Metabolic study of cafestol using in silico approach, zebrafish water tank experiments and liquid chromatography high-resolution mass spectrometry analyses

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2021-12, Vol.1186, p.123028-123028, Article 123028
• Main Authors: Andriolo, Cyrus Veiga, Novaes, Fábio Junior M., Pereira, Henrique Marcelo Gualberto, Sardela, Vinícius Figueiredo, Rezende, Claudia Moraes
• Format: Article
• Language: English
• Subjects: Animals; Biotransformation; Chromatography, High Pressure Liquid; Chromatography, Liquid; Coffee - chemistry; Coffee - metabolism; Coffee beverage; Computer Simulation; Diterpenes - chemistry; Diterpenes - metabolism; In silico analysis; In vivo analysis; LC-MS/HRMS; Mass Spectrometry; Metabolic pathways; Metabolic study; Models, Animal; Molecular Structure; Zebrafish - metabolism
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2021.123028

[Display omitted] •In silico approach proposed 25 structures as cafestol metabolites.•Zebrafish water tank model produced 5 structures of predicted cafestol metabolites.•MS2 fragmentation study of cafestol supports the identification of metabolites structure.•Cafestol consumption and metabolites formation were monitored by LC-MS during 9 h. Coffee is one of the most consumed beverages worldwide. Cafestol is an endogenous coffee diterpene present in raw coffee beans and also found in hot beverages, with several biological activities. However, there is still little information on this molecule after ingestion of coffee infusion. Zebrafish (Danio rerio) is a promising in vivo model for metabolic studies due to the annotation of mammalian orthologs to encode enzymes related to drug metabolism. Experiments using Zebrafish Water Tank (ZWT) model produce more significant number of metabolites for molecular investigation in a cleaner matrix than other classical models, such as purified hepatocytes. This work aimed to investigate the biotransformation of cafestol by the ZWT model using ultra-performance liquid chromatography coupled to hybrid quadrupole-orbitrap high-resolution mass spectrometry equipped with electrospray ionization (UPLC-HRMS) supported by in silico approach using SMARTCyp, Way2Drug and XenoSite Softwares. Twenty-five metabolites of cafestol were proposed by in silico analysis, in which 5 phase I metabolites were confirmed in the ZWT by UPLC and MS/HRMS investigation: 6-hydroxy-cafestol, 6,12-dihydroxy-cafestol, 2-oxo-cafestol, 6-oxo-cafestol and one isomer whose position in the carboxyl group was not determined. These metabolites were observed during 9 h of the experiment, whose contents were associated with the behavioral responses of the fish.