In vitro investigations of the metabolism of Victoria pure blue BO dye to identify main metabolites for food control in fish

Estelle Dubreil-Cheneau (1) , Luc Sczubelek , Viktoriia Burkina (2) , Vladimir Zlabek (2) , Sidika Sakalli (2) , Galia Zamaratskaia (3) , Dominique Hurtaud-Pessel (1) , Eric Verdon (1)

1 Laboratoire de Fougères - ANSES
2 Faculty of Fisheries and Protection of Waters [University of South Bohemia]
3 SLU - Swedish University of Agricultural Sciences = Sveriges lantbruksuniversitet

Although banned, dyes, such as Victoria pure blue BO (VPBO), are illicitly used in aquaculture to treat or prevent infections due to their therapeutic activities. The present study examined the formation of phase I and phase II metabolites derived from VPBO using trout liver microsomes and S9 proteins. The well-known malachite green (MG) dye was also studied as a positive control and to compare its metabolism with that of VPBO. First, we optimised the incubation conditions for the detection of VPBO and MG metabolites by studying the formation of cytochrome P450 (CYP) substrates. Using the determined conditions (2 h at 20 °C), we incubated VPBO with trout microsomal and S9 fractions induced with β-naphtoflavone, and analysed the supernatant in a LC-LTQ-Orbitrap-HRMS system. The in vitro assays led to the detection of 16 VPBO metabolites from Phase I reactions, arising in particular from reactions with CYP1A. No metabolites were detected from Phase II reactions. The main metabolite detected, deethyl-VPBO, was CID-fragmented to determine its chemical structure, and thus recommend a potential biomarker for the control of VPBO in farmed fish foodstuffs.

Mots clés: Victoria pure blue DO; dye; metabolite; analysis; residues; fish; food safety; LC-LTQ-Orbitrap-HRMS system; chemistry

Mots clés: Bleu Victoria; colorant; metabolite; analyse; résidus; poisson; sécurité des aliments; chimie; spectrométrie de masse Orbitrap

Domaines: Chimie analytique

Estelle Dubreil-Cheneau, Luc Sczubelek, Viktoriia Burkina, Vladimir Zlabek, Sidika Sakalli, et al.. In vitro investigations of the metabolism of Victoria pure blue BO dye to identify main metabolites for food control in fish. Chemosphere, 2020, 238, pp.1-12. ⟨10.1016/j.chemosphere.2019.124538⟩. ⟨anses-02337771⟩