Search Results (79)

Bamboo leaves contain structurally diverse flavonoid glycosides; whereas, compound-level information for Bambusa textilis remains limited. In this study, a flavonoid-enriched 40% ethanol fraction prepared from B. textilis leaves by petroleum-ether partitioning and AB-8 resin adsorption was analyzed by ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-ESI-Q-TOF-MS/MS) in negative-ion mode. Retention times and MS/MS spectra were compared with seven in-house reference standards, while the remaining constituents were annotated from accurate-mass measurements, diagnostic product ions, fragmentation behavior, and data from the literature. According to Metabolomics Standards Initiative criteria, seven compounds were assigned at Level 1 and twenty-four were reported as Level 3 annotations because their exact positional structures were not confirmed by additional standards or NMR. Thirty-one flavonoid-related constituents were annotated in the analyzed fraction, encompassing six glycoside classes; most annotations, by count, were apigenin- or luteolin-derived. These annotations add compound-level information for the analyzed B. textilis fraction and allow qualitative comparison with other bamboo flavonoid profiles. Because the sample was selectively enriched, the results do not describe the complete leaf metabolome or compound concentrations. Additional standards or NMR analysis are needed to confirm the glycosylation positions of tentatively annotated compounds. Full article

The high prevalence and incidence of neurodegenerative diseases pose a public health challenge and drive the search for alternative treatments. This study determined the chemical composition of hydroalcoholic extracts from the Antarctic species Polycauliona candelaria and Placopsis antarctica and evaluated their antioxidant and cholinesterase-inhibitory potential through in vitro assays and molecular docking. Using UHPLC/ESI/QToF/MS, 16 compounds were tentatively identified in P. candelaria and 11 in P. antarctica. P. antarctica exhibited greater antioxidant capacity (2.69 ± 0.15 mg GAE/g in TPC, and an IC₅₀ for DPPH and ABTS of 330.64 ± 0.02 and 63.33 ± 0.02 µg/mL, respectively) and inhibitory activity (IC₅₀ for AChE and BuChE of 654.42 ± 0.03 and 845.58 ± 0.01 µg/mL, respectively) similar to P. candelaria. Molecular docking analyses revealed that gyrophoric acid and stictic acid possess outstanding binding affinities, comparable to the drug galantamine, by effectively interacting with the catalytic sites of the enzymes. This is the first report on the chemical compounds present in extracts of P. antarctica and P. candelaria and contributes to the understanding of their therapeutic potential. Full article

Lichenized fungi are a source of secondary metabolites with multiple biological potential. The objective of the study was to determine the chemical composition, antioxidant, and anti-inflammatory activity of the hydroalcoholic extract of the Antarctic lichen Leptogium puberulum through metabolomic, in vitro, and in silico analyses. Seventeen compounds were tentatively identified using UHPLC-ESI-QToF-MS. The phenolic composition yielded 6.356 mg GAE/g, and antioxidant activity assays showed IC₅₀ values for DPPH• and ABTS•⁺ of 1187.149 and 207.00 µg/mL, respectively, along with 15.845 µmol Trolox/g for ORAC and 21.925 µmol Trolox/g for FRAP. The in silico evaluation was performed using OSIRIS Data Warrior, ProTox 3.0, and SwissTargetPrediction, identifying 9,10,12,13,14-pentahydroxytetracosanoic acid (PHTA), 9,10,12,13-tetrahydroxytricosanoic acid (THTA), 9,10,12,13-tetrahydroxyheneicosanoic acid (THHA), and 9,10,12,13-tetrahydroxydocosanoic acid (THDA) as the most promising compounds. These metabolites showed favorable pharmacokinetic properties, with no anticipated toxicological risks. Subsequently, their affinity for the cyclooxygenase-2 (COX-2) enzyme was evaluated by molecular docking with AutoDock Vina software version 1.2.3, and the most stable protein–ligand complexes were analyzed to characterize key interactions within the active site and subjected to molecular dynamics simulations with YASARA software version 19.1.27 for 100 ns. Overall, these results indicate that selected metabolites from L. puberulum may act as potential COX-2 inhibitors, supporting their relevance as lichen-derived anti-inflammatory agents and warranting further pharmacological investigation. Full article

