The role of biosynthesized zinc oxide nanoparticles in modulating the expression of virulence factors in multidrug-resistant Klebsiellapneumoniae.

This study investigates the valorization of two locally available Algerian plant biomasses, Malva sylvestris and Plantago ovata, as sustainable and eco-friendly biopolymer additives for water-based drilling fluids. Their effects on key rheological and filtration properties, including apparent viscosity, yield point, gel strength, and fluid loss control were comparatively evaluated at concentrations of 2, 4 and 6.6 g per 330 mL of base fluid. Fourier Transform infrared spectroscopy analysis confirmed the presence of polysaccharide-related functional groups, while Scanning Electron Microscopy observations revealed distinct microstructures, with M. sylvestris exhibiting fibrous cellulose-like morphologies and P. ovata showing porous textures favorable for water retention and gel formation. Rheological measurements indicated that all formulations displayed pseudo-plastic shear-thinning behavior, with apparent viscosity decreasing from above 2000 mPa.s. at low shear rates to below 200 mPa.s. at high shear rates. At the highest concentration, M. sylvestris achieved higher shear stress (85 Pa) and gel strength (9.6 Pa), while P. ovata reached 70 Pa. Both additives reduced API fluid loss to 9.5 mL, with M. sylvestris providing improved pH stability (11.5) and more consistent pressure control. Overall, the results highlight the potential of these renewable, locally sourced biomaterials as sustainable alternatives to synthetic drilling fluid additives contributing to cost effective and environmentally friendly drilling operations.

Antibiotic resistance is one of the greatest challenges posed to modern medicine. It impairs the effectiveness of treatment of bacterial infections and plays a role in the increase in mortality worldwide. Given this phenomenon, the interest in alternative therapeutic methods, including phytotherapy, is growing. The aim of this study is to review the antibacterial properties of common mallow (Malva sylvestris L.) extracts based on available scientific publications. The common mallow, which is a member of the Malvaceae family, has been used in natural medicine since ancient times. Due to its multifaceted mechanism of biological action, it is currently being investigated in numerous studies in terms of its therapeutic potential, e.g. the possible use of this plant in the treatment of bacterial infections. With its phytochemical composition, M. sylvestris effectively inhibits the growth and development of many pathogens (e.g. Staphylococcus aureus, Escherichia coli, Proteus vulgaris, and Enterobacter aerogenes), which suggests that, in the future, it may complement the conventional antibacterial treatment approaches and enhance their therapeutic potential.

Background/Objectives: Dry eye disease (DED) is a multifactorial condition affecting the ocular surface. It is characterized by tear film instability, hyperosmolarity, inflammation, and oxidative stress. First-line treatment for DED relies on lubricating and hydrating eye drops, usually containing hyaluronic acid (HA), which supports tear film stability and epithelial healing. However, HA alone cannot correct oxidative stress, a key driver of cellular damage and inflammation in DED. Accordingly, this study aimed to evaluate the antioxidant capacity of Malva sylvestris tincture (MalvaT) and its physicochemical properties in experimental eye-drop formulations containing HA. Methods: The antioxidant activity of reconstituted MalvaT lyophilisate (Malva) was assessed in cell-free assays against several oxygen radicals and in cell-based assays using the human HaCat keratinocyte cell line. The refractive index was measured in eye-drop formulations containing 0.15% or 0.3% HA and 0.5% MalvaT. Surface tension was assessed in eye-drop formulations containing 0.15% HA and increasing concentrations (0.25-2.0%) of MalvaT. Results: Malva showed potent oxygen radical scavenging activity in both cell-free and cell-based assays, indicating its antioxidant capacity and the efficient cellular uptake of antioxidant components. The refractive indices of experimental eye-drop solutions containing HA and MalvaT were close to that of tear fluid (1.334). The surface tension of the experimental eye-drop formulations, while not impacted by 0.15% HA, was significantly reduced by increasing concentrations of MalvaT (p < 0.0001). At the concentration of 0.5% MalvaT, the mean surface tension was reduced from 68.17 mN/m (HA control) to 59.80 mN/m (HA + MalvaT), thereby bringing it closer to that of tear fluid. Conclusions: This pre-clinical study suggests that combining the antioxidant properties of Malva tincture with the lubricating and hydrating effects of HA in eye-drop formulations exhibiting optimal rheological characteristics may offer a promising therapeutic approach for managing DED.

