© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 1
Manual Extraction of PFAS in Drinking Water
Following DIN 38407-42
Authors
William Jones, Biotage, Salem, NH, USA
Matthew Harden, Biotage, Salem, NH, USA
Application Note AN968
Introduction
Per- and polyfluorinated alkyl substances (PFAS) have been used abundantly since
their inception in the twentieth century and have become a closely monitored class of
compounds within environmental testing. This application note outlines a procedure for
those seeking to follow DIN 38407-42 for large volumes of water. The data presented
was generated using a Biotage® VacMaster™ vacuum manifold with a PFAS free Biotage®
VacMaster™ Large Volume Extraction (LVE) kit in conjunction with EVOLUTE® PFAS SPE
columns and a TurboVap® LV system.
Agilent
» InfinityLab PFC Delay Column, 4.6 x 30 mm, p/n 5062-8100
» ZORBAX RRHD Eclipse Plus C18, 95 Å,
2.1 x 50 mm, 1.8 µm, p/n 959757-902
Sigma-Aldrich
» Ammonium Acetate, ACS Reagent
Grade ≥ 97%, p/n 238074-25G
» Acetic Acid, Glacial, ReagentPlus®, ≥ 99% p/n A6283
Honeywell
» Water, ACS Certified, HPLC Grade, p/n AH365-4
» Methanol, Burdick & Jackson™, LC-MS Grade, p/n LC230-4
VWR
» 15 mL Polypropylene Centrifuge Tubes
with Caps, p/n 21008-670
» 250 mL Polypropylene Wide Mouth Bottles, p/n 414004-125
Equipment and Materials Used
Biotage
» Biotage® VacMaster™ 20 Sample Processing Station (with
15 mL rack), p/n 121-2015ML, fitted with polypropylene
(PFAS free) stopcocks (p/n 121-0009-PP)
» Biotage® VacMaster™ LVE Kit (PFAS)
for 1, 3, 6 mL SPE Columns (p/n 121-2190)
» EVOLUTE® PFAS 500 mg/6 mL SPE Columns, p/n 614-0050-CP
» EVOLUTE® PFAS 150 mg/6 mL SPE Columns, p/n 614-0015-CP
» TurboVap® LV Automated Solvent
Evaporation System, p/n 415000
» TurboVap® LV Multi Rack (48 Positions,
10–20 mm Tubes), p/n 414964
Wellington Laboratories
» ISO 21675:2019 Labelled Stock Solution,
1.2 mL, p/n ISO 21675-LSS
» ISO 21675:2019 Native Stock Solution,
1.2 mL, p/n ISO 21675-NSS
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 2
Analytes
Table 1. Listing of Target Analytes and Internal Standards.
Target Analyte
Acronym
CAS
Perfluorobutanoic acid
PFBA
375-22-4
Perfluoropentanoic acid
PFPeA
2706-90-3
Perfluorohexanoic acid
PFHxA
307-24-4
Perfluoroheptanoic acid
PFHpA
375-85-9
Perfluorooctanoic acid
PFOA
335-67-1
Perfluorononanoic acid
PFNA
375-95-1
Perfluorodecanoic acid
PFDA
335-76-2
Perfluorobutanesulfonic acid
PFBS
375-73-5
Perfluorohexanesulfonic acid
PFHxS
355-46-4
Perfluorooctanesulfonic acid
PFOS
1763-23-1
Perfluoroundecanoic acid1
PFUnA
2058-94-8
Perfluorododecanoic acid1
PFDoA
307-55-1
Perfluoroheptanesulfonic acid1
PFHpS
375-92-8
Perfluorodecanesulfonic acid1
PFDS
335-77-3
1H,1H,2H,2H-perfluorooctanesulfonic acid1
H4PFOS
27619-97-2
Internal Standard
Perfluoro-n-[1,2,3,4-13C4]butanoic acid
13
C4-PFBA
Perfluoro-n-[1,2,3,4,5-13C5]pentanoic acid
13
C5-PFPeA
Perfluoro-n-[1,2-13C2]hexanoic acid
13
C2-PFHxA
Sodium perfluoro-1-[1,2,3-13C3]hexanesulfonate
13
C3-PFHxS
Sodium perfluoro-1-[1,2,3-18O2]hexanesulfonate
18
O2-PFHxS
Perfluoro-n-[1,2,3,4-13C4]heptanoic acid
13
C4-PFHpA
Perfluoro-[1,2-13C4]octanoic acid
13
C4-PFOA
Sodium perfluoro-1-[1,2,3,4-13C4]octanesulfonate
13
C4-PFOS
Perfluoro-n-[13C5]nonanoic acid
13
C5-PFNA
Perfluoro-n-[1,2-13C2]decanoic acid
13
C2-PFDA
1
Target compounds are from the expanded list given in DIN 38407-42 Section 2.1.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 3
Solution Preparation
Ammonia/Methanol Solution
1. Add 400 μL of NH4OH for every 100 mL of methanol to
a clean beaker.
