DEQ-CFW_00002950

Supporting information
Legacy and Emerging Perfluoroalkyl Substances Are Important
Drinking Water Contaminants in the Cape Fear River Watershed
of North Carolina
Supporting information includes analytical method description, 6 tables, and 5 figures.
Mei Sun' 2- ", Elisa Areval02, Mark Strynar3, Andrew Lindstrom3, Michael Richardson 4, Ben
Kearns4, Adam Pickett5, Chris Smith6, and Detlef R.U. Knappe2
'Department of Civil and Environmental Engineering
University of North Carolina at Charlotte
Charlotte, North Carolina 28223, USA
2 Department of Civil, Construction, and Environmental Engineering
North Carolina State University
Raleigh, North Carolina 27695, USA
3 National Exposure Research Laboratory
U.S. Environmental Protection Agency
Research Triangle Park, North Carolina 27711, USA
4 Cape Fear Public Utility Authority
Wilmington, North Carolina 28403, USA
5 Town of Pittsboro
Pittsboro, North Carolina 27312, USA
6 Fayetteville Public Works Commission
Fayetteville, North Carolina 28301, USA
*Corresponding Author Email: [e-mail]; Phone: [phone]
Page 1 of 12
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Analytical standards: PFASs studied in this research are listed in Table S1. For legacy PFASs,
native and isotopically labeled standards were purchased from Wellington Laboratories
(Guelph, Ontario, Canada). Native PFPrOPrA was purchased from Thermo Fisher Scientific
(Waltham, MA). No analytical standards were available for other PFECAs.
PFAS quantification: PFAS concentrations in samples from DWTPs and adsorption tests were
determined by liquid chromatography tandem mass spectrometry (LC-MS/MS) using a large -
volume (0.9 mL) direct injection method. An Agilent 1100 Series LC pump and PE Sciex API
3000 LC-MS/MS system equipped with a 4.6 mm x 50 mm HPLC column (Kinetex C18 5µm
100A, Phenomenex Inc.) was used for PFAS analysis. The eluent gradient is shown in Table S4
in SI. All samples, calibration standards, and quality control samples were spiked with
isotopically labeled internal standards, filtered through 0.45-µm glass microfiber syringe filters,
and analyzed in duplicate. The MS transitions for PFAS analytes and internal standards are
shown in Table S5 in SI. The quantitation limit (QL) was 25 ng/L for PFOS and
perfluorodecanoic acid, and 10 ng/L for other legacy PFASs and PFPrOPrA. The QL was
defined as the first point of the standard curve, for which the regression equation yielded a
calculated value within ±30% error. For PFECAs without analytical standards, chromatographic
peak areas are reported.
PFAS concentrations along the treatment train of DWTP C were analyzed using a Waters
Acquity ultra performance liquid chromatograph interfaced with a Waters Quattro Premier XE
triple quadrupole mass spectrometer (Waters, Milford, MA, USA) after solid phase extraction.
Method details are described elsewhere.' The QL for all PFASs with analytical standards was
0.2 ng/L, and peak areas were recorded for PFECAs without standards.
Page 2 of 12
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Table S1. Perfluoroalkyl substances (PFASs) detected in the Cape Fear River (CFR) watershed
CompoundMolecular # of Chain length
. .• . •. (including all
carbonsweight
• and S)
Perfluorocarboxylic acids (PFCAs)
Perfluorobutanoic acid (PFBA)
214.0
C4HF7O2
375-22-4
3
4
Perfluoropentanoic acid (PFPeA)
264.0
C5HF9O2
2706-90-3
4
5
Perfluorohexanoic acid (PFHxA)
314.1
C6HF11O2
307-24-4
5
6
Perfluoroheptanoic acid (PFHpA)
364.1
C7HF13O2
375-85-9
6
7
Perfluorooctanoic acid (PFOA)
414.1
C8HF1502
335-67-1
7
8
Perfluorononanoic acid (PFNA)
464.1
C9HF17O2
375-95-1
8
9
Perfluorodecanoic acid (PFDA)
514.1
CloHF19O2
335-76-2
9
10
Perfluorosulfonic acids (PFSAs)
Perfluorobutane sulfonic acid (PFBS)
300.1
C4HF9SO3
375-73-5
4
5
Perfluorohexane sulfonic acid (PFHxS)
400.1
C6HF13SO3
355-46-4
6
7
Perfluorooctane sulfonic acid (PFOS)
500.1
C8HF17SO3
1763-23-1
8
9
Perfluoroalkyl ether carboxylic acids with one ether group (mono -ether PFECAs)
Perfluoro-2-methoxyacetic acid (PFMOAA)
180.0
C3HF503
674-13-5
2
4
Perfluoro-3-methoxypropanoic acid (PFMOPrA)
230.0
C4HF7O3
377-73-1
3
5
Perfluoro-4-methoxybutanoic acid (PFMOBA)
280.0
C5HF9O3
863090-89-5
4
6
Perfluoro-2-propoxypropanoic acid (PFPrOPrA)
330.1
C6HF11O3
13252-13-6
5
7
Perfluoroalkyl ether carboxylic acids with multiple ether group (multi -ether PFECAs)
Perfluoro(3,5-dioxahexanoic) acid (PFO2HxA)
246.0
C4HF7O4
39492-88-1
3
6
Perfluoro(3,5,7-trioxaoctanoic) acid (PFO3OA)
312.0
C5HF9O5
39492-89-2
4
8
Perfluoro(3,5,7,9-tetraoxadecanoic) acid (PFO4DA)
378.1
C6HF11O6
39492-90-5
5
10
Page 3 of 12

