André Santos Alkimim, Maria Luiza Nitzsche Morato, Caroline Roberta Leal da Mota, Marina Pereira Rocha, Juliana Rodrigues Vasconcelos, Claudinei da Silva Souza, Júlio César Dias Lopes, Wagner Campos Otoni, Cláudia Gontijo Silva, Vera Lucia Almeida, Priscilla Rodrigues Valadares Campana
The wild species Passiflora setacea DC., known as ‘maracujá-do-sono’ or ‘maracujá-sururuca’, has chemical and pharmacological properties that are important to the food and pharmaceutical industries. This study aimed to identify and quantify polyphenols and flavonoids in extracts from leaves of in vivo orchard-grown and greenhouse-derived P. setacea plants. The extracts were also evaluated for their antioxidant (AAO) and acetylcholinesterase (AChE) inhibitory activity. The polyphenols and flavonoids were quantified spectrophotometrically. Based on the results, the hydroalcoholic extract from the leaves of P. setacea (PSHF-01) plants grown in the orchard was selected for further studies. In this extract, the flavonoids were characterized and identified using spectrophotometric and chromatographic methods. In addition, its antioxidant potential was evaluated through DPPH free radical scavenging, the ß-carotene/linoleic acid lipid peroxidation method, and the liposome peroxidation assay (TBARS). Furthermore, in silico activity prediction for flavonoids from P. setacea was performed using the Active-IT system. The total polyphenol content of the extracts varied from 11.32 ± 0.33 to 40.59 ± 1.5 mg of gallic acid equivalents per gram of extract. For flavonoid quantification, the amounts ranged from 3.79 ± 1.10 to 33.65 ± 4.73 mg of total flavonoids per gram of extract, calculated as apigenin. The extracts were considered promising for antioxidant activity using the β-carotene/linoleic acid peroxidation method. Concerning AChE inhibition, the extracts did not show significant values (%I > 40%). In the selected extract, PSHF-01, isovitexin and isoorientin were identified by chromatographic techniques (TLC, HPLC-UV and UPLC-PDA-ESI-MS/MS). Furthermore, the compounds luteolin-(O -glucopyranosyl)- C -glucopyranoside, (iso)orientin-C-xyloside, vicenin II, (iso)vitexin-2”-O-xyloside, and (iso)orientin-C-rhamnoside were tentatively characterized by UPLC-PDA-ESI-MS/MS. HPLC-DAD method was developed and validated for the determination of isovitexin in PSHF-01. Antioxidant activity was detected only in the ß-carotene/linoleic acid assay, whereas no activity was observed in the DPPH or TBARS assays. These differences may be attributed to distinct mechanisms underlying each assay. In silico predictions indicated that the evaluated flavonoids have the potential to exhibit hepatic metabolic stability and antiparasitic, anti-inflammatory, antitumor, and immunomodulatory activities. These results enhance our understanding of the chemical composition and biological properties of P. setacea and open new avenues for future research that could support its pharmaceutical potential and agronomic applications.
Priscila Brasil Augusto de Souza, Davi Santos Oliveira, Estefane de Oliveira Picanço, Magno Perêa Muniz, Claudete Catanhede do Nascimento, Maria da Paz Lima
Ceiba pentandra is a pantropical species and one of the largest trees in Brazil, recognized for its rich diversity of secondary metabolites, especially flavonoids. In this study, phytochemical investigation of the methanolic extract obtained from the wood resulted in the isolation of seven flavonoids, including one new biflavonoid (7). The structures of compounds 1–7 were elucidated using 1D and 2D NMR spectroscopy and HRESI-MS. Comparison with literature data enabled the identification of the known compounds and supported the structural assignment of biflavonoid (7). The occurrence of this dimeric biflavonoid expands the phytochemical knowledge of Ceiba pentandra and reinforces the importance of Amazonian woody species as promising sources of structurally diverse phenolic metabolites.
Mustapha N. Abubakar, Vitor S. Mazucato, Paulo C. Vieira
Fungal communities mediate antagonism and emergent, interaction-specific inhibition through a dense chemical dialogue, yet linking ecological phenotypes to specific metabolites remains a central challenge in natural product discovery. Conventional bioassay-guided fractionation is iterative and often biased toward abundant constituents. We addressed this challenge using a consortium of protease-rich, fruit-derived fungi (Neofusicoccum ribis, Fusarium sacchari and F. verticillioides). A hypothesis-generating metabolomics workflow was applied, coupling controlled axenic, pairwise and three-way co-culture phenotyping with 1 H NMR-guided orthogonal projections to latent structures-discriminant analysis (OPLS-DA). An exploratory OPLS-DA model compared the 1 H NMR spectra of bioactive crude extracts with those of 31 purified metabolites to rank compounds by spectral similarity as a hypothesis-generating prioritization strategy. The five highest-ranked compounds were then screened in a preliminary antifungal assay at a single concentration of 50 μg/mL. Potent allelopathy of axenic F. verticillioides (V18) (∼70% inhibition against V6) was significantly attenuated in the V6/V18 duo co-culture (adjusted p < 0.0001, Cohen’s d = 3.1), whereas a unique three-way (trio) emergent inhibition arose against an otherwise resistant competitor. The trio culture produced the highest biomass, and the endophyte (V6) grew substantially faster than either pathogen. The OPLS-DA model (Q 2 = 0.748) prioritized several candidates, including a constitutively produced isocoumarin (Compound 5) and Compound 27 [7-hydroxy-3-(hydroxymethyl)-5-methyl-1H-isochromen-1-one], which was isolated exclusively from the trio co-culture—the only condition that inhibited F. sacchari. Preliminary testing confirmed statistically significant activity of the five highest-ranked compounds against at least one target fungus (Table S5). Compound 27 showed moderate activity against F. sacchari (20.9% inhibition), while Compounds 11, 5 and 6 exhibited comparable or higher activity against F. verticillioides. These findings generate chemically explicit hypotheses and provide initial experimental support for the prioritization workflow, but they do not establish causality or confirmed potency of individual compounds. Emergent bioactivities in this system are best interpreted as the combined outcome of ecological competition and interaction-triggered secondary metabolism. Dose-response assays are required before functional relevance can be claimed.
