Antiviral Activity and Chemical Characterization of Ficus citrifolia Mill. (Moraceae) Leaf Extract Against Emerging Arboviruses

Antiviral Activity and Chemical Characterization of Ficus citrifolia Mill. 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(Moraceae) Leaf Extract Against Emerging Arboviruses Horrana Acácio Mardegan, Allana Martins Ataíde, Loueny Celine Ribeiro Andrade, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7583932/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 24 Mar, 2026 Read the published version in Revista Brasileira de Farmacognosia → Version 1 posted 5 You are reading this latest preprint version Abstract Arboviral diseases pose a serious public health problem, particularly in tropical regions, where environmental conditions favor the proliferation of mosquito vectors, facilitating viral transmission and leading to debilitating clinical manifestations ranging from fever and joint pain to severe neurological complications. This study evaluated the in vitro antiviral activity and phytochemical profile of the ethanolic extract of Ficus citrifolia Mill. leaves (Moraceae). Using high-resolution LC-MS/MS analysis, 21 compounds were identified, including phenolic acids, flavonoids, lignans, terpenes, and steroids - many of which are known for their antiviral and anti-inflammatory properties. The extract exhibited significant antiviral activity against Mayaro virus (EC₅₀: 61.24 µg/mL, SI: 12.96), Chikungunya virus (EC₅₀: 117.92 µg/mL, SI: 6.73), and Zika virus (EC₅₀: 117.29 µg/mL, SI: 6.77), with no observed activity against Oropouche virus. The findings highlight the potential of F. citrifolia as a source of antiviral agents and support its traditional use in treating inflammatory and infectious diseases. Further studies, including bioassay-guided fractionation and mechanistic evaluations, are recommended to identify the active constituents and assess clinical applicability. LC-MS/MS Compound Annotation Antiviral Activity Moraceae Introduction Arboviruses stand out as infectious agents transmitted to humans by the bite of infected blood-feeding arthropods. Tropical and subtropical climates greatly facilitate the reproduction of these vectors, especially in countries such as Brazil, where climatic conditions favor the spread of arboviruses such as dengue, yellow fever, chikungunya, Zika, Mayaro, and Oropouche. Although these diseases are endemic, they continue to be neglected, with 1,645,678 cases reported in the Americas in 2020 based on PAHO and WHO (2020). In developing countries such as Brazil, unorganized urbanization and poor sanitation infrastructure increase the vulnerability of the population. In addition, the lack of effective vaccines and antivirals aggravates the situation, overburdening health systems and favoring outbreaks in the words Girard et al. ( 2020 ). The resurgence of Zika (ZIKV), Oropouche (OROV), Mayaro (MAYV), and Chikungunya (CHIKV) in urban centers has intensified these challenges. These viruses produce similar symptoms, such as fever, rashes, and severe joint pain, sometimes progressing to chronic arthritis. ZIKV is known to cause congenital malformations and vertical transmission, as noted by Pielnaa et al. ( 2020 ), and OROV has recently been linked to similar physical changes and vertical transmission as described by Schwartz, Dashraath & Baud ( 2024 ). MAYV can cause complications such as myocarditis, hemorrhagic syndromes, and neurological manifestations. Faced with these clinical challenges, the search for new therapeutic sources has increasingly turned to the ancestral knowledge of traditional communities about medicinal plants. The biodiversity of the Amazon offers several species with therapeutic potential, supporting traditional healing practices and providing an accessible alternative. Among these species, those of the genus Ficus are widely used in traditional medicine for their anti-inflammatory, analgesic, and healing properties, as stated by Lansky et al. (2008). These plants are known for their rich chemiodiversity, including organic acids, phenolic compounds, and terpenes reported in Cruz et al. (2022). Previous studies have demonstrated the therapeutic effects of species such as F. iteophylla , which reduced pain and inflammation in rodents, as discussed in Abdulmalik et al. ( 2011 )d amplissima , which exhibited anti-inflammatory, antioxidant, and wound-healing properties in vivo based on Arunachalam & Parimelazhagan (2012). Although these effects are not directly antiviral, viral pathogens induce inflammatory responses in the host as part of the infection process, in accordance with Pingen et al. ( 2016 ). Ethnopharmacological applications may therefore be linked to antiviral potential, as evidenced by F. sur and F. rubiginosa inhibiting herpes simplex virus type 1 (HSV-1), as described by Sieniawska et al. ( 2022 ) and Dell'Annunziata et al. (2022), F. deltoidea showing antiviral activity against dengue virus type 2, according to Wu et al. ( 2024 )d carica inhibiting SARS-CoV-2 replication, based on Hamed et al. ( 2023 ). Although several Ficus species have been studied, no research has evaluated the antiviral potential of F. citrifolia , commonly known as “fig tree” or “black gameleira.” Previous studies have highlighted the antimicobacterial effects of its ethanolic leaf extract against Mycobacterium tuberculosis , as well as the antiproliferative effect of three lignans isolated from the leaf extract with dichloromethane, as reported by Antoun et al. ( 2001 ) and Baetas et al. ( 2018 ). Furthermore, the isolated lignans show promising antiviral potential, in the words of Carvalho et al. (2023). Based on these findings and the ongoing search for new antiviral agents, we conducted a study to evaluate the in vitro antiviral properties of the ethanolic extract of F. citrifolia leaves against the MAYV, CHIKV, ZIKV, and OROV strains. In addition, we performed chemical profiling using high-resolution mass spectrometry (HR-LCMS) to support future phytopharmacological investigations of this species. Material and Methods Plant collection, identification and extraction Leaves of F. citrifolia (Moraceae) were collected in Belém, (Pará, Brazil) under the coordenates 01°02'28" latitude and 048°27'00" longitude, in the reserve of the Brazilian Agricultural Research Corporation (EMBRAPA-PA) and cataloged under voucher No. IAN202214. Access to genetic heritage was legalized before the National System for the Management of Genetic Heritage and Associated Traditional Knowledge (SISGEN) under the registration AF01766 (available at https://sisgen.gov.br ). The botanical material was washed, oven-dried at 45 ± 1°C. The dried material was ground in a knife mill and sieved through in mesh screens with an average nominal opening of 180 µm. Extraction was performed with ethanol (99.5%, w/w, TEDIA®; 1:10, m/v), using 300 g of dried and crushed material in two cycles of 24 hours each, at room temperature. The extract was vacuum filtered and dried in an oven (45 ± 1°C) until constant weight. The crude extract yielded (EEFC) 14 g. The sample (crude extract) was cleaned by solid phase extraction (SPE) using a 50 mg C18 cartridge (Phenomenex, Torrance, CA, USA). The cartridge was preconditioned according to the manufacturer. 10 mg of sample was dissolved in 1.0 mL of H 2 O:ACN (2:8, v/v) and eluted through the cartridge. The volume was collected, and a second volume was passed through the cartridge. The volumes were pooled and dried, yielding 2.3 mg, which was stored away from light until analysis. LC-MS analysis LC-MS analyses were performed on a Xevo G2-S QTof HRMS (Waters Corp., Milford, MA, USA) with an Electrospray source, using leucine enkephalin for accurate mass calibration. MassLynx 4.1 controlled the system and data acquisition. The extract (2 mg/mL) was injected (2 µL) into a BEH C18 column (50 × 2.1 mm, 1.7 µm) at 40°C, with Ultrapure water (A) and acetonitrile (B) as mobile phases at a 300 µL/min flow rate. The gradient was: 0–18 min (10–100% B), 18–20 min (100% B), 20–21 min (100–10% B), and 21–24 min (10% B). Positive and negative ionization modes were used across the m/z range 50–1200. Data Dependent Acquisition (Top 5) was applied, with collision energy from 15 to 65 eV and scan time of 0.1 s. Tolerances for charge states, peak extraction, and deisotoping were set at ± 0.2 Da, 2 Da, ± 3 Da, and 6 Da, respectively. Source/desolvation temperatures were 150°C/300°C, with gas flow rates of 20 L/h and 600 L/h, respectively. Data processing and annotation of chemical compounds The .raw data were converted to mzML format using MSConvert 3.0.2 (Chambers et al. 2012 ) (ProteoWizard, Palo Alto, CA, USA). Data processing was performed with MZmine 4.2.0 (Schmid et al. 2023 ). Ion detection thresholds were set at 1.0 x 10³ for MS 1 and 8.0 x 10¹ for MS 2 . Deconvolution was done using Local Minimum Resolver with a chromatographic threshold of 80%. Retention time and mobility intervals were set at 0.050, with a minimum absolute height of 1000 and peak-to-peak ratio of 1.7. Isotope detection had a 10 ppm error and 0.2 min retention time tolerance, for charges + 1 to + 2. Peak alignment used 3:1 m/z tolerance and 0.2 min retention time margin. Data were filtered to exclude features without MS 2 spectra and blank features, resulting in 256 features in negative and 353 in positive mode. Identification was performed via Global Natural Products Social Molecular Networking (GNPS), which is a MS/MS spectral library devised by Wang et al. ( 2016 ). The detailed analyses can be accessed via the IDs: for the negative mode, ID = 57c3e0b4cedf4f698ee422b761de598d, and for the positive mode, ID = 7f6942fd82484d0d9f2fd0ab8af95450. Antiviral activity of the extract Cell lines In the in vitro assays, Vero continuous lineage cells (CCL-81), which are fibroblast-like cells derived from the kidney of the African green monkey ( Cercopithecus aethiops ) obtained from the American Type Culture Collection (ATCC; USA) were used. The cells were cultured in Dulbecco's Modified Eagle's Minimal Essential Medium (DMEM; Sigma-Aldrich, USA), which was supplemented with 5% fetal bovine serum (FBS; Gibco, USA), penicillin/streptomycin (200U/mL) and amphotericin B (2.5g/mL) from Sigma-Aldrich (USA). Cell culture was conducted according to the guidelines established by the ATCC (2022), and the environment was maintained at 37°C with 5% CO 2 and humidity. The arboviruses Mayaro ( Alphavirus ), Chikungunya ( Alphavirus ), Zika ( Orthoflavivirus ) and Oropouche ( Orthobunyavirus ) were used in the antiviral trials. The Mayaro virus (MAYV), strain Acre27 (GenBank KM400591), was initially isolated during an epidemiological survey from a blood sample of a febrile, malaria-negative patient collected in Acrelândia, in the Amazon Basin, Acre, Brazil by Terzian et al. (2015). The Chikungunya virus (CHIKV) strain S27-African (GenBank AF369024) was initially isolated from a febrile patient and serves as a prototype for the African genotype authored by Khan et al. ( 2002 ). Zika virus (ZIKV) strain PE243/2015 (GenBank KX197192.1) was initially recovered from a patient in the state of Pernambuco, Brazil, who presented with the classic symptoms of Zika fever, but did not have any neurological complications during the 2015 outbreak reported by Donald et al. ( 2016 ). The Oropouche virus (OROV), strain BeAn19991 (GenBank KP052850-KP052852), which represents the prototype of the Brazilian genotype, was initially isolated from a “sloth” ( Bradypus tridactylus ) in Pará (Brazil) in 1960 made by Saeed et al. ( 2000 ). Cytotoxicity assays Vero cells were seeded in 96-well microplates at a density of 5 x 10 4 cells/well and incubated at 37°C overnight. After this period, the culture medium was replaced with various concentrations of F. citrifolia samples, serially diluted 1:2 in DMEM medium with 1% FBS. The experiment was carried out in triplicate. DMSO was used to solubilize the extracts, maintaining the same proportion as the vehicle control. Untreated cells were also kept as a control. The plates were incubated at 37°C for 48 hours. Cell viability was assessed using the MTT (methylthiazolyl diphenyl tetrazolium bromide) colorimetric assay, and absorbance was measured at 490 ƞm using a spectrophotometer. Using a regression analysis, the cytotoxic concentration for 50% of the cells (CC 50 ) of each extract was determined by comparing the viability of the treated cells with that of the untreated cells. Antiviral activity assays Vero cells were seeded in 96-well plates at a density of 5 x 10 4 cells/well and infected with MAYV, CHIKV, ZIKV or OROV at a multiplicity of infection (MOI) of 1. They were then treated with different concentrations of F. citrifolia extract (EEFC), starting with non-cytotoxic concentrations. The control groups, which include uninfected and untreated cells, received only the culture medium. In contrast, the virus control groups were infected without