. Moreover, relatively high chlorogenic acid and rutin content may suggest use of Ilex spp. to treat obesity, digestive problems, in chemoprevention, and as preservatives in the food industry.

Senga Sengana is a common strawberry (Fragaria x ananassa Duch.) cultivar growing in southern Poland. From leaves of this popular horticultural plant six triterpenoids were obtained including one glycoside nigaichigoside F1. All compounds were isolated by repeated silica gel column chromatography and preparative HPLC on Reverse phase RP-18. Their structures were evaluated by modern HR-MS and NMR techniques. This is the first report concerning triterpenoids constituents in Senga Sengana cultivar leaves and the only one that describes isolation of these compounds from the strawberry leaves. From all compounds nigaichigoside F1 is the dominant constituent. Compound T1 (3 beta, 19 alpha, 23 alpha-trihydroxy-2-oxoolean-12-en-28-oic acid) is a new triterpenoid, not previously described in nature. As triterpenoids are responsible for many biological effects the strawberry leaf extracts enriched in triterpenoid constituents may also be involved in antifungal, antitumor, antidiabetic and antinociceptive activities. The latter two activities could be especially linked to the presence of euscaphic acid and nigaichigoside F1, respectively. Overall, this study suggests that strawberry leaves are a promising waste and a valuable source of bioactive compounds for the food and pharmaceutical industry.

Therapeutic properties of Ilex species are widely used in natural medicine. Ilex × meserveae may become a potential substitute for Ilex paraguariensis (Yerba Mate). As a part of the preliminary safety verification of this European Ilex hybrid vs. Yerba Mate, an eight-week study concerning the impact of regular administration of leaves of both species on kidneys was conducted. The standard water infusion and three dominant fractions of Ilex × meserveae leaves’ constituents (polyphenols, saponins and less polar terpenoids) were separately tried on 96 male Wistar rats divided into 8-member groups. Animals were divided into two basic nutritional groups: the first one was rats fed standard feed and the second on was rats fed with high-cholesterol diet (20 g of cholesterol per kg of standard feed). Postmortem morphometric evaluation of stained kidney samples concerned the filtration barrier elements, which are crucial in proper diuresis. The results showed that saponins present in the hydroalcoholic dry extract (administered in a dose of 10 mg/kg of body weight/day) as well as in water infusions (1:20) from Ilex × meserveae and Ilex paraguariensis do not demonstrate nephrotoxicity but conversely, have a protective role on kidney status in animals fed with a normal diet and in a high-cholesterol diet.

Ethnopharmacological relevance: Theriac is considered the most popular cure-all multi-ingredient medicine and has been used for more than two millennia. It has also been used as one of the most important anti-epidemic drugs up to the 19th c., treated as an emergency medicine in case of e.g. bubonic plague. Aim of the study: Until now, no reliable information regarding the pharmacological effect of the treacle was available, including its possible toxic or narcotic properties. In order to change the state of knowledge in this matter we have selected the Theriac recipe that had been actually used for producing the treacle in 1630, which was confirmed by the official municipal documents of the time. Methods: The recipe was written in Latin, with the use of pre-Linnean nomenclature and then apothecary common names, which required translation into the modern scientific language in order to get reliable pharmacological conclusions. The information from historical sources has been compiled with the pharmacological data concerning the most potent compounds, which for the first time made it possible to calculate the amounts of active compounds in the doses taken by then patients. Results: Only two species included in Theriac can be harmful in humans: poppy and sea squill, but in both cases the calculated quantity of morphine and cardiac glycosides, respectively, were below toxic level. There are no indications, both from the historical and pharmacological point of view, for Theriac being toxic or narcotic in patients, when used as prescribed. Conclusions: As for now, the most probable is that the treacle owed its postulated efficacy in the main indications to the placebo effect. Still, the results should be further confirmed by reconstructing the actual Theriac and subjecting it to modern tests and analyses.

One of the modern nutritional trends is to enthusiastically look for natural products that can be considered functional food and be a source of ingredients with a health-promoting effect. Today, many food manufacturers offer Cistus × incanus leaves to prepare common self-preparations (e.g., infusions) or as ready-to-use dietary supplements. Cistus × incanus (rock rose, pink rock-rose, hoary rock-rose), belonging to the family Cistaceae, is widespread in Mediterranean countries. For many years, cistus extracts and its aromatic resin have been used in traditional Middle East medicine to treat, among others, colds, fever, stomach problems, and skin wounds. In past years, this plant was rediscovered by the public. Due to the growing popularity of Cistus products, the most recent scientific literature on this subject is reviewed here. This article aims to present the latest research results on the phytochemical composition of Cistus × incanus and the impact of its consumption on human health. Particular emphasis is put on antioxidant, anti-inflammatory, antibacterial, antiviral, and antiproliferative activities and support of digestive system functions. Studies have shown that the main active ingredients of Cistus × incanus are flavonoid compounds, including flavonol glycosides (myricetin, quercetin, kaempferol), flavan-3-ols, and tannins. It was demonstrated that the presence of these compounds determines the therapeutic and health-promoting properties of cistus leaves and its products. That applies primarily to a strong antioxidant effect, which may reduce the risk of noncommunicable diseases, including cardiovascular diseases, neurodegenerative diseases, and cancer. Cistus preparations are also recommended as immunostimulants, supporting the treatment of bacterial and viral infections. Labdanum oleoresin and essential oil are a valuable source of substances with strong antibacterial and anti-inflammatory properties, which can be used in the future in the production of pharmaceutical and cosmetic preparations, and also serve as a natural food preservative.

