a study of endocrine disrupting compounds in wastewater arthur ho spacegrant mentor: dr. bob arnold...
DESCRIPTION
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) x 10 3 (Excedrin Migraine) Ibuprofen(anti-inflammatory) (Motrin) 17α-ethinyl estradiol (oral contraceptive) (Alesse-21) 17β-estradiol (vertebrate hormone) (Phoenix)5.3 x 10 3 (Activella) 4-nonylphenol (surfactant metabolite) 40 (Tucson)4.6 x as estrogenic E 2, EE 2TRANSCRIPT
A Study of Endocrine A Study of Endocrine Disrupting Compounds in Disrupting Compounds in
WastewaterWastewaterArthur HoArthur Ho
Spacegrant Mentor: Dr. Bob ArnoldSpacegrant Mentor: Dr. Bob ArnoldChemical and Environmental Chemical and Environmental
EngineeringEngineeringUniversity of ArizonaUniversity of Arizona
Effects in AnimalsEffects in Animals
Kidd et al., 2007
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
Examples of Estrogenic Examples of Estrogenic CompoundsCompounds
Chemical Structure Molecular Weight
Log 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)
220.35 4.48 2×10-4
Octylphenol (OP)
206.33 4.12 3×10-4
Bisphenol A (BPA)
228.29 3.40 8×10-5
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%
Cytoplasm
E
R
R E
E
Cytoplasm
= Estrogen
= Receptor
Yeast Estrogen/Androgen Screen
R
hER/hAR
-gal
CPRGyellow
CPRred
RE
lac-z
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
A 570
nm -
A 630
nm
EC50=1.1 x 10-10 M
Cytoplasm
E
R
R E
E
Cytoplasm
= Estrogen
= Receptor
Yeast Estrogen/Androgen Screen
R
hER/hAR
-gal
CPRGyellow
CPRred
RE
lac-z
AgonismAgonism
Anti-estrogens: Mechanism of Anti-estrogens: Mechanism of ActionAction
AntagonismAntagonismR E R A
RE
RA
-gal
No protein production
Tamoxifen is an Anti-Tamoxifen is an Anti-estrogenestrogen
•Known anti-estrogens: (hydroxy) tamoxifen, chlorinated hormones,
•Known anti-androgens: 4-,4’-DDE, flutamide
0102030405060708090
100
1.00E-07 1.00E-06 1.00E-05
Tamoxifen concentration (M)
Perc
ent R
elat
ive
Est
roge
nic
Act
ivity
1 nM EE2TXFN + 1 nM EE2
MethodsMethods Obtaining Wastewater samplesObtaining Wastewater samples Extraction of OrganicsExtraction of Organics Preparation of organicsPreparation of organics Yeast Estrogen AssayYeast Estrogen Assay
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
% re
l B-g
alac
tosi
dase
20% MeOH eluent
50% MeOH eluent
80% MeOH eluent
NP w ater w / 1nM EE2
Anti-estrogen test: EXTRACT from RAW SLUDGE LIQUID
0102030405060708090
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
% re
l. 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
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
% re
l. 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
-gal
acto
sida
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
AcknowledgementsAcknowledgements
Dave Newman & Sondra TeskeDave Newman & Sondra Teske Susan BrewSusan Brew Matt Giardiana and AzSGC StaffMatt Giardiana and AzSGC Staff Bingfeng DongBingfeng Dong