Background: Malva sylvestris (the common mallow) is an herbaceous species widely used in ethnobotanical practices to treat gastrointestinal, hepatic and urinary inflammation. Objectives: Despite these beneficial effects on human health, the antineoplastic potential of this plant has not yet been fully explored. Thus, in the present study, two human colon cancer cell lines (i.e., HCT-116 and Caco-2) were treated with an extract obtained from M. sylvestris flowers (MFE), whose composition in terms of phytochemicals and microRNAs has been recently published by our research group, to explore its potential bioactivity. Methods/Results: MTT and Trypan blue assays demonstrated that MFE reduced tumour cell growth without causing significant cytotoxicity or apoptosis. Following the diphenylboric acid 2-aminoethyl ester-induced fluorescence of some plant metabolites, microscopy analysis proved that MFE components crossed the cell membranes, accumulating into nuclei. Wound assay and transwell tests documented that MFE was also able to reduce cell motility and invasiveness. In both cell lines qPCR experiments demonstrated that MFE caused the over-expression of factors, like VIMENTIN and E-CADHERIN, which negatively influence epithelial–mesenchymal transition in colon cancers. However, the effects of MFE appeared to be time-, dose- and cell type-dependent. In fact, the treatment induced senescence in P53-null Caco-2 cells (i.e., ROS, β-galactosidase and P21WAF1/Cip1) and a premise of differentiation (i.e., P27Kip1) in P53-wild-type HCT-116 cells, also via the CDK2/c-MYC/AKT axis, justifying its antiproliferative property. In parallel, the transfection of tumour cells with pure synthetic miR160b-5p—a microRNA identified in M. sylvestris flowers and predicted to target the human CDK2 transcript—resulted in gene silencing, thereby suggesting its central role in mediating the cross-kingdom effects of MFE on the investigated cancer models. Conclusions: Overall, these findings open new perspectives on the common mallow as a source of potential antimetastatic compounds and on the possible use of its plant microRNAs in the development of gene therapies.

This study investigated the clinical and immunological efficacy of combining Malva sylvestris L. extract with levamisole in calves naturally affected by bovine trichophytosis. Forty clinically diagnosed calves (8-11 months old) were randomly allocated into four groups: Control, Malva, Levamisole, and Combination. All animals received subcutaneous ivermectin (0.2 mg/kg) ten days before treatment and were maintained under uniform housing, feeding, and management conditions throughout the study. Treatments were applied for 21 days: Control (distilled water + saline), Malva (M. sylvestris extract + saline), Levamisole (distilled water + levamisole, 2.5 mg/kg), and Combination (both treatments). Lesion diameters were recorded on days 0 and 21. Blood samples collected on days 0, 7, 14, and 21 were analysed for leukocyte profiles and serum IgG, IL-6, and IFN-γ concentrations. The Combination group exhibited the most pronounced reduction in lesion size (p≤0.05) and marked elevations in leukocyte counts and IgG levels (p≤0.01). IL-6 concentrations significantly decreased in the Malva group by day 21 (p≤0.001), whereas IFN-γ levels showed notable increases in the Levamisole and Combination groups (p≤0.001). Overall, these findings underscore the therapeutic potential of integrating topical M. sylvestris with systemic levamisole as an effective complementary strategy for managing bovine dermatophytosis.

