Determination of Catechol Estrogen Adducts by High-Performance Liquid Chromatography: Establishing Biomarkers for the Early Detection of Breast Cancer

摘要

In order to better understand the role of estrogen metabolism as it relates to breast cancer etiology, a new analytical technique that can measure CE and CE-DNA adducts at low endogenous levels is being developed. This new technique is based on HPLC analysis of fluorescent probes specific for CE and CE-DNA adducts. An extraction procedure employing a solvent mixture of chloroform, dimethylforamide, and acetic acid was developed to extract CE-DNA adducts, CE, and MPEM from rat breast tissue. This extraction procedure involves homogenization of the tissue in 20 mL of the solvent mixture, removal of the solvent, followed by HPThC analysis. Reaction of alpha,alpha-dibromomalonamides occurs quickly with catechols, and this malonamide system is being developed to produce fluorescent probes for HPLC analysis. Reaction of N,N'-Bis- anthracen-9- ylmethyl-2,2-dibromo-malonamide with catechol results in ketal formation. The resulting product is highly fluorescent and can be detected at the low femtomole level. The aim of this work is to develop new biomarkers for the early detection of breast cancer.