Larrea ameghinoi Speg., an endemic species of Argentine Patagonia traditionally used in folk medicine to treat fever, stomach disorders, respiratory conditions, back pain, and as an emmenagogue, among others, still remains chemically and biologically underexplored compared to the other four members of the genus. This study aimed to perform a comprehensive metabolomic characterization of methanolic extracts from two populations (EMLaSAO and EMLaMAQ) using ultra-high-resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC–ESI–QTOF–MS) and to evaluate their antioxidant, antimicrobial, and enzyme-inhibitory activities of relevance to human health. Thirty-three compounds were tentatively identified by extensive UHPLC–MS analysis, including flavones, two major lignans, and oleanane-type triterpenes. Both extracts exhibited high phenolic content (215–239 mg of gallic acid equivalents (GAE)/g extract) and strong free radical scavenging activity, as evidenced by 2,2-diphenyl-1-picrylhydrazyl (DPPH, EC₅₀ ≈ 10 μg/mL), ferric-reducing antioxidant power (FRAP), and Trolox equivalent antioxidant activity (TEAC) assays. In addition, significant inhibition of butyrylcholinesterase (IC₅₀ ≈ 50 μg extract/mL) and α-glucosidase, together with selective antibacterial activity against methicillin-sensitive and resistant Staphylococcus aureus (MIC = 125 μg extract/mL), were recorded. These findings suggest that L. ameghinoi possesses a distinctive phytochemical composition conferring multitarget bioactivity, differing from other Larrea species dominated by lignans such as nordihydroguaiaretic acid (NDGA) and its derivatives. Overall, this work supports the potential of L. ameghinoi as a novel source of bioactive metabolites for managing oxidative stress-related disorders and opportunistic infections. This warrants future in vivo studies investigating biological activities associated with oxidative stress and their relevance to human health. Full article

Botrytis cinerea and Plasmopara viticola, the causal agents of grey mold and downy mildew, respectively, are two major grapevine pathogens whose control largely relies on synthetic fungicides, raising environmental and health concerns. Plant-derived secondary metabolites, particularly flavonoids involved in plant defense, represent promising sustainable alternatives. Among them, sakuranetin, a flavanone aglycone known for its antifungal activity in rice, remains poorly explored for grapevine protection. In this study, sakuranetin was purified from cherry branches (48 mg) and structurally characterized using UHPLC-ESI-QTOF-MS and NMR analyses. Its antifungal activity against B. cinerea and P. viticola was evaluated through in vitro, in vivo and in planta assays. For B. cinerea, our results showed a significant in vitro inhibition of mycelium growth, with EC₅₀ values of 16.43 mg·L⁻¹, while no protection of detached berries was observed. Against P. viticola, sakuranetin has no effect on the release of zoospores, but there is a total inhibition of spore germination at 1 mg·L⁻¹ in vitro, confirmed in vivo on a foliar disc. In planta, no significant protection is observed at 25 mg·L⁻¹, even if some targeted defense genes are induced. Further studies are needed to determine the best concentration of sakuranetin to use to manage B. cinerea and P. viticola in planta. Full article

Aeromonas hydrophila is a major seafood-borne pathogen capable of persisting under preservative-associated stress by entering a viable but non-culturable (VBNC) state, thereby evading culture-based detection. Here, untargeted metabolomics was applied as the primary analytical approach to elucidate metabolic reprogramming during VBNC formation under seafood-relevant preservation conditions. Cells were incubated at 4 °C for 30 days in sodium benzoate-supplemented saline, comparing 0.85% NaCl (culturable condition) and 4% NaCl (VBNC-inducing condition), with sampling every 6 days. Under 4% NaCl with sodium benzoate, culturability declined from 6.18 log CFU/mL at day 0 to undetectable levels by day 30, while cell viability was retained, confirming VBNC induction. UHPLC–ESI–QTOF–MS profiling detected over 893 intracellular metabolic features, of which 518 metabolites were significantly altered between VBNC and culturable states at day 30. Principal component analysis revealed clear, time-dependent metabolic divergence, with the VBNC trajectory explaining 34.4% (PC1) and 11.5% (PC2) of total variance. Pathway enrichment analysis demonstrated significant remodeling of alanine, aspartate and glutamate metabolism (8/28 hits, FDR = 5.7 × 10⁻⁴); arginine biosynthesis (5/14 hits, FDR = 5.44 × 10⁻³); purine metabolism (10/70 hits, FDR = 8.34 × 10⁻³); and pyrimidine metabolism (7/39 hits, FDR = 1.35 × 10⁻²), indicating nitrogen conservation and metabolic downshifting. A robust biomarker panel, including depleted cyclic AMP, aminoadipic acid, hypotaurine, O6-CM-dG, and betaine, and enriched urocanic acid, pipecolic acid, proline, azelaic acid, and orcinol perfectly discriminated VBNC from culturable cells. These findings demonstrate that sodium benzoate-based preservation can induce a metabolically reprogrammed VBNC state in A. hydrophila, highlighting a hidden food safety risk beyond culture-based assessment. Full article