BackgroundUnderstanding physician attitudes toward natural over the counter (OTC) products in respiratory medicine remains understudied in multinational contexts. This study represents the first multinational survey of physician perceptions regarding natural syrup formulations for cough and sore throat management.ObjectiveTo assess physician acceptance of a natural syrup containing Verbascum and Malva sylvestris extracts across diverse international healthcare settings by evaluating seven key product attributes.MethodsCross-sectional survey of 398 practicing physicians across 13 countries was conducted through the Sermo global physician network in May 2025. Participants evaluated clinical appropriateness, safety in vulnerable populations, co-prescription capabilities, throat protection properties, allergen-free composition, natural ingredient preference, and nighttime convenience compared to the leading lozenge in each country.Resultswhile regional variations were observed, overall physician acceptance was highest for absence of problematic pharmaceutical ingredients (86%), followed by safety in vulnerable populations and co-prescription safety (82%), with lower agreement for natural ingredients (77%), throat protection (73%), allergen-free composition (71%), and night-time convenience (63%). Regional patterns showed consistently higher acceptance in Middle Eastern countries and below-average values in most European countries (except Ireland), with Saudi Arabia highest and Finland and Italy lowest.ConclusionPhysicians across diverse healthcare systems reported favourable attitudes toward this natural syrup formulation, with particular emphasis on the perceived safety profile for vulnerable patient groups. The notable international variation suggests the potential influence of cultural and regional factors on prescribing attitudes. While these exploratory findings should be interpreted cautiously given the descriptive study design and industry sponsorship, they provide preliminary support for the continued investigation of natural formulations in respiratory care, particularly for patients with comorbidities who may require alternatives to conventional antitussive therapies.

This study examines the content of major groups of biologically active substances (BAS) in extracts of Malva sylvestris L., a wild medicinal plant widely distributed across temperate ecosystems. Using established chemical analysis methods, reliable data were obtained for 40% and 70% aqueous-ethanol extracts regarding their BAS composition. The total phenolic content was determined spectrophotometrically using the Folin–Ciocalteu method, while the total flavonoid content was quantified by the aluminum chloride colorimetric assay. Comparative analysis revealed that the 40% extract contained higher levels of phenolic compounds (1.28 ± 0.02 mg/mL), flavonoids (0.77 ± 0.03 mg/mL), and amino acids (10.7 ± 0.01 mg/mL) than the 70% extract (0.88 ± 0.03 mg/mL, 0.35 ± 0.02 mg/mL, and 6.8 ± 0.01 mg/mL, respectively). Thin-layer chromatography additionally confirmed the presence of polysaccharides, quercetin, ascorbic acid, and anthocyanins in the analyzed samples. It was found that a 40% ethanol solution is a more effective extractant for polar compounds, whereas increasing the ethanol concentration to 70% reduces their solubility. The results confirm the potential of M. sylvestris L. extracts as a promising source of BAS for the development of safe phytotherapeutic agents.

This study evaluated hot-water infusion as a practical and sustainable extraction method for functional flower petal teas. Six edible flowers—Tagetes erecta, Lonicera japonica, Celosia argentea var. cristata, Centaurea cyanus, Hibiscus sabdariffa, and Malva sylvestris—were compared under hot-water and 80% ethanol extraction. Hot-water extraction was performed at 100 °C for 15 min. Hot-water extracts showed 1.3–4.0 times higher total phenolic content (TPC) and stronger antioxidant activities (ABTS, DPPH, and FRAP) than 80% ethanol extracts, reflecting efficient extraction of hydrophilic phenolic acids. UPLC-ESI-Q-TOF-MS and GC–MS analyses of hot-water extracts revealed chlorogenic acid, caffeic acid, coumaric acid derivatives, flavonoid glycosides, and aroma volatiles such as hexanal and α-pinene. These findings confirm that simple hot-water infusion effectively recovers both bioactive and aroma-active compounds, supporting its application in developing safe, natural, and functional flower teas.

