Integrated analytical workflow for chromatographic profiling and metabolite annotation of a cytotoxic Phorbas amaranthus extract

Author(s)

do Amaral, Bruno S. ^{[1, 2]} ; da Silva, Fernanda B. ^[3] ; Leme, Gabriel Mazzi ^[2] ; Schmitz, Leticia S. S. ^[4] ; Jimenez, Paula C. ^[4] ; Campos Martins, Roberto Carlos ^[3] ; Cass, Quezia B. ^[2] ; Valverde, Alessandra L. ^{[2, 5]}

Affiliation

^[1] Inst Fed Educ Ciencia & Tecnol Sao Paulo, Campus Pirituba, BR-05110000 Sao Paulo, SP - Brazil

^[2] Univ Fed Sao Carlos, Dept Quim, Separare, Rodovia Washington Luis, Km 235, BR-13565905 Sao Carlos, SP - Brazil

^[3] Univ Fed Rio de Janeiro, Inst Pesquisas Prod Nat, BR-21941599 Rio De Janeiro, RJ - Brazil

^[4] Univ Fed Sao Paulo, Inst Mar, Dept Ciencias Mar, BR-11070100 Santos, SP - Brazil

^[5] Univ Fed Fluminense, Inst Quim, Lab Prod Nat, BR-24020141 Niteroi, RJ - Brazil

• Document type: Journal article
• Source: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES; v. 1174, JUN 1 2021.
• Web of Science Citations: 1

Abstract

Phorbas is a widely studied genus of marine sponge and produce structurally rich cytotoxic metabolites. Still, only few studies have assessed metabolites present in Brazilian species. To circumvent redundancy, in this work, we applied and herein report the use of a scouting liquid chromatographic system associate to the design of experiment produced by the DryLab (R) software to obtain a fast and efficient chromatographic separation of the active hexane fraction, further enabling untargeted high-resolution mass spectrometry (HRMS) data. To this end, a crude hydroalcoholic extract of the sponge Phorbas amaranthus collected in Brazilian coast was prepared and partitioned. The cytotoxicity of the crude extract and the fractions was evaluated using tumor cell culture models. Fragmentation pathways assembled from HRMS data allowed the annotation of 18 known Phorbas metabolites, while 17 metabolites were inferred based on Global Natural Product Social Molecular Networking (GNPS), matching with a further 29 metabolites annotated through molecular subnetwork. The workflow employed demonstrates that chromatographic method development can be accelerated by the use of automated scouting systems and DryLab (R), which is useful for profiling natural product libraries, as well as data curation by molecular clusters and should be incorporated to the tools of natural product chemists. (AU)

• FAPESP's process: 16/24957-0 - Models of binders rapid screening in extracts of natural products for bioconjugation tests and LC-HRMS
• Grantee: Gabriel Mazzi Leme
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50249-8 - Green chemistry: sustainable synthetic methods employing benign solvents, safer reagents, and bio-renewable feedstock
• Grantee: Arlene Gonçalves Corrêa
• Support Opportunities: Research Grants - Research Centers in Engineering Program