any treatment. All tests were carried out in triplicate and incubated for 48 hours at 37ºC for MAYV, CHIKV and OROV, and 72 hours for ZIKV. Cell viability was then determined using the MTT colorimetric assay after the incubation period. The effective concentration for 50% of the cells (EC 50 ) was determined using regression analysis and represented the concentration that protected 50% of the infected cells compared to the virus control. The antiviral selectivity of the extract was assessed using the selectivity index (SI), which is defined as the ratio of CC 50 to EC 50 . Results and Discussion Identification of Compounds LC-MS analysis in the negative [M–H] – and positive [M + H] + modes enabled the annotation of 21 chemical compounds belonging to different classes, such as: phenolic acids, flavonoids (flavones, flavanones, flavonols, isoflavones), lignans, fatty acids and terpenes. Of these, 10 were identified through an integrated approach, which combined high-precision experimental data in comparison with the fragments found in the literature and 11 through analysis carried out using the GNPS platform. The results obtained so far provide a solid basis for understanding the chemical composition of the species. Table 1 shows the compounds annotated with their respective experimental m/z values compared to the theoretical values calculated from the molecular formulas. The experimental error remained below 5 ppm for all the compounds, indicating high precision in the identification. Compounds such as caffeic acid ( 1 ), identified on the basis of its characteristic fragment m/z 135 by Sineiawska et al. (2022). Pinoresinol ( 13 ) with fragments m/z 342, 327, 311, 151 and 136 as done by Patyra, Dudek and Kiss (2022). Naringenin ( 14 ), with m/z 151, 119, 107 explained by Zeng et al. ( 2018 ). (Iso)orientin ( 5 ) with m/z 327, 297, 285 and (iso)vitexin ( 8 ) m/z 311, 283, 269, 134 by Amaral et al. (2019)., proved to be consistent with data previously reported in the literature, giving greater reliability to the annotation carried out. Table 1 Chemical compounds annotated for the ethanolic extract of F . citrifolia leaves. Compound RT (min) Molecular formula Experimental m/z Teorical m/z Error (ppm) Fragments (MS/MS) Putative name Chemical class Ionization mode 1 0.51 C 9 H 8 O 4 179.0346 179.0344 1.1 135 caffeic acid* Phenolic acid [M–H] – 2 2.43 C 21 H 20 O 11 447.0920 447.0927 1.5 357, 327, 297, 285 orientin Flavone [M–H] – 3 2.76 C 21 H 20 O 10 431.0973 431.0978 1.1 311, 284, 283, 269, 239, 161, 135 vitexin Flavone [M–H] – 4 2.89 C 11 H 16 O 3 197.1172 197.1178 3.0 197, 179, 161, 135, 107 (epi)loliolide Monoterpene [M + H] + 5 3.01 C 21 H 20 O 11 447.0920 447.0927 1.5 327, 297, 285 (iso)orientin* Flavone [M–H] – 6 3.29 C 11 H 16 O 3 197.1178 197.1178 0 197, 179, 161, 135, 107 loliolide Monoterpene [M + H] + 7 3.55 C 21 H 22 O 10 433.1122 433.1135 3.0 433, 313, 295, 272, 271 naringenin-7-O-glucoside Flavanone [M–H] – 8 3.56 C 21 H 20 O 10 431.0968 431.0978 2.3 311, 283, 269,134 (iso)vitexin* Flavone [M–H] – 9 4.59 C 15 H 12 O 6 287.0552 287.0556 1.3 287, 151, 135, 125, 107 eriodictyol Flavanone [M–H] – 10 4.78 C 15 H 10 O 6 285.0398 285.0399 0.3 285, 151, 133, 107 luteolin Flavone [M–H] – 11 4.88 C 15 H 10 O 7 301.0345 301.0348 0.9 178, 159, 151, 139, 121, 107 quercetin Flavonol [M–H] – 12 5.05 C 11 H 12 O 4 207.0654 207.0657 1.4 207, 179, 161, 135, 134 ethyl caffeate Phenolic acid [M–H] – 13 5.09 C 20 H 22 O 6 357.1335 357.1338 0.8 342, 327, 311, 151, 136 pinoresinol* Lignan [M–H] – 14 5.50 C 15 H 12 O 5 271.0607 271.0606 0.3 151, 119, 107 naringenin* Flavanone [M–H] – 15 5.60 C 15 H 10 O 5 269.0453 269.0450 1.1 159, 151, 117, 107 apigenin Flavone [M–H] – 16 7.71 C 15 H 12 O 4 255.0656 255.0657 0.3 255, 213, 187, 151, 125, 107 pinocembrin Flavanone [M–H] – 17 9.41 C 20 H 18 O 5 339.1222 339.1232 2.9 283, 271, 255, 153 isowighteone* Isoflavone [M + H] + 18 10.29 C 20 H 16 O 5 337.1070 337.1076 1.7 319, 295, 283, 271, 247 alpinumisoflavone* Isoflavone [M + H] + 19 10.71 C 20 H 16 O 5 337.1071 337.1076 1.4 319, 295, 283, 271, 247 (iso)alpinumisoflavone* Isoflavone [M + H] + 20 17.94 C 30 H 48 O 425.3763 425.3783 4.7 407, 231, 191, 177, 109 lupenone* Triterpene [M + H] + 21 19.04 C 29 H 48 O 413.3764 413.3783 4.5 311, 297, 283, 255 stigmasterol* Steroid [M + H] + *Notes made based on chemical prospecting of previous studies reported for the genus and comparative analysis of spectra available in the literature. Among the flavonoids, the isoflavones isowighteone ( 17 ), alpinumisoflavone ( 18 ) and (iso)alpinumisoflavone ( 19 ) were observed. Their characteristic fragmentations corroborate the proposed structures, including the loss of the prenyl group, resulting in the ion of m/z 283 and 271, common to all of them. Isowighteone ( 17 ) showed fragments at m/z 283, 271, 255 and 153 (Hostettmann; Marston and Wolfender 2005 ), and alpinumisoflavone ( 18 ) and (iso)alpinumisoflavone ( 19 ) showed fragments at m/z 319, 295, 283, 271 and 247 showed by Chen et al. (2024). The triterpene lupenone ( 20 ) with ions at m/z 407, 231, 191, 177 and 109 as described by Wang et al. ( 2020 ) and the steroid stigmasterol ( 21 ) with m/z at 311, 297, 283, 255 were also identified, reinforcing the chemical complexity of the species by Rozenberg et al. ( 2003 ). The compounds orientin ( 2 ), vitexin ( 3 ), (epi)loliolide ( 4 ), loliodide ( 6 ), narigerin-7-O-glucoside ( 7 ), eriodictiol ( 9 ), luteolin ( 10 ), quercetin ( 11 ), ethyl caffeate ( 12 ), apigenin ( 15 ) and pinocembrin ( 16 ) were identified using the GNPS platform. This tool made it possible to correlate the experimental mass spectra with data libraries of previously described compounds. Cytotoxicity and antiviral activity of the extract Table 2 presents the cytotoxicity (CC₅₀) and antiviral activity results for the ethanolic extract of F. citrifolia leaves. The extract showed a CC₅₀ of 793.92 µg/mL. The effective concentrations (EC₅₀) and selectivity indices (SI) were 61.24 µg/mL (SI 12.96) for MAYV, 117.92 µg/mL (SI 6.73) for CHIKV, and 117.29 µg/mL (SI 6.77) for ZIKV. No antiviral activity was observed against OROV. The lack of effect may be attributed to OROV’s segmented negative-sense RNA genome, which presents replication mechanisms distinct from unsegmented arboviruses like MAYV, CHIKV, and ZIKV (Files et al., 2022 ; Emmanuel et al., 2021 ). Table 2 – Antiviral activity and cytotoxicity threshold for the ethanolic extract of Ficus citrifolia leaves. Cytotoxic concentration (CC 50 , µg/mL) 793.92 Arbovirus Effective concentration (EC 50 , µg/mL) Selectivity index (SI) MAYV 61.24 ± 5.56 12.96 OROV nd nd CHIKV 117.92 ± 2.52 6.73 ZIKV 117.29 ± 37.16 6.77 Note: MAYV (Mayaro virus); OROV (Oropouche virus); CHIKV (Chikungunya virus); ZIKV (Zika virus); nd (not detected). (Brito, 2025; European Centre for Disease Prevention and Control, 2025). Ficus species are rich in diverse secondary metabolites such as flavonoids, terpenes, phenols, sterols, and lignans (Chandra et al., 2023 ), which may underlie the observed biological effects. The high SI values, particularly against MAYV (12.96), suggest a therapeutic potential with low cytotoxicity, reinforcing the extract’s promise against Orthoflavivirus and Alphavirus. LC-MS/MS analysis enabled the identification of compounds potentially linked to this antiviral activity. Caffeic acid ( 1 ) has shown activity against ILHV and ZIKV by binding to the viral envelope protein (Saivish et al., 2023 ). Its derivative, ethyl caffeate ( 12 ), also exhibited antiviral effects by inhibiting HIV protease (Wang et al., 2019 ). Among flavonoids, orientin ( 2 ) blocks SARS-CoV-2 entry into host cells (Bhowmik et al., 2021 ), while isoorientin ( 5 ) inhibits the spike-ACE2 interaction (Chen et al., 2023 ). Luteolin ( 10 ) is known for broad-spectrum antiviral action against influenza, herpesvirus, and coronaviruses (Lu et al., 2023 ). Vitexin ( 3 ) and isovitexin ( 8 ) inhibit H1N1 and offer protection against COVID-19 (Sadati et al., 2019; Ferdausi et al., 2022). Apigenin ( 15 ) interferes with replication of DNA and RNA viruses, including HSV, HCV, DENV, and SARS-CoV (Lee et al., 2023 ). Flavanones identified include eriodictyol ( 9 ), which modulates inflammatory cytokines relevant in CHIKV and MAYV infections (Deng et al., 2020 ). Naringenin ( 14 ) and its glycoside naringenin-7-O-glucoside ( 7 ), along with pinocembrin ( 16 ), reduce ZIKV RNA synthesis and affect viral proteins (Cataneo et al., 2021 ; Lee et al., 2019 ). Quercetin ( 11 ) disrupts MAYV entry and RNA replication (Santos et al., 2014 ), supporting the strong activity observed against this virus (EC₅₀ 61.24 µg/mL; SI 12.96). Pinoresinol ( 13 ), a lignan reported in F. citrifolia , has demonstrated anti-HBV effects (Wang et al., 2013). Prenylated isoflavones such as isowighteone ( 17 ), alpinumisoflavone ( 18 ), and its isomer, known from the genus (Bankeu et al., 2011), may also contribute due to increased lipophilicity and membrane interaction potential (Ateba et al., 2019). Monoterpenes like loliolide ( 6 ), identified in the genus (Zhang et al., 2024 ), inhibit HCV entry (Chung et al., 2016 ). Its epimer, (epi)loliolide ( 4 ), although not linked to antiviral activity, shows anti-hepatocellular carcinoma properties (Gangadhar et al., 2020 ). The triterpene lupenone ( 20 ) has activity against CHIKV, while stigmasterol ( 21 ) inhibits HSV replication (Bhakat & Soliman, 2015 ; Petrera, Níttolo & Alché, 2014). These results suggest that the antiviral effect may arise from synergistic interactions among m etabolites rather than isolated actions (Caesar & Cech, 2019 ). The chemical complexity of F. citrifolia supports its antiviral activity and points to its potential for therapeutic development. Further studies, including bioguided fractionation and molecular docking, are essential to elucidate specific mechanisms and identify active constituents. Declarations Authors’ contributions Horrana Acácio Mardegan designed the studies. Horrana Acácio Mardegan, Allana Martins dos Santos Ataíde, Loueny Celine Ribeiro Andrade and Johan Carlos Costa Santiago conducted LC-MS experiments. Cíntia Lopes de Brito Guimarães designed antiviral assays. Horrana Acácio Mardegan, Allana Martins dos Santos Ataíde and Johan Carlos Costa Santiago conducted MS/MS data processing. Horrana Acácio Mardegan and Johan Carlos Costa Santiago performed metabolite characterization. Horrana Acácio Mardegan, Allana Martins dos Santos Ataíde, Loueny Celine Ribeiro Andrade and Abraão de Jesus Barbosa Muribeca searched the pharmacological properties of the annotated metabolites. Ariane Coelho Ferraz, Marília Bueno da Silva Menegatto and Cíntia Lopes de Brito Magalhães performed and discussed in vitro antiviral assays with the extracts. Horrana Acácio Mardegan, Allana Martins dos Santos Ataíde, Johan Carlos Costa Santiago, Abraão de Jesus Barbosa Muribeca wrote the manuscript. Sônia das Graças Santa Rosa Pamplona, Cíntia Lopes de Brito Magalhães, Abraão de Jesus Barbosa Muribeca, Consuelo Yumiko Yoshioka e Silva and Milton Nascimento da Silva reviewed the whole manuscript and provided comments. All authors have read and agreed to the published version of the manuscript. Acknowledgements Horrana Acácio Mardegan would like to thank the Federal University of Pará (UFPA) and the Coordination for the Improvement of Higher Education Personnel (CAPES) for their support in awarding a master's degree scholarship. Supplementary Information Not applicable. Ethics approval Not applicable. This study did not involve human participants or animal experiments. Consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. References Abdulmalik I, Sule M, Yaro A, Abdullahi M, Abdulkadir M, Yusuf H (2011) Evaluation of analgesic and anti-inflammatory effects of ethanol extract of Ficus iteophylla leaves in rodents. Afr J Tradit Complement Altern Med. ;8(4) Andrzej Patyra, Dudek MK, Kiss AK (2022) LC-DAD–ESI-MS/MS and NMR Analysis of Conifer Wood Specialized Metabolites. Cells 11(20):3332–3332 Antoun MD, Ramos Z, Vazques J, Oquendo I, Proctor GR, Gerena L, Franzblau SG (2001) Evaluation of the flora of Puerto Rico for in vitro antiplasmodial and antimycobacterial activities. Phytotherapy Research: PTR 15(7):638–642 Arunachalam K, Parimelazhagan T (2013) Anti-inflammatory, wound healing and in-vivo antioxidant properties of the leaves of Ficus amplissima Smith. J Ethnopharmacol 145(1):139–145 Baetas AC, Fábio L, Paula A (2018) In vitro cytotoxicity of lignans isolated from Ficus citrifolia P. Miller (Moraceae) on C6 glioma cell line. Glo Adv Res J Med Med Sci 7:77–84 Bhakat S, Soliman MES (2015) Chikungunya virus (CHIKV) inhibitors from natural sources: a medicinal chemistry perspective. J Nat Med 69(4):451–462 Bhowmik A, Biswas S, Hajra S, Saha P (2021) In silico validation of potent phytochemical orientin as inhibitor of SARS-CoV-2 spike and host cell receptor GRP78 binding. Heliyon 7(1):e05923 Brito C, Falcão MB, Maria T, Cerqueira-Silva, Teixeira MG, Freitas R, Chikungunya (2025) From Hypothesis to Evidence of Increased Severe Disease and Fatalities. Viruses [Internet]. Jan 3 [cited 2025 May 28];17(1):62–2. Available from: https://www.mdpi.com/1999-4915/17/1/62 Caesar LK, Cech NB (2019) Synergy and antagonism in natural product extracts: when 1 + 1 does not equal 2. Natural Prod Reports [Internet] 36(6):869–888 Carolina A, Auguste AJ, Danila Vedovello, Ferreira MU et al (2015) Mônica da Silva-Nunes, Márcia Aparecida Sperança,. Isolation and Characterization of Mayaro Virus from a Human in Acre, Brazil. American Journal of Tropical Medicine and Hygiene [Internet]. ;92(2):401–4. