Identification of androstenedione in a river containing paper mill effluent

Effluent from a paper mill discharging into the Fenholloway River, Taylor C ounty, Florida, USA, contains chemicals that masculinize females of the res ident population of eastern mosquitofish (Gambusia holbrooki), as evidenced in females by elongated anal fins, which is normally a male-specific trait . To identify androgenic components in the effluent, water collected from t he Fenholloway River and a control tributary was fractionated using solid-p hase extraction and reverse-phase high-performance-liquid chromatography. T wo Fenholloway River fractions induced androgen receptor-dependent transcri ptional activity in transient transfection cell culture assays. Of these, a ndrostenedione was confirmed by liquid chromatography-mass spectrometry wit h multiple reaction monitoring.