Diabetic nephropathy is a major complication of diabetes mellitus, primarily driven by hyperglycemia-induced oxidative stress and renal tubular cell injury. Beetroot (Beta vulgaris L.) is rich in antioxidant phytochemicals, and its industrial processing generates large amounts of spent material that may retain significant bioactive compounds. This study evaluated the phytochemical profile, antioxidant capacity, and renoprotective potential of beetroot spent extracts under hyperglycemic conditions. Beetroot spent material was extracted using hot water and 70% ethanol. Total phenolic, flavonoid, and betalain contents were quantified, and antioxidant activity was assessed using the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. Phytochemical characterization was performed by ultra-high-pressure liquid chromatography–electrospray ionization–quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). Cytotoxicity was evaluated in peripheral blood mononuclear cells, SH-SY5Y, HEK-293, and MDA-MB-231 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Renoprotective effects were investigated in HEK-293 renal tubular cells cultured under normal (5.5 mM) and high-glucose (200 mM) conditions. UHPLC-ESI-QTOF-MS was used to identify over 80 phenolic and flavonoid compounds including quercetin, epicatechin, and epigallocatechin gallate. The hot water extract exhibited superior antioxidant activity, achieving approximately 90% ABTS radical inhibition. Beetroot spent extract showed no cytotoxicity at concentrations below 1 mg/mL and significantly restored HEK-293 cell viability (>90%) under high-glucose conditions at concentrations ≥31.25 µg/mL. In conclusion, beetroot spent water extract possesses strong antioxidant and renoprotective activities against hyperglycemia-induced renal cell damage, supporting its valorization as a sustainable functional food ingredient for diabetes-related health applications. Full article

Calophyllum spp. infusions are used to treat varicose veins, hemorrhoids, and hypertension. However, the chemical composition and mechanisms of action are poorly understood. Accordingly, the aim of this study was to investigate the phytochemical composition and vascular effects of hydroalcoholic extracts of Calophyllum longifolium. Phytochemical profiling was performed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF-MS). Extract effects on rat aortic rings and aortic vascular smooth muscle cells (VSMCs) were evaluated using wire myography and photometric measurement of intracellular Ca²⁺, respectively. UHPLC-ESI-Q-TOF-MS revealed the presence of coumarins, xanthones, flavonoids, triterpenes, and phenolic acids. Coumarin–resveratrol hybrids, such as gut-70 derivatives, were also abundant. In aortic rings from normotensive rats, C. longifolium induced a biphasic vascular response whereby low concentrations (1 μg/mL) produced significant vascular relaxation, whereas high concentrations (100 μg/mL) produced contraction. Blockade of ATP-sensitive (K_ATP) or voltage-gated (K_V) potassium channels attenuated these effects. Furthermore, effects were not observed in preparations preincubated with L-NG-Nitro-L-arginine methyl ester (L-NAME) or in endothelium-denuded rings. In aortic VSMCs, extracts (1 µg/mL) rapidly reduced sarcoplasmic reticulum (SR) Ca²⁺ content. This study provides the first UHPLC-ESI-Q-TOF-MS chemical profile of C. longifolium, revealing diverse bioactive metabolites. It is also the first to demonstrate that C. longifolium exerts an endothelium-dependent, nitric oxide- and Ca²⁺-mediated biphasic effect on vascular function. Taken together, these findings highlight C. longifolium as a potential novel source of vasculotropic phytopharmaceuticals. Full article

