SUPPLEMENTARY DATA
Role o me hanogenesis on he bio ans o ma ion o o ganic mic opollu an s
du ing anae obic diges ion
Lo ena Gonzalez-Gil *, a, Miguel Mau icio-Iglesias a, Denisse Se ano a, b, Juan M. Lema
a, Ma a Ca balla a
a Depa men o Chemical Enginee ing, School o Enginee ing, Uni e sidade de
San iago de Compos ela, Rúa Lope Gómez de Ma zoa, E-15782 San iago de
Compos ela, Spain
b Depa men o Wa e and En i onmen al Sciences, Ins i u o Tecnológico de Sono a, 5
de eb e o 818 su , Colonia Cen o, 85000 Ciudad Ob egón, México
*Co esponding Au ho
E-mail add esses: [email p o ec ed], [email p o ec ed],
[email p o ec ed], [email p o ec ed], ma[email p o ec ed]
1
Con en s
S1. Physicochemical cha ac e is ics o selec ed OMPs ........................................... 2
S2. Pa ame e es ima ion .......................................................................................... 3
S3. Analysis o o ganic mic opollu an s .................................................................. 4
S4. Me hanogenic eac o s ope a ion ....................................................................... 5
S5. So p ion o OMPs in diges ed sludge ................................................................ 7
S6. Bio ans o ma ion kine ics ................................................................................. 7
S7. Bio ans o ma ion a e cons an s in he me hanogenic s ep ............................ 14
S8. Bio ans o ma ion phase .................................................................................. 15
S9. Remo al o OMPs du ing sewage sludge AD ................................................. 16
Re e ences .................................................................................................................. 17
2
S1. Physicochemical cha ac e is ics o selec ed OMPs
Table S1. Applica ion and main physicochemical p ope ies o he selec ed OMPs.
OMP
Applica ion
MW
(g/mol)
s
(mg/L)
H
(a m m3/mol)
pKa
log Kow
ERY
An ibio ic
733.9
1.4
5.4·10-29
8.9
3.1
ROX
An ibio ic
837.1
0.02
5.0·10-31
9.1
2.8
SMX
An ibio ic
253.3
610
6.4·10-13
6.2
0.9
TMP
An ibio ic
290.3
400
2.4·10-14
7.1
0.9
FLX
An idep essan
309.3
60
8.9·10-8
9.8
4.1
CBZ
An icon ulsan
236.3
112
1.1·10-10
15.9
2.5
DZP
Anxioly ic
284.7
50
3.6·10-9
3.4
2.8
E1
Es ogen
270.4
30
3.8·10-10
10.3
3.1
E2
Es ogen
272.4
3.6
3.6·10-11
10.3
4.0
EE2
Con acep i e/
es ogen
296.4
11.3
7.9·10-12
10.3
3.7
ADBI
F ag ance
244.4
0.22
2.1·10-4
‒
5.9
HHCB
F ag ance
258.4
1.8
1.3·10-4
‒
5.9
AHTN
F ag ance
258.4
0.21
2.6·10-4
‒
5.8
IBP
An i-in lamma o y
206.3
21
1.5·10-6
4.9
4.0
NPX
An i-in lamma o y
230.3
15.9
3.4·10-10
4.2
3.2
DCF
An i-in lamma o y
296.2
2.4
4.7·10-12
4.2
4.2
OP
Su ac an
206.3
3.1
8.5·10-6
≈10
5.5
NP
Su ac an
220.4
7.0
3.4·10-5
≈10
5.8
TCS
An isep ic
289.5
10
5.0·10-9
7.7
4.7
BPA
Fungicide/plas icize
228.3
120
1.0·10-11
10.1
3.3
Molecula weigh (MW), Hen y’s law cons an (H), solubili y a 25 °C (s), acid dissocia ion cons an (pKa), oc anol-
wa e coe icien (Kow).
Da a ob ained om D ugBank and PhysP op da abases and om Va hanicko a e al. (1995).
3
S2. Pa ame e es ima ion
The pseudo- i s o de kine ic cons an was es ima ed by leas -squa e i ing o he esul s
o he kine ic expe imen s o equa ion S1.
