<Supplemental Materials>
Bacterial community compositions in sediment polluted by perfluoroalkyl
acids (PFAAs) using Illumina high-throughput sequencing
Yajun Sun a, b, Tieyu Wang a,*, Xiawei Peng a, b, Pei Wang a, Yonglong Lu
a
State Key Lab of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences,
Chinese Academy of Sciences, Beijing 100085, China
b
College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083,
China
* Corresponding author:
Tel: +86 10 62849466; Fax: +86 10 62918177
E-mail address: wangty@rcees.ac.cn (T. Wang)
1
Table S1 HPLC and ESI-MS Instrument Conditions.
Table S2 Target analytes of 12 PFASs measured in this study with QA/QC information.
QA/QC information including matrix spike recovery (MSR) and procedure
recovery (PR), which were presented as Mean ± SD (Mean: Arithmetic mean;
SD: Standard deviation; n indicates the number of samples analyzed.), limit of
detection (LOD), and limit of quantification (LOQ).
Table S3 Percentage of major classes in each sample (%)
Table S4 Spearman correlation coefficients of environmental factors (Spearman
correlation coefficients with p < 0.01 and p < 0.05 are shown in red). *p < 0.05; **p <
0.01
2
Table S1 HPLC and ESI-MS Instrument Conditions
HPLC conditions
Analytical column
Aglient ZORBAX Eclipse Plus C18, 2.1×100 mm, 3.5 μm
Guard column
Agilent 1290 Infinity In-line filter with 0.3 μm SS frit
Column temperature
40 °C
Injection volume
5 μL
Mobile phase
A = 2 mM ammonium acetate
B = 100% Acetonitrile
Run time
16 min + 4 min post time
Flow rate
0.3 mL/min
Gradient
Time (min)
Mobile phase
0
20% B
14
90% B
16
90% B
MS conditions
Acquisition parameters
ESI mode, negative ionization; MRM
Source gas temperature
350 °C
Source gas flow rate
9 L/min
Nebulizer pressure
40 psi
Capiliary
3500 V positive; 4000 V negative
Delta EMV(-)
200-400 V
3
Table S2 Target analytes of 12 PFAAs measured in this study with QA/QC information
Analyte
Acronym
Carbon
MS/MS
Frag
CE
%MSR
%PR
LOD
LOQ
number
transition
(V)
(V)
(n=4)
(n=4)
(ng/g
dw)
Perfluorocarboxylic acid
PFCAs
Perfluoro-butanoic acid
PFBA
C4
213.0 → 169.1
57
1
100±5
103±7
0.020
0.10
Perfluoro-pentanoic acid
PFPeA
C5
263.0 → 218.9
68
2
104±3
101±9
0.010
0.03
Perfluoro-hexanoic acid
PFHxA
C6
313.0 → 269.0
68
3
110±3
102±5
0.004
0.01
313.0 → 119.0
62
15
363.0 → 318.9
68
4
96±6
92±2
0.006
0.02
363.0 → 169.0
70
9
413.0 → 368.9
82
4
100±11
107±3
0.002
0.01
413.0 → 169.0
82
12
413.0 → 219.0
82
10
463.0 → 419.0
82
3
109±2
120±6
0.002
0.01
463.0 → 169.0
66
15
513.0 → 468.9
86
3
103±7
101±9
0.004
0.01
513.0 → 219.0
78
13
563.0 → 519.0
90
5
90±4
101±10
0.008
0.02
563.0 → 319.0
84
15
613.0 → 569.0
90
5
89±4
92±5
0.004
0.01
613.0 → 169.0
80
23
299.0 → 80.0
135
32
119±4
103±9
0.004
0.01
299.0 → 99.0
132
24
399.0 → 80.0
150
40
114±8
112±4
0.004
0.01
399.0 → 99.0
146
34
498.9 → 80.0
154
47
117±2
92±6
0.004