2. Agitate to homogenize.
3. Prepare new solution daily.
Acetate Buffer
1. Measure out 499.5 mL of reagent water in a clean beaker.
2. Add 0.193 g of NH4Ac.
3. Sonicate the solution for 5 minutes until the salt is
fully dissolved.
4. Add 570 μL of glacial acetic acid.
5. Agitate to homogenize the solution.
Working Spiking Solution
1. Dilute 100 μL of the native stock solution with 900 μL of
methanol to achieve a 10 ppt solution.
Summary of SPE Method
SPE Column Format
EVOLUTE® PFAS 500 mg/6 mL or EVOLUTE® PFAS 150 mg/6 mL
Sample Pre-Treatment
Adjust the pH of each sample to 3 using glacial acetic acid. Add
targets and internal standards.
Conditioning
Condition each column with 0.1 % NH4OH in methanol (10 mL)
followed by methanol (10 mL).
Equilibration
Equilibrate each column with reagent water (10 mL).
Sample Loading
Load sample at a flow rate of 5 mL/min.
Wash
Rinse the sample container with acetate buffer solution (10 mL)
and load onto the column. Repeat using reagent water (10 mL).
Dry
Dry the column for 5 minutes at a flow rate of 5 mL/min.
Elution
Rinse the sample container with methanol (5 mL) and use to
elute the analytes from the column at a flow rate of 2 mL/min.
Repeat using 0.1 % NH4OH in methanol (5 mL).
Post Extraction
Concentrate the extract to a volume of 1 mL. Add IS and mix
prior to analysis.
Note: EVOLUTE® PFAS 60 mg/3 mL columns may be used
however sample and solvent volumes should be adjusted.
Refer to DIN 38407-42 for the appropriate amounts.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 4
Sample Preparation Procedure
1. Clean all parts of the Biotage® VacMaster™ system per the
procedure given in Appendix A.
2. Set up and fill new sample containers with water;
250–500 mL are typical for this method.
3. Add glacial acetic acid to each of the sample containers to
reduce the pH to 3 (approximately 100 μL for 250 mL sample
volumes and 200 μL for 500 mL sample volumes).
4. Verify the pH of the sample is 3 using pH paper. To reduce
the possibility of contamination, a duplicate volume was
collected and adjusted to the appropriate pH and the same
volume of acid was added to the sample container.
5. Prepare for the determination of the initial sample volume by
either marking the level of the sample on the container or by
weighing the sample container.
6. Add 20 μL of the undiluted Labeled Stock Solution to each
of the sample containers. If desired, fortify a sample using
target analytes: the addition of 125 μL or 37.5 μL of the native
stock solution will yield either 50 ppt or 15 ppt concentra-
tions respectively, while the addition of 50 μL of the working
spiking solution will yield a 2 ppt concentration. If the mixes
used were different than the ones outlined in this note,
adjust the concentration or spiking amounts as needed.