𝑘𝑏𝑖𝑜𝑙 =a gmin ∑(𝐶𝑇(𝑡)−𝐶𝑇,𝑒𝑥𝑝)2
Equa ion S1
whe e CT was ob ained acco ding o equa ion 2 o he manusc ip :
𝐶𝑇= 𝐶0·𝑒(− 1
𝐻𝑅𝑇−𝑘𝑏𝑖𝑜𝑙·𝑋𝑉𝑆𝑆)·𝑡
To ob ain a obus es ima ion o kbiol a boo s ap p ocedu e was ollowed o de e mine he
expec ed alue and he con idence in e al o he pa ame e . The p ocedu e was done as
ollows:
i) Es ima e he esiduals (e) o equa ion S1 by equa ion S2, which a e used o simula e
he expe imen al e o in he measu emen s;
𝑒 =𝐶𝑇(𝑡)−𝐶𝑇,𝑒𝑥𝑝
Equa ion S2
ii) Simula e new expe imen al esul s (C*T,exp) by andomly adding some o he esiduals
o he expe imen al da a;
𝐶∗𝑇,𝑒𝑥𝑝 =𝐶𝑇,𝑒𝑥𝑝 +𝑒𝑗 whe e ej is a andom a iable ej
∈
e
Equa ion S3
iii) Ca y ou he leas squa e es ima ion o k*biol o he new simula ed da a by using
equa ion S1.
i ) I e a e h ough s eps ii) and iii) un il con e gence (numbe o i e a ions = 1000) on a
dis ibu ion o kbiol and use his dis ibu ion o ind he expec ed alue and i s con idence
in e al.
4
S3. Analysis o o ganic mic opollu an s
Table S2. Limi s o quan i ica ion (LOQ) and eco e y anges (n=3) o OMPs in he
liquid and solid phase o he me hanogenic sludge.
LOQ
Reco e y ange (%)
OMP
Liquid
(ng/L)
Solid
(ng/g)
Liquid
Solid
ERY
3
0.6
90-110
65-80
ROX
3
0.6
80-90
55-65
SMX
15
3
85-90
55-60
TMP
15
3
70-75
55-60
FLX
3
0.6
30-35
45-50
CBZ
15
3
65-75
70-80
DZP
15
3
50-60
60-70
E1
30
6
40-50
60-70
E2
30
6
60-70
60-70
EE2
30
6
60-80
60-75
ADBI
150
30
45-55
120-150
HHCB
150
30
40-60
120-150
AHTN
150
30
35-50
130-160
IBP
60
12
130-150
120-140
NPX
75
15
110-120
100-120
DCF
300
60
120-140
130-140
OP
60
12
60-70
120-140
NP
60
12
60-70
120-140
TCS
150
30
55-65
110-130
BPA
75
15
110-120
70-80

5
S4. Me hanogenic eac o s ope a ion
Figu e S1. Pe o mance o he me hanogenic eac o s (MR1 le side and MR2 igh side) du ing he whole ope a ion. O ganic loading a e (OLR)
and me hane p oduc ion (CH4) a e ep esen ed in he uppe g aphs, while pH, ola ile suspended solids (VSS) and in e media e/ o al alkalini y a io
(IA/TA) a e depic ed in he bo om g aphs. G ey a eas highligh he pe iods when he kine ic expe imen s wi h OMPs we e conduc ed.
6
Table S3. A e age eeding cha ac e is ics o me hanogenic eac o s du ing bo h s eady-
s a e ope a ional s ages (OLR=1 and 2 g COD/L d).
OLR=1 g COD/L d
OLR=2 g COD/L d
pH
6.3 ± 0.7
6.4 ± 0.6
Ace ic acid (HAc, g/L)
4.7 ± 0.3
9.5 ± 0.4
P opionic acid (g/L)
1.7 ± 0.1
3.3 ± 0.1
Bu y ic acid (g/L)
1.4 ± 0.1
2.7 ± 0.1
To al VFA (g HAceq/L)
7.9 ± 0.7
15.9 ± 0.5
COD (g/L)
9.5 ± 1.0
18.8 ± 1.0
NH4Cl (g/L)
0.6
0.6
KH2PO4 (g/L)
0.4
0.4
T ace concen a ions o mic o-nu ien s (Fe, Ca, Mg, C , Co, Cu, Mn, Mo,
Ni, Se, Zn, B) we e added acco ding o Angelidaki and Sande s (2004).
7
S5. So p ion o OMPs in diges ed sludge
Table S4. Pa i ioning coe icien s (log Kd) epo ed o anae obically diges ed sludge.
