title: Quantification of bufadienolides in the poisons of Rhinella marina and Rhaebo guttatus by HPLC-UV.
abstract: Bufadienolides are the main active compounds in the Bufonidae family of frogs. Recent studies have demonstrated cytotoxic and/or antitumor activity in these molecules. A HPLC-DAD method was developed and validated to quantify three bufadienolides (telocinobufagin, marinobufagin and bufalin) in ethyl acetate extracts of the cane toad poison frogs and smooth-sided toad. The chromatographic analysis was performed on Phenomenex Luna C18 (250.0 × 4.6 mm, 5 μm), using gradient elution with acetonitrile and water, at a flow rate of 1.0 mL min(-1) and detection at 296 nm. The method showed linearity (r > 0.999) and adequate recovery values (86%-111%). The limits of quantification of bufadienolides were 7.4 μg mL(-1) for telocinobufagin, 4.2 μg mL(-1) for marinobufagin and 4.0 μg mL(-1) for bufalin. Intraday and interday values of the method were evaluated and presented standard deviation values lower than 5%. The method was successfully applied to quantify the bufadienolides in the venom extract of the cane toad, which showed a content of 60% of marinobufagin. The same method was not selective for the venom extract of the Rhaebo guttatus, despite being linear, accurate and precise, requiring the development of a technique that presents a greater selectivity.
authors.affiliation: Institute of Natural, Humanities and Social Sciences, Federal University of Mato Grosso, 78.577-267, Sinop, MT, Brazil.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Kerkhoff
authors.affiliation: Institute of Natural, Humanities and Social Sciences, Federal University of Mato Grosso, 78.577-267, Sinop, MT, Brazil.
authors.forename: Janaina da C
authors.initials: Jda C
authors.lastname: Noronha
authors.affiliation: Faculty of Pharmaceutical Sciences, Federal University of Alfenas, 37.130-000, Alfenas, MG, Brazil.
authors.forename: Rudy
authors.initials: R
authors.lastname: Bonfilio
authors.affiliation: Institute of Natural, Humanities and Social Sciences, Federal University of Mato Grosso, 78.577-267, Sinop, MT, Brazil.
authors.forename: Adilson P
authors.initials: AP
authors.lastname: Sinhorin
authors.affiliation: Institute of Natural, Humanities and Social Sciences, Federal University of Mato Grosso, 78.577-267, Sinop, MT, Brazil.
authors.forename: Domingos de J
authors.initials: Dde J
authors.lastname: Rodrigues
authors.affiliation: Department of Chemistry, Federal University of Piauí, 64.049-550, Teresina, PI, Brazil.
authors.forename: Mariana H
authors.initials: MH
authors.lastname: Chaves
authors.affiliation: Department of Chemistry, Federal University of Piauí, 64.049-550, Teresina, PI, Brazil. Electronic address: gerardovieira@yahoo.com.br.
authors.forename: Gerardo Magela
authors.initials: GM
authors.lastname: Vieira
mesh_terms: D000664:Amphibian Venoms / Q000737:chemistry*; D000818:Animals; D002018:Bufanolides / Q000032:analysis*; D002024:Bufo marinus / Q000378:metabolism*; D002025:Bufonidae / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
chemical_list: D000664:Amphibian Venoms; D002018:Bufanolides
keywords: Bufadienolides; Method validation; Rhaebo guttatus; Rhinella marina