Degradation of Hormone

Contaminants in Waters by •OH

OxidationKaty

SwancuttStephen Mezyk

Background• What are current water treatment

methods?– Preliminary Treatment (screens)

Background• What are current water treatment

methods?– Preliminary Treatment (screens)– Primary Treatment (Sedimentation,

addition of alum/lime/iron salts/polymers)

Background• What are current water treatment

methods?– Preliminary Treatment (screens)– Primary Treatment (Sedimentation,

addition of alum/lime/iron salts/polymers)

– Secondary Treatment (aeration, bacteria)

Background• What are current water treatment

methods?

Background• What are current water treatment

methods?– Disinfection (still Secondary Treatment)

• Chlorine (dangerous by-products)• Ozone (high energy cost of formation from

O2)

• Ultraviolet Radiation (bacteria may survive low doses)

Background• What are current water treatment

methods?– Disinfection (still Secondary Treatment)

• Chlorine (dangerous by-products)

• Ozone (high energy cost of formation from O2)

• Ultraviolet Radiation (bacteria may survive low doses)

– Advanced Treatment• Membrane filtration• Reverse osmosis• Ion exchange• Carbon absorption

Background

• What are the results of ineffective treatment?– Trace contaminants are hard to remove from water!

Background

• What are the results of ineffective treatment?– Trace contaminants are hard to remove from water!

Pharmaceuticals

Hormones

Pesticides

Fragrances

Chlorinated hydrocarbons

Antibiotics

Background

• What are the results of ineffective treatment?– Trace contaminants are hard to remove from water!

Pharmaceuticals

Hormones

Fish devastated by sex-changing chemicals in municipal wastewaterAuthor: Natural Sciences and Engineering Research Council CanadaPublished on Feb 16, 2008 - 7:31:49 AM

Pesticides

Fragrances

Chlorinated hydrocarbons

Antibiotics

Male fish becoming female?Researchers worry about estrogen

and pollutants in the water By Tom Costello

CorrespondentNBC News Nov. 9, 2004

What's In The Water? Estrogen-like Chemicals Found In Fish

Caught In Pittsburgh's Rivers, USA

ScienceDaily (Apr. 17, 2007)

Background

• What are Advanced Oxidation Processes?

•OH

Electron BeamsNon-Thermal Plasmas

O3/UV

H2O2/O3

H2O2/UV

H2O2/O3/UV

Photocatalytic Redox Processes (TiO2/UV)

Electrohydraulic Cavitation & Sonolysis

Supercritical Water Oxidation

Gamma Radiation

Graphic adapted from the Journal of Advanced Oxidation Technologies at http://www.jaots.net/

Background• How effective are AOPs for treating hormones?

– Ethinylestradiol: studied along with many other contaminants by Huber, Canonica, Park, and von Gunten (ES&T 2008, 37(5): 1016-1024)

Background• How effective are AOPs for treating

hormones?– Ethinylestradiol: studied along with many other

contaminants by Huber, Canonica, Park, and von Gunten (ES&T 2008, 37(5): 1016-1024)• kozone is approximately 3 x 106 M-1s-1 and k•OH was

guessed to range from 3.3 to 9.8 x 109 M-1s-1

– •OH is faster than other methods, but not well understood.

•OH + hormones → productsk

Background• How effective are AOPs for treating

hormones?– Ethinylestradiol: studied along with many

other contaminants by Huber, Canonica, Park, and von Gunten (ES&T 2008, 37(5): 1016-1024)• kozone is approximately 3 x 106 M-1s-1 and k•OH was

guessed to range from 3.3 to 9.8 x 109 M-1s-1

– •OH is faster than other methods, but not well understood.

• Why don’t we know much about •OH reactions with hormones?– Insolubility

•OH + hormones → productsk

Goals

To evaluate •OH as a method of degradation of the following hormone compounds:

HO

H H

H

OHestradiol

HO

H H

H

OH

OH

estriol

HO

H H

H

O

estrone

HO

H H

H

OH

ethinylestradiol

O

H H

H

O

progesterone

Specific Aims

1. Measure fundamental rate constants

2. Analyze oxidation products3. Elucidate mechanisms4. Quantify removal efficiencies5. Evaluate loss of estrogenic or

endocrine disrupting activity

1. Rate Constants

How do we make free radicals?

H2O 0.28OH + 0.27eaq- + 0.06H

+ 0.07H2O2 + 0.05H2 + 0.27H+

Coefficients are relative yields in μmol/Joule

Buxton et al, (1988) J. Phys. Chem. Ref. Data, Vol. 17, pp. 513-886

1. Rate Constants

How do we make free radicals?

How do we isolate •OH?