Sustainable protein production from mallow leaves by lactic acid bacteria as a green alternative protein

Abstract Background: Wild edible plants (WEPs) have long underpinned subsistence strategies and cultural traditions in rural Europe, yet regional syntheses remain scarce. Therefore, this research aims to provide a comprehensive synthesis of Piedmont ethnobotanical heritage and investigate the abundance of WEPs along a gradient of anthropogenic influence. Methods: This study presents an in-depth literature review of WEPs in Piedmont (NW Italy), integrating a large body of grey Italian-language monographs and published papers, grouping the collected data in a database. Results: We recorded 527 species across 78 families, classifying each one according to its (i) typical environment based on three levels of anthropogenic influence (low, intermediate and high), (ii) harvested organ(s) (seven categories) and (iii) traditional culinary use(s) (eight categories). Our findings reveal that highly anthropized habitats – namely annual and perennial ruderal communities and nutrient-rich grasslands – yield the highest WEPs species richness (208 species), whereas low-disturbed forests and alpine grasslands contribute to organ and use diversity. Aerial organs (leaves, young shoots and flowers) are most commonly collected, facilitating raw and cooked applications, while hypogeal organs (tubers, bulbs) and niche uses (oils, coffee substitutes) are comparatively rare. Among all taxa, 12 species ( Foeniculum vulgare , Tanacetum vulgare, Borago officinalis , Carum carvi, Taraxacum officinale, Tilia cordata, Malva sylvestris , Myrrhis odorata , Rubus fruticosus , Rubus idaeus , Sambucus nigra and Thymus vulgaris ) stand out for their combination of high organ versatility and multiple culinary roles, highlighting their potential as flagship WEPs for gastronomic valorization. Conclusions: The persistence of traditional knowledge in the face of modernization underscores both the resilience of cultural practices and the urgency of conservation. Landscape dynamics, including both land-use intensification and abandonment, exert complex effects on WEP diversity; we advocate tailored land-management strategies that preserve habitat heterogeneity and support the WEP populations. By systematizing Piedmont ethnobotanical heritage and defining the influence of the anthropogenic gradient over it, this work establishes a foundation for future ecological, nutritional and socio-economic research. Moreover, it provides a roadmap for integrating underutilized WEPs into sustainable food systems, promoting the conservation of biodiversity and revitalizing local food traditions in the Western Alps.

Arbuscular mycorrhizal fungi (AMF) are ubiquitous in arid ecosystems, yet their distribution and community structure along spatial and ecological gradients remains insufficiently explored at regional scales. Here, we employed Malva sylvestris L., a native spontaneous plant species, to investigate the distribution patterns, phylogenetic structure, and community interactions of AMF and the associated root microbiome in dryland ecosystems. Sampling was conducted along a 700km transect extending from the Atlantic coast to inland Morocco, encompassing predominantly semi-arid ecosystems. Amplicon sequencing of the LSU rDNA region of roots and soil samples revealed a highly diverse AMF assemblage spanning ten families, including Domikaceae, Diversisporaceae, Entrophosporaceae, Sclerocystaceae, and Septoglomeraceae, while the most frequent taxa belonged to the genera Dominikia, Entrophospora, Funneliformis, and Rhizophagus. Phylogenetic alpha diversity declined with increasing soil phosphorus (P) and nitrogen (N) but increased with soil potassium, precipitation, and distance from the coastline. AMF community dissimilarity in the rhizosphere was primarily explained by distance from the coastline, MAT, and precipitation together with soil P, N, whereas AMF communities in roots were mainly structured by soil P, N, and carbon. Community assembly processes among root-associated AMF were mainly shaped by total soil N and P: total N drove local AMF community structure (positive Nearest Taxon Index) while variation in soil P increased community turnover among locations (positive beta Nearest Taxon Index). Consequently, network topology was negatively correlated with soil P, and temperature, but positively with precipitation. Specialized AMF taxa, particularly Septoglomus and Funneliformis, acted as hubs in the root fungal network, whereas generalists such as Rhizophagus and Entrophospora drove cross-kingdom associations, interacting strongly with Rhizobium, Sphingomonas, and Caulobacter. Overall, this study advances our understanding of AMF ecology in dryland ecosystems and introduces an innovative bioinformatic workflow that provides new opportunities for exploring mycorrhizal diversity and functions.