7. Load the desired EVOLUTE® PFAS columns onto the Biotage®
VacMaster™. Seal any unused positions using VacMaster Port
Sealing Plugs (p/n 121-0005)
8. Rinse each column with 10 mL of 0.1 % NH4OH in methanol
and apply vacuum at 10 mL/min to pull it to waste. Do not
allow the sorbent to go dry.
9. Rinse each column with 10 mL of methanol and apply
vacuum at 10 mL/min to pull it to waste. Do not allow the
sorbent to go dry.
10. Rinse each column with 10 mL of reagent water and apply
vacuum at 10 mL/min to send it to waste. Do not allow the
water level to drop below the top of the packing.
11. Using the Biotage® VacMaster™ LVE Kit, place one end of
the cleaned tubing into the bottom of each of the sample
containers, and secure in position using the clips provided.
12. Load the samples onto the columns using a flow rate of
5 mL/min.
13. Once the sample has been fully loaded, rinse the sample
containers using 10 mL of acetate buffer solution, swirl to
ensure the full rinsing of the container, and load the aliquot
onto the column at a rate of 5 mL/min.
14. Rinse the sample containers using 10 mL of reagent water,
swirl to ensure the full rinsing of the container, and load the
aliquot onto the column at a rate of 5 mL/min.
15. Dry the column for 5 minutes at a rate of 5 mL/min.
16. Load 15 mL centrifuge tubes into the rack corresponding to
each of the column positions and load into the VacMaster.
17. Rinse each sample container using 5 mL of methanol and
swirl to ensure the full rinsing of the container. Load the
aliquot through the appropriate column and collect at a
dropwise rate.
18. Rinse each sample container using 5 mL of 0.1% NH4OH
in methanol and swirl to ensure the full rinsing of the
container. Load the aliquot through the appropriate column
and collect at a dropwise rate.
19. Determine the initial sample volume by either using a gradu-
ated cylinder and filling the sample container to the original
mark or by taking an additional weight of the container.
20. Transfer the centrifuge tubes to the TurboVap® LV system and
concentrate the samples to just under 1 mL using nitrogen
according to the parameters in Table 2.
21. Bring the final extract to 1 mL and transfer to an
autosampler vial.
22. Load the extract onto a calibrated LC-MS/MS system and
process using the conditions given in the below sections.
Table 2. TurboVap® LV Concentration Protocol.
Bath Temp:
60 ˚C
Evaporation Mode
Method (Ramp Gradient)
Manifold Setup
48 positions
Rack Row Height
120 mm*
Step 1:
1.5 L/min for 20 min
Step 2:
3.0 L/min for 15 min
Step 3:
3.5 L/min for 45 min
*The nozzle position was adjusted such that it was as far to the right as
possible to give the user a clear view of the vortex within the tube.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 5
LC-MS/MS Conditions
Agilent 1290 Infinity II LC System
» 1290 Infinity II Multicolumn Thermostat, G7116B
» 1290 Infinity II Multisampler, G7167B
» 1290 Infinity II High Speed Pump, G7120A
» InfinityLab PFC-free HPLC Conversion Kit, 5004-0006
Columns
» InfinityLab PFC Delay Column, 4.6 x 30 mm, p/n 5062-8100
» ZORBAX RRHD Eclipse Plus C18, 95 Å,
2.1 x 50 mm, 1.8 µm, p/n 959757-902
Mobile Phases
» A: 20 mM Ammonium Acetate in Water
» B: Methanol
Table 3. LC Gradient.
Time (min)
%A
%B
0.50
95.00
5.00
3.00
60.00
40.00
16.00
20.00
80.00
18.00
20.00
80.00
20.00
5.00
95.00
20.50
0.00
100.00
25.00
0.00
100.00
26.00
5.00
95.00
» Flow Rate: 0.2 mL/min
» Injection Volume: 5 μL
» Column Temperature: 50 ˚C
Agilent 6470 MS/MS, G6470B
» Gas Temperature: 230 ˚C
» Gas Flow: 4 L/min
» Nebulizer: 20 psi
» Sheath Gas Temperature: 375 ˚C
» Sheath Gas Flow: 12 L/min
» Capillary Voltage (Positive): 3500 V
» Capillary Voltage (Negative): 3500 V
» Nozzle Voltage (Positive): 500 V
» Nozzle Voltage (Negative): 0 V
For a complete listing of MRM Transitions, see Appendix B
Results
System Calibration
For the work being done here, a total of six points were used
in the calibration covering a range of 0.2-20 ppt. The lowest
three points were below the calculated MRL. The curve was
forced through zero and achieved excellent linearity across the
calibration range.
