1. The University of Arizona
Assessment of Chemical and Biological Techniques to Evaluate the Quality of Water from Pilot Scale Direct Potable Reuse Treatment Facilities
Kevin Daniels, Christiane Hoppe-Jones, Shawn Beitel, Israel Lopez, Minkyu Park, Erica Clevenger, Shane Snyder
The University of Arizona

2. The Status of Water
783 million people do not have access to clean drinking water
6-8 million people die per year from a water related disease
By the year 2025, 1.8 billion people will experience absolute water scarcity and 66% of the world population experiencing "stress conditions
Reliable sources of fresh water are becoming increasingly limited in the southwestern United States and globally. Growing populations and often unpredictable climatic events have strained already fragile water systems, adding urgency to securing safe and sustainable water sources (Grant et al., 2012). Potential restrictions on potable water withdrawals due to drought, coupled with rising energy costs, increase the viability of alternative water sources worldwide.
Fresh water is one of the most limited and important resources needed for all human life. Although the earth has approximately 3% drinking water, only 0.5% is currently unfrozen and available for consumption. Today, 783 million people do not have access to clean drinking water and as a consequence, 6-8 million people die per year from a water related disease. With climate change on the rise, weather patterns have altered the and water availability in many parts of the world. Furthermore, it is projected that by the year 2025, 1.8 billion people; 22% of the projected population (UN), will experience absolute water scarcity and 66% of the world population experiencing "stress conditions". As a result, alternate sources of water are being investigated for their practicality and effectiveness they could prove to be. Such options are: Desalination, Direct Potable Ruse (DPR), and Indirect Potable Reuse (IPR)

3. Alternative sources Most commonly applied: Desalination Water Reuse
Conservation, desalination, and water reuse are the primary means to augment dwindling freshwater supplies.2 There aren’t any other practices at the large scale.
Mayes, A. M., Shannon, M. A., Georgiadis, J. G., Elimelech, M., Mariñas, B. J., & Bohn, P. W. (2008). Science and technology for water purification in the coming decades. Nature, 452(7185),
Shannon et al., 2008
WateReuse, 2015

4. Conventional Wastewater Treatment
Composed of 3 parts, primary, secondary, and tertiary.
Primary consists of screens, grates, and sedimentation
Secondary treatment normally consists of a biological treatment
Lastly, tertiary treatment consist of disinfection
Many compounds remain
Monitoring does not exist

5. Source Contaminants
Approximately 15,000 new chemicals discovered daily
Most substances have propensity to enter WWTPs
Infeasible to evaluate each substance individually
Current paradigm does not consider mixture effects
Goal: The purpose of our research is to provide insight about methods to characterize water quality from source, drinking, and reuse waters at multiple different locations throughout the United States that represent a diverse geography and source organic quality. Methods discussed include the use of bulk organics parameters (i.e. EEM/TOC), Non-target Analysis (NTA), size exclusion chromatography (SEC)-TOC, and targeted analysis of Contaminants of Emerging Concern (CECs) using a Direct Water Injection (DWI) and Online-Solid Phase Extraction (O-SPE) method. In addition to the chemical analysis, a variety of bioassays were applied to evaluate the effectiveness of the advanced treatment processes in removing specific classes of compounds without creating more harmful disinfection byproducts. Data indicates that both RO and UV are effective at reducing estrogen, glucocorticoid, and P53 activity below limits of detection. TOC analysis exhibited 94% removal and fluorescence demonstrated 98.9% removal across the advanced treatment processes evaluated (MF to RO to UV). Each method performed can be useful tools in demonstrating treatment effectiveness.

6. Purpose
The purpose of this research is to provide insight about methods to characterize and treat secondary wastewater effluent for direct potable reuse applications
Goal
1. Determine a monitoring framework
2. Identify the most efficient treatment train
Goal: The purpose of our research is to provide insight about methods to characterize water quality from source, drinking, and reuse waters at multiple different locations throughout the United States that represent a diverse geography and source organic quality. Methods discussed include the use of bulk organics parameters (i.e. EEM/TOC), Non-target Analysis (NTA), size exclusion chromatography (SEC)-TOC, and targeted analysis of Contaminants of Emerging Concern (CECs) using a Direct Water Injection (DWI) and Online-Solid Phase Extraction (O-SPE) method. In addition to the chemical analysis, a variety of bioassays were applied to evaluate the effectiveness of the advanced treatment processes in removing specific classes of compounds without creating more harmful disinfection byproducts. Data indicates that both RO and UV are effective at reducing estrogen, glucocorticoid, and P53 activity below limits of detection. TOC analysis exhibited 94% removal and fluorescence demonstrated 98.9% removal across the advanced treatment processes evaluated (MF to RO to UV). Each method performed can be useful tools in demonstrating treatment effectiveness.

7. Analytical Methods Bulk Organic Parameters Targeted Analysis Bioassays
Total Organic Carbon
Organic Carbon Distribution
UV absorbance
Total Fluorescence
Excitation Emission Matrix
Targeted Analysis
Bioassays
Non-Targeted Analysis
Goal: The purpose of our research is to provide insight about methods to characterize water quality from source, drinking, and reuse waters at multiple different locations throughout the United States that represent a diverse geography and source organic quality. Methods discussed include the use of bulk organics parameters (i.e. EEM/TOC), Non-target Analysis (NTA), size exclusion chromatography (SEC)-TOC, and targeted analysis of Contaminants of Emerging Concern (CECs) using a Direct Water Injection (DWI) and Online-Solid Phase Extraction (O-SPE) method. In addition to the chemical analysis, a variety of bioassays were applied to evaluate the effectiveness of the advanced treatment processes in removing specific classes of compounds without creating more harmful disinfection byproducts. Data indicates that both RO and UV are effective at reducing estrogen, glucocorticoid, and P53 activity below limits of detection. TOC analysis exhibited 94% removal and fluorescence demonstrated 98.9% removal across the advanced treatment processes evaluated (MF to RO to UV). Each method performed can be useful tools in demonstrating treatment effectiveness.