Common mallow (Malva sylvestris) is an important medicinal plant in both traditional and modern medicine. Various parts of this plant, such as the leaves, flowers, roots, and seeds, produce important biological compounds used in conventional medicine and the food industry. Despite its medicinal value, little is known about how its populations respond to drought stress at the physiological and biochemical levels. This research aimed to investigate the diversity and response of M. sylvestris populations to different physio-biochemical traits under drought stress. A field experiment was conducted with 10 populations of M. sylvestris using a randomized complete block design with three replications under normal irrigation up to 80% FC (field capacity) and drought stress 60% FC in the research institute forest and rangeland in Tehran, Iran, in 2023. Leaf samples were collected during the flowering stage, and some physio-biochemical traits and antioxidant activities were measured. Results indicated that drought stress decreased leaf area index (LAI), relative water content (RWC), chlorophyll levels, and stomatal density, while increasing osmolyte content, phenolics, anthocyanins, and antioxidant enzyme activity. These adjustments promoted osmoprotectant accumulation and strengthened antioxidant defenses, indicating an adaptive response to oxidative stress under water deficit. Genotype G9 (code 38767, Shirvan) showed the highest solute accumulation and drought tolerance, making it a promising candidate for breeding drought-resistant cultivars. Further research is recommended to explore the pharmacological properties of its secondary metabolites and to develop drought-resilient cultivars for use in herbal medicine and phytopharmaceuticals industries.

Oilfield-produced water is a major environmental concern due to elevated concentrations of dissolved metals, notably Sr, which can form insoluble scales as SrSO₄ and facilitate the co-precipitation of naturally occurring radioactive materials. In this study, for the first time, a biosurfactant extracted from Malva sylvestris leaves, rich in saponins, was explored as a green and biodegradable agent for Sr removal from highly saline produced water collected in western Iran. Batch experiments investigated the effect of temperature (25°C and 60°C) and biosurfactant concentration (0.006, 0.015, and 0.024g/mL) on removal efficiency. The removal mechanism is primarily attributed to coordination between Sr ions and the deprotonated carboxylate groups in saponin molecules, forming stable Sr-saponin complexes. Elevated temperature enhances this process through increased molecular mobility, partial dehydration of Sr2⁺ hydration shells, and favorable thermodynamic shifts, as indicated by Van't Hoff analysis. Increasing biosurfactant concentration supplies more active binding sites and improves selectivity over competing cations such as Ca2⁺ and Mg2⁺. Time-dependent experiments further revealed that complexation starts after 2 days, and complete sedimentation was observed at the end of the test (day 5). The absence of further Sr reduction between days 5 and 10 indicates the long-term stability of the complexes and their resistance to re-dissolution. A peak Sr removal efficiency of 63.6% was achieved at 60°C and 0.024g/mL with no significant change in the pH of the aqueous phase, demonstrating the potential of M. sylvestris for stable and efficient Sr removal from real oilfield produced water. This approach provides a novel, sustainable, and cost-effective alternative to conventional synthetic chemicals for produced water treatment.