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4347347 Cataneo AHD, Ávila EP, Mendes LA, de O VG, Ferraz CR, de Almeida MV et al (2021) Flavonoids as Molecules With Anti-Zika virus Activity. Front Microbiol. ;12 Chambers MC, Maclean B, Burke R, Amodei D, Ruderman DL, Neumann S et al (2012) A cross-platform toolkit for mass spectrometry and proteomics. Nature Biotechnology [Internet]. ;30(10):918–20. Available from: https://www.nature.com/articles/nbt.2377 Chandra S, Kala K, Negi MS, Anuj, Nautiyal (2023) The Genus Ficus : a review of its phytochemicals and pharmacological studies. ;7(1):230. Available from: https://www.researchgate.net/publication/371874948_The_Genus_Ficus_a_review_of_its_phytochemicals_and_pharmacological_studies Chen G, Li X, Saleri FD, Guo M (2016) Analysis of Flavonoids in Rhamnus davurica and Its Antiproliferative Activities. Molecules 21(10):1275–1275 Chen G, Sun Y, Yushan D, Shaerbayi N, Zhang H, He H et al (2023) Identification and Characterization of Chemical Constituents from Ammopiptanthus nanus Stem and Their Metabolites in Rats by UHPLC-Q-TOF-MS/MS. Planta Med 90(02):138–153 Chen YL, Chen CY, Lai KH, Chang YC, Hwang TL (2023) Anti-inflammatory and antiviral activities of flavone C-glycosides of Lophatherum gracil e for COVID-19. J Funct Foods 101:105407 Chung CY, Liu CH, Thierry Burnouf, Wang GH, Chang SP, Alagie Jassey et al (2016) Activity-based and fraction-guided analysis of Phyllanthus urinaria identifies loliolide as a potent inhibitor of hepatitis C virus entry. Antiviral Res 130:58–68 Cruz RF, Corrêa (2022) Carlos Victor Lamarão, Edgar Aparecido Sanches, Pedro Henrique Campelo, Jaqueline. Ficus spp .: Phytochemical composition and medicinal potential. Res Soc Dev 11(12):e265111234135–e265111234135 Darbour N, Bayet C, Rodin-Bercion S, Elkhomsi Z, Lurel F, Chaboud A et al (2007) Isoflavones from Ficus nymphaefolia . Nat Prod Res 21(5):461–464 Deng Z, Hassan S, Rafiq M, Li H, He Y, Cai Y et al (2020) Pharmacological Activity of Eriodictyol: The Major Natural Polyphenolic Flavanone. Wu H, editor. Evidence-Based Complementary and Alternative Medicine . ;2020:1–11 Diagne CT, Bengue M, Choumet V, Hamel R, Pompon J, Missé D (2020) Mayaro Virus Pathogenesis and Transmission Mechanisms. Pathogens 9(9):738 Donald CL, Brennan B, Cumberworth SL, Rezelj VV, Clark JJ, Cordeiro MT et al (2016) Full Genome Sequence and sfRNA Interferon Antagonist Activity of Zika Virus from Recife, Brazil. PLoS neglected tropical diseases [Internet]. ;10(10):e0005048. Available from: https://pubmed.ncbi.nlm.nih.gov/27706161/ dos Santos AE, Kuster RM, Yamamoto KA, Salles TS, Campos R, de Meneses MD et al (2014) Quercetin and quercetin 3-O-glycosides from Bauhinia longifolia (Bong.) Steud. show anti-Mayaro virus activity. Parasites Vectors. ;7(1) Emmanuel V, Otávio JB, Brustolini CL, Filipe, Castro F, Guimarães AP et al (2021) Common Dysregulation of Innate Immunity Pathways in Human Primary Astrocytes Infected With Chikungunya, Mayaro, Oropouche, and Zika Viruses. Front Cell Infect Microbiol. ;11 Epidemiological Update Dengue and other Arboviruses – 10 June 2020 (em inglês e espanhol) [Internet]. Paho.org. 2020 . Available from: https://www.paho.org/pt/documents/epidemiological-update-dengue-and-other-arboviruses-10-june-2020 Erina Petrera (2014) Analía Gabriela Níttolo, Alché LE. Antiviral Action of Synthetic Stigmasterol Derivatives on Herpes Simplex Virus Replication in Nervous Cells In Vitro . Biomed Res Int 2014:1–9 European Centre for Disease Prevention and Control. Chikungunya worldwide overview [Internet]. European Centre for Disease Prevention and Control (2025) Available from: https://www.ecdc.europa.eu/en/chikungunya-monthly Federica Dell’Annunziata, Sellitto C, Franci G, Marcotullio MC, Piovan A, Marca RD et al (2022) Antiviral Activity of Ficus rubiginosa Leaf Extracts against HSV-1, HCoV-229E and PV-1. Viruses 14(10):2257–2257 Ferdausi N, Islam S, Rimti F, Quayum S, Arshad E, Ibnat A et al Point-specific interactions of isovitexin with the neighboring amino acid residues of the hACE2 receptor as a targeted therapeutic agent in suppressing the SARS-CoV-2 influx mechanism. Journal of Advanced Veterinary and Animal Research [Internet]. 2022 [cited 2023 Mar 8];9(2):230. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9298103/ Ficus spp . (fig): Ethnobotany and potential as anticancer and anti-inflammatory agents. Journal of Ethnopharmacology [Internet]. (2008) ;119(2):195–213 Files MA, Hansen CA, Herrera VC, Schindewolf C, Barrett ADT, Beasley DWC et al (2022) Baseline mapping of Oropouche virology, epidemiology, therapeutics, and vaccine research and development. Vaccines. ;7(1) Gangadhar KN, Rodrigues MJ, Pereira H, Gaspar H, Malcata FX, Barreira L et al (2020) Anti-Hepatocellular Carcinoma (HepG2) Activities of Monoterpene Hydroxy Lactones Isolated from the Marine Microalga Tisochrysis lutea . Mar Drugs 18(11):567 Girard M, Nelson CB, Picot V, Gubler DJ (2020) Arboviruses: A global public health threat. Vaccine 38(24):3989–3994 Hamed M, Khalifa M, Mahmoud, Mohammed OA, Kamaly, Alanazi AS et al (2023) The Potential Roles of Ficus carica Extract in the Management of COVID-19 Viral Infections: A Computer-aided Drug Design Study. Curr Comput - Aided Drug Des 20(6):974–986 Hostettmann K, Marston A, Wolfender JL (2005) Strategy in the Search for New Lead Compounds and Drugs from Plants. CHIMIA Int J Chem 59(6):291–294 Hostettmann K, Marston A, Wolfender JL (2005) Strategy in the search for new lead compounds and drugs from plants. Chimia 59(6):291–291 Jean R, Khayala, Lenta BN, Noungoué DT, Ngouela SA, Mustafa SA et al (2011) Isoflavone Dimers and Other Bioactive Constituents from the Figs of Ficus mucuso . J Nat Prod 74(6):1370–1378 Jean R, Khayala, Lenta BN, Noungoué DT, Ngouela SA, Mustafa SA et al (2011) Isoflavone Dimers and Other Bioactive Constituents from the Figs of Ficus mucuso . J Nat Prod 74(6):1370–1378 Khan AH, Morita K, Parquet M del, Hasebe C, Mathenge F, Igarashi EGM (2002) Complete nucleotide sequence of chikungunya virus and evidence for an internal polyadenylation site. J Gen Virol 83(12):3075–3084 Lee IG, Lee J, Hong SH, Seo YJ (2023) Apigenin’s Therapeutic Potential Against Viral Infection. Front Bioscience-Landmark 28(10):237–237 Lee JL, Loe MWC, Lee RCH, Chu JJH (2019) Antiviral activity of pinocembrin against Zika virus replication. Antiviral Res 167:13–24 Lu P, Zhang T, Ren Y, Rao H, Lei J, Zhao G et al (2023) Literature Rev Antiviral Mechanism Luteolin 18(4):1934578X2311715–1934578X2311715 Saivish MV, Pacca CC, Vivaldo, Gabriela, Alves R, Liliane, Nebo et al (2023) Caffeic Acid Has Antiviral Activity against Ilhéus Virus In Vitro . Viruses 15(2):494–494 Paixão ES, Teixeira MG, Rodrigues LC (2017) Zika, chikungunya and dengue: the causes and threats of new and re-emerging arboviral diseases. BMJ Global Health [Internet]. ;3(Suppl 1):e000530. Available from: https://gh.bmj.com/content/3/Suppl_1/e000530 Pearson JL, Lee S, Suresh H, Low M, Nang M, Singh S et al (2014) The Liquid Chromatographic Determination of Chlorogenic and Caffeic Acids in Xu Duan ( Dipsacus asperoides ) Raw Herb. ISRN Anal Chem 2014:1–6 Pielnaa P, Al-Saadawe M, Saro A, Dama MF, Zhou M, Huang Y et al (2020) Zika virus-spread, epidemiology, genome, transmission cycle, clinical manifestation, associated challenges, vaccine and antiviral drug development. Virology [Internet]. ;543(0042-6822):34–42. Available from: https://pubmed.ncbi.nlm.nih.gov/32056845/ Pingen M, Bryden Steven R, Pondeville E, Schnettler E, Kohl A, Merits A et al (2016) Host Inflammatory Response to Mosquito Bites Enhances the Severity of Arbovirus Infection. Immunity [Internet]. ;44(6):1455–69. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4920956/ Qi C, Fu YH, Chen W, Chen G, Dai C, Song X et al (2017) A new isoflavone from the roots of Ficus auriculata . Nat Prod Res 32(1):43–47 Rhelly A, Diogo J, Paulo AC, Ferraz, Mardegan HA, Menegatto S et al (2023) Untargeted-based metabolomics analysis and in vitro/in silico antiviral activity of extracts from Phyllanthus brasiliensis (Aubl.) Poir. Phytochem Anal 34(7):869–883 Ricardo Guimarães Amaral, Vieira S, Claudia M, do C, Andrade LN, Severino P et al (2019) Cytotoxic potential of 14 Passiflora species against cancer cells. J Med Plants Res 13(7):157–166 Rozenberg R, Ruibal-Mendieta NL, Petitjean G, Cani PD, Delacroix DL, Delzenne NM et al (2003) Phytosterol analysis and characterization in spelt (Triticum aestivum ssp. spelta L.) and wheat ( T. aestivum L.) lipids by LC/APCI-MS. J Cereal Sci 38(2):189–197 Sadati S, Gheibi N, Ranjbar S, Hashemzadeh M (2018) Docking study of flavonoid derivatives as potent inhibitors of influenza H1N1 virus neuraminidase. Biomedical Rep. Nov 23 Saeed MF, Watts DM, Shope RE, Barrett ADT, Tesh RB, Nunes M et al (2000) Nucleotide sequences and phylogeny of the nucleocapsid gene of Oropouche virus. J Gen Virol 81(3):743–748 Schmid R, Heuckeroth S, Korf A, Smirnov A, Myers O, Dyrlund TS et al (2023) Integrative analysis of multimodal mass spectrometry data in MZmine 3. Nature Biotechnology [Internet]. ;41(4):447–9. Available from: https://www.nature.com/articles/s41587-023-01690-2 Schwartz DA, Dashraath P, Baud D (2024) Oropouche Virus (OROV) in Pregnancy: An Emerging Cause of Placental and Fetal Infection Associated with Stillbirth and Microcephaly following Vertical Transmission. Viruses [Internet]. ;16(9):1435. Available from: https://www.mdpi.com/1999-4915/16/9/1435 Sieniawska E, Świątek Ł, Sinan KI, Zengin G, Boguszewska A, Polz-Dacewicz M et al (2022) Phytochemical Insights into Ficus sur Extracts and Their Biological Activity. Molecules (Basel, Switzerland) [Internet]. ;27(6):1863. Available from: https://pubmed.ncbi.nlm.nih.gov/35335228/ Mvondo SBAMA, Séfirin, Djiogue Stéphane, Zingue, Krenn L (2019) Dieudonné Njamen. A Pharmacological Overview of Alpinumisoflavone, a Natural Prenylated Isoflavonoid. Frontiers in Pharmacology . ;10 VALLI M, RUSSO HM, BOLZANI VS (2018) The potential contribution of the natural products from Brazilian biodiversity to bioeconomy. An Acad Bras Cienc 90(1 suppl 1):763–778 Wang GK, Lin BB, Rao R, Zhu K, Qin XY, Xie GY et al (2012) A new lignan with anti-HBV activity from the roots of Bombax ceiba . Nat Prod Res 27(15):1348–1352 Wang M, Carver JJ, Phelan VV, Sanchez LM, Garg N, Peng Y et al (2016) Sharing and community curation of mass spectrometry data with Global Natural Products Social Molecular Networking. Nat Biotechnol 34(8):828–837 Wang X, Wei Y, Tian WY, Sakharkar MK, Liu Q, Yang X et al (2019) Characterization of Nine Compounds Isolated from the Acid Hydrolysate of Lonicera fulvotomentosa Hsu et S. C. Cheng and Evaluation of Their In Vitro Activity towards HIV Protease. Molecules [Internet]. Dec 11 [cited 2023 Sep 17];24(24):4526. Available from: http://www.ncbi.nlm.nih.gov/pmc/articles/pmc6943440/ Wang X, Zhong XJ, Zhou N, Cai N, Xu JH, Wang QB et al (2020) Rapid Characterizaiton of Chemical Constituents of the Tubers of Gymnadenia conopsea by UPLC–Orbitrap–MS/MS Analysis. Molecules 25(4):898–898 Wei L, Ma R, Fu Y (2022) Differences in Chemical Constituents between Dalbergia oliveri Heartwood and Sapwood and Their Effect on Wood Color. Molecules 27(22):7978–7978 World Health Organization. WHO Coronavirus (COVID-19) [Internet]. World Health Organization (2023) Available from: https://covid19.who.int/ Wu YS, Afzal S, Appalaraju V, Wei TQ, Manap ASA, Albokhadaim I (2024) Evaluation for Antiviral Potential of Ficus deltoidea against Dengue Virus Type-2 (Penilaian Potensi Antiviral Ficus deltoidea terhadap Virus Denggi Jenis-2). Sains Malaysiana 53(2):383–396 Zeng X, Su W, Zheng Y, Liu H, Li P, Zhang W et al (2018) UFLC-Q-TOF-MS/MS-Based Screening and Identification of Flavonoids and Derived Metabolites in Human Urine after Oral Administration of Exocarpium Citri Grandis Extract. Molecules 23(4):895–895 Zhang N, Guo M, Zhou B, Zhao X, Fu M, Shen X et al (2024) Chemical Components and Their α-Glucosidase Inhibitory Activity from the Leaves of Ficus carica Linn. Records Nat Prod. ;(1):165–170 Zhou N, Zheng D, You Q, Chen T, Jiang J, Shen W et al (2023) Therapeutic Potential of Biochanin A in Herpes Simplex Keratitis. Pharmaceuticals 16(9):1240–1240 Supplementary Files Graphicalabstract.pdf Cite Share Download PDF Status: Published Journal Publication published 24 Mar, 2026 Read the published version in Revista Brasileira de Farmacognosia → Version 1 posted Reviewers agreed at journal 03 Nov, 2025 Reviewers invited by journal 03 Nov, 2025 Editor invited by journal 22 Oct, 2025 Editor assigned by journal 08 Oct, 2025 First submitted to journal 08 Oct, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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16:28:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1087835,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7583932/v1/2264701c-ff14-4d7d-a9f8-2955cc86ddaf.pdf"},{"id":95810094,"identity":"5ec2ccf6-fbe1-4a67-955d-4cffa604155c","added_by":"auto","created_at":"2025-11-13 08:51:46","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":937325,"visible":true,"origin":"","legend":"","description":"","filename":"Graphicalabstract.