Rayssa Ribeiro

Oleoresins are complex natural lipophilic matrices traditionally analyzed using chromatographic techniques that require extensive sample preparation, derivatization, and authentic standards. Amazonian oleoresins from Copaifera and Eperua species (Fabaceae) represent valuable bioresources with recognized pharmacological potential, largely attributed to diterpenoids such as copalic and hardwickiic acids, as well as bioactive sesquiterpenes, including the cannabinoid β-caryophyllene. In this study, we present a proof-of-concept application of Direct Analysis in Real Time coupled with High-Resolution Mass Spectrometry (DART-HRMS) as a rapid, direct, and environmentally friendly approach for chemical fingerprinting and semi-targeted screening of the two most important amazonian oleoresins from these two genera: Eperua oleifera and Copaifera multijuga. Analyses were performed using a Q Exactive Orbitrap coupled to a DART ion source under optimized conditions. Hardwickiic acid was used as a model compound for method optimization, with optimal performance achieved at 200 °C and 100 V, yielding stable signal intensities (CV < 10%) and high mass accuracy (<1 ppm). The method enabled reproducible detection of diterpenic acids in both oleoresins, allowing differentiation of their chemical profiles and assessment of short-term stability under ambient conditions. In addition to diterpenes, free fatty acids were also detected, expanding the compositional characterization of these matrices. Compound annotation was performed based on accurate mass measurements and literature comparison, corresponding to Level 5 confidence according to established metabolomics criteria. Although the absence of chromatographic separation limits isomer discrimination and absolute quantification, DART-HRMS provides a rapid and solvent-free strategy for chemical fingerprinting and preliminary characterization of oleoresins. This approach aligns with Green Chemistry principles and shows strong potential as a screening and triage tool for quality control, chemotaxonomic studies, and sustainable valorization of Amazonian natural products.

Oleoresins are complex natural lipophilic matrices traditionally analyzed using chromatographic techniques that require extensive sample preparation, derivatization, and authentic standards. Amazonian oleoresins from Copaifera and Eperua species (Fabaceae) represent valuable bioresources with recognized pharmacological potential, largely attributed to diterpenoids such as copalic and hardwickiic acids, as well as bioactive sesquiterpenes, including the cannabinoid b-caryophyllene. In this study, we present a proof-of-concept application of Direct Analysis in Real Time coupled with High-Resolution Mass Spectrometry (DART-HRMS) as a rapid, direct, and environmentally friendly approach for chemical fingerprinting and semi-targeted screening of the two most important amazonian oleoresins from these two genera: Eperua oleifera and Copaifera multijuga. Analyses were performed using a Q Exactive Orbitrap coupled to a DART ion source under after conditions optimization. Hardwickiic acid was used as a model compound for method optimization, with optimal performance achieved at 200 °C and 100 V, yielding stable signal intensities (CV < 10%) and high mass accuracy (< 1 ppm). The method enabled reproducible detection of diterpenic acids in both oleoresins, allowing differentiation of their chemical profiles and assessment of short-term stability under ambient conditions. In addition to diterpenes, free fatty acids were also detected, expanding the compositional characterization of these matrices. Compound annotation was performed based on accurate mass measurements and literature comparison, corresponding to Level 5 confidence according to established metabolomics criteria. Although the absence of chromatographic separation limits isomer discrimination and absolute quantification, DART-HRMS provides a rapid and solvent-free strategy for chemical fingerprinting and preliminary characterization of oleoresins. This approach aligns with Green Chemistry principles and shows strong potential as a screening and triage tool for quality control, chemotaxonomic studies, and sustainable valorization of Amazonian natural products.

Eperua oleifera Ducke, commonly known as “copaíba‐jacaré,” produces oilresins traditionally used in folk medicine for wound healing and as antifungal and antibacterial agents, similar to those of Copaifera species. However, its chemical composition remains poorly characterized. This study investigated the oilresin of E. oleifera collected in the Brazilian Amazon region. After derivatization, the sample was analyzed by GC–MS, with data compared to literature, mass spectral libraries, and standards previously isolated. In addition, neutral and acidic fractions were separated using ion‐exchange open‐column chromatography with silica modified with KOH‐impregnated silica as the stationary phase. Twelve diterpenes were identified, including nine carboxylic acids and three alcohols. Hardwickiic acid and copalic acid were the major components, fully characterized by NMR. The most abundant compound in the neutral fraction was isolated and identified by HRMS and NMR as the methyl ester of hardwickiic acid—a compound not previously reported in oilresins. The application of GC–MS enabled the identification of diterpenes based on known fragmentation patterns. In addition, the direct infusion experiment detected a methyl ester that, to our knowledge, has not been previously reported in oilresins.