PFBS
PFHxS
Figure 1. Calibration curves for PFBS and PFHxS. Calibration curves for
the remaining target analytes in Table 1 are shown in Appendix C.
Determination of the Minimum Reporting
Level (MRL) and Detection Limits (DL)
A target MRL of 2 ng/L was selected and at least seven replicate
laboratory fortified blanks (LFBs) were created and run at that
concentration. Figure 2 below illustrates the results of this test
for both the 150 mg and 500 mg columns using 250 mL sample
volumes; all compounds were recovered within 15% of the
spiked amount and had less than 10% CV.
Figure 2. MRL and DL Recoveries. Those compounds with an asterisk were
used in salt form.
The data for individual compounds is shown in Appendix D.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 6
Demonstration of Low System Background
An investigation into the background of the complete process
was done in three steps. The first step was to run blank
injections of methanol on the analytical system (system blank).
The second step was to load centrifuge tubes containing a
similar volume of methanol as would result from the extraction
process onto the evaporation system, allowing them to
concentrate and then run on the analytical system (evaporation
blank). The third and final step was to create a full Laboratory
Reagent Blank (LRB), extract and concentrate it, and run it on
the analytical system. By separating the process into three
steps it becomes easier to determine what, if any, contribution
to the overall background each of the steps has. The result of
these tests are given in Appendix E and selected data are shown
below in Figures 3–5.
Figure 3. Contribution of the TurboVap® LV to the PFAS Background.
Those compounds with an asterisk were used in salt form.
Figure 4. PFAS Background for full LRB using EVOLUTE® PFAS 500 mg/6 mL
columns. Those compounds with an asterisk were used in salt form.
Figure 6. Initial Demonstration of Accuracy (15 ng/L, n=4).
Those compounds with an asterisk were used in salt form.
Figure 7. Initial Demonstration of Precision (15 ng/L, n=4). Those
compounds with an asterisk were used in salt form.
Figure 5. PFAS Background for full LRB using EVOLUTE® PFAS 150 mg/6 mL
columns. Those compounds with an asterisk were used in salt form.
For those results which were generated using only the analytical
system, all target analytes were N.D. (unable to be separated
from the noise in the baseline) and so were not listed out in the
previous tables.
When examining the data resulting for both the TurboVap® LV
and the full LRB tests (which includes the Biotage® VacMaster™
manifold, PFAS Free Large Volume Loading Kit, and the
EVOLUTE® cartridges as well as the TurboVap® LV) there are clear
indications of the presence of a PFAS background. However,
even at the highest concentrations detected, all levels are much
lower than the 1/3 MRL limit indicating that the background is
acceptable and will not interfere with future sample runs.
Initial Demonstration of Precision
and Accuracy (IDP, IDA)
To determine the precision and accuracy of the sample prepara-
tion process, four LFB samples were prepared at concentrations
of 15 ppt. The data is given in Appendix F and illustrated in
Figures 6 and 7.
The results show that the average recovery for each target
analyte was within 15% of the nominal value and that the
coefficient of variation (CV) for each analyte fell under 10%
on average.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 7
Examination of System Carryover
To simulate an influent sample, four LFB samples were
created with concentrations which were above the range of
the calibration curve. These samples were extracted, and
the cleanup procedure given in Appendix A was run three
times. To ensure that the system background was adequately
reduced, a set of four LRB samples were extracted immediately
afterwards and analyzed. The LRB data is presented in
Appendix G and illustrated in Figure 8.
Figure 8. Results of carryover study following four, 50 ng/L LFB samples
using EVOLUTE® PFAS 500 mg/6 mL columns. Those compounds with an
asterisk were used in salt form.
