Separation of substrates and closely related glucuronide metabolites using various chromatographic modes.

摘要: The aim of this study was to assess the retention and selectivity a cocktail 10 substrates uridine diphosphate glucuronosyltransferase enzymes (UGTs) their respective glucuronides using four chromatographic approaches. For purpose, seven different stationary phases were employed in reversed phase liquid chromatography (RPLC), two hydrophilic interaction (HILIC), one aqueous normal (ANPC) subcritical fluid (SFC). Highly orthogonal separations achieved with these modes. Hydrophobic interactions mainly governed polar RPLC despite use chemical bonding, involving additional possible interactions. In ANPC, atypical poor peak shapes observed selected compounds. HILIC SFC conditions, metabolites more retained than because polarity increase related glucuronic acid moiety. latter, very high proportion organic solvent (up 80%) required elute that often displayed shapes. Finally, nine systems compared for separation isomeric diastereoisomeric used mode selective towards positional isomers morphine since they possess distinct lipophilicity. columns found be promising critical diastereoisomers pair, namely epitestosterone-glucuronide testosterone-glucuronide.