Antinociceptive in vivo activity and chemical profiling by UHPLC-MS/MS of stem bark and leaves extracts of Ficus maxima Mill. (Moraceae)

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Ethnopharmacological relevanceFicus maxima is a medicinal plant extensively used in traditional medicine by Indigenous peoples across Central and South America. It is a member of the family Moraceae, subgenus Pharmacosycea, employed in treating various conditions, including intestinal parasites, gingivitis, internal inflammations, and snake bites. Despite its significant pharmacological potential, the species remains underrepresented in scientific literature. Aim of the studyThis study aimed to evaluate the in vivo antinociceptive properties of leaf (ELFM) and stem bark (EBFM) extracts from Ficus maxima. Additionally, the chemical composition of these extracts was determined using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Materials and methodsPlant material was collected in Abaetetuba, Pará, Brazil, in October 2013 and subjected to static maceration to obtain crude ELFM and EBFM. Bio-guided fractionation was performed by sequential liquid-liquid partitioning with hexane (Hex), dichloromethane (DCM), and ethyl acetate (EtOAc), yielding the following fractions: ELFM-Hex and EBFM-Hex, ELFM-DCM and EBFM-DCM, and ELFM-EtOAc and EBFM-EtOAc. The biological activity of EBFM, ELFM, and their respective fractions were evaluated using the formalin-induced pain test and the hot plate test, followed by an assessment of their mechanisms of action. The UHPLC-MS/MS analysis was conducted using electrospray ionization (ESI) in both positive and negative modes. Metabolite annotation was facilitated by MS/MS libraries and molecular networks constructed on the GNPS platform. ResultsThe reactivity time to formalin in the neurogenic phase was reduced from 84.7 ± 7.6 s (100%) to 37.3 ± 4.7 s (44%), 33.1 ± 6.3 s (39%), 40.7 ± 7.4 s (48%), 57.2 ± 2.6 s (77%), 49.7 ± 4.1 s (58%), 46.8 ± 8.1 s (55%), and 52.4 ± 5.3 s (61%) after treatment with ASA, morphine, EBFM, ELFM, ELFM-Hex, ELFM-DCM, and ELFM-EtOAc at doses of 30 mg/kg, respectively. In the inflammatory phase, the reactivity time to formalin was reduced from 124.3 ± 25.9 s (100%) to 49.7 ± 4.7 s (40%), 9.8 ± 4.3 s (8%), 32.5 ± 8.5 s (26%), 59.8 ± 16.8 s (48%), and 54.4 ± 7.3 s (44%) after treatment with ASA, morphine, EBFM, ELFM, and ELFM-Hex at doses of 30 mg/kg, respectively. A reversal of the antinociceptive action of EBFM and ELFM was observed in the inflammatory phase after treatment with atropine, a muscarinic antagonist, and naloxone, an opioid antagonist, respectively. In the hot plate test, EBFM showed Antinociceptive Activity (AA) of 62.6 ± 9.2% after 90 min; however, there was a reversal of AA to 8.6 ± 2.8% when naloxone was used. The UHPLC-MS/MS metabolite analysis revealed the presence of loliolide (3), luteolin (13), lupeol (14), gallic acid (15), chlorogenic acid (16), pygenic acid A (17), and other metabolites from the alkaloids and fatty acids classes.