1. A Study of Endocrine Disrupting Compounds in Wastewater Arthur Ho Spacegrant Mentor: Dr. Bob Arnold Chemical and Environmental Engineering University of Arizona

2. Effects in Animals Kidd et al., 2007

3. USGS Surface Water Reconnaissance (Kolpin et al., 2002) 139 Sites 95 Compounds Comparison of 4 Compounds: Compound (Use and Effect)C MAX (µg/L)Days to Med Dose Caffeine (stimulant) 5.75.7 x 10 3 (Excedrin Migraine) Ibuprofen(anti-inflammatory) 1.010 5 (Motrin) 17α-ethinyl estradiol (oral contraceptive) 0.27 37 (Alesse-21) 17β-estradiol (vertebrate hormone) 0.093 (Phoenix)5.3 x 10 3 (Activella) 4-nonylphenol (surfactant metabolite) 40 (Tucson)4.6 x 10 -4 as estrogenic E 2, EE 2

4. Examples of Estrogenic Compounds ChemicalStructure Molecular Weight Log K ow Relative Estrogenic Activity (YES bioassay) 17  -estradiol (E 2 ) 272.39 3.94 1.0 17  -ethinyl estradiol (EE 2 ) 296.404.151.32 Estrone (E 1 ) 270.373.430.20 Estriol (E 3 ) 288.392.810.01 Nonylphenol (NP) 220.35 4.48 2×10 -4 Octylphenol (OP) 206.33 4.123×10 -4 Bisphenol A (BPA) 228.293.408×10 -5

5. Estrogenic Activity at JWPCP Effluent: 0.12 mols EE2 equivalents/day Dewatered Cake: 0.26 mols EE 2 equivalents /day Influent: 1.66 mols EE 2 equivalents/day Removal of estrogenic activity through the whole treatment plant = [1.656 – (0.1178 + 0.26)] / 1.656 = 77%

7. Cytoplasm ERREE = Estrogen = Receptor Yeast Estrogen/ Androgen Screen R hER/hAR  -gal CPRG yellow CPR red R E lac-z

8. Yeast Estrogen Screen (YES) Assay

9. Cytoplasm ERREE = Estrogen = Receptor Yeast Estrogen/ Androgen Screen R hER/hAR  -gal CPRG yellow CPR red R E lac-z

10. Agonism Agonism Anti-estrogens: Mechanism of Action Antagonism Antagonism RE R A R E R A  -gal No protein production

11. Tamoxifen is an Anti-estrogen Known anti-estrogens: (hydroxy) tamoxifen, chlorinated hormones, Known anti-androgens: 4-,4’-DDE, flutamide

12. Methods Obtaining Wastewater samples Obtaining Wastewater samples Extraction of Organics Extraction of Organics Preparation of organics Preparation of organics Yeast Estrogen Assay Yeast Estrogen Assay

13. RESULTS RAW SLUDGE SOLID EXTRACTS ARE HIGHLY ESTROGENIC RAW SLUDGE LIQUID EXTRACTS ARE ANTI- ESTROGENIC

14. RESULTS DIGESTED SLUDGE SOLIDS LOST ESTROGENIC ACTIVITY DIGESTED SLUDGE LIQUIDS HIGHLY ANTI- ESTROGENIC

15. Acknowledgements Dave Newman & Sondra Teske Dave Newman & Sondra Teske Susan Brew Susan Brew Matt Giardiana and AzSGC Staff Matt Giardiana and AzSGC Staff Bingfeng Dong Bingfeng Dong