Eperua oleifera Ducke (Fabaceae), commonly known as copaíba-jacaré, is traditionally used for therapeutic purposes, like Copaifera oleoresins. Previous GC-MS studies reported its chemical composition as mainly composed of diterpenic acids, consistent with species of the same genus. Although GC-MS remains widely used for comparing compound retention times and fragmentation patterns, its application to diterpenic acids requires a derivatization step to form methyl esters due to the poor chromatographic performance of carboxylic acids on methyl silicone stationary phases. This step may lead to misinterpretations, especially considering recent findings of naturally occurring methyl esters in oleoresins that may co-elute with derivatized acids. This study aimed to apply more sensitive analytical techniques to identify both target and untargeted compounds. The resin of E. oleifera was analyzed by GC-MS to assess the presence of volatile components. Additionally, UHPLC-HRMS was employed using full-scan MS, data-dependent acquisition (DDA), and parallel reaction monitoring (PRM) in both positive and negative ESI modes. GC-MS confirmed the absence of volatile sesquiterpenes, classifying E. oleifera as a resin. Targeted UHPLC-HRMS detected natural methyl esters of diterpenic acids, while untargeted analysis using Compound Discoverer 3.3 software revealed flavonoids and phenolic compounds not previously reported. These findings support the application of UHPLC-HRMS as a powerful tool in phytochemical studies.

Eperua oleifera Ducke (Fabaceae), commonly known as copaíba-jacaré, is traditionally used for therapeutic purposes, like Copaifera oleoresins. Previous GC-MS studies reported its chemical composition as mainly composed of diterpenic acids, consistent with species of the same genus. Although GC-MS remains widely used for comparing compound retention times and fragmentation patterns, its application to diterpenic acids requires a derivatization step to form methyl esters due to the poor chromatographic performance of carboxylic acids on methyl silicone stationary phases. This step may lead to misinterpretations, especially considering recent findings of naturally occurring methyl esters in oleoresins that may co-elute with derivatized acids. This study aimed to apply more sensitive analytical techniques to identify both target and untargeted compounds. The resin of E. oleifera was analyzed by GC-MS to assess the presence of volatile components. Additionally, UHPLC-HRMS was employed using full-scan MS, data-dependent acquisition (DDA), and parallel reaction monitoring (PRM) in both positive and negative ESI modes. GC-MS confirmed the absence of volatile sesquiterpenes, classifying E. oleifera as a resin. Targeted UHPLC-HRMS detected natural methyl esters of diterpenic acids, while untargeted analysis using Compound Discoverer software revealed flavonoids and phenolic compounds not previously reported. These findings support the application of UHPLC-HRMS as a powerful tool in phytochemical studies.

Trema micranthum (L.) Blume is a species from the Cannabaceae botanical family, present all over the tropics. A recent study conducted by our group identified the presence of cannabidiol (CBD), Δ⁹‐tetrahydrocannabinol, cannabidiolic acid, and tetrahydrocannabinolic acid‐A in T. micranthum extracts. Their presence in the plant's fruit, inflorescence, and leaf extracts was detected and quantified. T. micranthum has been identified as a potential new source of cannabinoids such as CBD. The present work focuses on a new Cannabinoid Profiling System (CPS) to present a wide view of cannabinoid chemical composition with several compassionate and modern mass spectrometry (MS) tools. Extracts from fruits, leaves, and inflorescences were obtained using a methanol/hexane (9:1, v/v) solvent mixture. Ultra high‐performance liquid chromatography coupled with high‐resolution tandem MS using targeted and untargeted approaches were used in these approaches. CPS allowed the identification of the 26 cannabinoids in T. micranthum described for the first time in this plant.

