Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions

Abstract Background: Bufadienolides are the main secondary metabolites found in the paratoid gland secretions (PGS) of toads of the Bufonidae family. These compounds are considered the main bioactive components of PGS. The aim of this study was to develop and validate the first method for the quan...

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Main Authors: Elcio Daniel Sousa Barros, Evaldo dos Santos Monção Filho, Mariana Helena Chaves, Cícero Alves Lopes Júnior, Gerardo Magela Vieira Júnior
Format: Article
Language:English
Published: SciELO 2025-05-01
Series:Journal of Venomous Animals and Toxins including Tropical Diseases
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Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992025000100309&lng=en&tlng=en
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author Elcio Daniel Sousa Barros
Evaldo dos Santos Monção Filho
Mariana Helena Chaves
Cícero Alves Lopes Júnior
Gerardo Magela Vieira Júnior
author_facet Elcio Daniel Sousa Barros
Evaldo dos Santos Monção Filho
Mariana Helena Chaves
Cícero Alves Lopes Júnior
Gerardo Magela Vieira Júnior
author_sort Elcio Daniel Sousa Barros
collection DOAJ
description Abstract Background: Bufadienolides are the main secondary metabolites found in the paratoid gland secretions (PGS) of toads of the Bufonidae family. These compounds are considered the main bioactive components of PGS. The aim of this study was to develop and validate the first method for the quantification of total bufadienolides (free and esterified) in samples of paratoid secretions from toads, using the UV-Vis absorption spectrophotometry technique. Methods: The proposed method was based on the bathochromic shift induced by the reaction of the α-pyrone group of bufadienolides (296 nm) with a 5% (w:v) aqueous solution of sodium hydroxide and detection at 356 nm, after 60 min (time defined based on the evaluation of kinetic assays). Results: The proposed method showed wide linearity (r = 0.9999), low LOD (1.3 × 10-4 µg/mL) and LOQ (3.9 × 10-4 µg/mL), recovery (84%-99%), repeatability (%RSD ≤ 5), reproducibility and robustness (p > 0.05). The total bufadienolide content in PGS extracts from 12 samples of R. diptycha ranged from 478 to 801 mg of EqMB/g of extract, while the R. granulosa sample presented 661 mg of EqMB/g of extract. Conclusion: The new developed method is innovative, simple, fast, accurate, robust, low cost, and can contribute to future research focused on the quantification of total bufadienolides in samples of toad glandular secretions. In addition to serving as a strategic tool in the selection of work matrices, optimizing time, and minimizing costs.
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spelling doaj-art-d4b6ee675f54462a80931ceaec2b6c5d2025-08-20T03:07:44ZengSciELOJournal of Venomous Animals and Toxins including Tropical Diseases1678-91992025-05-013110.1590/1678-9199-jvatitd-2024-0064Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretionsElcio Daniel Sousa Barroshttps://orcid.org/0000-0001-9837-2248Evaldo dos Santos Monção Filhohttps://orcid.org/0000-0001-6184-706XMariana Helena Chaveshttps://orcid.org/0000-0003-1541-6757Cícero Alves Lopes Júniorhttps://orcid.org/0000-0001-8379-8174Gerardo Magela Vieira Júniorhttps://orcid.org/0000-0001-8113-8957Abstract Background: Bufadienolides are the main secondary metabolites found in the paratoid gland secretions (PGS) of toads of the Bufonidae family. These compounds are considered the main bioactive components of PGS. The aim of this study was to develop and validate the first method for the quantification of total bufadienolides (free and esterified) in samples of paratoid secretions from toads, using the UV-Vis absorption spectrophotometry technique. Methods: The proposed method was based on the bathochromic shift induced by the reaction of the α-pyrone group of bufadienolides (296 nm) with a 5% (w:v) aqueous solution of sodium hydroxide and detection at 356 nm, after 60 min (time defined based on the evaluation of kinetic assays). Results: The proposed method showed wide linearity (r = 0.9999), low LOD (1.3 × 10-4 µg/mL) and LOQ (3.9 × 10-4 µg/mL), recovery (84%-99%), repeatability (%RSD ≤ 5), reproducibility and robustness (p > 0.05). The total bufadienolide content in PGS extracts from 12 samples of R. diptycha ranged from 478 to 801 mg of EqMB/g of extract, while the R. granulosa sample presented 661 mg of EqMB/g of extract. Conclusion: The new developed method is innovative, simple, fast, accurate, robust, low cost, and can contribute to future research focused on the quantification of total bufadienolides in samples of toad glandular secretions. In addition to serving as a strategic tool in the selection of work matrices, optimizing time, and minimizing costs.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992025000100309&lng=en&tlng=enGlandular secretionBufadienolidesRhinella genusUV-Vis methodQuantification
spellingShingle Elcio Daniel Sousa Barros
Evaldo dos Santos Monção Filho
Mariana Helena Chaves
Cícero Alves Lopes Júnior
Gerardo Magela Vieira Júnior
Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
Journal of Venomous Animals and Toxins including Tropical Diseases
Glandular secretion
Bufadienolides
Rhinella genus
UV-Vis method
Quantification
title Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
title_full Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
title_fullStr Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
title_full_unstemmed Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
title_short Development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
title_sort development and validation of a spectrophotometric method for the quantification of total bufadienolides in samples of toad glandular secretions
topic Glandular secretion
Bufadienolides
Rhinella genus
UV-Vis method
Quantification
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992025000100309&lng=en&tlng=en
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