The a e age ange ep esen s he minimum and he maximum alues om li e a u e.
log Kd
OMP
Gonzalez-Gil
e al. (2016)
Ca balla
e al. (2007)
Na umiya
e al. (2013)
Cla a
e al. (2011)
I ashechkin e
al. (2004)
A e age
ange
TCS
3.56-4.35
3.56-4.35
AHTN
4.22-4.86
3.7-4.43
2.60-2.90
2.60-4.86
HHCB
2.58-5.14
3.45-4.3
2.58-5.14
FLX
2.44-3.44
2.44-3.44
E2
2.22-3.34
2.30-2.83
2.22-3.34
E1
1.76-2.91
2.18-2.77
1.76-2.91
BPA
2.09-2.30
2.09-2.30
TMP
1.92-2.86
1.92-2.86
ERY
1.60-3.10
1.60-3.10
ROX
2.52-3.97
1.14-1.92
1.90-3.30
1.14-3.97
EE2
1.21-1.40
2.08-2.85
1.21-2.85
DZP
1.85-1.88
1.85-1.88
DCF
1.26-2.18
1.90-2.20
1.26-2.20
CBZ
1.60-2.27
1.31-1.83
1.60-2.00
1.31-2.27
SMX
0.77-1.79
0.77-1.79
IBP
1.03-1.76
1.00-1.78
1.00-1.78
NPX
1.03-1.71
1.03-1.71
S6. Bio ans o ma ion kine ics
The concen a ions measu ed in he liquid and solid phase o he me hanogenic e luen
du ing he h ee kine ic expe imen s a e displayed in Figu es S2. An a e age
me hanogenic kine ics ob ained om he h ee expe imen s is shown as well. Excep o
NP, EE2, SMX, TMP, and NPX, he esul ing a e age alues p esen ed a low de ia ion,
meaning ha he concen a ions o OMPs measu ed in h ee kine ic expe imen s we e
qui e simila . Mo eo e , con inuous lines depic he concen a ion pa e ns o OMPs in
bo h phases i solely washou was he esponsible o he emo al. The e o e, he
di e ence be ween he washou line and he expe imen al poin co esponds o
bio ans o ma ion.
8
Fig. S2a. Measu ed concen a ions in he liquid (blue diamonds) and solid (o ange squa es) phases o hyd ophobic compounds (G oup 3).
Con inuous lines show he emo al o he OMP by washou in he liquid (blue) and solid phase (o ange).
OLR=1 (Kine ic I)
OLR=1 (Kine ic II)
OLR=2 (Kine ic III)
AVERAGE
15
S8. Bio ans o ma ion phase
Fig. S3. Con ibu ion o each phase o he o al me hanogenic bio ans o ma ion o OMPs. The
appa en bio ans o ma ions measu ed in he solid (BS) and liquid phases (BL) a e ep esen ed by he
o ange, lowe ba s and by he blue, uppe ba s, espec i ely.

16
S9. Remo al o OMPs du ing sewage sludge AD
Table S6. Summa y o epo ed OMPs emo al du ing AD o sewage sludge unde simila ope a ional condi ions. The a e age emo al was
calcula ed wi h he mean alue o each e e ence and o hose cases whe e wo comple ely di e en alues we e epo ed bo h we e conside ed.
OMP
AD emo al (%)
Gonzalez-Gil
e al. (2016)
Ca balla e
al. (2007)
Na umiya e
al. (2013)
Sama as e
al. (2014)
Malmbo g &
Magné
(2015)
Cla a e al.
(2011)
Pa e akis e
al. (2012)
Be ge sen e
al. (2012)
Yang e al.
