1. Patterns of Estrogenic Activity in Treated Wastewater
A study from Switzerland
NICHT vergessen:
Paper Sebastian & io 2017
Workshop planar-YES in Wädenswil Feb. 2018
Andreas Schönborn, Tamara Mainetti, Andrea A. Grimmer
July 5, 2017 // HPTLC 2017, Berlin

2. Ø 1-3 ng/l EE2 in domestic wastewater Sewage Treatment Plant (STP)
Effects observed from > 0,1 ng/l EE2
Photo Michel Roggo,

3. EE2 interferes with reproduction of fish!
Fathead minnow (Pimephales promelas)
3 generations, exposed to 0-20 ng/L EE2
In mesocosms and aquariums over 1 year
Male survival in the F0 generation was significantly reduced by EE2
F1 parents exposed early in life produced almost no F2 eggs and no larvae
Relevanz des Themas:
We need affordable and reliable test systems that are able to depict the situation in which the fish are in today
The planar-YES is such a system
Schwindt et al., Journal of Applied Ecology 2014, 51,

4. planar-YES (state of 2017) LLE 138 x
Schoenborn, Mainetti, Grimmer, Durisch-Kaiser, Sennhauser 2017, Aqua & Gas (5), p

5. Fingerprinting estrogenic activity of STPs
Estrogen standards [pg] MeOH STPin STPin MeOH STPout STPout MeOH Proc. EtOH
Key message: the development has reached a point where we can reliably, reproducibly and very sensitively measure profiles of estrogenic activity in water samples

6. Comparison of activity fingerprints
M-A3 P1, AW
STP 1
M-A2
P3, AW
STP 2
M-A3
P1, AW
STP 3
M-A2 P1, AW
STP 4
V104
P3,
4.2.16
STP 5
M-A2
P5, AW
STP 6
Type 1
Type 2
Type 3 E1
EE2 E2
Evtl. BPA
3 distinguishable types.
Type 1 (rarest): no activity -> small STPs
Type 2: 1 or several of the estrogens E1, E2, EE2
Type 3: multiple activities
ARA 1 = Horgen
ARA 2 = Dübendorf
ARA 3 = Thalwil
ARA 4 = Bassersdorf
ARA 5 = BfG
ARA 6 = Werdhölzli ? E3
E1: estrone
EE2: 17α-ethinylestradiol
E2: 17β-estradiol
E3: estriol
BPA: Bisphenole-A

7. Different days – similar fingerprint
EE2
DB day 1
DB day 2
Typ 2: Nur Östrogene
Schoenborn, Mainetti, Grimmer, Durisch-Kaiser, Sennhauser 2017, Aqua & Gas (5), p

8. Multiple origin of estrogenicity possibleA
EE2 E1 E2 C D
Typ 3 -> Typ 2 + multiple unknown substances
We started collecting their Rf values
This is where it gets interesting for monitoring
Schoenborn, Mainetti, Grimmer, Durisch-Kaiser, Sennhauser 2017, Aqua & Gas (5), p

9. Estrogenicity removal by treatment steps
EE2 E1 E2 A L B F
Effluent pretreatment C
Efluent filter after ozonation
Effluent ozonation
Effluent clarifier D
Overall EA removal > 95% (up to 99.5%)
We can follow specific activities through the process
In this case it was an experimental STP, with a fourth treatment step using ozonation and subsequent filtartion
Substances A and C survived the fourth treatment step and still show activity
Keep in mind that the planar-YES is an activity test. A and C are not steroids, they must be something else that binds to the estrogen receptor

10. Identification and quantification
n=73 n=69 n=75
Mobile phase: chloroform, acetone, petroleum fraction 55:20:25
Identification of known substances is quite reliable (+/- 7-8% of Rf-value)
However, we can never be 100% sure if a detected signal belongs to a certain substance

11. STP Nr. Code Sampling location Nr of samples Activity bands
EEQ (ng/l) sum 1 RT
Filter -
< LOD 2 WL
< LOQmin 3 DR 4a DB
Clarifier
0.7 4b
0.9 5 BS
2.0 6 KO
2.6 7 WR
2.3 8 TA 4
0.8 9 HR 10 IL
0.4 11 WN
0.1 12 RG 13 NE 14 DE
1.4 15 EG 16 GS 17 WT 18 HN
0.2 19 FH
0.6
20 a
BL 1
Pretreatment
Max. 8
>LOQmax
20 b
BL 2
Max. 5
20 c
BL 3*
Ozonation
20 d
BL 4*
Ozon./Filtration
20 different STPs, 30 composite 24-hr samples
3 STPs with no activity (Type 1)
50% of all samples > 0.4 ng/l EEQ
In 1 case, 6 different active peaks were found

12. Summary: Canton of Zurich
Person equivalents
> 100’000 p.e.
50’-100’000 p.e.
10’-50’000 p.e.
>100 – 10’000 p.e.
Sampling site
Filtration
Clarifier
Estrogenic activity
Schoenborn, Mainetti, Grimmer, Durisch-Kaiser, Sennhauser 2017, Aqua & Gas (5), p

13. Challenges for the future (apart from technical details…)
Certification of the planar-YES procedure
Wanted: more labs that join in (please contact C. Weins, last speaker of this session)
Planned: planar-YES Workshop, Wädenswil, 2/2018 (please leave your name with me)

14. In the name of the fish: Thank you for your attention!
Planar-YES well suited for monitoring wastewater on estrogenicity
In the name of the fish: Thank you for your attention!