The all-d-enantiomeric-peptide RD2 was developed for the treatment of Alzheimer’s disease. This study aimed to develop a specific and highly sensitive liquid chromatography-mass-spectrometric (UHPLC-ESI-QTOF) method for quantifying RD2 in the mouse brain and to validate it according to the ICH M10 guideline to investigate the pharmacokinetic profile of RD2 in its target organ. Sample preparation, chromatographic separation and quantification were very challenging due to RD2’s highly hydrophilic properties, the complex matrix and the required lower limit of quantification (LLOQ). Chromatographic separation was performed on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm particle size) within 5 min at 50 °C with a flow rate of 0.5 mL·min⁻¹. Mobile phases consisted of water and acetonitrile with 0.2% formic acid and 0.015% heptafluorobutyric acid. Ions were generated by electrospray ionization in the positive mode, and RD2 was quantified by QTOF-MS. The developed extraction method revealed complete recovery. The linearity of the calibration curve was in the range of 2 ng·mL⁻¹ to 500 ng·mL⁻¹ (R² > 0.99) with a LLOQ of 5 ng·mL⁻¹. The intraday and interday accuracy and precision ranged from 0.4% to 12.2% and from 1.0% to 12.0%. RD2 remained stable in the freshly homogenized brain even after several freeze–thaw cycles, but stability decreased over time during long-term storage at −80 °C. Using this validated method, RD2-spiked brain homogenate samples and samples of a pharmacokinetic study with RD2 in mice were analyzed. Full article

Breast cancer remains one of the most prevalent and lethal malignancies affecting women worldwide, underscoring the need for safer and more effective therapeutic strategies. This study investigated the phytochemical composition, antioxidant activity, and antiproliferative potential of POW9™, a proprietary botanical blend formulated from nine medicinal plant extracts. Comprehensive phytochemical profiling was performed using ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) in both positive and negative ionization modes. A total of 34 compounds were identified in negative mode and 27 compounds in positive mode, comprising flavonoids, terpenoids, steroids, organic acids, peptides, glycosides, and lipids. POW9™ exhibited high total phenolic content (190.3 ± 3.5 mg gallic acid equivalents/g) and total flavonoid content (115.2 ± 1.5 mg quercetin equivalents/g), along with strong antioxidant activity, demonstrated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging with a half-maximal inhibitory concentration (IC₅₀) of 1.66 mg/mL (33.73 mg Trolox equivalents/g). Cytotoxicity assessment revealed minimal toxicity toward normal Vero cells. In contrast, POW9™ significantly inhibited the proliferation of human breast cancer cell lines in a concentration- and time-dependent manner. The IC₅₀ values were 6.75 mg/mL for MCF-7 cells and 18.08 mg/mL for MDA-MB-231 cells after 72 h of treatment, while prolonged exposure (96 h) further enhanced antiproliferative efficacy, reducing the IC₅₀ to 2.34 mg/mL. These findings demonstrate that POW9™ is a chemically diverse herbal formulation with potent antioxidant and selective anti-breast cancer activities, supporting its potential development as a complementary therapeutic or nutraceutical agent for breast cancer management. Full article