Table S2. Operational conditions of DWTP C on sampling day (August 18, 2014)
Parameter
Raw water ozone dose
Value
Raw water total organic carbon concentration
6.0 m /L
Aluminum sulfate coagulant dose
43 m /L
Coagulation pH
5.70
Settled water ozone dose
1.3 m /L
Settled water total organic carbon concentration
1.90 m /L
Empty bed contact time in
biological activated carbon filters
9.4 minutes for granular activated
carbon layer
2.3 minutes for sand layer
Medium pressure UV dose
25 mJ/cm2
Free chlorine dose
1.26 mg/L as C12
Free chlorine contact time
17.2 hours
Table S3. Water quality characteristics of surface water used in adsorption tests
Table S4. LC gradient method for PFAS analysis
Time (min) Mobile Phase A% (v/v) Mobile Phase B% Flow Rate (mL/min)
Mobile phase A: 2 mM ammonium acetate in ultrapure water with 5% methanol
Mobile phase B: 2 mM ammonium acetate in acetonitrile with 5% ultrapure water
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DEQ-CFW 00002953
Mobile phase A: 2 mM ammonium acetate in ultrapure water with 5% methanol
Mobile phase B: 2 mM ammonium acetate in acetonitrile with 5% ultrapure water
Page 4 of 12
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Table S5. MS transitions for PFAS Analysis
Legacy PFASs
PFECAs
Internal standards
PFBA
PFPeA
PFHxA
PFHpA
PFOA
PFNA
PFDA
PFBS
PFHxS
PFOS
PFMOAA
PFMOPrA
PFMOBA
PFPrOPrA
PFO2HxA o ^,5 L
PFO3OA 7,z
PFO4DA
Perfluoro-n-[1,2,3,4-13C4]butanoic acid
(13C4-PFBA)
Perfluoro-n-[1,2-13C2]hexanoic acid
(13C2-PFHxA)
Perfluoro-n-[1,2,3,4-13C2]octanoic acid
(13C4-PFOA)
Perfluoro-n-[1,2-13C2]decanoic acid
(13C2-PFDA)
Sodium perfluoro-1-
hexane[1802]sulfonate (1802-PFHxS)
Sodium perfluoro-l-[1,2,3,413C4]octane
sulfonate (13C4-PFOS)
L12.8 168.8
262.9 218.8
313.6 268.8
362.9 318.8
413.0 --> 368.8
463.0 418.8
513.1 68.8
299.1 --> 98.8
399.1 ---> 98.8
498.9 - 98.8
180.0 - 85.0
229.1 184.9
279.0 234.8
329.0 284.7
245.1 85.0
311. 84.9
377.1 - 85.0
217.0 - 172
315.1 -� 269.8
417.0 372.0
515.1 469.8
403.1 --+ 83.8
502.9 - 79.9
13C4-PFBA
13C2- PFHxA
13C2- PFHxA
13C4- PFOA
13C4- PFOA
13C4- PFOA
13C2-PFDA
1802-PFHxS
1802-PFHxS
13C4-PFOS
N/A
N/A
N/A
13C2- PFHxA
N/A
N/A
N/A
Not applicable
Page 5 of 12
DEQ-CFW 00002954