Background and objectivesStomatitis is among the most debilitating complications of cancer treatment, particularly chemotherapy. The use of medicinal plants has been suggested as a potential therapeutic approach to mitigate this condition. This study aimed to evaluate the effect of Malva sylvestris (mallow) mouthwash on chemotherapy-induced stomatitis in cancer patients.Materials and methodsThis triple-blind randomized clinical trial was conducted in 2024 on 70 cancer patients with chemotherapy-induced stomatitis who attended medical centers in an urban area of Iran. Participants were randomly allocated to two groups using the minimization method. The intervention group received 15 ml of Malva sylvestris mouthwash three times daily for 14 days, while the control group received standard chlorhexidine mouthwash. The severity of stomatitis and associated pain were assessed at baseline and on days 3, 7, and 14 using the World Health Organization (WHO) Mucositis Scale and the Visual Analog Scale (VAS) for pain, respectively. Data were analyzed using SPSS version 22.ResultsBefore the intervention, no statistically significant differences were observed in mean stomatitis or pain scores between the two groups. By day 7, the intervention group demonstrated significantly lower mean scores for both stomatitis and pain compared with the control group (P < 0.001). Moreover, the reduction in stomatitis severity over time was greater in the intervention group than in the control group (P < 0.05). On day 14, although the intervention group maintained lower mean stomatitis scores, the difference between the two groups was not statistically significant (P = 0.08).ConclusionThe findings suggest that Malva sylvestris mouthwash has a beneficial effect in reducing the severity of stomatitis and associated pain in patients undergoing chemotherapy. Therefore, its use may be recommended as a supportive intervention for patients, caregivers, and healthcare professionals involved in cancer care.Trial registrationTrial registration number: IRCT20150713023190N15.Date of registration: 2024/07/20.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-025-15158-w.

Urinary tract infection (UTI) is a common infection, particularly among reproductive- age and elderly women. While antibiotics remain the primary treatment, antimicrobial resistance is a growing global concern, necessitating effective non-antibiotic alternatives. This study determines the safety and efficacy of herbal formulation compared to nitrofurantoin in treating uncomplicated UTI and its impact on health-related quality of life (HRQoL) in women. A randomized, standard-controlled study was conducted involving 66 women diagnosed with uUTI. Participants were randomly assigned to either the Unani Herbal Formulations group (UHF) or the nitrofurantoin group (NG). The UHF group (n=33) received a Unani herbal formulation (3.5 g powder of Cucumis sativus L., Lagenaria siceraria, Portulaca oleracea L., Malva sylvestris L., and Adiantum capillus-veneris L.) along with 25 ml of Viola odorata L. syrup, administered twice daily for 8 days. The NG (n=33) received nitrofurantoin (100 mg) twice daily for 7 days. The primary outcome was the Urinary Tract Infection Symptom Assessment (UTISA) questionnaire. The secondary outcomes were symptom severity, urine dipstick test, urine culture and sensitivity, SF-12 HRQoL survey, and safety evaluation. Standardization, in vitro, and phytochemical analyses of the herbal formulation were also conducted. UTISA scores significantly improved within both groups (p < 0.001). The UTISA total score from 14.21 ± 3.43 and 14.18 ± 3.36 was reduced to 3.12 ± 4.65 and 1.61 ± 3.10 in the UHF and NG, respectively, on the 9th day. On the 9th day, 81.81% (UHF) and 87.87% (NG) had negative cultures. HRQoL scores showed significant within-group improvements (p < 0.001), with no significant difference between groups (p > 0.05). No adverse effects were reported. HPLC analysis confirmed the presence of Luteolin, a bioactive antimicrobial metabolite. The herbal formulations were as effective as nitrofurantoin in treating bacterial uUTIs, with no adverse effects and enhanced HRQoL. The presence of Luteolin supports its antimicrobial potential, offering a favourable non-antibiotic alternative for UTI management. The research project is registered at the Clinical Trial Registry of India, ICMR with No: CTRI/2021/09/036138 dated 01/09/2021. The protocol can be retrieved from CTRI.