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7583932/v1/1528d606c8568adb325b1198.pdf"}],"financialInterests":"","formattedTitle":"Antiviral Activity and Chemical Characterization of Ficus citrifolia Mill. (Moraceae) Leaf Extract Against Emerging Arboviruses","fulltext":[{"header":"Introduction","content":"\u003cp\u003eArboviruses stand out as infectious agents transmitted to humans by the bite of infected blood-feeding arthropods. Tropical and subtropical climates greatly facilitate the reproduction of these vectors, especially in countries such as Brazil, where climatic conditions favor the spread of arboviruses such as dengue, yellow fever, chikungunya, Zika, Mayaro, and Oropouche. Although these diseases are endemic, they continue to be neglected, with 1,645,678 cases reported in the Americas in 2020 based on PAHO and WHO (2020).\u003c/p\u003e\u003cp\u003eIn developing countries such as Brazil, unorganized urbanization and poor sanitation infrastructure increase the vulnerability of the population. In addition, the lack of effective vaccines and antivirals aggravates the situation, overburdening health systems and favoring outbreaks in the words Girard et al. (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The resurgence of Zika (ZIKV), Oropouche (OROV), Mayaro (MAYV), and Chikungunya (CHIKV) in urban centers has intensified these challenges. These viruses produce similar symptoms, such as fever, rashes, and severe joint pain, sometimes progressing to chronic arthritis. ZIKV is known to cause congenital malformations and vertical transmission, as noted by Pielnaa et al. (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and OROV has recently been linked to similar physical changes and vertical transmission as described by Schwartz, Dashraath \u0026amp; Baud (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). MAYV can cause complications such as myocarditis, hemorrhagic syndromes, and neurological manifestations.\u003c/p\u003e\u003cp\u003eFaced with these clinical challenges, the search for new therapeutic sources has increasingly turned to the ancestral knowledge of traditional communities about medicinal plants. The biodiversity of the Amazon offers several species with therapeutic potential, supporting traditional healing practices and providing an accessible alternative.\u003c/p\u003e\u003cp\u003eAmong these species, those of the genus Ficus are widely used in traditional medicine for their anti-inflammatory, analgesic, and healing properties, as stated by Lansky et al. (2008). These plants are known for their rich chemiodiversity, including organic acids, phenolic compounds, and terpenes reported in Cruz et al. (2022). Previous studies have demonstrated the therapeutic effects of species such as \u003cem\u003eF. iteophylla\u003c/em\u003e, which reduced pain and inflammation in rodents, as discussed in Abdulmalik et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2011\u003c/span\u003e)d \u003cem\u003eamplissima\u003c/em\u003e, which exhibited anti-inflammatory, antioxidant, and wound-healing properties \u003cem\u003ein vivo\u003c/em\u003e based on Arunachalam \u0026amp; Parimelazhagan (2012).\u003c/p\u003e\u003cp\u003eAlthough these effects are not directly antiviral, viral pathogens induce inflammatory responses in the host as part of the infection process, in accordance with Pingen et al. (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Ethnopharmacological applications may therefore be linked to antiviral potential, as evidenced by \u003cem\u003eF. sur\u003c/em\u003e and \u003cem\u003eF. rubiginosa\u003c/em\u003e inhibiting herpes simplex virus type 1 (HSV-1), as described by Sieniawska et al. (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) and Dell'Annunziata et al. (2022), \u003cem\u003eF. deltoidea\u003c/em\u003e showing antiviral activity against dengue virus type 2, according to Wu et al. (\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)d \u003cem\u003ecarica\u003c/em\u003e inhibiting SARS-CoV-2 replication, based on Hamed et al. (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAlthough several Ficus species have been studied, no research has evaluated the antiviral potential of \u003cem\u003eF. citrifolia\u003c/em\u003e, commonly known as \u0026ldquo;fig tree\u0026rdquo; or \u0026ldquo;black gameleira.\u0026rdquo; Previous studies have highlighted the antimicobacterial effects of its ethanolic leaf extract against \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e, as well as the antiproliferative effect of three lignans isolated from the leaf extract with dichloromethane, as reported by Antoun et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2001\u003c/span\u003e) and Baetas et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, the isolated lignans show promising antiviral potential, in the words of Carvalho et al. (2023).\u003c/p\u003e\u003cp\u003eBased on these findings and the ongoing search for new antiviral agents, we conducted a study to evaluate the \u003cem\u003ein vitro\u003c/em\u003e antiviral properties of the ethanolic extract of \u003cem\u003eF. citrifolia\u003c/em\u003e leaves against the MAYV, CHIKV, ZIKV, and OROV strains. In addition, we performed chemical profiling using high-resolution mass spectrometry (HR-LCMS) to support future phytopharmacological investigations of this species.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003ePlant collection, identification and extraction\u003c/h2\u003e\u003cp\u003eLeaves of \u003cem\u003eF. citrifolia\u003c/em\u003e (Moraceae) were collected in Bel\u0026eacute;m, (Par\u0026aacute;, Brazil) under the coordenates 01\u0026deg;02'28\" latitude and 048\u0026deg;27'00\" longitude, in the reserve of the Brazilian Agricultural Research Corporation (EMBRAPA-PA) and cataloged under voucher No. IAN202214. Access to genetic heritage was legalized before the National System for the Management of Genetic Heritage and Associated Traditional Knowledge (SISGEN) under the registration AF01766 (available at \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://sisgen.gov.br\u003c/span\u003e\u003cspan address=\"https://sisgen.gov.br\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe botanical material was washed, oven-dried at 45\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C. The dried material was ground in a knife mill and sieved through in mesh screens with an average nominal opening of 180 \u0026micro;m.\u003c/p\u003e\u003cp\u003eExtraction was performed with ethanol (99.5%, w/w, TEDIA\u0026reg;; 1:10, m/v), using 300 g of dried and crushed material in two cycles of 24 hours each, at room temperature. The extract was vacuum filtered and dried in an oven (45\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C) until constant weight. The crude extract yielded (EEFC) 14 g.\u003c/p\u003e\u003cp\u003eThe sample (crude extract) was cleaned by solid phase extraction (SPE) using a 50 mg C18 cartridge (Phenomenex, Torrance, CA, USA). The cartridge was preconditioned according to the manufacturer. 10 mg of sample was dissolved in 1.0 mL of H\u003csub\u003e2\u003c/sub\u003eO:ACN (2:8, v/v) and eluted through the cartridge. The volume was collected, and a second volume was passed through the cartridge. The volumes were pooled and dried, yielding 2.3 mg, which was stored away from light until analysis.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eLC-MS analysis\u003c/h3\u003e\n\u003cp\u003eLC-MS analyses were performed on a Xevo G2-S QTof HRMS (Waters Corp., Milford, MA, USA) with an Electrospray source, using leucine enkephalin for accurate mass calibration. MassLynx 4.1 controlled the system and data acquisition. The extract (2 mg/mL) was injected (2 \u0026micro;L) into a BEH C18 column (50 \u0026times; 2.1 mm, 1.7 \u0026micro;m) at 40\u0026deg;C, with Ultrapure water (A) and acetonitrile (B) as mobile phases at a 300 \u0026micro;L/min flow rate. The gradient was: 0\u0026ndash;18 min (10\u0026ndash;100% B), 18\u0026ndash;20 min (100% B), 20\u0026ndash;21 min (100\u0026ndash;10% B), and 21\u0026ndash;24 min (10% B). Positive and negative ionization modes were used across the m/z range 50\u0026ndash;1200. Data Dependent Acquisition (Top 5) was applied, with collision energy from 15 to 65 eV and scan time of 0.1 s. Tolerances for charge states, peak extraction, and deisotoping were set at \u0026plusmn;\u0026thinsp;0.2 Da, 2 Da, \u0026plusmn;\u0026thinsp;3 Da, and 6 Da, respectively. Source/desolvation temperatures were 150\u0026deg;C/300\u0026deg;C, with gas flow rates of 20 L/h and 600 L/h, respectively.\u003c/p\u003e\n\u003ch3\u003eData processing and annotation of chemical compounds\u003c/h3\u003e\n\u003cp\u003eThe .raw data were converted to mzML format using MSConvert 3.0.2 (Chambers et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) (ProteoWizard, Palo Alto, CA, USA). Data processing was performed with MZmine 4.2.0 (Schmid et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Ion detection thresholds were set at 1.0 x 10\u0026sup3; for MS\u003csup\u003e1\u003c/sup\u003e and 8.0 x 10\u0026sup1; for MS\u003csup\u003e2\u003c/sup\u003e. Deconvolution was done using Local Minimum Resolver with a chromatographic threshold of 80%. Retention time and mobility intervals were set at 0.050, with a minimum absolute height of 1000 and peak-to-peak ratio of 1.7. Isotope detection had a 10 ppm error and 0.2 min retention time tolerance, for charges\u0026thinsp;+\u0026thinsp;1 to +\u0026thinsp;2. Peak alignment used 3:1 m/z tolerance and 0.2 min retention time margin. Data were filtered to exclude features without MS\u003csup\u003e2\u003c/sup\u003e spectra and blank features, resulting in 256 features in negative and 353 in positive mode. Identification was performed via Global Natural Products Social Molecular Networking (GNPS), which is a MS/MS spectral library devised by Wang et al. (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The detailed analyses can be accessed via the IDs: for the negative mode, ID\u0026thinsp;=\u0026thinsp;57c3e0b4cedf4f698ee422b761de598d, and for the positive mode, ID\u0026thinsp;=\u0026thinsp;7f6942fd82484d0d9f2fd0ab8af95450.\u003c/p\u003e\n\u003ch3\u003eAntiviral activity of the extract\u003c/h3\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003eCell lines\u003c/h2\u003e\u003cp\u003eIn the \u003cem\u003ein vitro\u003c/em\u003e assays, Vero continuous lineage cells (CCL-81), which are fibroblast-like cells derived from the kidney of the African green monkey (\u003cem\u003eCercopithecus aethiops\u003c/em\u003e) obtained from the American Type Culture Collection (ATCC; USA) were used. The cells were cultured in Dulbecco's Modified Eagle's Minimal Essential Medium (DMEM; Sigma-Aldrich, USA), which was supplemented with 5% fetal bovine serum (FBS; Gibco, USA), penicillin/streptomycin (200U/mL) and amphotericin B (2.5g/mL) from Sigma-Aldrich (USA). Cell culture was conducted according to the guidelines established by the ATCC (2022), and the environment was maintained at 37\u0026deg;C with 5% CO\u003csub\u003e2\u003c/sub\u003e and humidity.