Phytochemical Profiling and Biological Activities of Rhododendron Subsect. Ledum: Discovering the Medicinal Potential of Labrador Tea Species in the Northern Hemisphere.

Rhododendron subsect. Ledum is a distinct taxonomic subdivision within the genus Rhododendron, comprising a group of evergreen shrubs and small trees. This review will comprehensively analyse the phytochemical profiles and biological properties of the Rhododendron subsect. Ledum species subsect. Ledum consists of eight plant species indigenous to temperate and subarctic regions of the Northern Hemisphere, collectively known as Labrador tea. Recent investigations have concentrated on the phytochemical constituents of these plants due to limited data, emphasizing their evergreen nature and potential industrial significance. This review summarizes their major phytochemical constituents, including flavonoids, phenolic acids, and terpenoids, and discusses their potential biological activities, such as antioxidant, anti-inflammatory, antimicrobial, antitumor, hypoglycemic, hepatoprotective, neuroprotective, and cardioprotective effects. Traditional uses of these plant species align with scientific findings, emphasizing the significance of these plants in traditional medicine. However, despite promising results, gaps exist in our understanding of specific compounds' therapeutic effects, necessitating further research for comprehensive validation. This review serves as a valuable resource for researchers, identifying current knowledge, uncertainties, and emerging trends in the study of the Rhododendron subsect. Ledum species.

Comparative Phytoprofiling of Achillea millefolium Morphotypes: Assessing Antioxidant Activity, Phenolic and Triterpenic Compounds Variation across Different Plant Parts.

Achillea millefolium L., commonly known as yarrow, is a versatile and widely distributed plant species with a rich history of ethnopharmacological significance. This study aimed to evaluate the comparative differences of A. millefolium inflorescence morphotypes. The phytochemical profile of white and pink inflorescence morphotypes was characterised by a complex of thirty-four phenolic and triterpene compounds. The species has distinct morphotypes of white and pink inflorescence. Phenolic and triterpenic profiles were determined, and individual compounds were quantified in inflorescence, leaf, and stem samples of two morphotypes tested. The antioxidant activity of plant extracts was evaluated by free radical scavenging (ABTS) and ferric-reducing antioxidant power (FRAP) assays. Caffeoylquinic acids predominated in all parts of the plant tested. Chlorogenic acid and 3,5-dicaffeoylquinic acid were the principal compounds in the phenolic profile. Betulin, betulinic acid, and α-amyrin were the prevailing triterpenic components in the triterpenic profiles of Achillea millefolium morphotypes. The predominant flavonoids in inflorescences were flavones, while in leaves, flavonols were the organ-specific compounds. The quantitative differences were observed between plant parts of morphotypes. Leaves consistently displayed the highest amounts of identified compounds and have been testified as the main source of antioxidant activity. Overall, white inflorescences accumulated a higher total amount of compounds compared to pink ones. The observed differences between morphotypes derived from the same population reflect the differences in specialised metabolites and their chemotypes. This study addresses gaps in knowledge, particularly in phenolic and triterpenic profiling of coloured inflorescence morphotypes, enhancing our understanding of chemotypes and morphotypes within the species.

Phytoprofiling of Sorbus L. Inflorescences: A Valuable and Promising Resource for Phenolics.

The leaves and fruits of various Sorbus L. genotypes have long ethnopharmacological and food-usage histories, but inflorescences are still underutilized and neglected materials with scarce phytochemical scientific evidence. The aim of this study was to determine the phenolic profiles of inflorescence extracts of 26 Sorbus species, genotypes, and cultivars. HPLC and UPLS with MS detection were applied, and coupled data revealed unique phytochemical phenolic profiles. Neochlorogenic and chlorogenic acids were the key compounds, reaching up to 5.8 mg/g of dw. Rutin, isoquercitrin, quercetin 3-O-malonylglucoside, isorhamnetin 3-O-rutinoside, sexangularetin derivative, and kaempferol acetyl hexoside were detected in all Sorbus inflorescence samples. Overall, high quantitative heterogeneity across the various Sorbus genotypes was found by profiling. Phenolic fingerprint profiles and sexangularetin derivatives could serve as markers in authenticity studies and quality control schemes. The species S. amurensis, S. arranensis, S. commixta, and S. discolor and the cultivars 'Chamsis Louing', 'Coral Beauty', and 'Edulis' could be used as target genotypes for production of smart and innovative inflorescence matrix-based ingredients.