8. Bulk Organic Parameters 1
DOC
Organic carbon remaining in a sample after filtering the sample using a 0.45 µm filter
Used to measure the DOC in water samples from 4 µg/L to 30,000 mg/L
Size exclusion chromatography (SEC) hyphenated with organic carbon detector (OCD)
Is employed to identify the size distribution of natural organic matter (NOM)
Shimadzu TOC Analyzer

9. Bulk Organic Parameters 2
UV and Total Fluorescence
UVA254 is an indicator of aromatic organic constituents
Many organic and anthropogenic compounds are fluorescent
Excitation Emission Matrix (EEM)
Used to characterize dissolved organic matter (DOM), natural organic matter (NOM) and effluent organic matter (EfOM) from wastewater treatment plants and natural waters
Horiba Aqualog

10. Targeted Analysis Used to identify the targeted CECs, including:
Personal Care Products - Household/Commercial Chemicals
Pharmaceuticals - Perfluorinated Compounds
Pesticides - X-ray Contrast Media
Two methods are applied,
Direct water injection – applying sample directly to the separation column
Online Solid Phase Extraction – sample gets extracted by a solid phase before separation
Collected Sample
Filter Sample
Online SPE
LC Column
QQQ
DWI
OSPE 10

11. In-Vitro Bioassays
Used to identify the overall toxicity of known and unknown contaminants in a mixture with the use of a biological system
5 Bioassays tested:
Estrogen Receptor (ER)
Glucocorticoid Receptor (GR)
Aryl hydrocarbon Receptor (AhR)
p53 pathway
HepG2 (Cytotoxicity)
Jia et al., 2016

12. Non-Targeted Analysis (NTA)
Provide information regarding the formation of transformation products and occurrence/fate of unknown contaminants
Principle Component Analysis (PCA), Heat Maps and Cluster Analysis will be used to reveal compounds of interest (that are removed or recalcitrant or transformation products)
Combining the results with water treatment variables and toxicity analysis will provide information on the compounds or group of compounds that correlate statistically to the biological observations

13. Treatments Investigated
Ozonation
Biologically Activated Carbon
Granular Activated Carbon
Microfiltration
Reverse Osmosis
UV based technologies
Ozone – oxidizes
BAC utilizes metabolzism
GAC – adsorption
Microfiltration – size exclusion
RO-
UV-

14. Sites Site 1 Site 2 Site 3 Site 4 Site 5
Sampling campaign is from oct 2015 to jan 2018
Full data set is not complete as we have additional sampling campaign underway
I will discuss a brief overview of site one due to time constraints

15. TOC Results: Site 1
Make bar graphs and show two compounds from each class
Used the word removed, not resistant

16. SEC-OCD Results: Site 1
Make bar graphs and show two compounds from each class
Used the word removed, not resistant
Huber, S.A et al., 2011

17. UVA/TF Results: Site 1

18. EEM Results: Site 1

19. Targeted Analysis Results: Site 1
Most Prevalent CECs
Concentration (ng/L)

20. Targeted Analysis Results: Site 1
All other CECs
Concentration (ng/L)

21. Bioassay Results: Site 1

22. Bioassay Results: Site 1

23. Bioassay Results: Site 1
Cytotoxicity (HepG2):
All samples from the sampling campaigns tested did not show cytotoxicity at a final concentration of 12.5x the original sample
p53 Bioassay:
All samples from the sampling campaigns tested did not show any interaction within this pathway (were <LOQ)
Aryl hydrocarbon Receptor Bioassay:
Still underway

24. NTA Results: Site 1

25. NTA Results: Site 1

26. Summary
For site 1, MF to RO to UV was more effective at removing TOC, aromatic double bonds, etc. based on initial sampling campaigns
All monitoring techniques showed similar trends with decreased responses throughout the treatment train
Bulk organic parameters might be suitable as a surrogate to monitor water quality for DPR purposes
Additional campaings will be collected for further confirmation
When compared to the other sites, the prelimary data suggest the same results
Each method performed can be useful tools in demonstrating treatment effectiveness.

27. Future Work Finish remaining sampling campaigns
Confirm most efficient treatment train
Determine which analytical techniques are the most appropriate to monitor water quality for DPR purposes
Discuss p53 assays

28. Acknowledgements
Many thanks to Dr. Armando Durazo, Guillermo Flores, and Juliana Ordine

29. References
Shannon, M. A., Bohn, P. W., Elimelech, M., Georgiadis, J. G., Mariñas, B. J., & Mayes, A. M. (2008). Science and technology for water purification in the coming decades. Nature, 452(7185),
Grant, S., et al. Taking the “Waste” Out of “Wastewater” for Human Water Security and Ecosystem Sustainability. Science 2012, 337 (6095),
Huber, S.A., Balz, A., Abert, M. and Pronk, W. (2011), Characterisation of aquatic humic and non-humic matter with size-exclusion chromatography--organic carbon detection--organic nitrogen detection (LC-OCD-OND).Water Research 45(2),
Watereuse, A. (2015). Framework for Direct Potable Reuse.

30. Preliminary CEC Data: Overview
TOrC Removal
% Remaining
Site 1A
Site 1B
Site 2
Site 3
Site 4
Site 5
= Removal from previous treatment
= Removal from WWE