Optimum Separation and Compound Class Separation of the Metabolites of Benzo[a]pyrene-DNA Adducts with Reversed-Phase Liquid Chromatography

Abstract

Abstract Mobile phases were optimized for the reversed-phase liquid chromatographic separation of a complex mixtures of fourteen metabolites of benzo[a]pyrene (B[a]P). The metabolites constituted groups of isomers that were difficult to separate. The groups of isomers were tetrols, dihydrodiols, diones, and monohydroxyl-benzo[a]pyrenes. The window diagram optimization approach was used to initially optimize the binary mobile phases. Based on the data obtained from the optimum binary mobile phases, a solubility parameter optimization method was employed to obtain an optimum ternary mobile phase. Both the binary and ternary mobile phases were very effective in separating the metabolites. However, complete baseline resolution of the complex mixture of the metabolites was not achieved under the conditions investigated. Nevertheless, it was possible to obtain a separation of all fourteen of the metabolites with some overlap of the chromatographic bands. Also, compound class separation was obtained with the cla...