Table S6. Maximum,
minimum, mean and median concentrations
(ng/L) of PFASs at three drinking
water intakes. *
min
medianmax
•. ian
mean
PFBA
99
<10
26
33
38
<10
12
12
104
<10
12
22
PFPeA
191
14
44
62
38
<10
19
19
116
<10
30
36
PFHxA
318
<10
48
78
42
<10
<10
11
24
<10
<10
<10
PFHpA
324
<10
39
67
85
<10
<10
11
24
<10
<10
<10
PFOA
137
<10
34
46
32
<10
<10
<10
17
<10
<10
<10
PFNA
38
<10
<10
<10
<10
<10
<10
<10
<10
<10
<10
<10
PFDA
35
<25
<25
<25
<25
<25
<25
<25
<25
<25
<25
<25
PFBS
80
<10
<10
<10
11
<10
<10
<10
<10
<10
<10
<10
PFHxS
193
<10
10
14
14
<10
<10
<10
14
<10
<10
<10
PFOS
346
<25
29
44
43
<25
<25
<25
40
<25
<25
<25
PFPrOPrA
<10
<10
<10
<10
10
<10
<10
<10
4560
55
304
631
PFOA+PFOS
447
0
64
90
59
0
0
9
55
<10
<10
<10
E PFASs**
1502
18
212
355
189
0
47
62
4696
55
345
710
* Concentrations less than quantitation limits were considered as zero to calculate means and Y- PFASs.
** Other PFECAs were present in water samples from community C but could not be quantified and were therefore not included in
PFASs
Page 6 of 12

F O
O
F
i4OH
F
F
PFMOAA
F F F O
O
F
F F F OH
F
F
PFMOBA
F F O
O
F
F F OH
F
F
PFMOPrA
PFPrOPrA
PFO2HxA PF03vA
F F F F F
p O O
OH
O O
F F F F F
PFO4DA O
Figure S1. Molecular structures of PFECAs evaluated in this study
Page 7 of 12
DEQ-CFW 00002956

Cape Fear River watershed
Haw
River
Commt ' A N
DWTP
Deep River
r
Flow direction
Community S DWTP
North Carolina ` �- Cape Fear River
v .
P1.AS Surface Neater sampling site
� •
Cape Fear river basin manufacturing for PAC test
plant
w� -- No Community CWIT �-.f'
MN MI _ DWTP
YV
r
OR Y SO - Yh MI NY } NN
NE IA__ _. 1 t MA
I
__ IL IN ON FA �►LI CT •� �,J
NV VT CO -' KS---- '--MO TOE
KY VA
CA --z
OK TN NC
AR
AZ NM SC S
MS AL OA
rx LA
FL d
f 100 km
Figure S2. Sampling sites in the Cape Fear River watershed, North Carolina. The scale is for the
Cape Fear River watershed map.
Page 8 of 12
DEQ-CFW 00002957

1200
1000
J 800
a"
c
c
0 600
ca
L
c
400
0
A
200
0
QFQtCPcP PF0 PFPe PF* Q 01.9PpP PF�P PF�P PF6 PP��S QF� PF �PxPFC�
PF
200
Community B
150
J
c
0 100
cB
c
O
U
0 50
U
0
4
PFQ�oPtP PF6 PFQe 1?0 PQ�QPPPpP PF�P PF�P QF� PP��S QFOlb
PF OP ,011b
PF
Page 9 of 12
DEQ-CFW 00002958