(2016)
A e age
SMX
80
100
100
‒
‒
‒
‒
‒
‒
93 ± 12
NPX
100
85
‒
85
85
‒
‒
‒
90
89 ± 7
TMP
75
‒
100
‒
100
‒
‒
‒
90
91 ± 12
NP
‒
‒
‒
35
‒
‒
0/100
‒
‒
45 ± 51
FLX
70
‒
‒
‒
0
‒
‒
30
30
33 ± 29
EE2
65
40/95
‒
‒
0
‒
20
‒
‒
44 ± 37
TCS
15
‒
30
65
‒
‒
‒
‒
50
40 ± 22
Musks
10
60
‒
‒
‒
0/45
‒
‒
‒
29 ± 28
BPA
‒
‒
‒
80
‒
‒
‒
‒
0
40 ± 40
E1+E2
0
80
‒
‒
0
‒
50
‒
‒
33 ± 39
CBZ
40
5
0
‒
15
‒
‒
‒
0
11 ± 15
DZP
50
30
‒
‒
‒
‒
‒
‒
‒
40 ± 10
ERY
‒
‒
45
‒
‒
‒
‒
‒
‒
45
ROX
85
95
65
‒
‒
‒
‒
‒
‒
82 ± 15
DCF
‒
0/80
25
95
25
‒
‒
‒
0
38 ± 41
IBP
30
45
‒
95
30
‒
‒
‒
10
42 ± 32
OP
‒
‒
‒
‒
‒
‒
‒
‒
‒
‒
Ope a ional condi ions
Con inuous diges e
Lab-scale
Lab-scale
Full-scale
Lab-scale
Lab-scale
Full-scale
Lab-scale
Lab-scale
Full-scale
OMPs spike
No
Yes
No
Yes
Yes
No
No
Yes
No
Subs a e*
MixS
MixS
MixS
MixS
MixS
SS
MixS & PS
SS
PS
Tempe a u e (°C)
37
37
30-55
37
37
37
35
37
35
HRT (d)
20-30
10-30
20-30
20
20
15-20
30
20
15-30
*MixS: mixed p ima y and seconda y sewage sludge; PS: p ima y sludge; SS: sewage sludge ( ype no speci ied).
17
Re e ences
Be ge sen, O., Øs nes, K., Vasskog, T., 2012. Anae obic ea men o sewage sludge
con aining selec i e se o onin eup ake inhibi o s. Bio esou . Technol. 117, 325–332.
Ca balla, M., Omil, F., Te nes, T., Lema, J.M., 2007. Fa e o pha maceu ical and pe sonal
ca e p oduc s (PPCPs) du ing anae obic diges ion o sewage sludge. Wa e Res. 41,
2139–50.
Cla a, M., Gans, O., Windho e , G., K enn, U., Ha l, W., B aun, K., Scha , S.,
Sche knech , C., 2011. Occu ence o polycyclic musks in was ewa e and ecei ing
wa e bodies and a e du ing was ewa e ea men . Chemosphe e 82, 1116–1123.
D ugBank da abase; h p://www.d ugbank.ca.
Gonzalez-Gil, L., Papa, M., Fe e i, D., Ce e i, E., Mazzoleni, G., S eimbe g, N.,
Ped azzani, R., Be anza, G., Lema, J.M., Ca balla, M., 2016. Is anae obic diges ion
e ec i e o he emo al o o ganic mic opollu an s and biological ac i i ies om
sewage sludge? Wa e Res. 102, 211–220.
I ashechkin, P., Co ini, P.-X., Dohmann, M., 2004. Beha iou o endoc ine dis up ing
chemicals du ing he ea men o municipal sewage sludge. Wa e Sci. Technol. 50,
133–140.
Malmbo g, J., Magné , J., 2015. Pha maceu ical esidues in sewage sludge: E ec o
sani iza ion and anae obic diges ion. J. En i on. Manage. 153, 1–10.
Na umiya, M., Nakada, N., Yamashi a, N., Tanaka, H., 2013. Phase dis ibu ion and
emo al o pha maceu icals and pe sonal ca e p oduc s du ing anae obic sludge
diges ion. J. Haza d. Ma e . 260, 305–12.
Pa e akis, N., Chiu, T.Y., Koh, Y.K.K., Les e , J.N., McAdam, E.J., Sc imshaw, M.D.,
Soa es, A., Ca mell, E., 2012. The e ec i eness o anae obic diges ion in emo ing
es ogens and nonylphenol e hoxyla es. J. Haza d. Ma e . 199–200, 88–95.
Sama as, V.G., S asinakis, A.S., Thomaidis, N.S., Mamais, D., Lekkas, T.D., 2014. Fa e
o selec ed eme ging mic opollu an s du ing mesophilic, he mophilic and
empe a u e co-phased anae obic diges ion o sewage sludge. Bio esou . Technol.
162, 365–72.
Sy acuse Resea ch Co po a ion (SRC) PhysP op da abase; h p://www.s cinc.com.
Va hanicko a, D., Shiu, W.-Y., Mackay, D., 1995. Aqueous solubili ies o alkylphenols
and me hoxyphenols a 25 oC. J. Chem. Eng. Da a 40, 448–451.
Yang, S., Hai, F.I., P ice, W.E., McDonald, J., Khan, S.J., Nghiem, L.D., 2016.
Occu ence o ace o ganic con aminan s in was ewa e sludge and hei emo als by
anae obic diges ion. Bio esou . Technol. 210, 153–159.