Phytochemical Diversity and Antioxidant Potential of Wild Heather (Calluna vulgaris L.) Aboveground Parts.

Calluna vulgaris L. (heather) is a traditional medicinal plant with anti-inflammatory and calming activities that are determined by the notable amounts of phytochemicals. The evaluation of different populations of plants that accumulate great amounts of bioactive compounds are requisite for the quality determination of plant materials and medicinal and nutritional products. The assessment of natural resources from a phytogeographic point of view is relevant. Phytochemical analysis of heather samples was carried out using spectrophotometric methods and HPLC-PDA techniques, while antioxidant activity was determined using ABTS and FRAP assays. A significant diversification of phenolic and triterpenic compounds and antioxidant activity was determined in the heather samples collected in distinct habitats. Natural habitats, due to their characteristic chemical heterogeneity, lead to the diversity of indicators characterizing the quality of plant raw materials. Chlorogenic acid and hyperoside were found to be predominant among the phenolic compounds, while ursolic, oleanolic acids, and uvaol prevailed among the triterpenic compounds. Thus, these compounds can be suggested as phytochemical markers, characteristic of the heather raw material from central Europe.

Development and Validation of the UPLC-DAD Methodology for the Detection of Triterpenoids and Phytosterols in Fruit Samples of Vaccinium macrocarpon Aiton and Vaccinium oxycoccos L.

Cranberries are used in the production of medicinal preparations and food supplements, which highlights the importance of triterpene compounds determination in cranberry fruit raw material. The aim of our study was to develop and validate for routine testing suitable UPLC-DAD methodology for the evaluation of triterpene acids, neutral triterpenoids, phytosterols, and squalene content in cranberry samples. The developed and optimized UPLC-DAD methodology was validated according to the guidelines of the International Council for Harmonization (ICH), evaluating the following parameters: range, specificity, linearity (R2 > 0.999), precision, LOD (0.27−1.86 µg/mL), LOQ (0.90−6.18 µg/mL), and recovery (80−110%). The developed and validated technique was used for the evaluation of triterpenic compounds in samples of Vaccinium macrocarpon and Vaccinium oxycoccos fruits, and their peels, pulp and seeds. The studied chromatogram profiles of Vaccinium macrocarpon and Vaccinium oxycoccos were identical but differed in the areas of the analytical peaks. Ursolic acid was the dominant compound in fruit samples of Vaccinium macrocarpon and Vaccinium oxycoccos. The highest amounts of triterpenic compounds were detected in the cranberry peels samples. The developed method for the detection of triterpene compounds can be applied in further studies for routine testing on the qualitative and quantitative composition of fruit samples of Vaccinium macrocarpon and Vaccinium oxycoccos species and cultivars.

Exploring Vaccinium vitis-idaea L. as a potential source of therapeutic agents: antimicrobial, antioxidant, and anti-inflammatory activities of extracts and fractions.

ETHNOPHARMACOLOGICAL RELEVANCE: Vaccinium vitis-idaea L. (lingonberry) leaves and fruits have traditionally been used in Asian and European countries as a natural solution for urinary tract infections, gastrointestinal distress, neurodegenerative diseases, and related inflammatory disorders, which are overall associated with free radical damage and presence of triggering pathogenic strains in the human body. Considering growing attention to natural products, there are not enough scientific data to confirm predominant specialized metabolites, responsible for the traditional therapeutic use of lingonberries. AIM OF THE STUDY: The present study aimed at an in-depth study of specialized metabolite profiling and biological activity evaluation of lingonberry crude extracts and isolated fractions. MATERIALS AND METHODS: Crude dry extracts and fractions from lingonberry leaves and fruits were analyzed by the UPLC-MS method. Potential inhibiting properties against different bacterial strains and hyaluronidase, ability to scavenge hydrogen peroxide, and effect on its production in a macrophage culture J774 were examined. RESULTS: Findings suggested the tentative presence of 59 compounds, mainly phenolics, displayed higher bioactivities of particular fractions than that of crude extracts and elucidated particular compounds as candidates in pharmaceuticals. Trimeric and dimeric proanthocyanidins from lingonberry leaves and fruits were shown to have the strongest antimicrobial, antioxidant, and anti-inflammatory potential. CONCLUSIONS: This study revealed specialized metabolites responsible for the traditional medicinal properties of lingonberries and pointed out demand for further purification and new research directions of proanthocyanidins in the frame of their multipharmacological perspectives.

