New derivatization method using ferrocene reagents for the determination of steroid glucuronides by high-performance liquid chromatography with electrochemical detection.
New derivatization methods using ferrocene reagents have been developed for the determination of steroid glucuronides by high-performance liquid chromatography with electrochemical detection. Condensation of glucuronides with 2-ferrocenylethylamine is effected in the presence of water-soluble carbodiimide and 1-hydroxybenzotriazole. The resulting amides show the satisfactory sensitivity at +0.45 V versus Ag/AgCl reference electrode, with a detection limit of 0.5 pmole. The present method is found to be applicable to the estimation of estrogen glucuronyltransferase activities and estrogen glucuronides in pregnancy urine.