Carolina Paula de Souza Moreira, Natasha Reynaud, Marcos Paulo Gomes Mol, Gilson Faria, Sérgio Caldas
Arboviral diseases such as dengue, Zika, and chikungunya remain major public health challenges worldwide. In the absence of broadly effective antiviral therapies and considering the limitations of current vaccination strategies, vector control remains the primary approach for reducing disease transmission. Essential oils have emerged as promising alternatives to synthetic insecticides because of their renewable origin, biodegradability, and multiple mechanisms of action. Nevertheless, their practical application is often limited by poor water solubility and physicochemical instability. This study investigated the chemical composition, larvicidal activity, and microemulsion-based formulation of Mentha piperita L. essential oil against Aedes aegypti larvae. The chemical composition of the essential oil was determined by gas chromatography–mass spectrometry (GC–MS). Larvicidal activity against third-instar A. aegypti larvae was evaluated by determining mortality rates and the median lethal concentration (LC 50). A pseudoternary phase diagram was constructed to identify suitable microemulsion regions, and selected formulations were characterized according to droplet size, polydispersity index (PDI), and short-term physical stability. The larvicidal activity of the resulting microemulsions was subsequently evaluated at different essential-oil concentrations. A blank microemulsion containing the formulation components without M. piperita essential oil was also evaluated as a control. GC–MS analysis identified 14 compounds representing nearly 100% of the essential oil composition, with menthol (50.17%) and menthone (23.02%) as the predominant constituents. The free essential oil exhibited concentration-dependent larvicidal activity against A. aegypti, with an LC 50 of 175.7 μg/mL and complete larval mortality at 250 μg/mL. Among the tested formulations, PME-05 was selected as the most promising formulation for further development because it achieved complete larval mortality while containing only 5% peppermint essential oil and exhibited the lowest PDI (0.06 ± 0.007), indicating a highly homogeneous dispersed system. The results support the use of microemulsion technology as a suitable delivery system for M. piperita essential oil, facilitating its dispersion and application in aqueous environments while retaining its larvicidal activity. The developed formulation represents a promising platform for the application of botanical larvicides in mosquito control. Further studies are needed to evaluate long-term physicochemical stability, residual activity, effects on non-target organisms, and performance under semi-field and field conditions.
Alberto Araujo de Caux, José Cláudio Klier Monteiro Filho, Ruan Barbosa da Silva e Medeiros, Marili Villa Nova Rodrigues, Marcos Nopper Alves, Marcus Bruno Soares Forte, Maria Isabel Rodrigues, Rodney Alexandre Ferreira Rodrigues
Monteverdia ilicifolia (Mart. Ex Reiss.) Biral, popularly known as espinheira-santa, is a medicinal plant recommended in Brazilian primary healthcare due to its well-recognized antidyspeptic, antacid, and gastric protective properties. Despite its clinical relevance, the development of herbal medicine from this species remains challenging, particularly due to the intrinsic variability of plant derived materials. However, most studies focus on isolated stages of the process, and there is a lack of integrated approaches that evaluate the entire production chain. In this context, Quality by Design (QbD) emerges as a promising framework to systematically understand, control, and optimize this process. Thus, the objective of this study was to use QbD in the evaluation of the production processes of an herbal medicine made from M. ilicifolia, aiming to ensure its quality. The production processes were investigated through process mapping, risk assessment, design of experiments (DoE), and process validation. The initial stages involved hazard identification and risk assignment to identify potentially critical factors, which were subsequently challenged by DoE. The processes were compared based on physicochemical, operational, and technological responses related to Critical Quality Attributes (CQAs), which impact product efficacy and safety. Finally, the optimized condition was subjected to process validation to ensure reproducibility under routine manufacturing conditions. The stages of harvesting, drying, milling, extraction, concentration, spray drying, and granulation were evaluated. Among the nine factors assessed during lab-scale extraction, “drug-to-extract ratio” (1% and 2%) and “degree of fragmentation of the herbal drug” (shredded and powdered) showed consistent significant effects. Upon scale-up of the extraction process, “agitation” (0–400 rpm) also proved to be a determining factor at larger scale. For the spray drying and granulation stages, the addition of maltodextrin (0%–60%), feed rate (3.9–8.1 mL/min), and inlet temperature (120°C–200 °C) were significant for at least one CQA. In conclusion, the application of the QbD approach enabled the identification of the main elements related to the production of herbal medicines from espinheira-santa, satisfying the process capability.
