Structure and Oxidation of Pyrrole Adducts Formed between Aflatoxin B 2a and Biological Amines

• Published in: Chemical research in toxicology Vol. 30; no. 6; pp. 1275 - 1285
• Main Authors: Rushing, Blake R, Selim, Mustafa I
• Format: Journal Article
• Language: English
• Published: United States 19.06.2017
• Subjects: Aflatoxins - chemistry; Aflatoxins - metabolism; Amines - chemistry; Amines - metabolism; Binding Sites; Chromatography, High Pressure Liquid; DNA Adducts - chemical synthesis; DNA Adducts - chemistry; DNA Adducts - metabolism; Kinetics; Mass Spectrometry; Molecular Structure; Oxidation-Reduction; Pyrroles - chemical synthesis; Pyrroles - chemistry; Pyrroles - metabolism
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/acs.chemrestox.7b00002

Aflatoxin B has been shown to bind to proteins through a dialdehyde intermediate under physiological conditions. The proposed structure of this adduct has been published showing a Schiff base interaction, but adequate verification using structural elucidation instrumental techniques has not been performed. In this work, we synthesized the aflatoxin B amino acid adduct under alkaline conditions, and the formation of a new product was determined using high performance liquid chromatography-time-of-flight mass spectrometry. The resulting accurate mass was used to generate a novel proposed chemical structure of the adduct in which the dialdehyde forms a pyrrole ring with primary amines rather than the previously proposed Schiff base interaction. The pyrrole structure was confirmed using H, C, correlation spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond correlation NMR and tandem mass spectrometry. Reaction kinetics show that the reaction is overall second order and that the rate increases as pH increases. Additionally, this study shows for the first time that aflatoxin B dialdehyde forms adducts with phosphatidylethanolamines and does so through pyrrole ring formation, which makes it the first aflatoxin-lipid adduct to be structurally identified. Furthermore, oxidation of the pyrrole adduct produced a product that was 16 m/z heavier. When the aflatoxin B -lysine (ε) adduct was oxidized, it gave a product with an accurate mass, mass fragmentation pattern, and H NMR spectrum that match aflatoxin B -lysine, which suggest the transformation of the pyrrole ring to a pyrrolin-2-one ring. These data give new insight into the fate and chemical properties of biological adducts formed from aflatoxin B as well as possible interferences with known aflatoxin B exposure biomarkers.