0.01
498.9 → 99.0
150
42
Perfluoro-heptanoic acid
Perfluoro-octanoic acid
Perfluoro-nonanoic acid
Perfluoro-decanoic acid
Perfluoro-undecanoic acid
Perfluoro-dodecanoic acid
PFHpA
PFOA
PFNA
PFDA
PFUdA
PFDoA
Perfluorinated sulfonic acid
PFSAs
Perfluoro-butanesulfonate
PFBS
Perfluoro-hexanesulfonate
Perfluoro-octanesulfonate
PFHxS
PFOS
C7
C8
C9
C10
C11
C12
C4
C6
C8
Internal standards
13
13
-
217.0 → 172.0
57
1
-
-
-
-
13
13
-
315.0 → 270.0
68
3
-
-
-
-
13
13
-
417.0 → 372.0
82
4
-
-
-
-
13
13
-
468.0 → 423.0
82
3
-
-
-
-
13
13
-
515.0 → 470.0
86
3
-
-
-
-
13
13
-
565.0 → 520.0
90
5
-
-
-
-
13
13
-
615.0 → 570.0
90
5
-
-
-
-
18
18
-
403.0 → 103.0
150
40
-
-
-
-
13
13
-
503.0 → 99.0
154
47
-
-
-
-
C4 Perfluoro-butanoic acid
C4 Perfluoro-hexanoic acid
C4 Perfluoro-octanoic acid
C4 Perfluoro-nonanoic acid
C4 Perfluoro-decanoic acid
C4 Perfluoro-undecanoic acid
C2 Perfluoro-dodecanoic acid
O2 Perfluoro-hexanesulfonate
C4 Perfluoro-octanesulfonate
C4 PFBA
C4 PFHxA
C4 PFOA
C4 PFNA
C4 PFDA
C4 PFUdA
C2 PFDoA
O2 PFHxS
C4 PFOS
4
Table S3 Percentage of the major classes in each sample (%)
Class
XQ1
XQ2
XQ3
XQ4
XQ5
XQ6
XQ7
XQ8
XQ9
XQ10
β-Proteobacteria
18.9
20.7
36.7
31.9
39.1
26.0
21.5
17.1
22.5
7.3
γ-Proteobacteria
17.3
19.5
15.7
24.5
16.4
15.7
17.7
17.0
19.8
21.1
δ-Proteobacteria
8.9
6.9
7.3
6.0
3.7
8.9
9.7
9.2
10.9
6.2
Gemmatimonadetes-1
8.2
6.6
4.2
2.6
1.3
5.1
6.6
4.6
3.8
2.0
α-Proteobacteria
4.2
4.1
1.9
4.0
2.2
3.7
5.0
5.8
5.3
13.9
Sphingobacteria
2.9
2.4
2.7
2.5
0.7
3.3
2.4
2.7
1.4
1.7
Pedosphaerae
2.3
4.0
4.1
1.4
0.6
2.9
1.9
2.3
1.1
0.8
Anaerolineae
2.2
2.2
1.3
1.3
0.6
2.6
2.1
3.2
4.1
3.0
Acidobacteria-6
1.9
4.1
1.3
1.3
0.5
1.6
3.4
4.8
2.2
1.4
Nitrospira
1.9
1.6
1.8
1.1
0.6
1.9
2.7
1.6
2.6
0.8
Verrucomicrobiae
1.3
2.8
1.1
1.3
1.4
1.4
1.1
0.9
1.1
2.2
Flavobacteria
1.3
1.8
2.2
3.1
0.9
1.9
2.8
1.9
1.3
15.8
Bacteroidia
1.0
0.7
3.1
1.9
0.7
2.8
1.2
1.1
1.1
1.1
ε-Proteobacteria
0.6
0.8
1.2
1.7
23.4
0.9
0.7
0.7
1.2
3.7
Others
27.1
21.8
15.4
15.4
7.9
21.3
21.2
27.1
21.6
19
5
Table S4 Spearman correlation coefficients of environmental factors (Spearman correlation coefficients with p < 0.01 and p < 0.05 are shown in red).
T
DO
Conductiv
ity
Salinity
pH
TOC
TN
TP
Moistur
e
Clay
Powder
Sand
T
DO
Conducti
-0.110
-0.006
0.742*
Salinity
-0.107
0.591
0.936**
pH
-0.040
0.362
-0.278
-0.427
TOC
-0.316
0.018
-0.188
-0.231
-0.062
TN
-0.003
-0.141
-0.263
-0.408
0.078
0.850**
TP
-0.024
-0.231
-0.600
-0.523
0.247
0.515
0.239
Moisture
0.055
-0.134
-0.285
-0.347
-0.086
0.733*
0.771**
0.442
Clay
0.030
-0.383
-0.103
-0.164
-0.358
0.358
0.587
-0.164
0.455
Powder
-0.310
-0.231
-0.212
-0.255
-0.204
0.564
0.440
0.394
-0.212
0.079
Sand
0.134
-0.353
0.248
0.298
0.210
-0.697*
-0.679*
-0.345
-0.370
-0.527
-0.818**
PFOA
0.158
-0.073
0.200
0.219
-0.401
0.430
0.575
-0.200
0.600
0.624
-0.188
vity
*p < 0.05; **p < 0.01
6
-0.164
PFO
A