Background and Rationale: Tinospora crispa (L.) Hook.f. & Thomson (T. crispa) is a climbing medicinal plant with long-standing ethnopharmacological use, particularly in inflammatory and hepatic disorders and cancer-related conditions. There is a knowledge gap regarding how wild versus cultivated ecotypes differ in chemotype, bioactivity, and safety, and how this might support or refine traditional use. Study Objectives: This study aimed to compare wild and cultivated ecotypes of T. crispa from the Nile Delta (Egypt) in terms of quantitative and qualitative phytochemical profiles; selected in vitro biological activities (especially antioxidant and cytotoxic actions); genetic markers potentially associated with metabolic variation; and short-term oral safety in an animal model. Core Methodology: Standardized extraction of plant material from wild and cultivated ecotypes. Determination of total phenolics, total flavonoids, and major phytochemical classes (alkaloids, tannins, terpenoids). Metabolomic characterization using UHPLC-ESI-QTOF-MS, supported by NMR, to confirm key compounds such as berberine, palmatine, chlorogenic acid, rutin, and borapetoside C. In vitro bioassays including: Antioxidant activity (e.g., radical-scavenging assay with EC₅₀ determination). Cytotoxicity against human cancer cell lines, with emphasis on HepG2 hepatoma cells and calculation of IC₅₀ values. Targeted genetic analysis to detect single-nucleotide polymorphisms (SNPs) in the gen1 locus that differentiate ecotypes. A 14-day oral toxicity study in rats, assessing liver and kidney function markers and performing histopathology of liver and kidney tissues. Principal Results: The wild ecotype showed a 43–65% increase in total flavonoid and polyphenol content compared with the cultivated ecotype, as well as substantially higher levels of key alkaloids, particularly berberine (around 12.5 ± 0.8 mg/g), along with elevated chlorogenic acid and borapetoside C. UHPLC-MS and NMR analyses confirmed the identity of the main bioactive constituents and defined a distinct chemical fingerprint for the wild chemotype. Bioassays demonstrated stronger antioxidant activity of the wild extract than the cultivated one and selective cytotoxicity of the wild extract against HepG2 cells (IC₅₀ ≈ 85 µg/mL), being clearly more potent than extracts from cultivated plants. Genetic profiling detected a C → T SNP within the gen1 region that differentiates the wild ecotype and may be linked to altered biosynthetic regulation. The 14-day oral toxicity study (up to 600 mg/kg) revealed no evidence of hepatic or renal toxicity, with biochemical markers remaining within physiological limits and normal liver and kidney histology. Conclusions and Future Perspectives: The wild Nile-Delta ecotype of T. crispa appears to be a stress-adapted chemotype characterized by enriched levels of multiple bioactive metabolites, superior in vitro bioactivity, and an encouraging preliminary safety margin. These findings support further evaluation of wild T. crispa as a candidate source for standardized botanical preparations targeting oxidative stress-related and hepatic pathologies, while emphasizing the need for: More comprehensive in vivo efficacy studies. Cultivation strategies that deliberately maintain or mimic beneficial stress conditions to preserve phytochemical richness. Broader geographical and genetic sampling to assess how generalizable the present chemotypic and bioactivity patterns are across the species. Full article

Bacterial outer-membrane vesicles (OMVs) mediate stress tolerance, biofilm formation, and interkingdom communication, but their role in beneficial endophytes remains underexplored. We isolated 11 non-redundant isolates associated with Bacillus, Enterococcus, Kosakonia and Kocuria from Agave tequilana seeds, identified by MALDI-TOF MS and 16S rRNA gene sequencing. We focused on the catalase-negative Enterobacter cloacae SEA01, which exhibits plant-promoting traits and support agave growth under nutrient-poor microcosms. In addition, this endophyte produces OMVs. Time-resolved SEM documented OMV release and cell aggregation within 9 h, followed by mature biofilms at 24 h with continued vesiculation. Purified OMVs (≈80–300 nm) contained extracellular DNA and were characterized by dynamic light scattering and UHPLC–ESI–QTOF-MS lipidomics. The OMV lipidome was dominated by phosphatidylethanolamine (~80%) and was enriched in monounsaturated fatty acids (16:1, 18:1), while the stress-associated cyclopropane fatty acids (17:1, 19:1) were comparatively retained in the whole-cell membranes; OMVs also exhibited reduced ubiquinone-8. SEA01 is catalase-negative, uncommon among plant-associated Enterobacter, suggesting a testable model in which oxidative factors modulate OMV output and biofilm assembly. These may have implications for recognition and redox signaling at the root interface. Future works should combine targeted proteomics/genomics with genetic or chemical disruption of catalase/OMV pathways. Full article

