M. Tech (Department of Chemistry, Faculty of Applied and Computer Sciences) Vaal University of Technology. / A simple and rapid method for determination of estrone (E1) and β-estradiol (E2) was developed and validated using high performance liquid chromatography (HPLC). The solutions of standards and sample were prepared with distilled water. HPLC separation was performed in isocratic method 50/50 (water/methanol) using 4.6 mm x 250 mm id film thickness 5 µm) XDB-C18 capillary column, detector DAD, UV on 254 nm, temperature 20 ºC with flow rate of 2 mL/min, sample volume 20 µL and run time of 10 min. Calibration curves were linear between concentration range 1.0 - 15.0 ppm. The method was validated for limit of detection and quantification, linearity, precision, trueness and specificity. Also the method was applied to directly and easily to the analysis of the E1 and E2. Adsorption experiments were carried out in batch mode using multistirrer in a series of Erlenmeyer flasks of 50 ml capacity covered to prevent contamination having concentration ranges of E1 and E2 from 1 to 10 mg/L with adsorbent dose range 0.01 to 1 g at pH range 1 to 10 and temperature range 15°C to 35°C, placed on multistirrer. The results of the batch studies showed that simultaneous adsorption shows the maximum percent (91%) removal of E1 and (86 %) E2 at optimum temperature 25 °C of adsorbent dose 0.1 g, and pH 7. The mechanism, isotherms and kinetics of removal of two endocrine disrupting chemicals, estrone (E1) and β-estradiol (E2) by activated carbon adsorption were investigated in an agitated non-flow batch adsorption studies. Mathematical models were used to describe the adsorption phenomenon with the kinetic and thermodynamic parameters evaluated using the adsorption equilibrium data at varying temperatures. Higher adsorption rates were achieved at acidic to neutral pH ranges, with the sorption kinetic data showing a good fit to the pseudo second order rate equation and the Langmuir adsorption isotherm model for both E1 and E2. The Gibbs free energy were –16.68 kJ/mol and –17.34 kJ/mol for E1 and E2 respectively. The values of enthalpy for both E1 (84.50 kJ/mol) and E2 (90 kJ/mol) indicated a chemical nature of the sorption process. Both the isotherm and thermodynamic data obtained all supported the mechanism of adsorption of E1 and E2 to be mainly chemisorption’s supported by some physical attractions.

The disposal of waste from agricultural activities has been recognised as a source of environmental contamination by endocrine disrupting chemicals (EDCs). The New Zealand dairy industry produces a large volume of dairy farm effluent, which contains EDCs in the form of estrogens. Most of this dairy farm effluent is applied onto the land for disposal. Groundwater and soil contamination by estrogens following waste application on the land have been reported overseas, but our understanding of the processes and factors governing the fate of estrogens in the soil is poor. Therefore the main goal of the present study was to better understand the fate and transport of estrogens, in particular 17β-estradiol (E2) and estrone (E1) in soil. In order to quantify E1 and E2 in drainage water and soil samples, chemical analysis by gas-chromatography mass-spectrometry (GC-MS) was carried out. This included sample extraction, sample clean-up through silica gel and gel permeation chromatography, and sample extract derivatisation prior to analysis. In order to develop a reliable method to extract estrogens from soil, research was conducted to optimise E1 and E2 extraction conditions by adjusting the number of sonication and shaking events, as well as the volume and type of solvent. Among five solvents and solvent mixtures tested, the best recovery on spiked and aged soil was obtained using an isopropanol/water (1:1) mix. A microcosm experiment was carried out to determine the dissipation rates of E2 and E1, at 8°C and at field capacity, in the Templeton soil sampled at two different depths (5-10 cm and 30-35 cm). The dissipation rates decreased with time and half-life values of 0.6-0.8 d for E1 and 0.3-0.4 d for E2 were found for the two depths studied. A field transport experiment was also carried out in winter, over three months, by applying dairy farm effluent spiked with estrogens onto undisturbed Templeton soil lysimeters (50 cm in diameter and 70 cm deep). The hormones were applied in dairy farm effluent at 120 mg m⁻² for E2 and 137 mg m⁻² for E1. The results of the transport experiment showed that in the presence of preferential/macropore flow pathways 0.3-0.7% of E2 and 8-13% of E1 was recovered in the leachate at the bottom of the lysimeters after 3 months, and 1-7% of the recovered E2 and 3-54% of the recovered E1 was leached within 2 days of application. These results suggest that leaching of estrogens via preferential/macropore flow pathways is the greatest concern for groundwater contamination. In the absence of preferential/macropore flow pathways, a significant amount (> 99.94%) of both hormones dissipated in the top 70 cm of soil, due to sorption and rapid biodegradation. Surprisingly, in all cases, estrogen breakthrough occurred before that of an inert tracer (bromide). This could not be explained by the advection-dispersion transport of estrogens, nor by their presence as antecedent concentrations in the soil. It was therefore suggested that colloidal enhanced transport of estrogens was responsible for the earlier breakthrough of estrogens and caused the leaching of a fraction of the applied estrogens to a soil depth of 70 cm. A two-phase model, adapted from a state-space mixing cell model, was built to describe the observed estrogen transport processes under transient flow. The model takes into account 3 transport processes namely, advection-dispersion, preferential/macropore flow and colloidal enhanced transport. This model was able to successfully describe the estrogen transport observed from the lysimeters.