a study of endocrine disrupting compounds in wastewater
DESCRIPTION
A Study of Endocrine Disrupting Compounds in Wastewater. Arthur Ho Spacegrant Mentor: Dr. Bob Arnold Chemical and Environmental Engineering University of Arizona. Effects in Animals. Kidd et al ., 2007. 139 Sites 95 Compounds. Comparison of 4 Compounds: - PowerPoint PPT PresentationTRANSCRIPT
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A Study of Endocrine A Study of Endocrine Disrupting Compounds in Disrupting Compounds in
WastewaterWastewater
Arthur HoArthur HoSpacegrant Mentor: Dr. Bob ArnoldSpacegrant Mentor: Dr. Bob Arnold
Chemical and Environmental Chemical and Environmental EngineeringEngineering
University of ArizonaUniversity of Arizona
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Effects in AnimalsEffects in Animals
Kidd et al., 2007
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USGS Surface Water ReconnaissanceUSGS Surface Water Reconnaissance(Kolpin (Kolpin et alet al., 2002)., 2002)
• 139 Sites
• 95 Compounds
Comparison of 4 Compounds:
Compound (Use and Effect) CMAX (µg/L) Days to Med Dose
• Caffeine (stimulant) 5.7 5.7 x 103 (Excedrin Migraine)
• Ibuprofen (anti-inflammatory) 1.0 105 (Motrin)
• 17α-ethinyl estradiol (oral contraceptive) 0.27 37 (Alesse-21)
• 17β-estradiol (vertebrate hormone) 0.093 (Phoenix) 5.3 x 103 (Activella)
• 4-nonylphenol (surfactant metabolite) 40 (Tucson) 4.6 x 10-4 as estrogenic E2, EE2
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Examples of Estrogenic Examples of Estrogenic CompoundsCompounds
Chemical StructureMolecular
WeightLog Kow
Relative Estrogenic Activity
(YES bioassay)
17-estradiol (E2)
272.39
3.94
1.0
17-ethinyl estradiol (EE2)
296.40 4.15 1.32
Estrone (E1)270.37 3.43 0.20
Estriol (E3)288.39 2.81 0.01
Nonylphenol (NP)
220.354.48
2×10-4
Octylphenol (OP)
206.33 4.12 3×10-4
Bisphenol A (BPA)
228.29 3.40 8×10-5
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Estrogenic Activity at JWPCPEstrogenic Activity at JWPCP
Effluent: 0.12 mols EE2 equivalents/day
Dewatered Cake: 0.26 mols EE2
equivalents /day
Influent:1.66 mols EE2
equivalents/day
Removal of estrogenic activity through the whole treatment plant = [1.656 – (0.1178 + 0.26)] / 1.656 = 77%
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Cytoplasm
E
R
R E
E
Cytoplasm
= Estrogen
= Receptor
Yeast Estrogen/Androgen Screen
R
hER/hAR
-gal
CPRGyellow
CPRred
RE
lac-z
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Yeast Estrogen Screen (YES) AssayYeast Estrogen Screen (YES) Assay
17- Ethinylestradiol
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
1.E-13 1.E-12 1.E-11 1.E-10 1.E-09 1.E-08 1.E-07 1.E-06
Molar concentration in well
A570 n
m - A
630 n
m
EC50=1.1 x 10-10 M
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Cytoplasm
E
R
R E
E
Cytoplasm
= Estrogen
= Receptor
Yeast Estrogen/Androgen Screen
R
hER/hAR
-gal
CPRGyellow
CPRred
RE
lac-z
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AgonismAgonism
Anti-estrogens: Mechanism of Anti-estrogens: Mechanism of ActionAction
AntagonismAntagonismR E R A
RE
RA
-gal
No protein production
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Tamoxifen is an Anti-Tamoxifen is an Anti-estrogenestrogen
•Known anti-estrogens: (hydroxy) tamoxifen, chlorinated hormones,
•Known anti-androgens: 4-,4’-DDE, flutamide
01020304050
60708090
100
1.00E-07 1.00E-06 1.00E-05
Tamoxifen concentration (M)
Per
cent
Rel
ativ
e E
stro
geni
c A
ctiv
ity
1 nM EE2
TXFN + 1 nM EE2
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MethodsMethods Obtaining Wastewater samplesObtaining Wastewater samples Extraction of OrganicsExtraction of Organics Preparation of organicsPreparation of organics Yeast Estrogen AssayYeast Estrogen Assay
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RESULTSRESULTS Anti-Estrogen Test: EXTRACT from RAW SLUDGE SOLIDS
0
10
20
30
40
50
60
70
80
90
1.00E-09 1.00E-08 1.00E-07 1.00E-06 1.00E-05 1.00E-04 1.00E-03 1.00E-02 1.00E-01 1.00E+00
Fraction of extract in well
% r
el B
-gala
cto
sid
ase
20% MeOH eluent
50% MeOH eluent
80% MeOH eluent
NP w ater w / 1nM EE2
Anti-estrogen test: EXTRACT from RAW SLUDGE LIQUID
0
10
20
30
40
50
60
70
80
90
100
1.00E-09 1.00E-08 1.00E-07 1.00E-06 1.00E-05 1.00E-04 1.00E-03 1.00E-02 1.00E-01 1.00E+00
Fraction of extract in well
% r
el. B
-gal
acto
sida
se
20% MeOH eluent
50% MeOH eluent
80% MeOH eluent
NP water w/ 1nM EE2
RAW SLUDGE SOLID EXTRACTS ARE HIGHLY ESTROGENIC
RAW SLUDGE LIQUID EXTRACTS ARE ANTI-ESTROGENIC
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RESULTSRESULTSAnti-estrogen test: EXTRACT from DIGESTED SLUDGE SOLID
0
10
20
30
40
50
60
70
80
90
100
1.00E-09 1.00E-08 1.00E-07 1.00E-06 1.00E-05 1.00E-04 1.00E-03 1.00E-02 1.00E-01 1.00E+00
Fraction of extract in well
% r
el. B
-gal
acto
sida
se
20% MeOH eluent
50% MeOH eluent
80% MeOH eluent
NP water w/ 1nM EE2
Anti-estrogen test: EXTRACT from DIGESTER SLUDGE LIQUID
0
10
20
30
40
50
60
70
80
1.00E-09 1.00E-08 1.00E-07 1.00E-06 1.00E-05 1.00E-04 1.00E-03 1.00E-02 1.00E-01 1.00E+00
Fraction of extract in well
% B
-ga
lac
tos
ida
se
20% MeOH eluent
50% MeOH eluent
80% MeOH eluent
NP water w/ 1nM EE2
DIGESTED SLUDGE SOLIDS LOST ESTROGENIC ACTIVITY
DIGESTED SLUDGE LIQUIDS HIGHLY ANTI-ESTROGENIC
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AcknowledgementsAcknowledgements
Dave Newman & Sondra TeskeDave Newman & Sondra Teske
Susan BrewSusan Brew
Matt Giardiana and AzSGC StaffMatt Giardiana and AzSGC Staff
Bingfeng DongBingfeng Dong