This study aimed at the evaluation of the antioxidant and cognitive-enhancing effect of methanol–aqueous extract from Helichrysum italicum ssp. italicum aerial parts. Significant radical scavenging activity (110.33 ± 3.47 and 234.70 ± 5.21 mg TE/g for DPPH and ABTS) and reducing power (354.23 ± 17.51 and 210.24 ± 8.68 mg TE/g for CUPRAC and FRAP) were observed. The extract showed average acetylcholinesterase and low butyrylcholinesterase inhibitory potential. H. italicum extract (200 mg/kg/po) administered in combination with galantamine (3 mg/kg/po) for 12 days significantly improved the memory and learning process compared with galantamine alone in the passive avoidance test. The effect was comparable to that of Ginkgo biloba extract (100 mg/kg/po). In deep secondary metabolite annotation of the extract by UHPLC-HRMS, more than 90 hydroxybenzoic and hydroxicinnamic acid-glycosides, phenylethanoid glycosides, a series of acylquinic and caffeoylhexaric acids, methoxylated derivatives of scutellarein, quercetagetin and 6-hydroxyluteolin, and prenylated phloroglucinol-α-pyrones were reported for the first time in H. italicum. Fragmentation patterns of four subclasses of heterodimer-pyrones were proposed. In-depth profiling of the pyrones revealed 23 compounds undescribed in the literature. Pyrones and acylphloroglucinols together with acylquinic acids could account for memory improvement. The presented research advanced our knowledge of H. italicum, highlighting the species as a rich source of secondary metabolites with cognitive-enhancing potential.

Oxidative stress is a common phenomenon of many liver disorders; it both affects patient survival and directly influences the applicability, effectiveness, and toxicity of drugs. In the pursuit of reliable natural remedies for hepatoprotection, this study reports on the complete phytochemical characterization, antioxidant, and hepatoprotective activities of the Prenanthes purpurea methanol-aqueous extract in an in vitro model of diclofenac-induced liver injury (DILI). An ultra high-performance liquid chromatography–high-resolution mass spectrometry analysis (UHPLC-HRMS) was conducted, delineating more than 100 secondary metabolites for the first time in the species, including a series of phenolic acid-hexosides, acylquinic, acylhydroxyquinic and acyltartaric acids, and flavonoids. Quinic acid, chlorogenic, 3,5-dicaffeoylquinic and 5-feruloylhydroxyquinic acid, caffeoyltartaric and cichoric acids, eryodictiol-O-hexuronide, and luteolin O-hexuronide dominated the phytochemical profile and most likely contributed to the observed hepatoprotective activity of the studied P. purpurea leaf extract. The potency and molecular basis of cellular protection were investigated in parallel with pure caffeoylquinic acids in a series of pretreatment experiments that verified the antiapoptotic and antioxidant properties of the natural products.

Erica spiculifolia Salisb. (formerly Bruckenthalia spiculifolia Benth.) (Balkan heath) is renowned for its traditional usage as a diuretic, anti-inflammatory and antioxidant agent. For the first time, acylquinic acids, flavonoids and numerous proanthocyanidin oligomers were annotated/dereplicated by liquid chromatography–high-resolution mass spectrometry in methanol–aqueous extracts from E. spiculifolia aerial parts harvested at the early and full flowering stage. Chlorogenic acid and proanthocyanidin tetra- and trimer A, B-type together with quercitrin and (+) catechin were the predominant compounds in the semi-quantitative analysis. Neutral triterpenoids, triterpenoid acids and phytosterols were determined in apolar extracts by gas chromatography–mass spectrometry. Triterpenoid acids accounted for 80% of the total triterpenoid content, dominated by ursolic and oleanolic acid, reaching up to 32.2 and 6.1 mg/g dw, respectively. Ursa/olean-2,12-dien-28-oic acids and 3-keto-derivatives together with α-amyrin acetate as a chemotaxonomic marker, α-amyrenone, α- and β-amyrin were evaluated. Total phenolic and flavonoid contents were 83.85 ± 0.89 mg gallic acid equivalents/g and 78.91 ± 0.41 mg rutin equivalents/g, respectively. The extract actively scavenged DPPH and ABTS radicals (540.01 and 639.11 mg Trolox equivalents (TE)/g), possessed high potential to reduce copper and iron ions (660.32 and 869.22 mg TE/g, respectively), and demonstrated high metal chelating capacity (15.57 Ethylenediaminetetraacetic acid equivalents/g). It exhibited prominent anti-lipase (18.32 mg orlistat equivalents/g) and anti-tyrosinase (71.90 mg kojic acid equivalents/g) activity. The extract inhibited α-glucoside (1.35 mmol acarbose equivalents/g) and acetylcholinesterase (2.56 mg galanthamin equivalents/g), and had moderate effects on α-amylase, elastase, collagenase and hyaluronidase. Balkan heath could be recommended for raw material production with antioxidant and enzyme inhibitory properties.

Ostericum palustre (Besser) Besser (synonym Angelica pancicii Vandas ex. Velen.) is a Eurasian species from the Apiaceae family, previously related to the Balkan endemic species A. pancicii. The study aims to provide a thorough profiling of methanol-aqueous extracts from O. palustre leaves, roots, and inflorescences integrated with an evaluation of antioxidant potential and enzyme inhibitory activity towards some therapeutic targets. For the first time, a series of simple coumarins and furanocoumarins alongside phenolic and acylquinic acids, and flavonoids were annotated/dereplicated in the O. palustre of Bulgarian origin by liquid chromatography coupled with quadrupole—Orbitrap high resolution mass spectrometry acquisition platform. According to the discriminant analysis (sPLS-DA) of the biological potential, radical scavenging activity (47.9 mg TE/g in DPPH and 61.8 mg TE/g in ABTS), reducing power (102.2 mg TE/g in CUPRAC and 57.4 mg TE/g in FRAP), and metal-chelating capacity (20.1 mg EDTAE/g) accounted mainly for the stronger antioxidant activity of inflorescences extract than roots and leaves. Root extracts exhibited anti-collagenase, anti-elastase, and anti-hyaluronidase effects with lower IC₅₀ values (IC₅₀ 37.22, 42.47 and 32.09 μg/mL, respectively). Pearson relationship analysis revealed potent antioxidants including furanocoumarins (oxypeucedanin hydrate, xanthotoxol/bergaptol, byakangelicin/isobyakangelicin, ostruthol) and phenolic acids, while a series of angelols alongside feruloylquinic and dicaffeoylquinic acids, and flavonol glycosides hold significance for the neuroprotective activity of the leaves extract. The enzyme inhibitory activity of the root extracts towards collagenase, elastase and hyaluronidase, related to the anti-aging activity, was ascribed to simple hydroxylated/methoxylated coumarins. The study suggests the potential health benefits of O. palustre extracts as antioxidant, anti-aging, and neuroprotective agents.