Characterisation of B(a)P metabolites formed in an ex vivo pig skin model using three complementary analytical methods.
An original method was developed to separate, identify and quantify the different benzo(a)pyrene (B(a)P) metabolites formed through oxidative and conjugative pathways. All B(a)P metabolites were separated by an improved high-performance liquid chromatography method, then detected and quantified relatively by online radioactivity detection. At the same time, metabolite structures were characterised by tandem mass spectrometry using two complementary ionisation modes: electrospray ionisation in the negative mode and atmospheric pressure chemical ionisation in the positive mode. This method was successfully applied to the analysis of B(a)P metabolites, produced by incubation of B(a)P with the ex vivo pig ear skin model. These include glucuronic acid and sulphate conjugates of B(a)P-OHs and B(a)P-diols, as well as direct phase I metabolites: B(a)P-tetrol, B(a)P-diones, B(a)P-catechols, B(a)P-diols and B(a)P-OHs.
Jacques
Carine
C
INRA, UMR 1089 Xénobiotiques INRA-ENVT, 180 chemin de Tournefeuille, BP 93173, Toulouse, 31027, France.
Jamin
Emilien L
EL
Perdu
Elisabeth
E
Duplan
Hélène
H
Mavon
Alain
A
Zalko
Daniel
D
Debrauwer
Laurent
L
D000818:Animals; D001564:Benzo(a)pyrene / Q000031:analogs & derivatives / Q000032:analysis* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004431:Ear, External; D005260:Female; D020723:Glucuronic Acid / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D023421:Models, Animal; D015394:Molecular Structure; D010084:Oxidation-Reduction; D012867:Skin / Q000378:metabolism*; D013237:Stereoisomerism; D013431:Sulfates / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D013552:Swine; D053719:Tandem Mass Spectrometry
D013431:Sulfates; D001564:Benzo(a)pyrene; D020723:Glucuronic Acid