In the first part of this thesis the principles of capillary electrophoresis (CE) are presented from the aspects of steroid and sterol analysis focusing mainly on two separation techniques: micellar electrokinetic chromatography (MEKC) and capillary electrochromatography (CEC). Analytes are delineated in steroids, corticosteroids, phytosterols, and cholesterol. Conventional chromatographic methods: gas chromatography (GC) and high-pressure liquid chromatography (HPLC) are somewhat challenging for steroid and sterol analysis, since direct analysis of steroids/sterols and their conjugates is rarely feasible. Hence, alternative separation and analysis methods need to be approached. MEKC and CEC have provided intriguing new opportunities for steroids and sterols, respectively.
The experimental part covers the study on finding out the steroid composition and concentrations of wastewater samples collected from wastewater treatment plants (WWTP) around Finland. In addition, the efficiencies of the WWTPs were resolved.
There were two types of wastewater samples: influent and effluent. Influent is the unclean water and effluent is the cleaned water that has passed through the process steps. The sample pretreatment includes filtering (glass fiber and membrane filters), solid phase extraction (SPE) (with C18 (Strata-X) and quaternary amine (N+) sorbents), and liquid-liquid extraction (LLE) (with diethyl ether). SPE was effective in purifying and concentrating the water samples, with a concentration factor of 20,000.
The analysis was performed with partial filling-micellar electrokinetic chromatography, utilizing UV detection. It was found that the method was suitable for both qualitative and quantitative analysis of endogenous steroids and their corresponding metabolites. Androstenedione, testosterone glucuronide, and progesterone were found from the samples. Some notable results are that biological treatment most likely increases the amount of androstenedione, whereas enzymatic processes remove efficiently progesterone. Overall, the lowest steroid concentrations were obtained from the samples of Espoo, Pori, and Uusikaupunki. On the contrary, highest concentrations were in Kajaani, Mikkeli, and Porvoo.