Distribution of Phenolic Compounds and Antioxidant Activity in Plant Parts and Populations of Seven Underutilized Wild Achillea Species.

Evaluation of phytochemical composition of underutilized Achillea species provides the primary selection of germplasms with the desired quality of raw material for their further applications. The aim of the study was to evaluate the comprehensive distribution patterns of phenolic compounds in seven wild Achillea spp. and their plant parts, and to assess their antioxidant activity. Plant material was collected from different sites in Turkey. A complex of hydroxycinnamic acids, flavonols and flavones was identified and quantified in methanolic extracts using HPLC-PDA method. Antioxidant activity was assessed by radical scavenging assay. The results showed that qualitative and qualitative profiles of caffeoylquinic acids and flavonoids were species-specific, explaining the characteristic patterns of their variation in the corresponding species and plant parts. The highest total amount of caffeoylquinic acids was detected in A. setacea. A. arabica exposed the highest accumulation of mono-caffeoylquinic acids and flavonoids with the greatest levels of quercetin and luteolin derivatives and the flavonol santin. Santin was detected in all plant parts of A. cappadocica, A. setacea, A. santolinoides subsp. wilhelmsii, and A. arabica. A notable antiradical capacity was confirmed in A. arabica, A. setacea and A. cappadocica plant extracts. The leaves of all studied species were found to have priority over inflorescences and stems in terms of radical scavenging activity. The new data complemented the information that may be relevant for the continuation of chemophenetic studies in the heterogeneous genus Achillea.

Development, Validation, and Application of the UPLC-DAD Methodology for the Evaluation of the Qualitative and Quantitative Composition of Phenolic Compounds in the Fruit of American Cranberry (Vaccinium macrocarpon Aiton).

Phenolic compounds in the fruit of American cranberry (Vaccinium macrocarpon Aiton) determine the antioxidant, anti-inflammatory, anticancer, and other biological effects. The berries are used in the production of medicinal preparations and food supplements, which highlights the importance of qualitative and quantitative analysis of phenolic compounds in cranberry fruit raw material. The aim of our study was to develop and validate an efficient, cost-effective, reproducible, and fast UPLC-DAD methodology for the evaluation of the qualitative and quantitative composition of phenolic compounds in raw material and preparations of American cranberry fruit. During the development of the methodology, chlorogenic acid and the following flavonols were identified in cranberry fruit samples: myricetin-3-galactoside, quercetin-3-galactoside, quercetin-3-glucoside, quercetin-3-α-L-arabinopyranoside, quercetin-3-α-L-arabinofuranoside, quercetin-3-rhamnoside, myricetin, and quercetin. The developed and optimized UPLC-DAD methodology was validated according to the guidelines of the International Council for Harmonization (ICH), evaluating the following parameters: range, specificity, linearity (R2 > 0.999), precision (%RSD < 2%), LOD (0.38-1.01 µg/mL), LOQ (0.54-3.06 µg/mL), and recovery (80-110%). The developed methodology was applied to evaluate the qualitative and quantitative composition of phenolic compounds in fruit samples of cranberry cultivars 'Baifay', 'Bergman', 'Prolific', and 'Searles', as well as 'Bain-MC' and 'BL-12' clones. In the tested samples, the majority (about 70%) of the identified flavonols were quercetin derivatives. The greatest amount of quercetin-3-galactoside (1035.35 ± 4.26 µg/g DW) was found in fruit samples of the 'Searles' cultivar, and the greatest amount of myricetin-3-galactoside (940.06 ± 24.91 µg/g DW) was detected in fruit samples of the 'Woolman' cultivar.

Sea Buckthorn Leaf Powders: The Impact of Cultivar and Drying Mode on Antioxidant, Phytochemical, and Chromatic Profile of Valuable Resource.

