| Pharmaceutical substances such as 17alpha-ethinylestradiol (EE2), the active component of contraceptive pill, are often not eliminated during wastewater treatment and are not biodegraded in the environment. In this Master's project, analytical techniques for analyzing the low concentration of EE2 during biodegradation were developed and the biodegradability of 17alpha-ethinylestradiol was studied using five bacteria. A preliminary identification of the produced metabolites was also completed.;A low cost solid phase extraction (SPE) procedure was developed and optimized to obtain reproducible analyte recoveries in order to pre-concentrate the EE2 samples prior to HPLC analysis. The limit of analysis (LOA) was significantly lowered from 1.73 ppm to 0.173 ppm upon implementation of the combined SPE/HPLC method. R. erythropolis degraded EE2 up to 47% of its initial concentration in 13 h, while after 65 h, R. equi degraded EE2 up to 39% of its initial concentration. No significant EE2 degradation was observed using Rhodococcus rhodochrous and Rhodococcus zopfii. Preliminary analysis of EE2 degradation products confirmed the presence of phenol among other possible metabolites of high molecular weight. |
