The fresh sponge was kept in methanol for three days, after which a remaining solution was ltered, concentrated under reduced pressure at temperature of up to 40°C to obtain the crude extract. The Ag extract was suspended in 6:1 (v/v) MeOH: H 2 O and subjected to liquid/liquid partition with the increasing polarity solvents n-hexane, chloroform, ethyl acetate and n-butanol. Subsequently, the marine natural product was dried under reduced pressure in rotoevaporator (FISATOM® ) ) and stored for partition and chromatographic fractionation, obtaining fraction 2 (Ag2). The chemical characterization of the fractions was obtained by mass spectrum coupled with liquid chromatography (HPLC-MS-MS) using the Dionex Ultimate 3000 liquid chromatography system. Thus, the constituents were separated into a Phenomenex Hydro C-18 column (2.0 x 50 mm, 4 µm), with an equivalent precolumn, at a ow rate of 600 µl/min. The mobile phase was a gradient of 0.1% formic acid in water and 0.1% formic acid in methanol.
The mass spectra were recorded on an AB Sciex triple linear quadrupole ion capture spectrometer (3200 QTRAP® LC-MS/MS), type ion source (AB Sciex TurboIonSpray®). For chromatograms and spectra acquisition, the software PeakView® version 2.2 was used. The spectral data of Ag2 fraction were similar to those found in the literature and thus used as the object of study on this research [9, 13] .
The mass spectra were recorded on an AB Sciex triple linear quadrupole ion capture spectrometer (3200 QTRAP® LC-MS/MS), type ion source (AB Sciex TurboIonSpray®). For chromatograms and spectra acquisition, the software PeakView® version 2.2 was used. The spectral data of Ag2 fraction were similar to those found in the literature and thus used as the object of study on this research [9, 13] .