Girly Grass Dana Devin-Clarke. Sally Brown, Michael Muramoto*, and Michael Doubrava*

1. Girly Grass Dana Devin-Clarke. Sally Brown, Michael Muramoto*, and Michael Doubrava* University of Washington and *King County Environmental Laboratory Methods of Extraction Study Design Background • The leachate is collected and transferred to King County for extraction and analysis. • Ibuprofen and Triclosan: • Liquid/ Liquid extraction • Concentration on Turbovap • Analysis on GC/MS • The Estrogens: • ZHE Filtration • Vacuum Degassing • C-18 SPE loading • NH2 Cleanup • Silica Cleanup • Alumina Cleanup • Derivatizing the Extracts • Analysis on the GC/MS • * The soil extraction and analysis methods are currently being researched. Many pharmaceuticals and personal care products or PCPPs enter and leave wastewater treatment plants (WWTP) unaltered or incompletely removed and are subsequently released into the environment through wastewater outflows. Fish exposed to PPCPs in wastewater outflows have been shown to exhibit intersex characteristics. Instead of releasing effluent into streams, wastewater can be further treated and beneficially reused for irrigation of golf courses, farmland or forestry plantations. However, the persistence of PCPPs once applied to a soil system, such as a golf course, is yet to be researched. Subsequently, this study was created to assess the benefits and potential for exposure to PCPPs by using reclaimed water produced by Renton Treatment Plant to grow turf grass from the Foster Links Golf Course in Tukwila. The PCPPs that will be monitored for mobility as well as presence in the soils include: ibuprofen, triclosan and the natural and synthetic estrogens: estriol, 17β-estradiol and ethinylestradiol 6’’ X 6’’ squares of Fairway and Tee turf grass were transplanted from the Foster Links Golf Course to pots in the CUH greenhouse. • These grasses were then assigned one of five treatments including: • T1 =Control (Tap water + 100%Fert) • T2 = Reclaimed water (RW) only • T3 = RW + 50% Fertilizer • T4 = RW + 100% Fertilizer • T5 = Biosolids (Tap water only) • Each pot is watered with 500 mL of water and received fertilizer amendments of 50% and 100% actual course application rates Compounds of Interest Preliminary Conclusions Ibuprofen: an anti-inflammatory drug - Concentrations as low as 1 μg/L affect the heat shock response in trout fry -USGS stream surveys in thirty states revealed a detected stream concentrations: Median = 0.20 µg/L Max =1.0 µg/L Triclosan: anti-microbial compound in household products - Lethal conc. (LC50 rainbow trout = 0.35 μg/L) - The degradation byproducts: methyl-TCS, which bioaccumulates in fish and dioxin, a known carcinogen -Stream conc: Median(0.80 g/L) and Max. (40 g/L) Natural and Synthetic estrogens: -Fish exposed to concentrations as low as 1 ng/L begin to develop intersex characteristics -Stream conc.: a median and maximum 0.073 and 0.831 μg/L for ethinylestradiol (EE2) 0.009 and 0.2 μg/L for estradiol (E2) 0.027 and 0.112 μg/L for estrone (E1) • The source water and leachate is collected every other week and tested at the King County Environmental Laboratory. • These are tested to determine: • the total concentration of compounds being loaded into the system • the amount of compound that may potentially pass through the system in the leachate • The soil and plant samples will be collected and tested at the end of the study to determine plant uptake as well as potential accumulation or persistence of compounds in a sediment system. • * ND = Not Detected, < MDL = less than methods detection limit • ** Note these are the preliminary numbers from the first extraction. More data is required to fully characterize the fate and concentration of compounds within this system • Initial results indicate: • The estrogens, added with the reclaimed water, do not pass through the soil system.