B-Estradiol is an estrogen steroid hormone. It is the major estrogen secreted by the premenopausal ovary. It is synthesized from testosterone primarily in the ovarian granulosa cells and placenta. B-estradiol is used as a medication, for instance in menopausal hormone therapy. Primesep 200, a reverse phase column, contains embedded acidic ionizable groups and can retain B-Estradiol. The method is UV compatible and can be used as a general approach for analyzing similar compounds.