H2O 0.28OH + 0.27eaq- + 0.06H

+ 0.07H2O2 + 0.05H2 + 0.27H+

Coefficients are relative yields in μmol/Joule

Buxton et al, (1988) J. Phys. Chem. Ref. Data, Vol. 17, pp. 513-886

eaq- + N2O + H2O → N2 + OH- + •OH

•H + N2O → •OH + N2

1. Rate Constants

1. Rate Constants

1 2

3 4

1. Rate Constants

• How to measure absorbance:– Directly– Competition Kinetics– Vary the concentration of the compound

(steroid)

1. Rate Constants

• How to measure absorbance:– Directly– Competition Kinetics– Vary the concentration of the compound (steroid)

• Steroids in water? Only about 10 μM…– Must come up with a new way to measure

steroids- one that works around insolubility!

– Instead of changing the steroid concentration, use competition kinetics and alter the [SCN-]

Competition Kinetics•OH + SCN- (+SCN-) → OH- + (SCN)2

•-

•OH + X → products

1. Rate Constants

][

][1

SCNk

Xk

Abs

Abs

SCN

X

SCN

o

SCN

][

1][*

11

SCNAbs

X

k

k

AbsAbsSCNSCN

xo

SCNSCN

Old Method: Vary the compound of interest (steroids, etc.)

New Method: Vary the thiocyanate, [steroid] can be constant

kSCN-

kX

• The new method works!– Suwannee River fulvic acid + •OH kinetics

(in M-1s-1)

1. Rate Constants

• The new method works!– Suwannee River fulvic acid + •OH kinetics

(in M-1s-1)

1. Rate Constants

Direct Absorption

Measurement1

(1.39 ± 0.16) x 108

at 400nm

(1.87 ± 0.07) x 108

at 272nm

1Westerhoff et al, (2007) Environmental Science & Technology, 41: 4640-4646

2Rosario-Ortiz et al, recently accepted for publication in Environmental Science & Technology

• The new method works!– Suwannee River fulvic acid + •OH kinetics

(in M-1s-1)

1. Rate Constants

Direct Absorption

Measurement1

(1.39 ± 0.16) x 108

at 400nm

(1.87 ± 0.07) x 108

at 272nm

Old Competition Kinetics1

(1.55 ± 0.04) x 108

1Westerhoff et al, (2007) Environmental Science & Technology, 41: 4640-4646

2Rosario-Ortiz et al, recently accepted for publication in Environmental Science & Technology

• The new method works!– Suwannee River fulvic acid + •OH kinetics

(in M-1s-1)

1. Rate Constants

Direct Absorption

Measurement1

(1.39 ± 0.16) x 108

at 400nm

(1.87 ± 0.07) x 108

at 272nm

Old Competition Kinetics1

(1.55 ± 0.04) x 108

New Competition Kinetics2

(1.61 ± 0.06) x 108

1Westerhoff et al, (2007) Environmental Science & Technology, 41: 4640-4646

2Rosario-Ortiz et al, recently accepted for publication in Environmental Science & Technology

• Desired information…2. Identifying products3. Elucidating mechanisms4. Calculating degradation efficiencies

2,3 and 4. Product Analysis

• Desired information…2. Identifying products3. Elucidating mechanisms4. Calculating degradation efficiencies

• How to…– Step One: Saturate with N2O

– Step Two: Irradiate with 60Co– Step Three: Analyze products using

LCMS

2,3 and 4. Product Analysis

2,3 and 4. Product Analysis

HO

H H

H

OH

? ? ?

• Test irradiation products for remaining estrogen activity– Yeast Estrogen Screen (YES)

• lac-Z• beta-galactosidase• chlorophenol red-beta-D-galactopyranoside

(CPRG)

5. Estrogen Activity

• Test irradiation products for remaining estrogen activity– Yeast Estrogen Screen (YES)

• lac-Z• beta-galactosidase• chlorophenol red-beta-D-galactopyranoside

(CPRG)

5. Estrogen Activity

Yellow = no remaining estrogen activity

Red = estrogen activity intact

Anticipated Costs

From 8/1/2008 to 8/1/2011 (a three-year project)

• Total: $202,450

• Personnel: only a two person project = less spent on salaries: $97,787

• General lab supplies and chemicals: cheap and dirt cheap: $12,000

Anticipated Costs

• Equipment– Tetronix oscilloscope: $21,973– UV/VIS Spectrophotometer: $7,090– Millipore MilliQ System: $7,200

• Travel: $22,500 (airfare is not cheap)• Facility use

– Notre Dame Rad Lab: $200 hr-1 x ~120 hr =$24,000

– UCI Mass Spec Lab: $40 sample-1 x ~200 samples =$8,000

The Big Picture

• There are deficiencies in standard wastewater treatment- steroids in particular pose an environmental health threat and must be degraded

The Big Picture

• There are deficiencies in standard wastewater treatment- steroids in particular pose an environmental health threat and must be degraded

• AOPs may lead to improved wastewater treatment practices

The Big Picture

• There are deficiencies in standard wastewater treatment- steroids in particular pose an environmental health threat and must be degraded

• AOPs may lead to improved wastewater treatment practices

• It is necessary to understand how steroids react with •OH. No one has been able to do so due to solubility issues… until now.

THANK YOU