Sea buckthorn (Hippophae rhamnoides L. (HR)) leaf powders are the underutilized, promising resource of valuable compounds. Genotype and processing methods are key factors in the preparation of homogenous, stable, and quantified ingredients. The aim of this study was to evaluate the phenolic, triterpenic, antioxidant profiles, carotenoid and chlorophyll content, and chromatic characteristics of convection-dried and freeze-dried HR leaf powders obtained from ten different female cultivars, namely 'Avgustinka', 'Botaniceskaja Liubitelskaja', 'Botaniceskaja', 'Hibrid Percika', 'Julia', 'Nivelena', 'Otradnaja', 'Podarok Sadu', 'Trofimovskaja', and 'Vorobjovskaja'. The chromatic characteristics were determined using the CIELAB scale. The phytochemical profiles were determined using HPLC-PDA (high performance liquid chromatography with photodiode array detector) analysis; spectrophotometric assays and antioxidant activities were investigated using ABTS (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (ferric ion reducing antioxidant power) assays. The sea buckthorn leaf powders had a yellowish-green appearance. The drying mode had a significant impact on the total antioxidant activity, chlorophyll content, and chromatic characteristics of the samples; the freeze-dried samples were superior in antioxidant activity, chlorophyll, carotenoid content, and chromatic profile, compared to convection-dried leaf powder samples. The determined triterpenic and phenolic profiles strongly depend on the cultivar, and the drying technique had no impact on qualitative and quantitative composition. Catechin, epigallocatechin, procyanidin B3, ursolic acid, α-amyrin, and ß-sitosterol could be used as quantitative markers in the phenolic and triterpenic profiles. The cultivars 'Avgustinka', 'Nivelena', and 'Botaniceskaja' were superior to other tested cultivars, with the phytochemical composition and antioxidant activity.

Optimization, Validation and Application of HPLC-PDA Methods for Quantification of Triterpenoids in Vaccinium vitis-idaea L.

Triterpenoids have regained much attention as promising multi-targeting bioactive agents of natural origin in the treatment of numerous disorders. Due to the high potential for phytopharmaceutical development, accurate qualitative and quantitative analysis of triterpenoids for screening and quality control is required. Vaccinium vitis-idaea L. (lingonberry) raw materials have aroused interest as a rich source of triterpenoids. However, currently, no validated, rapid, and easy-to-perform quantification method is available for the routine control of these compounds in lingonberries. This research aimed at developing and validating HPLC-PDA methods for the determination and screening of triterpenoids in extracts of lingonberry leaves, fruits, and flowers. The developed methods were deemed satisfactory by validation, which revealed acceptable analytical specificity, linearity (r2 > 0.9999), precision (RSD < 2%), trueness (94.70-105.81%), and sensitivity (LOD: 0.08-0.65 µg/mL). The real sample analysis demonstrated established methods applicability for quantification of 13 triterpenoids in lingonberries and emphasized differences between raw materials. Lingonberry fruits were distinguished by the richness of ursolic acid; lingonberry flowers by similar profile to fruits, but low content of neutral triterpenoids; whereas lingonberry leaves by the particularly high level of α-amyrin. Thus, the proposed methods proved to be reliable and applicable for quantification and routine analysis of triterpenoids in lingonberry samples.

Investigation of Antibacterial and Antiinflammatory Activities of Proanthocyanidins from Pelargonium sidoides DC Root Extract.