611I1ZI
4000
J
� 3000
c
O
m
2000
a�
U
C
O
U 1000
M
• Community C
•
•
•
el
PFP�pPtP PF6 QFPe QF� QP�Qp Popp PF�P PF�P PF� QQ�� P �oPkP�pS
P
Figure S3. PFAS concentration distributions in the CFR watershed at three drinking water
intakes. Concentrations less than quantitation limits were considered as zero. Upper and lower
edges of a box represent the 751h and 25th percentile, respectively; the middle line represents the
median; upper and lower bars represent the 90th and 101h percentile, respectively; and dots
represent outliers (>901h or <101h percentile).
Page 10 of 12
DEQ-CFW 00002959

2.5E+07
•
2.0E+07
a
•
m
1.5E+07
•
3
•
o
••
C
1.0E+07
•�
oft
5.0E+06
� i• • 'A-0
S• At
2500
7Mean
2000
t�
1500
e °
e
1000
a
° e ee
e
w
e e
A
Qe o A
500
eY
0
0.0E+00
6/15/13 7/30/13 9/13/13 10/28/13 12/12/13
5. E+07
4. E+07
-B
E
3. E+07
3
0
2.E+07
1. E+07
0 E+00
Community B
s Mean flow
61
PFASs
e
e °
ebb
°
500
400
J
300
in
200 a
w
100
0
6/15/13 7/30/13 9/13/13 10/28/13 12/12/13
6. E+07
•t Community C
5.E+07F-
4.E+07 • • • Mean flow
•
e PFASs
E•
n
0 3.E+07 •
C L,
v 2. E+07 °
e
1. E+07 , �
0.E+00 �� ao
6/1/13 7/1/13 7/31/13 8/30/13 9/29/13
6000
5000
4000
ao
V
3000 cn
Q
2000 w
1000
0
Figure S4. Total PFAS concentrations in the source water and stream flow at the three studied
DWTPs. Stream flow data were acquired from US Geological Survey stream gage records
Page 11 of 12
DEQ-CFW 00002960

1W7o
a
80%
60%-
m
40% —�1C
E• o--- .� _ _.`a
20%
0%
-20% r i i
0 20 40 60 80 100 120 140
time (min)
--HPFBA-*-PFPeA -A- PFHxA -*-PFHpA-W-PFOA
-- PFNA -0-PFDA --c)--PFBS -0- PFHxS - -PFOS
100% _
c
80%
601%
'm
0 40%
E
d
Cr 20%
0%
-20%
0 20 40 60 80 100 120 140
time (min)
-�-PFBA -4&-PFPeA-*-PFHxA-E-PFHpA-W-PFOA
-,--PFNA-O-PFDA -a-PFBS-O-PFHS--*-PFOS
100 % _
80%
a^ 60%.�.�---�"�
A
'0 40%
E
a
°C 201%
0% e
-20%
0 20 40 60 80 100 120 140
time (min)
-I-PFBA-+-PFPeA -& PFHxA-*-PFHpA fit(-PFOA
-�-•PFNA-O-PFDA-o--PFBS-0-PFHxS-*w-PFOS
1W7o
80%
60%
m
E40%
20%
0%
-20%
IUU7a
80%
60%
'm
0 40%
E
L
Cr 20%
0%
-20%
1W7o
80%
60%
m
'0 40%
E
w
20%
0%
-20%
b
0 20 40 60 80 100 120 140
time (min)
-+-PFMOPrA-If-PFMOBA- -PFPrOPrA
-0-PF02HxA-0-PF030A •;r-PF04DA
d
0 20 40 60 80 100 120 140
time (min)
--PFMOPrA-f-PFMOBA--PFPrOPrA
-O-PF02HxA-D-PF030A •-C�--PF04DA
f
0 20 40 60 80 100 120 140
time (min)
-#-PFMOPrA-fPFMOBA --PFPrOPrA
-O-PF02HxA-0-PF030A o.-PF04DA
Figure S5. PFAS adsorption at powdered activated carbon doses of (a, b) 30 mg/L, (c, d) 60 mg/L
and (e, f) 100 mg/L. Figures show average PFAS removal percentages of duplicate tests.
Reference
1. Nakayama, S.; Strynar, M. J.; Helfant, L.; Egeghy, P.; Ye, X.; Lindstrom, A. B.,
Perfluorinated compounds in the Cape Fear drainage basin in North Carolina. Environ. Sci.
Technol. 2007, 41, (15), 5271-5276.
Page 12 of 12
DEQ-CFW 00002961