Background/Objectives: Andiroba oil, extracted from Carapa guianensis seeds, possesses therapeutic properties including anti-inflammatory and wound healing effects. This study aimed to develop and characterize a nanoemulsion formulation containing andiroba oil (NeAnd) and to evaluate its cytotoxicity and wound healing potential in vitro. Methods: The oil was evaluated for acidity, antioxidant activity, and fatty acid composition. NeAnd was produced by ultrasonication and characterized using FTIR (Fourier transform infrared spectroscopy), Raman spectroscopy, dynamic light scattering, and transmission electron microscopy. Results: NeAnd exhibited a spherical shape and stable physicochemical properties, with an average hydrodynamic diameter (HD) of 205.7 ± 3.9 nm, a polydispersity index (PdI) of 0.295 ± 0.05, a negative zeta potential of −4.16 ± 0.414 mV, and pH of approximately 6.5. These nanodroplets remained stable for 120 days when stored at 4 °C and maintained their parameters even under pH variations. FTIR and Raman analyses confirmed the presence of functional groups and the organization of fatty acid chains in NeAnd. Cell viability assays revealed no statistically significant differences in cytotoxicity at various concentrations (90–360 µg/mL) after 24 and 48 h. In scratch wound healing assays, NeAnd significantly enhanced wound closure (88.9%) compared to the PBS control (38%) and free andiroba oil (68.6%) in keratinocytes (p < 0.05). Conclusions: These promising findings indicate NeAnd as a potential nanophytomedicine for wound healing and tissue regeneration treatments.

Trema micranthum (Cannabaceae) has emerged as a promising new source of cannabinoids, including cannabidiol (CBD). Given the substantial medicinal demand for cannabinoids and the regulatory challenges associated with Cannabis sativa due to the presence of Δ⁹-tetrahydrocannabinol (THC), this study sought to explore the presence of CBD, THC, and their precursors, Δ⁹-tetrahydrocannabinolic acid A (THCA A) and cannabidiolic acid (CBDA), in various parts of Trema micranthum using UHPLC-HRMS/MS (Orbitrap). Extracts from fruits, leaves, inflorescences, and stems were obtained using a methanol/hexane (9:1, v/v) solvent mixture. UHPLC coupled with an Orbitrap mass spectrometer was employed for cannabinoid identification and quantification, with standard mixtures prepared in methanol. The extracts yielded significant quantities, such as 6.6%/g from leaves and 3%/g from fruits. Cannabinoids were detected in fruits, leaves, and inflorescences, with acidic forms (CBDA and THCA A) present in higher concentrations than their neutral counterparts. Notably, leaves contained 4.43 × 10⁻³ µg/g of CBD and 1.05 × 10⁻³ µg/g of THC. These findings, facilitated by high-resolution analytical methods, underscore the potential of Trema micranthum as an alternative source for cannabinoids, guiding future research in this area.

Copaiba oil is a natural product commonly employed in traditional medicine due to qualities as anti-inflammatory and antimicrobial mainly. On dentistry, studies have focused on the use of copaiba in cavity varnishes, endodontic cements, bacterial biofilm control and even in the composition of local anesthetics. The aim of this study was to investigate on the use of copaiba oil within dental specialties. The search was conducted on databases using the primary keywords ‘Copaifera’ and ‘Dentistry’ resulting in 687 studies. Was observed that Copaifera multijuga was the most prevalent species in the evaluated articles, endodontics was the most presente dental specialty in publications. Antimicrobial tests and biocompatibility tests was most prevalents. Based on the results of this study, despite the limited number of evaluated works, it can be concluded that over the years, copaiba has been increasingly used in dentistry, yielding promising results such as the development of new bioproducts that combine copaiba's pharmacological characteristics without compromising the necessary physical attributes for clinical use.

The current era witnesses a remarkable advancement in biomass utilization, guided by the principles of green chemistry and biorefinery and the comprehensive exploitation of plant-based raw materials. Predominantly, large-scale production methods have been pursued, akin to approaches in the oil industry, enabling the incorporation of novel products into energy and petrochemical markets. However, the viability of such systems on a small and medium scale is hindered by logistical challenges and the constraints of economies of scale. For small agricultural producers and food processing companies, the complete utilization of biomass transcends environmental responsibility, evolving into a strategy for survival through the diversification of by-products with enhanced value. The state of Rio de Janeiro in Brazil presents a range of population dynamics, geographical features, climate conditions, and agricultural production patterns that closely resemble those found in various tropical countries and agricultural regions worldwide. This region, sustaining a green belt supporting 17 million people, provides an apt case study for investigating chemical compounds with potential value among agro-industrial residues, which can motivate the creation of a lucrative biotechnological industry. Examples include naringenin and hesperidin from oranges and lemons, epi-gallo-catechin gallate from bananas, caffeic acids from coffee, and the bromelain enzyme from pineapples. This study addresses the challenges associated with developing biotechnological alternatives within the agroindustry, considering economic, technological, logistical, and market-related aspects. The insights from examining the Brazilian state of Rio de Janeiro will contribute to the broader discourse on sustainable biomass utilization and the creation of value-added by-products.