Abstract This study offers a green, sustainable route to synthesize copper oxide (CuO) and nickel‐doped CuO (Ni‐doped CuO) nanoparticles using Malva sylvestris leaf extract as a natural reducing and stabilizing agent. Nickel content was varied from 1 to 10 mol%, and the materials were characterized by FE‐SEM, EDX, XRD, FTIR, and UV–vis spectroscopy. The 7.5 mol% Ni‐doped CuO exhibited the most favorable characteristics, uniform crystallite size, and an optimal optical band gap of ∼3.0 eV, resulting in excellent photocatalytic activity, achieving 98% degradation of methylene blue (MB) under visible light within 3 h. Band‐structure estimates for CuO and 7.5 mol% Ni‐doped CuO place the conduction band minimum and valence band maximum at −0.95 and −2.02 eV, respectively, providing sufficient driving force for reactive oxygen species generation and efficient dye degradation. The nanoparticles also demonstrated good stability and reusability over four successive cycles. Overall, this work highlights the potential of locally available M. sylvestris leaf extract for eco‐friendly, reproducible synthesis of high‐performance Ni‐doped CuO photocatalysts, offering a promising route for sustainable wastewater treatment.

Potential benefits of Malva sylvestris in treating dry-eye pathology in vitro

This study developed edible bilayer films from Malva sylvestris gum (MSG) and chitosan (CH), with a focus on incorporating Salvia abrotanoides essential oil (EO) as a bioactive additive. EO addition remarkably decreased moisture content from 0.096 ± 0.028 to 26.08 ± 0.15, solubility from 65.84 ± 0.54 to 31.45 ± 0.19, while thickness increased significantly from 0.096 ± 0.028 to 0.118 ± 0.09 for MSG/CH and MSG/CH-2 %EO, respectively. Color and optical characteristics were determined. SEM images revealed film's microstructure and FTIR analysis evaluated chemical interactions within polymer matrix. Differential scanning calorimetry revealed a reduction in Tg, from 27 °C (MSG/CH) to 10.8 °C (MSG/CH-2 %EO), indicating enhanced chain mobility. Mechanical evaluation showed a decrease in tensile strength (12.5 to 5.1 MPa) but an improvement in elongation at break (12.2 % to 18.6 %) (p < 0.05). Antioxidant evaluation demonstrated a significant rise in DPPH scavenging activity at 2 % EO in bilayer films. These findings highlight EO-incorporated MSG/CH films as a promising biodegradable packaging material with enhanced functional properties.

Introduction: A Polyherbal Unani Powder Formulation for Urinary Tract Infections (PUFP-UTI) comprises Adiantum capillus-veneris L., Cucumis sativus L., Lagenaria siceraria, Malva sylvestris L., and Portulaca oleracea L. This study aimed to evaluate the physicochemical characteristics, phytochemical composition, quantification of luteolin using HPLC, microbial safety, and in vitro antibacterial activity to assess the quality, efficacy, and safety of the formulation. Methods: Physicochemical analysis included organoleptic properties, ash value, density, moisture content, and extractive values. Phytochemical screening detected phenols, tannins, flavonoids, alkaloids, glycosides, saponins, terpenoids, and sterols. Quantitative analysis measured total phenols, tannins, flavonoids, and alkaloids. Luteolin content was quantified by HPLC. Microbial contamination was assessed following WHO guidelines. Antibacterial activity was tested against E. coli, S. aureus, K. pneumoniae, E. faecalis, P. mirabilis, S. saprophyticus, and P. aeruginosa using the agar well diffusion method. Results: PUFP-UTI exhibited acceptable physicochemical properties and contained significant amounts of phenols, tannins, flavonoids, alkaloids, and luteolin. The formulation met WHO safety standards for microbial contamination. It also showed substantial antibacterial activity against all tested pathogens. These results indicate that PUFP-UTI is a microbiologically safe herbal formulation with notable antimicrobial efficacy. Discussion: The presence of multiple phytoconstituents, such as flavonoids and tannins, along with measurable luteolin content, may contribute to the observed antibacterial effect of PUFP-UTI. Its broad-spectrum activity against common uropathogens supports its pharmacological relevance. The formulation also complies with basic quality and safety benchmarks, reinforcing its potential as a safe traditional remedy. Conclusion: PUFP-UTI's antibacterial properties support its traditional use in treating urinary tract infections. Further stability studies and clinical trials are recommended to confirm its therapeutic potential.