This study aimed to characterize the chemical composition and evaluate the biological activities of the aerial parts of Alkanna corcyrensis Hayek (AC), an endemic Greek species not previously studied. Phytochemical analysis of the methanolic extract was performed using UHPLC-ESI-Q-TOF–MS/MS combined with molecular networking analysis. Additionally, the total phenolic content (TPC) and total flavonoid content (TFC) were determined. Chromatographic separations were carried out to isolate major compounds, and the antioxidant capacity, along with enzyme inhibitory activity, was assessed. The analysis led to the tentative identification of 86 compounds, including 67 phenolic compounds (mainly caffeic acid derivatives and flavonoid glycosides), 10 pyrrolizidine alkaloids of trachelanthamidine, platynecine, and retronecine types, and nine organic and fatty acid derivatives. Among these, one flavonol glycoside (kaempferol-O-malonyl methyl ester hexoside) and three pyrrolizidine alkaloids (9-sarracinoyl-trachelanthamidine/isoretronecanol, retronecine-pentoside, and trachelanthamidine/isoretronecanol-hexoside) were reported for the first time. The extract exhibited high TPC (74.45 mg GAE/g extract) and TFC (46.66 mg GAE/g extract). Chromatographic separations resulted in the isolation of five major metabolites, namely rosmarinic acid, danshensu, kaempferol-3-O-glucoside, kaempferol-3-O-galactoside, and quercetin-3-O-glycoside. Biological evaluation revealed considerable antioxidant activity and inhibitory effects against α-glucosidase (6.65 mmol ACAE/g extract). Overall, this study highlights the remarkable phytochemical diversity and richness of AC among alkanet species and demonstrates its promising antioxidant potential, laying the foundation for further investigations towards its future exploitation. Full article

American ginseng (Panax quinquefolius L.) is a rare and valuable plant utilized for medicinal and culinary purposes, with its geographic origin and cultivation significantly affecting its quality and efficacy. However, the metabolic differences between cultivated and wild American ginseng are not well understood. An accurate and reliable method for tracing the origin and evaluating the quality of American ginseng is therefore urgently required. This study introduces a UHPLC-Q/TOF-MS-based comparative metabolomics and machine learning strategy for the rapid identification of wild and cultivated American ginseng. Both principal component analysis and hierarchical cluster analysis revealed distinct metabolic phenotypes between wild and cultivated American ginseng. Furthermore, the integration of univariate and multivariate statistical analyses identified eight differential metabolites in the ESI⁺ mode and three in the ESI^- mode, including seven ginsenosides. A potential ginsenosides marker panel was used to construct five machine learning models to assist in diagnosing the metabolic phenotypes of American ginseng. The Random Forest model, based on the eight differential metabolites in the ESI⁺ mode, achieved a 100% classification rate in both test and validation sets for distinguishing between wild and cultivated American ginseng. This study highlights the feasibility and application of our artificial intelligence-driven comparative metabolomics strategy for cultivar identification and geographic tracing of American ginseng, offering new insights into the molecular basis of metabolic variation in cultivated American ginseng. Full article

Anemone baicalensis, a plant abundant in Northeast China, has garnered attention for its potential medicinal properties. While its aerial parts (ABAP) have demonstrated significant antioxidant activity, the rhizome (ABR) remains less explored, particularly in comparison to the well-documented Anemones raddeanae Rhizoma, a valuable traditional Chinese medicine. This study investigates the chemical composition and bioactivity of ABR, comparing it with ABAP and evaluating its potential as a substitute for Anemones raddeanae Rhizoma. Phytochemical analyses, including qualitative and quantitative assessments, were conducted on ABR extracts using various solvents. Antioxidant activity was evaluated through multiple assays, and stability studies were performed on methanol and 80% ethanol extracts. UHPLC-ESI-Q-TOF-MS was employed to identify chemical constituents. Acute toxicity and hepatoprotective effects were assessed in vivo using a rat model. The results revealed that ABR and ABAP share nearly half of their chemical components, with ABR containing a higher diversity of triterpenoid saponins. The 80% ethanol extract of ABR exhibited the highest extraction yield, rich in phenolics and triterpenoids, and demonstrated superior antioxidant activity and stability. In vivo studies showed that ABR significantly reduced liver injury markers (ALT, AST, $\gamma$-GT, and MDA), enhanced antioxidant enzyme activity (CAT), and increased albumin concentration, comparable to the effects of Anemones raddeanae Rhizoma. Acute toxicity tests indicated low toxicity, supporting its safety for medicinal use. ABR shows significant potential as a substitute for Anemones raddeanae Rhizoma, particularly due to its rich triterpenoid content and hepatoprotective effects. While ABAP outperforms ABR in terms of antioxidant activity, ABR’s unique chemical profile and liver-protective capabilities highlight its value in drug development. This study provides a scientific foundation for the further exploration of ABR as a sustainable alternative in traditional medicine. Full article