Hydrophilic interaction chromatography coupled to high resolution mass spectrometry (HILIC-LC-HRMS)
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Journal of Chromatography, A
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Although proteins in snake venoms have been extensively studied and characterized, low-mass molecules remain
relatively unexplored, mainly due to their low abundance, secondary role in envenomation, and some analytical
technique limitations. However, these small molecules can provide new important data related to venom toxins’
molecular structure, functions, and evolutionary relationships. This research aimed to characterize molecules
below 10 kDa in the venoms of snakes from the Viperidae families (Bothrops, Agkistrodon, and Bitis) and compare
two chromatographic approaches: reverse-phase chromatography (RP), a classic technique, and hydrophilic
interaction liquid chromatography (HILIC), an alternative technique, both coupled with high-resolution mass
spectrometry (HRMS). The results showed that the separation of the HILIC column provided a more efficient
evenly distributed ion profile than RP, contributing to a 25.6% increase in the sequences identified. Homologous
sequences for Bradykinin-potentiating peptides (BPPs) and fragments of major venom proteins, possibly cryptids,
were found. In addition, BPP 13a, peptides rich in histidine and glycine (pHpG), and spacer sequences were
identified in all snakes analyzed, especially with HILIC separation, suggesting that these sequences may be
conserved within Viperidae. These findings indicate that the use of the HILIC column, compared to RP, is a
promising approach for characterizing peptides in snake venom obtained by the ultrafiltration process. It contributes
to the study of these still poorly understood molecules and is also a good option for studying other
complex protein/peptide mixtures.
Como referenciar
VALLA, MONICA V.; LEBRUN, IVO; PUDENZI, MARCOS A.; OLIVEIRA, LAUDICEIA A.; ALMEIDA, HELOISA F.; SANTOS, NATHALIA G.; RODRIGUES, MARIANA S.; SPENCER, PATRICK J.; ROCHA, MARISA M.; PIMENTA, DANIEL C.; COELHO, GUILHERME R. Hydrophilic interaction chromatography coupled to high resolution mass spectrometry (HILIC-LC-HRMS): an approach to study natural peptides in Viperidae snake venom. Journal of Chromatography, A, v. 1743, p. 1-10, 2025. DOI: 10.1016/j.chroma.2025.465715. Disponível em: https://repositorio.ipen.br/handle/123456789/49850. Acesso em: 04 May 2026.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.