\u003c/p\u003e\u003cp\u003eThe arboviruses Mayaro (\u003cem\u003eAlphavirus\u003c/em\u003e), Chikungunya (\u003cem\u003eAlphavirus\u003c/em\u003e), Zika (\u003cem\u003eOrthoflavivirus\u003c/em\u003e) and Oropouche (\u003cem\u003eOrthobunyavirus\u003c/em\u003e) were used in the antiviral trials. The Mayaro virus (MAYV), strain Acre27 (GenBank KM400591), was initially isolated during an epidemiological survey from a blood sample of a febrile, malaria-negative patient collected in Acrel\u0026acirc;ndia, in the Amazon Basin, Acre, Brazil by Terzian et al. (2015). The Chikungunya virus (CHIKV) strain S27-African (GenBank AF369024) was initially isolated from a febrile patient and serves as a prototype for the African genotype authored by Khan et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Zika virus (ZIKV) strain PE243/2015 (GenBank KX197192.1) was initially recovered from a patient in the state of Pernambuco, Brazil, who presented with the classic symptoms of Zika fever, but did not have any neurological complications during the 2015 outbreak reported by Donald et al. (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The Oropouche virus (OROV), strain BeAn19991 (GenBank KP052850-KP052852), which represents the prototype of the Brazilian genotype, was initially isolated from a \u0026ldquo;sloth\u0026rdquo; (\u003cem\u003eBradypus tridactylus\u003c/em\u003e) in Par\u0026aacute; (Brazil) in 1960 made by Saeed et al. (\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eCytotoxicity assays\u003c/h2\u003e\u003cp\u003eVero cells were seeded in 96-well microplates at a density of 5 x 10\u003csup\u003e4\u003c/sup\u003e cells/well and incubated at 37\u0026deg;C overnight. After this period, the culture medium was replaced with various concentrations of \u003cem\u003eF. citrifolia\u003c/em\u003e samples, serially diluted 1:2 in DMEM medium with 1% FBS. The experiment was carried out in triplicate. DMSO was used to solubilize the extracts, maintaining the same proportion as the vehicle control. Untreated cells were also kept as a control. The plates were incubated at 37\u0026deg;C for 48 hours. Cell viability was assessed using the MTT (methylthiazolyl diphenyl tetrazolium bromide) colorimetric assay, and absorbance was measured at 490 ƞm using a spectrophotometer. Using a regression analysis, the cytotoxic concentration for 50% of the cells (CC\u003csub\u003e50\u003c/sub\u003e) of each extract was determined by comparing the viability of the treated cells with that of the untreated cells.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eAntiviral activity assays\u003c/h3\u003e\n\u003cp\u003eVero cells were seeded in 96-well plates at a density of 5 x 10\u003csup\u003e4\u003c/sup\u003e cells/well and infected with MAYV, CHIKV, ZIKV or OROV at a multiplicity of infection (MOI) of 1. They were then treated with different concentrations of \u003cem\u003eF. citrifolia\u003c/em\u003e extract (EEFC), starting with non-cytotoxic concentrations. The control groups, which include uninfected and untreated cells, received only the culture medium. In contrast, the virus control groups were infected without any treatment. All tests were carried out in triplicate and incubated for 48 hours at 37\u0026ordm;C for MAYV, CHIKV and OROV, and 72 hours for ZIKV. Cell viability was then determined using the MTT colorimetric assay after the incubation period. The effective concentration for 50% of the cells (EC\u003csub\u003e50\u003c/sub\u003e) was determined using regression analysis and represented the concentration that protected 50% of the infected cells compared to the virus control. The antiviral selectivity of the extract was assessed using the selectivity index (SI), which is defined as the ratio of CC\u003csub\u003e50\u003c/sub\u003e to EC\u003csub\u003e50\u003c/sub\u003e.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eIdentification of Compounds\u003c/h2\u003e\u003cp\u003eLC-MS analysis in the negative [M\u0026ndash;H]\u003csup\u003e\u0026ndash;\u003c/sup\u003e and positive [M\u0026thinsp;+\u0026thinsp;H]\u003csup\u003e+\u003c/sup\u003e modes enabled the annotation of 21 chemical compounds belonging to different classes, such as: phenolic acids, flavonoids (flavones, flavanones, flavonols, isoflavones), lignans, fatty acids and terpenes. Of these, 10 were identified through an integrated approach, which combined high-precision experimental data in comparison with the fragments found in the literature and 11 through analysis carried out using the GNPS platform. The results obtained so far provide a solid basis for understanding the chemical composition of the species.\u003c/p\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the compounds annotated with their respective experimental \u003cem\u003em/z\u003c/em\u003e values compared to the theoretical values calculated from the molecular formulas. The experimental error remained below 5 ppm for all the compounds, indicating high precision in the identification. Compounds such as caffeic acid (\u003cb\u003e1\u003c/b\u003e), identified on the basis of its characteristic fragment \u003cem\u003em/z\u003c/em\u003e 135 by Sineiawska et al. (2022). Pinoresinol (\u003cb\u003e13\u003c/b\u003e) with fragments \u003cem\u003em/z\u003c/em\u003e 342, 327, 311, 151 and 136 as done by Patyra, Dudek and Kiss (2022). Naringenin (\u003cb\u003e14\u003c/b\u003e), with \u003cem\u003em/z\u003c/em\u003e 151, 119, 107 explained by Zeng et al. (\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). (Iso)orientin (\u003cb\u003e5\u003c/b\u003e) with \u003cem\u003em/z\u003c/em\u003e 327, 297, 285 and (iso)vitexin (\u003cb\u003e8\u003c/b\u003e) \u003cem\u003em/z\u003c/em\u003e 311, 283, 269, 134 by Amaral et al. (2019)., proved to be consistent with data previously reported in the literature, giving greater reliability to the annotation carried out.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eChemical compounds annotated for the ethanolic extract of \u003cem\u003eF\u003c/em\u003e. \u003cem\u003ecitrifolia\u003c/em\u003e leaves.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"10\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCompound\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRT (min)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMolecular formula\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eExperimental\u003c/p\u003e\u003cp\u003e\u003cem\u003em/z\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTeorical\u003c/p\u003e\u003cp\u003e\u003cem\u003em/z\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eError (ppm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eFragments (MS/MS)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003ePutative name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eChemical class\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eIonization mode\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e8\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e179.0346\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e179.0344\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e135\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003ecaffeic acid*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ePhenolic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e11\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e447.0920\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e447.0927\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e357, 327, 297, 285\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eorientin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e3\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e431.0973\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e431.0978\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e311, 284, 283, 269, 239, 161, 135\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003evitexin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e4\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e197.1172\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e197.1178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e3.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e197, 179, 161, 135, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e(epi)loliolide\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eMonoterpene\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e5\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e11\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e447.0920\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e447.0927\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e327, 297, 285\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e(iso)orientin*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e197.1178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e197.1178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e197, 179, 161, 135, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eloliolide\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eMonoterpene\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e7\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e433.1122\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e433.1135\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e3.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e433, 313, 295, 272, 271\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003enaringenin-7-O-glucoside\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavanone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e8\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e21\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e10\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e431.0968\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e431.0978\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e311, 283, 269,134\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e(iso)vitexin*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e9\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e4.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e287.0552\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e287.0556\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e287, 151, 135, 125, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eeriodictyol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavanone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e10\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e4.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e285.0398\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e285.0399\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e285, 151, 133, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eluteolin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e11\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e4.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e301.0345\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e301.0348\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e178, 159, 151, 139, 121, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003equercetin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavonol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e12\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e207.0654\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e207.0657\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e207, 179, 161, 135, 134\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eethyl caffeate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ePhenolic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e22\u003c/sub\u003eO\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e357.1335\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e357.1338\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e342, 327, 311, 151, 136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003epinoresinol*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eLignan\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e14\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e271.0607\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e271.0606\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e151, 119, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003enaringenin*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavanone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e15\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5.60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e269.0453\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e269.0450\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e159, 151, 117, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eapigenin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e16\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e7.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e15\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e255.0656\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e255.0657\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e255, 213, 187, 151, 125, 107\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003epinocembrin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eFlavanone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026ndash;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e9.