Conclusion
With the scrutiny being given to the presence of PFAS
compounds in the environment, it is essential to find reliable
products which can meet the requirements of DIN 38407-42.
This application note has shown that the VacMaster™ vacuum
manifold with PFAS free accessories, EVOLUTE® PFAS SPE
columns and the TurboVap® LV can be used to easily meet and
exceed the demands of the method.
Ordering Information
Part Number
Description
Qty
121-2015ML
Biotage® VacMaster™ 20 Sample Processing
Station With 15 mm Rack
1
121-2190
Biotage® VacMaster™ LVE Kit (PFAS) for 1, 3,
6 mL SPE Columns
1
121-0009-PP
Polypropylene (PFAS) Stopcocks
10
614-0050-CP EVOLUTE® PFAS 500 mg/6 mL columns
30
614-0015-CP
EVOLUTE® PFAS 150 mg/6 mL columns
30
614-0006-BP EVOLUTE® PFAS 60 mg/3 mL columns
50
415000
TurboVap® LV Automated Solvent
Evaporation System
1
414964
TurboVap® LV Multi Rack (48 Positions,
10–20 mm Tubes)
1
The graph shown in Figure 8 shows a clear indication that the
cleaning procedure in Appendix A was successful in reducing
the background of PFAS compounds to below the 1/3 MRL limit.
For further reductions in the background, additional cleaning
steps could be employed.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 8
Appendix A
Biotage® VacMaster™ Cleaning Procedure
For the best results, it is recommended that this procedure be
completed before the use of the VacMaster™ each day and at the
end of each extraction prior to proceeding with the next set of
samples.
1. Ensure that a column and column adapter is installed onto
each VacMaster™ position slated to be cleaned.
2. Fill a clean beaker with 50 mL of methanol and place no more
than four of the LVE Kit lines into the beaker.
3. Apply vacuum to the manifold and pull the methanol through
the positions into the waste container.
4. Remove the column and discard.
5. Using methanol in a squeeze bottle, clean the exterior of the
LVE Kit’s lines, the column adapters, the stopcock, and the
metal cannula. Discard all rinsate.
6. Repeat this up to three times for all positions which
require cleaning.
Note: In situations where the previous sample was highly
concentrated, the above cleaning procedure may need to be
repeated multiple times. If there is concern regarding potential
carryover contamination regardless of the cleaning procedure,
a laboratory reagent blank should be run in that position to
ensure its cleanliness.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 9
Appendix B
MRM Transitions
Table 4. MRM Transitions for Agilent 6470 MS/MS.
Cpd Name
ISTD?
Prec
Ion
MS1 Res
Prod
Ion
MS2 Res
Frag
(V)
CE
(V)
Cell Acc
(V)
Ret Time
(min)
Ret
Window
Polarity
H4PFOS
No
427
Unit/Enh (6490)
406.8
Unit/Enh (6490)
125
24
5
13.3
1.37
Negative
H4PFOS
No
427
Unit/Enh (6490)
80.9
Unit/Enh (6490)
125
40
5
13.3
1.37
Negative
