The Liquid Chromatographic Separation of Metabolites of Benzo[a]pyrene with γ-Cyclodextrin as a Mobile Phase Additive

Abstract
The effects of γ-cyclodextrin as a mobile phase additive on the retention characteristics of fourteen metabolites of benzo[a]pyrene were investigated with reversed-phase liquid chromatography. It was possible to separate dihydrodiols, diones, and monohydroxyl-benzo[a]pyrenes as individual classes of metabolites. Also, with γ-cyclodextrin in the mobile phase, monohydroxyl-benzo[a]pyrene isomers that were very difficult to separate were readily separated. For example, 6-hydroxyl-benzo- [a]pyrene was separated from 12-hydroxyl-benzo[a]pyrene and 9-hydroxyl-benzo[a]pyrene was separated from 2-hydroxyl-benzo- [a]pyrene. The four tetrols were readily separated with shorter retention times using 4.0 mM of γ-cyclodextrin in a methanol-water (55:45) mobile phase. Overall, the results showed that γ-cyclodextrin was more effective in separating the metabolites than was β-cyclodextrin.

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