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Bioassay-Guided Fractionation of Siparuna glycycarpa n-Butanol Extract with Inhibitory Activity against Influenza A(H1N1)pdm09 Virus by Centrifugal Partition Chromatography (CPC)
Authors:Carla Monteiro Leal  Suzana Guimares Leito  Leonardo Luiz Oliveira de Mello  Isabel de Castro Rangel  Carlos Vinicius Azevedo da Silva  Milene Dias Miranda  Amanda Resende Tucci  Camilla Blanco de Assis  Carolina de Queiroz Sacramento  Natalia Fintelman-Rodrigues  Hector Henrique Ferreira Koolen  Boniek Gontijo Vaz  Rosineide Costa Simas  Gilda Guimares Leito
Abstract:Siparuna glycycarpa occurs in the Amazon region, and some species of this genus are used in Brazilian folk medicine. A recent study showed the inhibitory effect of this species against influenza A(H1N1)pdm09 virus, and in order to acquire active fractions, a polar solvent system n-butanol-methanol-water (9:1:10, v/v) was selected and used for bioassay-guided fractionation of n-butanol extract by centrifugal partition chromatography (CPC). The upper phase was used as stationary phase and the lower phase as mobile (descending mode). Among the collected fractions, the ones coded SGA, SGC, SGD, and SGO showed the highest antiviral inhibition levels (above 74%) at 100 µg·mL−1 after 24 h of infection. The bioactive fractions chemical profiles were investigated by LC-HRMS/MS data in positive and negative ionization modes exploring the Global Natural Products Social Molecular Networking (GNPS) platform to build a molecular network. Benzylisoquinoline alkaloids were annotated in the fractions coded SGA, SGC, and SGD collected during elution step. Aporphine alkaloids, O-glycosylated flavonoids, and dihydrochalcones in SGO were acquired with the change of mobile phase from lower aqueous to upper organic. Benzylisoquinolinic and aporphine alkaloids as well as glycosylated flavonoids were annotated in the most bioactive fractions suggesting this group of compounds as responsible for antiviral activity.
Keywords:centrifugal partition chromatography  influenza virus  mass spectrometry  off-line LC-MS/MS  Siparunaceae  solvent system
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