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e339.1222\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e339.1232\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e283, 271, 255, 153\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eisowighteone*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIsoflavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e18\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10.29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e337.1070\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e337.1076\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e319, 295, 283, 271, 247\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003ealpinumisoflavone*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIsoflavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e19\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e20\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e337.1071\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e337.1076\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e319, 295, 283, 271, 247\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e(iso)alpinumisoflavone*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIsoflavone\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e20\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e17.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e30\u003c/sub\u003eH\u003csub\u003e48\u003c/sub\u003eO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e425.3763\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e425.3783\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e407, 231, 191, 177, 109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003elupenone*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eTriterpene\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003e21\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e19.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC\u003csub\u003e29\u003c/sub\u003eH\u003csub\u003e48\u003c/sub\u003eO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e413.3764\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e413.3783\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e311, 297, 283, 255\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003estigmasterol*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eSteroid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e[M\u0026thinsp;+\u0026thinsp;H]\u003c/b\u003e\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"10\"\u003e*Notes made based on chemical prospecting of previous studies reported for the genus and comparative analysis of spectra available in the literature.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eAmong the flavonoids, the isoflavones isowighteone (\u003cb\u003e17\u003c/b\u003e), alpinumisoflavone (\u003cb\u003e18\u003c/b\u003e) and (iso)alpinumisoflavone (\u003cb\u003e19\u003c/b\u003e) were observed. Their characteristic fragmentations corroborate the proposed structures, including the loss of the prenyl group, resulting in the ion of \u003cem\u003em/z\u003c/em\u003e 283 and 271, common to all of them. Isowighteone (\u003cb\u003e17\u003c/b\u003e) showed fragments at \u003cem\u003em/z\u003c/em\u003e 283, 271, 255 and 153 (Hostettmann; Marston and Wolfender \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2005\u003c/span\u003e), and alpinumisoflavone (\u003cb\u003e18\u003c/b\u003e) and (iso)alpinumisoflavone (\u003cb\u003e19\u003c/b\u003e) showed fragments at \u003cem\u003em/z\u003c/em\u003e 319, 295, 283, 271 and 247 showed by Chen et al. (2024). The triterpene lupenone (\u003cb\u003e20\u003c/b\u003e) with ions at \u003cem\u003em/z\u003c/em\u003e 407, 231, 191, 177 and 109 as described by Wang et al. (\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and the steroid stigmasterol (\u003cb\u003e21\u003c/b\u003e) with \u003cem\u003em/z\u003c/em\u003e at 311, 297, 283, 255 were also identified, reinforcing the chemical complexity of the species by Rozenberg et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). The compounds orientin (\u003cb\u003e2\u003c/b\u003e), vitexin (\u003cb\u003e3\u003c/b\u003e), (epi)loliolide (\u003cb\u003e4\u003c/b\u003e), loliodide (\u003cb\u003e6\u003c/b\u003e), narigerin-7-O-glucoside (\u003cb\u003e7\u003c/b\u003e), eriodictiol (\u003cb\u003e9\u003c/b\u003e), luteolin (\u003cb\u003e10\u003c/b\u003e), quercetin (\u003cb\u003e11\u003c/b\u003e), ethyl caffeate (\u003cb\u003e12\u003c/b\u003e), apigenin (\u003cb\u003e15\u003c/b\u003e) and pinocembrin (\u003cb\u003e16\u003c/b\u003e) were identified using the GNPS platform. This tool made it possible to correlate the experimental mass spectra with data libraries of previously described compounds.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eCytotoxicity and antiviral activity of the extract\u003c/h2\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e presents the cytotoxicity (CC₅₀) and antiviral activity results for the ethanolic extract of \u003cem\u003eF. citrifolia\u003c/em\u003e leaves. The extract showed a CC₅₀ of 793.92 \u0026micro;g/mL. The effective concentrations (EC₅₀) and selectivity indices (SI) were 61.24 \u0026micro;g/mL (SI 12.96) for MAYV, 117.92 \u0026micro;g/mL (SI 6.73) for CHIKV, and 117.29 \u0026micro;g/mL (SI 6.77) for ZIKV. No antiviral activity was observed against OROV. The lack of effect may be attributed to OROV\u0026rsquo;s segmented negative-sense RNA genome, which presents replication mechanisms distinct from unsegmented arboviruses like MAYV, CHIKV, and ZIKV (Files et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Emmanuel et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003e\u0026ndash; Antiviral activity and cytotoxicity threshold for the ethanolic extract of \u003cem\u003eFicus citrifolia\u003c/em\u003e leaves.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCytotoxic concentration (CC\u003csub\u003e50\u003c/sub\u003e, \u0026micro;g/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\u003cp\u003e793.92\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eArbovirus\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eEffective concentration (EC\u003csub\u003e50\u003c/sub\u003e, \u0026micro;g/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSelectivity index (SI)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eMAYV\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e61.24\u0026thinsp;\u0026plusmn;\u0026thinsp;5.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.96\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eOROV\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003end\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003end\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eCHIKV\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e117.92\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.73\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eZIKV\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e117.29\u0026thinsp;\u0026plusmn;\u0026thinsp;37.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.77\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003eNote: MAYV (Mayaro virus); OROV (Oropouche virus); CHIKV (Chikungunya virus); ZIKV (Zika virus); nd (not detected).\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003e(Brito, 2025; European Centre for Disease Prevention and Control, 2025).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eFicus\u003c/em\u003e species are rich in diverse secondary metabolites such as flavonoids, terpenes, phenols, sterols, and lignans (Chandra et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), which may underlie the observed biological effects. The high SI values, particularly against MAYV (12.96), suggest a therapeutic potential with low cytotoxicity, reinforcing the extract\u0026rsquo;s promise against Orthoflavivirus and Alphavirus.\u003c/p\u003e\u003cp\u003eLC-MS/MS analysis enabled the identification of compounds potentially linked to this antiviral activity. Caffeic acid (\u003cb\u003e1\u003c/b\u003e) has shown activity against ILHV and ZIKV by binding to the viral envelope protein (Saivish et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Its derivative, ethyl caffeate (\u003cb\u003e12\u003c/b\u003e), also exhibited antiviral effects by inhibiting HIV protease (Wang et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eAmong flavonoids, orientin (\u003cb\u003e2\u003c/b\u003e) blocks SARS-CoV-2 entry into host cells (Bhowmik et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), while isoorientin (\u003cb\u003e5\u003c/b\u003e) inhibits the spike-ACE2 interaction (Chen et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Luteolin (\u003cb\u003e10\u003c/b\u003e) is known for broad-spectrum antiviral action against influenza, herpesvirus, and coronaviruses (Lu et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Vitexin (\u003cb\u003e3\u003c/b\u003e) and isovitexin (\u003cb\u003e8\u003c/b\u003e) inhibit H1N1 and offer protection against COVID-19 (Sadati et al., 2019; Ferdausi et al., 2022). Apigenin (\u003cb\u003e15\u003c/b\u003e) interferes with replication of DNA and RNA viruses, including HSV, HCV, DENV, and SARS-CoV (Lee et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFlavanones identified include eriodictyol (\u003cb\u003e9\u003c/b\u003e), which modulates inflammatory cytokines relevant in CHIKV and MAYV infections (Deng et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Naringenin (\u003cb\u003e14\u003c/b\u003e) and its glycoside naringenin-7-O-glucoside (\u003cb\u003e7\u003c/b\u003e), along with pinocembrin (\u003cb\u003e16\u003c/b\u003e), reduce ZIKV RNA synthesis and affect viral proteins (Cataneo et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Lee et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Quercetin (\u003cb\u003e11\u003c/b\u003e) disrupts MAYV entry and RNA replication (Santos et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), supporting the strong activity observed against this virus (EC₅₀ 61.24 \u0026micro;g/mL; SI 12.96).\u003c/p\u003e\u003cp\u003ePinoresinol (\u003cb\u003e13\u003c/b\u003e), a lignan reported in \u003cem\u003eF. citrifolia\u003c/em\u003e, has demonstrated anti-HBV effects (Wang et al., 2013). Prenylated isoflavones such as isowighteone (\u003cb\u003e17\u003c/b\u003e), alpinumisoflavone (\u003cb\u003e18\u003c/b\u003e), and its isomer, known from the genus (Bankeu et al., 2011), may also contribute due to increased lipophilicity and membrane interaction potential (Ateba et al., 2019).\u003c/p\u003e\u003cp\u003eMonoterpenes like loliolide (\u003cb\u003e6\u003c/b\u003e), identified in the genus (Zhang et al., \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), inhibit HCV entry (Chung et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Its epimer, (epi)loliolide (\u003cb\u003e4\u003c/b\u003e), although not linked to antiviral activity, shows anti-hepatocellular carcinoma properties (Gangadhar et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The triterpene lupenone (\u003cb\u003e20\u003c/b\u003e) has activity against CHIKV, while stigmasterol (\u003cb\u003e21\u003c/b\u003e) inhibits HSV replication (Bhakat \u0026amp; Soliman, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Petrera, N\u0026iacute;ttolo \u0026amp; Alch\u0026eacute;, 2014).\u003c/p\u003e\u003cp\u003eThese results suggest that the antiviral effect may arise from synergistic interactions among m etabolites rather than isolated actions (Caesar \u0026amp; Cech, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The chemical complexity of \u003cem\u003eF. citrifolia\u003c/em\u003e supports its antiviral activity and points to its potential for therapeutic development. Further studies, including bioguided fractionation and molecular docking, are essential to elucidate specific mechanisms and identify active constituents.\u003c/p\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHorrana Ac\u0026aacute;cio Mardegan designed the studies. Horrana Ac\u0026aacute;cio Mardegan, Allana Martins dos Santos Ata\u0026iacute;de, Loueny Celine Ribeiro Andrade and Johan Carlos Costa Santiago conducted LC-MS experiments. C\u0026iacute;ntia Lopes de Brito Guimar\u0026atilde;es designed antiviral assays. Horrana Ac\u0026aacute;cio Mardegan, Allana Martins dos Santos Ata\u0026iacute;de and Johan Carlos Costa Santiago conducted MS/MS data processing. Horrana Ac\u0026aacute;cio Mardegan and Johan Carlos Costa Santiago performed metabolite characterization. Horrana Ac\u0026aacute;cio Mardegan, Allana Martins dos Santos Ata\u0026iacute;de, Loueny Celine Ribeiro Andrade and Abra\u0026atilde;o de Jesus Barbosa Muribeca searched the pharmacological properties of the annotated metabolites. Ariane Coelho Ferraz, Mar\u0026iacute;lia Bueno da Silva Menegatto and C\u0026iacute;ntia Lopes de Brito Magalh\u0026atilde;es performed and discussed \u003cem\u003ein vitro\u003c/em\u003e antiviral assays with the extracts. Horrana Ac\u0026aacute;cio Mardegan, Allana Martins dos Santos Ata\u0026iacute;de, Johan Carlos Costa Santiago, Abra\u0026atilde;o de Jesus Barbosa Muribeca wrote the manuscript. S\u0026ocirc;nia das Gra\u0026ccedil;as Santa Rosa Pamplona, C\u0026iacute;ntia Lopes de Brito Magalh\u0026atilde;es, Abra\u0026atilde;o de Jesus Barbosa Muribeca, Consuelo Yumiko Yoshioka e Silva and Milton Nascimento da Silva reviewed the whole manuscript and provided comments. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHorrana Ac\u0026aacute;cio Mardegan would like to thank the Federal University of Par\u0026aacute; (UFPA) and the Coordination for the Improvement of Higher Education Personnel (CAPES) for their support in awarding a master\u0026apos;s degree scholarship. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSupplementary Information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This study did not involve human participants or animal experiments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdulmalik I, Sule M, Yaro A, Abdullahi M, Abdulkadir M, Yusuf H (2011) Evaluation of analgesic and anti-inflammatory effects of ethanol extract of \u003cem\u003eFicus iteophylla\u003c/em\u003e leaves in rodents. Afr J Tradit Complement Altern Med. ;8(4)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAndrzej Patyra, Dudek MK, Kiss AK (2022) LC-DAD\u0026ndash;ESI-MS/MS and NMR Analysis of Conifer Wood Specialized Metabolites. Cells 11(20):3332\u0026ndash;3332\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAntoun MD, Ramos Z, Vazques J, Oquendo I, Proctor GR, Gerena L, Franzblau SG (2001) Evaluation of the flora of Puerto Rico for in vitro antiplasmodial and antimycobacterial activities. Phytotherapy Research: PTR 15(7):638\u0026ndash;642\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eArunachalam K, Parimelazhagan T (2013) Anti-inflammatory, wound healing and in-vivo antioxidant properties of the leaves of \u003cem\u003eFicus amplissima\u003c/em\u003e Smith. J Ethnopharmacol 145(1):139\u0026ndash;145\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBaetas AC, F\u0026aacute;bio L, Paula A (2018) In vitro cytotoxicity of lignans isolated from \u003cem\u003eFicus citrifolia\u003c/em\u003e P. Miller (Moraceae) on C6 glioma cell line. Glo Adv Res J Med Med Sci 7:77\u0026ndash;84\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBhakat S, Soliman MES (2015) Chikungunya virus (CHIKV) inhibitors from natural sources: a medicinal chemistry perspective. J Nat Med 69(4):451\u0026ndash;462\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBhowmik A, Biswas S, Hajra S, Saha P (2021) In silico validation of potent phytochemical orientin as inhibitor of SARS-CoV-2 spike and host cell receptor GRP78 binding. Heliyon 7(1):e05923\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBrito C, Falc\u0026atilde;o MB, Maria T, Cerqueira-Silva, Teixeira MG, Freitas R, Chikungunya (2025) From Hypothesis to Evidence of Increased Severe Disease and Fatalities. \u003cem\u003eViruses\u003c/em\u003e [Internet]. Jan 3 [cited 2025 May 28];17(1):62\u0026ndash;2. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.mdpi.com/1999-4915/17/1/62\u003c/span\u003e\u003cspan address=\"https://www.mdpi.com/1999-4915/17/1/62\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCaesar LK, Cech NB (2019) Synergy and antagonism in natural product extracts: when 1\u0026thinsp;+\u0026thinsp;1 does not equal 2. Natural Prod Reports [Internet] 36(6):869\u0026ndash;888\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCarolina A, Auguste AJ, Danila Vedovello, Ferreira MU et al (2015) M\u0026ocirc;nica da Silva-Nunes, M\u0026aacute;rcia Aparecida Speran\u0026ccedil;a,. Isolation and Characterization of Mayaro Virus from a Human in Acre, Brazil. \u003cem\u003eAmerican Journal of Tropical Medicine and Hygiene\u003c/em\u003e [Internet]. ;92(2):401\u0026ndash;4. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4347347\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4347347\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCataneo AHD, \u0026Aacute;vila EP, Mendes LA, de O VG, Ferraz CR, de Almeida MV et al (2021) Flavonoids as Molecules With Anti-Zika virus Activity. Front Microbiol. ;12\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChambers MC, Maclean B, Burke R, Amodei D, Ruderman DL, Neumann S et al (2012) A cross-platform toolkit for mass spectrometry and proteomics. \u003cem\u003eNature Biotechnology\u003c/em\u003e [Internet]. ;30(10):918\u0026ndash;20. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.nature.com/articles/nbt.2377\u003c/span\u003e\u003cspan address=\"https://www.nature.com/articles/nbt.2377\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChandra S, Kala K, Negi MS, Anuj, Nautiyal (2023) The Genus \u003cem\u003eFicus\u003c/em\u003e: a review of its phytochemicals and pharmacological studies. ;7(1):230. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.researchgate.net/publication/371874948_The_Genus_Ficus_a_review_of_its_phytochemicals_and_pharmacological_studies\u003c/span\u003e\u003cspan address=\"https://www.researchgate.net/publication/371874948_The_Genus_Ficus_a_review_of_its_phytochemicals_and_pharmacological_studies\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen G, Li X, Saleri FD, Guo M (2016) Analysis of Flavonoids in Rhamnus davurica and Its Antiproliferative Activities. Molecules 21(10):1275\u0026ndash;1275\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen G, Sun Y, Yushan D, Shaerbayi N, Zhang H, He H et al (2023) Identification and Characterization of Chemical Constituents from \u003cem\u003eAmmopiptanthus nanus\u003c/em\u003e Stem and Their Metabolites in Rats by UHPLC-Q-TOF-MS/MS. Planta Med 90(02):138\u0026ndash;153\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen YL, Chen CY, Lai KH, Chang YC, Hwang TL (2023) Anti-inflammatory and antiviral activities of flavone C-glycosides of \u003cem\u003eLophatherum gracil\u003c/em\u003ee for COVID-19. J Funct Foods 101:105407\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChung CY, Liu CH, Thierry Burnouf, Wang GH, Chang SP, Alagie Jassey et al (2016) Activity-based and fraction-guided analysis of \u003cem\u003ePhyllanthus urinaria\u003c/em\u003e identifies loliolide as a potent inhibitor of hepatitis C virus entry. Antiviral Res 130:58\u0026ndash;68\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCruz RF, Corr\u0026ecirc;a (2022) Carlos Victor Lamar\u0026atilde;o, Edgar Aparecido Sanches, Pedro Henrique Campelo, Jaqueline. \u003cem\u003eFicus spp\u003c/em\u003e.: Phytochemical composition and medicinal potential. Res Soc Dev 11(12):e265111234135\u0026ndash;e265111234135\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDarbour N, Bayet C, Rodin-Bercion S, Elkhomsi Z, Lurel F, Chaboud A et al (2007) Isoflavones from \u003cem\u003eFicus nymphaefolia\u003c/em\u003e. Nat Prod Res 21(5):461\u0026ndash;464\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDeng Z, Hassan S, Rafiq M, Li H, He Y, Cai Y et al (2020) Pharmacological Activity of Eriodictyol: The Major Natural Polyphenolic Flavanone. Wu H, editor. \u003cem\u003eEvidence-Based Complementary and Alternative Medicine\u003c/em\u003e. ;2020:1\u0026ndash;11\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDiagne CT, Bengue M, Choumet V, Hamel R, Pompon J, Miss\u0026eacute; D (2020) Mayaro Virus Pathogenesis and Transmission Mechanisms. Pathogens 9(9):738\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDonald CL, Brennan B, Cumberworth SL, Rezelj VV, Clark JJ, Cordeiro MT et al (2016) Full Genome Sequence and sfRNA Interferon Antagonist Activity of Zika Virus from Recife, Brazil. \u003cem\u003ePLoS neglected tropical diseases\u003c/em\u003e [Internet]. ;10(10):e0005048. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/27706161/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/27706161/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003edos Santos AE, Kuster RM, Yamamoto KA, Salles TS, Campos R, de Meneses MD et al (2014) Quercetin and quercetin 3-O-glycosides from Bauhinia longifolia (Bong.) Steud. show anti-Mayaro virus activity. Parasites Vectors. ;7(1)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEmmanuel V, Ot\u0026aacute;vio JB, Brustolini CL, Filipe, Castro F, Guimar\u0026atilde;es AP et al (2021) Common Dysregulation of Innate Immunity Pathways in Human Primary Astrocytes Infected With Chikungunya, Mayaro, Oropouche, and Zika Viruses. Front Cell Infect Microbiol. ;11\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEpidemiological Update Dengue and other Arboviruses \u0026ndash;\u0026thinsp;10 June 2020 (em ingl\u0026ecirc;s e espanhol) [Internet]. \u003cem\u003ePaho.org. 2020\u003c/em\u003e. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.paho.org/pt/documents/epidemiological-update-dengue-and-other-arboviruses-10-june-2020\u003c/span\u003e\u003cspan address=\"https://www.paho.org/pt/documents/epidemiological-update-dengue-and-other-arboviruses-10-june-2020\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eErina Petrera (2014) Anal\u0026iacute;a Gabriela N\u0026iacute;ttolo, Alch\u0026eacute; LE. Antiviral Action of Synthetic Stigmasterol Derivatives on Herpes Simplex Virus Replication in Nervous Cells \u003cem\u003eIn Vitro\u003c/em\u003e. Biomed Res Int 2014:1\u0026ndash;9\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEuropean Centre for Disease Prevention and Control. Chikungunya worldwide overview [Internet]. European Centre for Disease Prevention and Control (2025) Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ecdc.europa.eu/en/chikungunya-monthly\u003c/span\u003e\u003cspan address=\"https://www.ecdc.europa.eu/en/chikungunya-monthly\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFederica Dell\u0026rsquo;Annunziata, Sellitto C, Franci G, Marcotullio MC, Piovan A, Marca RD et al (2022) Antiviral Activity of \u003cem\u003eFicus rubiginosa\u003c/em\u003e Leaf Extracts against HSV-1, HCoV-229E and PV-1. Viruses 14(10):2257\u0026ndash;2257\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFerdausi N, Islam S, Rimti F, Quayum S, Arshad E, Ibnat A et al Point-specific interactions of isovitexin with the neighboring amino acid residues of the hACE2 receptor as a targeted therapeutic agent in suppressing the SARS-CoV-2 influx mechanism. \u003cem\u003eJournal of Advanced Veterinary and Animal Research\u003c/em\u003e [Internet]. 2022 [cited 2023 Mar 8];9(2):230. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9298103/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9298103/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003e\u003cem\u003eFicus spp\u003c/em\u003e. (fig): Ethnobotany and potential as anticancer and anti-inflammatory agents. \u003cem\u003eJournal of Ethnopharmacology\u003c/em\u003e [Internet]. (2008) ;119(2):195\u0026ndash;213\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFiles MA, Hansen CA, Herrera VC, Schindewolf C, Barrett ADT, Beasley DWC et al (2022) Baseline mapping of Oropouche virology, epidemiology, therapeutics, and vaccine research and development. Vaccines. ;7(1)\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGangadhar KN, Rodrigues MJ, Pereira H, Gaspar H, Malcata FX, Barreira L et al (2020) Anti-Hepatocellular Carcinoma (HepG2) Activities of Monoterpene Hydroxy Lactones Isolated from the Marine Microalga \u003cem\u003eTisochrysis lutea\u003c/em\u003e. Mar Drugs 18(11):567\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGirard M, Nelson CB, Picot V, Gubler DJ (2020) Arboviruses: A global public health threat. Vaccine 38(24):3989\u0026ndash;3994\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHamed M, Khalifa M, Mahmoud, Mohammed OA, Kamaly, Alanazi AS et al (2023) The Potential Roles of \u003cem\u003eFicus carica\u003c/em\u003e Extract in the Management of COVID-19 Viral Infections: A Computer-aided Drug Design Study. Curr Comput - Aided Drug Des 20(6):974\u0026ndash;986\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHostettmann K, Marston A, Wolfender JL (2005) Strategy in the Search for New Lead Compounds and Drugs from Plants. CHIMIA Int J Chem 59(6):291\u0026ndash;294\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHostettmann K, Marston A, Wolfender JL (2005) Strategy in the search for new lead compounds and drugs from plants. Chimia 59(6):291\u0026ndash;291\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJean R, Khayala, Lenta BN, Noungou\u0026eacute; DT, Ngouela SA, Mustafa SA et al (2011) Isoflavone Dimers and Other Bioactive Constituents from the Figs of \u003cem\u003eFicus mucuso\u003c/em\u003e. J Nat Prod 74(6):1370\u0026ndash;1378\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJean R, Khayala, Lenta BN, Noungou\u0026eacute; DT, Ngouela SA, Mustafa SA et al (2011) Isoflavone Dimers and Other Bioactive Constituents from the Figs of \u003cem\u003eFicus mucuso\u003c/em\u003e. J Nat Prod 74(6):1370\u0026ndash;1378\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKhan AH, Morita K, Parquet M del, Hasebe C, Mathenge F, Igarashi EGM (2002) Complete nucleotide sequence of chikungunya virus and evidence for an internal polyadenylation site. J Gen Virol 83(12):3075\u0026ndash;3084\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLee IG, Lee J, Hong SH, Seo YJ (2023) Apigenin\u0026rsquo;s Therapeutic Potential Against Viral Infection. Front Bioscience-Landmark 28(10):237\u0026ndash;237\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLee JL, Loe MWC, Lee RCH, Chu JJH (2019) Antiviral activity of pinocembrin against Zika virus replication. Antiviral Res 167:13\u0026ndash;24\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLu P, Zhang T, Ren Y, Rao H, Lei J, Zhao G et al (2023) Literature Rev Antiviral Mechanism Luteolin 18(4):1934578X2311715\u0026ndash;1934578X2311715\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSaivish MV, Pacca CC, Vivaldo, Gabriela, Alves R, Liliane, Nebo et al (2023) Caffeic Acid Has Antiviral Activity against Ilh\u0026eacute;us Virus \u003cem\u003eIn Vitro\u003c/em\u003e. Viruses 15(2):494\u0026ndash;494\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePaix\u0026atilde;o ES, Teixeira MG, Rodrigues LC (2017) Zika, chikungunya and dengue: the causes and threats of new and re-emerging arboviral diseases. \u003cem\u003eBMJ Global Health\u003c/em\u003e [Internet]. ;3(Suppl 1):e000530. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://gh.bmj.com/content/3/Suppl_1/e000530\u003c/span\u003e\u003cspan address=\"https://gh.bmj.com/content/3/Suppl_1/e000530\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePearson JL, Lee S, Suresh H, Low M, Nang M, Singh S et al (2014) The Liquid Chromatographic Determination of Chlorogenic and Caffeic Acids in Xu Duan (\u003cem\u003eDipsacus asperoides\u003c/em\u003e) Raw Herb. ISRN Anal Chem 2014:1\u0026ndash;6\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePielnaa P, Al-Saadawe M, Saro A, Dama MF, Zhou M, Huang Y et al (2020) Zika virus-spread, epidemiology, genome, transmission cycle, clinical manifestation, associated challenges, vaccine and antiviral drug development. \u003cem\u003eVirology\u003c/em\u003e [Internet]. ;543(0042-6822):34\u0026ndash;42. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/32056845/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/32056845/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePingen M, Bryden Steven R, Pondeville E, Schnettler E, Kohl A, Merits A et al (2016) Host Inflammatory Response to Mosquito Bites Enhances the Severity of Arbovirus Infection. \u003cem\u003eImmunity\u003c/em\u003e [Internet]. ;44(6):1455\u0026ndash;69. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4920956/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4920956/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eQi C, Fu YH, Chen W, Chen G, Dai C, Song X et al (2017) A new isoflavone from the roots of \u003cem\u003eFicus auriculata\u003c/em\u003e. Nat Prod Res 32(1):43\u0026ndash;47\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRhelly A, Diogo J, Paulo AC, Ferraz, Mardegan HA, Menegatto S et al (2023) Untargeted-based metabolomics analysis and in vitro/in silico antiviral activity of extracts from \u003cem\u003ePhyllanthus brasiliensis\u003c/em\u003e (Aubl.) Poir. Phytochem Anal 34(7):869\u0026ndash;883\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRicardo Guimar\u0026atilde;es Amaral, Vieira S, Claudia M, do C, Andrade LN, Severino P et al (2019) Cytotoxic potential of 14 \u003cem\u003ePassiflora\u003c/em\u003e species against cancer cells. J Med Plants Res 13(7):157\u0026ndash;166\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRozenberg R, Ruibal-Mendieta NL, Petitjean G, Cani PD, Delacroix DL, Delzenne NM et al (2003) Phytosterol analysis and characterization in spelt \u003cem\u003e(Triticum aestivum\u003c/em\u003e ssp. spelta L.) and wheat (\u003cem\u003eT. aestivum\u003c/em\u003e L.) lipids by LC/APCI-MS. J Cereal Sci 38(2):189\u0026ndash;197\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSadati S, Gheibi N, Ranjbar S, Hashemzadeh M (2018) Docking study of flavonoid derivatives as potent inhibitors of influenza H1N1 virus neuraminidase. Biomedical Rep. Nov 23\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSaeed MF, Watts DM, Shope RE, Barrett ADT, Tesh RB, Nunes M et al (2000) Nucleotide sequences and phylogeny of the nucleocapsid gene of Oropouche virus. J Gen Virol 81(3):743\u0026ndash;748\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSchmid R, Heuckeroth S, Korf A, Smirnov A, Myers O, Dyrlund TS et al (2023) Integrative analysis of multimodal mass spectrometry data in MZmine 3. \u003cem\u003eNature Biotechnology\u003c/em\u003e [Internet]. ;41(4):447\u0026ndash;9. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.nature.com/articles/s41587-023-01690-2\u003c/span\u003e\u003cspan address=\"https://www.nature.com/articles/s41587-023-01690-2\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSchwartz DA, Dashraath P, Baud D (2024) Oropouche Virus (OROV) in Pregnancy: An Emerging Cause of Placental and Fetal Infection Associated with Stillbirth and Microcephaly following Vertical Transmission. \u003cem\u003eViruses\u003c/em\u003e [Internet]. ;16(9):1435. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.mdpi.com/1999-4915/16/9/1435\u003c/span\u003e\u003cspan address=\"https://www.mdpi.com/1999-4915/16/9/1435\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSieniawska E, Świątek Ł, Sinan KI, Zengin G, Boguszewska A, Polz-Dacewicz M et al (2022) Phytochemical Insights into \u003cem\u003eFicus sur\u003c/em\u003e Extracts and Their Biological Activity. \u003cem\u003eMolecules\u003c/em\u003e (Basel, Switzerland) [Internet]. ;27(6):1863. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://pubmed.ncbi.nlm.nih.gov/35335228/\u003c/span\u003e\u003cspan address=\"https://pubmed.ncbi.nlm.nih.gov/35335228/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMvondo SBAMA, S\u0026eacute;firin, Djiogue St\u0026eacute;phane, Zingue, Krenn L (2019) Dieudonn\u0026eacute; Njamen. A Pharmacological Overview of Alpinumisoflavone, a Natural Prenylated Isoflavonoid. \u003cem\u003eFrontiers in Pharmacology\u003c/em\u003e. ;10\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVALLI M, RUSSO HM, BOLZANI VS (2018) The potential contribution of the natural products from Brazilian biodiversity to bioeconomy. An Acad Bras Cienc 90(1 suppl 1):763\u0026ndash;778\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang GK, Lin BB, Rao R, Zhu K, Qin XY, Xie GY et al (2012) A new lignan with anti-HBV activity from the roots of \u003cem\u003eBombax ceiba\u003c/em\u003e. Nat Prod Res 27(15):1348\u0026ndash;1352\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang M, Carver JJ, Phelan VV, Sanchez LM, Garg N, Peng Y et al (2016) Sharing and community curation of mass spectrometry data with Global Natural Products Social Molecular Networking. Nat Biotechnol 34(8):828\u0026ndash;837\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang X, Wei Y, Tian WY, Sakharkar MK, Liu Q, Yang X et al (2019) Characterization of Nine Compounds Isolated from the Acid Hydrolysate of \u003cem\u003eLonicera fulvotomentosa\u003c/em\u003e Hsu et S. C. Cheng and Evaluation of Their In Vitro Activity towards HIV Protease. \u003cem\u003eMolecules\u003c/em\u003e [Internet]. Dec 11 [cited 2023 Sep 17];24(24):4526. Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.ncbi.nlm.nih.gov/pmc/articles/pmc6943440/\u003c/span\u003e\u003cspan address=\"http://www.ncbi.nlm.nih.gov/pmc/articles/pmc6943440/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang X, Zhong XJ, Zhou N, Cai N, Xu JH, Wang QB et al (2020) Rapid Characterizaiton of Chemical Constituents of the Tubers of \u003cem\u003eGymnadenia conopsea\u003c/em\u003e by UPLC\u0026ndash;Orbitrap\u0026ndash;MS/MS Analysis. Molecules 25(4):898\u0026ndash;898\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWei L, Ma R, Fu Y (2022) Differences in Chemical Constituents between \u003cem\u003eDalbergia oliveri\u003c/em\u003e Heartwood and Sapwood and Their Effect on Wood Color. Molecules 27(22):7978\u0026ndash;7978\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWorld Health Organization. WHO Coronavirus (COVID-19) [Internet]. World Health Organization (2023) Available from: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://covid19.who.int/\u003c/span\u003e\u003cspan address=\"https://covid19.who.int/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu YS, Afzal S, Appalaraju V, Wei TQ, Manap ASA, Albokhadaim I (2024) Evaluation for Antiviral Potential of \u003cem\u003eFicus deltoidea\u003c/em\u003e against Dengue Virus Type-2 (Penilaian Potensi Antiviral \u003cem\u003eFicus deltoidea\u003c/em\u003e terhadap Virus Denggi Jenis-2). Sains Malaysiana 53(2):383\u0026ndash;396\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZeng X, Su W, Zheng Y, Liu H, Li P, Zhang W et al (2018) UFLC-Q-TOF-MS/MS-Based Screening and Identification of Flavonoids and Derived Metabolites in Human Urine after Oral Administration of Exocarpium Citri Grandis Extract. Molecules 23(4):895\u0026ndash;895\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang N, Guo M, Zhou B, Zhao X, Fu M, Shen X et al (2024) Chemical Components and Their α-Glucosidase Inhibitory Activity from the Leaves of \u003cem\u003eFicus carica\u003c/em\u003e Linn. Records Nat Prod. ;(1):165\u0026ndash;170\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou N, Zheng D, You Q, Chen T, Jiang J, Shen W et al (2023) Therapeutic Potential of Biochanin A in Herpes Simplex Keratitis. Pharmaceuticals 16(9):1240\u0026ndash;1240\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"revista-brasileira-de-farmacognosia","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"rbfa","sideBox":"Learn more about [Revista Brasileira de Farmacognosia](https://www.springer.com/journal/43450)","snPcode":"43450","submissionUrl":"https://www.editorialmanager.com/rbfa/default2.aspx","title":"Revista Brasileira de Farmacognosia","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"LC-MS/MS, Compound Annotation, Antiviral Activity, Moraceae","lastPublishedDoi":"10.21203/rs.3.rs-7583932/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7583932/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eArboviral diseases pose a serious public health problem, particularly in tropical regions, where environmental conditions favor the proliferation of mosquito vectors, facilitating viral transmission and leading to debilitating clinical manifestations ranging from fever and joint pain to severe neurological complications. This study evaluated the \u003cem\u003ein vitro\u003c/em\u003e antiviral activity and phytochemical profile of the ethanolic extract of \u003cem\u003eFicus citrifolia\u003c/em\u003e Mill. leaves (Moraceae). Using high-resolution LC-MS/MS analysis, 21 compounds were identified, including phenolic acids, flavonoids, lignans, terpenes, and steroids - many of which are known for their antiviral and anti-inflammatory properties. The extract exhibited significant antiviral activity against Mayaro virus (EC₅₀: 61.24 µg/mL, SI: 12.96), Chikungunya virus (EC₅₀: 117.92 µg/mL, SI: 6.73), and Zika virus (EC₅₀: 117.29 µg/mL, SI: 6.77), with no observed activity against Oropouche virus. The findings highlight the potential of \u003cem\u003eF. citrifolia\u003c/em\u003e as a source of antiviral agents and support its traditional use in treating inflammatory and infectious diseases. Further studies, including bioassay-guided fractionation and mechanistic evaluations, are recommended to identify the active constituents and assess clinical applicability.\u003c/p\u003e","manuscriptTitle":"Antiviral Activity and Chemical Characterization of Ficus citrifolia Mill. (Moraceae) Leaf Extract Against Emerging Arboviruses","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-13 08:32:18","doi":"10.21203/rs.3.rs-7583932/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-11-03T15:11:59+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-03T14:20:27+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Revista Brasileira de Farmacognosia","date":"2025-10-22T12:28:17+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-09T02:17:16+00:00","index":"","fulltext":""},{"type":"submitted","content":"Revista Brasileira de Farmacognosia","date":"2025-10-08T16:19:07+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"revista-brasileira-de-farmacognosia","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"rbfa","sideBox":"Learn more about [Revista Brasileira de Farmacognosia](https://www.springer.com/journal/43450)","snPcode":"43450","submissionUrl":"https://www.editorialmanager.com/rbfa/default2.aspx","title":"Revista Brasileira de Farmacognosia","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"12be0281-c357-4745-ad98-265fd16aade6","owner":[],"postedDate":"November 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-30T16:25:02+00:00","versionOfRecord":{"articleIdentity":"rs-7583932","link":"https://doi.org/10.1007/s43450-026-00752-1","journal":{"identity":"revista-brasileira-de-farmacognosia","isVorOnly":false,"title":"Revista Brasileira de Farmacognosia"},"publishedOn":"2026-03-24 16:12:47","publishedOnDateReadable":"March 24th, 2026"},"versionCreatedAt":"2025-11-13 08:32:18","video":"","vorDoi":"10.1007/s43450-026-00752-1","vorDoiUrl":"https://doi.org/10.1007/s43450-026-00752-1","workflowStages":[]},"version":"v1","identity":"rs-7583932","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7583932","identity":"rs-7583932","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}