The leaves of Cistus x incanus L. (pink rock-rose) are used as plant material rich in antioxidants by the pharmaceutical and food industry. The main components which occur in pink rock-rose are polyphenols, especially flavonoids (glycosides of flavonols) and tannins (proanthocyanidins and ellagitannins). Derivatives of flavonols and flavan-3-ols as bioactive compounds of this species were identified previously. However, the full composition of ellagitannins has not been characterized yet. In this context, the study aimed to characterize the chemical composition of pink rock-rose ellagitannins and to identify and elucidate structures of major compounds from this group. In conclusion, the new ellagitannin cistusin together with well-known terflavin A and punicalagin from C. incanus leaves were isolated. Their structures, including the gallagyl, flavogallonyl, valoneoyl, and hexahydroxydiphenoyl acyl groups, were revealed by spectroscopic evidence (HR-ESI-MS, 1D and 2D-NMR).

So far, only a few primrose species have been analyzed regarding their saponin composition and content. Moreover, the roots of only two of them are defined by the European Union (EU) Pharmacopoeia monograph and commercially utilized by the pharmaceutical industry. Thus, this study intended to find some new sources of main triterpene saponins from Primulae radix, namely primulasaponins I and II together with the closely related sakurasosaponin. Using isolated standards, UHPLC-ESI-HRMS served to assess over 155 Primulaceae members qualitatively and quantitatively. Nine examples of plants accumulating over 5% of primulasaponin I in their roots were found. Among them, in one case, it was found as the almost sole secondary metabolite with the concentration of 15–20% (Primula grandis L.). A reasonable content of primulasaponin II was found to be typical for Primula vulgaris Huds. and P. megaseifolia Boiss. & Bal. The sakurasosaponin level was found in seven species to exceed 5%. The finding of new, single and rich sources of the abovementioned biomolecules among species that were never analyzed phytochemically is important for future research and economic benefit. The chemotaxonomic significance of the occurrence of these three saponins in Primulaceae is discussed.

A magistral preparation based on the sage leaf decoction or infusion is one of the most popular herbal prescriptions prepared in pharmacies in Lower Silesia (Poland) for gargling in the case of pharyngitis or mouth infections. Other components of this preparation are boric acid, aluminum(III) acetotartrate or Burowis solution, and glycerol. The study aimed to investigate the polyphenolic composition and relations between herbal and chemical ingredients that are present in this mixture in comparison with the traditional aqueous galenic forms - infusion and decoction. The analysis was performed using high-performance liquid chromatography with diode-array detector (HPLC-DAD) and electrospray ionization mass spectrometry (ESI-MS) methods. The main polyphenolic ingredients of sage leaf were rosmarinic acid (RA) and luteolin 7-O-beta-glucuronide. The RA predominance was observed in the infusion, decoction and magistral preparation. Comparing the contents of analyzed polyphenols, it was confirmed that their levels in the decoction were higher than in the infusion. Thus, the pharmacological activity of the examined magistral preparation results from the presence of boric acid, aluminum(III) salts and polyphenolic components of sage leaf hot water extract. The succeeding study showed that in the presence of excess salts of aluminum(III) and boric acid, sage polyphenols are partially soluble in an aqueous medium (mainly phenolic acids) and complexed as precipitated sediment (especially flavonoids). The aqueous solutions of this preparation obtained according to the recommendation are cloudy, which suggests limited solubility of the formed complexes. Therefore, the therapeutic activity should be associated with the presence of water-soluble caffeic acid esters like rosmarinic acid. The influence of luteolin glycosides coordination complexes is not evident. The obtained results also confirm the stability of the examined prescription formulation in the conditions of cold storage (4-6 degrees C) within 7 days from its production.

The phytochemistry of the genera Androsace, Cortusa, Soldanella, and Vitaliana, belonging to the Primulaceae family is not well studied so far. Hence, in this paper, we present the results of UHPLC-MS/MS analysis of several primrose family members as well as isolation and structure determination of two new saponins from Vitaliana primuliflora subsp. praetutiana. These two nor-triterpenoid saponins were characterized as (23S)-17α,23-epoxy-29-hydroxy-3β-[(O-β-d-glucopyranosyl-(1→2)-O-α-l-rhamnopyranosyl-(1→2)-O-β-d-glucopyranosyl-(1→2)-O-α-l-arabinopyranosyl-(1→6)-β-d-glucopyranosyl)oxy]-27-nor-lanost-8-en-25-one and (23S)-17α,23-epoxy-29-hydroxy-3β-[(O-α-l-rhamnopyranosyl-(1→2)-O-β-d-glucopyranosyl-(1→2)-O-α-l-arabinopyranosyl-(1→6)-β-d-glucopyranosyl)oxy]-27-nor-lanost-8-en-25-one, respectively. Their structures were determined by high resolution mass spectrometry (HRMS), tandem mass spectrometry (MS/MS), one- and two-dimensional nuclear magnetic resonance spectroscopy (1D-, and 2D-NMR) analyses. So far, the 27-nor-lanostane monodesmosides were rarely found in dicotyledon plants. Therefore their presence in Vitaliana and also in Androsace species belonging to the Aretia section is unique and reported here for the first time. Additionally, eleven other saponins were determined by HRMS and MS/MS spectra. The isolated lanostane saponins can be considered as chemotaxonomic markers of the family Primulaceae.