The study explores antibacterial, antiinflammatory and cytoprotective capacity of Pelargonium sidoides DC root extract (PSRE) and proanthocyanidin fraction from PSRE (PACN) under conditions characteristic for periodontal disease. Following previous finding that PACN exerts stronger suppression of Porphyromonas gingivalis compared to the effect on commensal Streptococcus salivarius, the current work continues antibacterial investigation on Staphylococcus aureus, Staphylococcus epidermidis, Aggregatibacter actinomycetemcomitans and Escherichia coli. PSRE and PACN are also studied for their ability to prevent gingival fibroblast cell death in the presence of bacteria or bacterial lipopolysaccharide (LPS), to block LPS- or LPS + IFNγ-induced release of inflammatory mediators, gene expression and surface antigen presentation. Both PSRE and PACN were more efficient in suppressing Staphylococcus and Aggregatibacter compared to Escherichia, prevented A. actinomycetemcomitans- and LPS-induced death of fibroblasts, decreased LPS-induced release of interleukin-8 and prostaglandin E2 from fibroblasts and IL-6 from leukocytes, blocked expression of IL-1ß, iNOS, and surface presentation of CD80 and CD86 in LPS + IFNγ-treated macrophages, and IL-1ß and COX-2 expression in LPS-treated leukocytes. None of the investigated substances affected either the level of secretion or expression of TNFα. In conclusion, PSRE, and especially PACN, possess strong antibacterial, antiinflammatory and gingival tissue protecting properties under periodontitis-mimicking conditions and are suggestable candidates for treatment of the disease.

Antioxidant Activities of Vaccinium vitis-idaea L. Leaves within Cultivars and Their Phenolic Compounds.

Lingonberry leaves are the subject of numerous studies because of antioxidant properties, positive influence on the health and potential use in the prevention and treatment of chronic diseases. In this work, the radical scavenging, reducing, chelating activities, and phenolic composition of ten lingonberry leaves cultivars, one subspecies, and one variety were investigated. Furthermore, the antioxidant activity of individual phenolic compounds, that can be found in lingonberry leaves, were analyzed, and structure-activity relationship was determined. Wide diversity for phenolic profile and antioxidant properties of lingonberry leaves has been observed in the present material. Cultivars 'Kostromskaja rozovaja', 'Rubin', and Vaccinium vitis-idaea var. leucocarpum surpassed all others tested cultivars and lower taxa by contents of phenolic compounds and antioxidant activity. Leaves of lingonberry cultivars and lower taxa are rich in arbutin, flavonol glycosides, proanthocyanidins, and the latter were considered to be the major contributor to antioxidant properties of lingonberry leaves.

EFFECT OF PERILLA FRUTESCENS EXTRACTS AND ROSMARINIC ACID ON RAT HEART MITOCHONDRIAL FUNCTIONS.

Perilla frutescens L. due to its aromatic, antibacterial, anti-inflammatory and antioxidant traits has been traditionally used as medicinal plant in Eastern Asia. Alterations of mitochondria are interconnected with many chronic diseases. Bioactives of herbal extracts can modulate mitochondrial effects and be beneficial in prevention of mitochondrial related chronic diseases. Direct effects of the red-leaf form P. frutescens extract (PFE) and the green-leaf form P. frutescens var. crispa f. viridis extract (PCE) were evaluated investigating activities on the oxidative phosphorylation and antioxidant activity in the rat heart mitochondria in vitro. HPLC-MS analysis was applied for the identification of phenolic compounds. Cell with a Clark-type oxygen electrode was used for mitochondrial respiration measurement. The generation of reactive oxygen species was estimated in isolated rat heart mitochondria and determined fluorimetrically. State 3 respiration rate was not affected by lower concentrations, however, it was inhibited at higher concentrations by 22-70% for PFE and by 45-55% for PCE. PFE containing anthocyanins induced the concentration-dependent stimulation (by 23-76%) of the State 4 respiration rate after addition of cytochrome c due to reducing properties. Significant reduction of H2O2 pro- duction was observed with investigated concentrations of rosmarinic acid and both perilla extracts. Our results demonstrate that the effect of PFE and PCE extracts on rat heart mitochondria depend on the qualitative characteristics of complex of biologically active compounds. Selective effects on mitochondrial function could enable the regulation of apoptosis or another mechanisms occurring in cells.

Phenolic Profiles and Contribution of Individual Compounds to Antioxidant Activity of Apple Powders.

Apples (Malus domestica L.) are the most common source of phenolic compounds in northern European diet. Besides pectins, dietary fibers, vitamins, and oligosaccharides they contain phenolic compounds of different classes. Apple powders are convenient functional forms retaining significant amounts of phenolic antioxidants. In this study reducing and radical scavenging profiles of freeze-dried powders of "Aldas,Ë® "Auksis,Ë® "Connel Red,Ë® "Ligol,Ë® "Lodel,Ë® and "RajkaË® were determined and phenolic constituents were identified using ultra high-performance liquid chromatography coupled to quadrupole and time-of-flight mass spectrometers. A negative ionization mode was applied and seventeen compounds: phenolic acids (coumaroylquinic, chlorogenic), flavonoids (quercetin derivatives), and procyanidin derivatives (B1, B2, and C1) were identified in all tested apple samples. Total values of Trolox equivalents varied from 7.72 ± 0.32 up to 20.02 ± 0.52 and from 11.10 ± 0.57 up to 21.42 ± 0.75 µmol/g of dry weight of apple powder in FRAP (ferric reducing antioxidant power) and ABTS (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) postcolumn assays, respectively. The greatest Trolox equivalent values were determined for apples of "AldasË® cultivar. Chlorogenic acid and procyanidin C1 were the most significant contributors to total reducing and radical scavenging activity in all apple cultivars tested, therefore they could be considered as markers of antioxidant activity.

Phytochemical and antioxidant profiles of leaves from different Sorbus L. species.

Leaves of Sorbus L. have been used in various traditional medicine systems. Phenolic compounds determine the main pharmacological effects of Sorbus L. In this study, phytochemical and antioxidant profiles of Sorbus anglica, Sorbus aria, Sorbus arranensis, Sorbus aucuparia, Sorbus austriaca, Sorbus caucasica, Sorbus commixta, Sorbus discolor, Sorbus gracilis, Sorbus hostii, Sorbus semi-incisa and Sorbus tianschanica were determined. Twenty four constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers. Post-column FRAP assay identified compounds with reducing activity and revealed significantly greatest total antioxidant activity of 175.30 µmol TE/g DW, 169.20 µmol TE/g DW and 148.11 µmol TE/g DW in S. commixta, S. discolor and S. gracilis leaf samples, respectively, with neochlorogenic and chlorogenic acids being most significant contributors. Characteristic fingerprints of phytochemical and antioxidant profiles could be applied for the quality evaluation of various raw materials of Sorbus L. species.

Quality evaluation of different preparations of dry extracts of birch (Betula pendula Roth) leaves.

Qualitatively prepared dry extracts are advantaged over conventional fluid extracts. Extracts of Betula pendula Roth leaves possess mild diuretic effects and anti-inflammatory activity. In this work, the spray-drying and lyophilisation processes were evaluated for producing dry extracts of birch leaves. The purpose was to identify the best drying method that provides the highest content of phenolic compounds and possesses greatest antioxidant activity. Predominant compounds hyperoside and chlorogenic acid were chosen as markers for the quantitative control of phenolic compounds. Their determined amounts in hydroalcoholic birch leaf extract were 15.85 and 8.77 mg/g of dry residue, respectively. Insignificant differences were determined in spray- and freeze-dried extracts. In the antioxidant profile, hyperoside (28.9 µmol TE/g dw) and chlorogenic acid (22.8 µmol TE/g dw) possessed strongest ferric-reducing activity. Also, no significant differences of antioxidant activity were determined in the spray- and freeze-dried extracts.

Effects of raspberry fruit extracts and ellagic acid on respiratory burst in murine macrophages.

The mechanism of action of polyphenolic compounds is attributed to their antioxidant, anti-inflammatory, and anti-proliferative properties and their effects on subcellular signal transduction, cell cycle impairment and apoptosis. A raspberry (Rubus idaeus L.) fruit extract contains various antioxidant active compounds, particularly ellagic acid (EA); however the exact intracellular mechanism of their action is not fully understood. The aim of the study was to evaluate the antioxidant effect of raspberry extracts, and that of ellagic acid by assessment of the production of the reactive oxygen species (ROS) by murine macrophage J774 cells. Raspberry extracts and their active compound EA did not affect or had very minor effects on cell viability. No significant difference in the ROS generation in arachidonic acid stimulated macrophages was determined for raspberry extracts and EA whereas in the phorbol-12 myristate-13 acetate model ROS generation was significantly (p < 0.05) reduced. Our observation that raspberry pomace extracts in vitro reduce ROS production in a J774 macrophage culture suggests that raspberry extract and ellagic acid mediated antioxidant effects may be due to the regulation of NADPH oxidase activity.