C3-PFHxS
No
402
Unit/Enh (6490)
80
Unit/Enh (6490)
100
49
5
11.7
1.5
Negative
C4-PFBA
No
217
Unit/Enh (6490)
172
Unit/Enh (6490)
60
8
5
5.1
1.45
Negative
C4-PFHpA
No
367
Unit/Enh (6490)
322
Unit/Enh (6490)
72
0
5
11.6
1.2
Negative
C5-PFHxA
No
318
Unit/Enh (6490)
273
Unit/Enh (6490)
70
8
5
9.7
1.14
Negative
C5-PFPeA
No
268
Unit/Enh (6490)
223
Unit/Enh (6490)
60
20
5
7.4
1.55
Negative
C6-PFDA
No
519
Unit/Enh (6490)
474
Unit/Enh (6490)
81
4
5
16.2
1.65
Negative
C8-PFOA
No
421
Unit/Enh (6490)
376
Unit/Enh (6490)
80
8
5
13.4
1.38
Negative
C8-PFOS
No
507
Unit/Enh (6490)
80
Unit/Enh (6490)
100
50
5
15
1.53
Negative
C9-PFNA
No
472
Unit/Enh (6490)
427
Unit/Enh (6490)
66
4
5
14.9
1.52
Negative
PFBA
No
213
Unit/Enh (6490)
168.9
Unit/Enh (6490)
60
8
5
5.1
1.48
Negative
PFBS
No
298.9
Unit/Enh (6490)
98.9
Unit/Enh (6490)
100
29
5
7.9
1.41
Negative
PFBS
No
298.9
Unit/Enh (6490)
80
Unit/Enh (6490)
100
45
5
7.9
1.41
Negative
PFDA
No
513
Unit/Enh (6490)
469
Unit/Enh (6490)
81
4
5
16.2
1.65
Negative
PFDA
No
513
Unit/Enh (6490)
218.7
Unit/Enh (6490)
100
16
5
16.2
1.65
Negative
PFDoA
No
613
Unit/Enh (6490)
569
Unit/Enh (6490)
79
5
5
18.2
1.84
Negative
PFDoA
No
613
Unit/Enh (6490)
268.7
Unit/Enh (6490)
100
20
5
18.2
1.84
Negative
PFDS
No
599
Unit/Enh (6490)
99
Unit/Enh (6490)
100
40
5
17.15
1.75
Negative
PFDS
No
599
Unit/Enh (6490)
80
Unit/Enh (6490)
100
40
5
17.15
1.75
Negative
PFHpA
No
362.9
Unit/Enh (6490)
319
Unit/Enh (6490)
72
0
5
11.6
1.2
Negative
PFHpA
No
362.9
Unit/Enh (6490)
169
Unit/Enh (6490)
72
12
5
11.6
1.2
Negative
PFHpS
No
448.9
Unit/Enh (6490)
98.7
Unit/Enh (6490)
100
44
5
13.5
1.39
Negative
PFHpS
No
448.9
Unit/Enh (6490)
79.7
Unit/Enh (6490)
100
52
5
13.5
1.39
Negative
PFHxA
No
313
Unit/Enh (6490)
268.9
Unit/Enh (6490)
70
8
5
9.7
1.12
Negative
PFHxA
No
313
Unit/Enh (6490)
119
Unit/Enh (6490)
70
18
5
9.7
1.12
Negative
PFHxS
No
398.9
Unit/Enh (6490)
99
Unit/Enh (6490)
100
45
5
11.7
1.5
Negative
PFHxS
No
398.9
Unit/Enh (6490)
80
Unit/Enh (6490)
100
49
5
11.7
1.5
Negative
PFNA
No
463
Unit/Enh (6490)
419
Unit/Enh (6490)
66
4
5
14.9
1.52
Negative
PFNA
No
463
Unit/Enh (6490)
219
Unit/Enh (6490)
66
17
5
14.9
1.52
Negative
PFOA
No
413
Unit/Enh (6490)
369
Unit/Enh (6490)
69
4
5
13.4
1.38
Negative
PFOA
No
413
Unit/Enh (6490)
169
Unit/Enh (6490)
69
12
5
13.4
1.38
Negative
PFOS
No
498.9
Unit/Enh (6490)
99
Unit/Enh (6490)
100
50
5
15
1.53
Negative
PFOS
No
498.9
Unit/Enh (6490)
80
Unit/Enh (6490)
100
50
5
15
1.53
Negative
PFPeA
No
263
Unit/Enh (6490)
218.9
Unit/Enh (6490)
60
8
5
7.4
1.77
Negative
PFUnA
No
563
Unit/Enh (6490)
519
Unit/Enh (6490)
73
5
5
17.2
1.75
Negative
PFUnA
No
563
Unit/Enh (6490)
269
Unit/Enh (6490)
100
20
5
17.2
1.75
Negative
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 10
Appendix C
Calibration Curves
Figure 9. Calibration curves for the target analytes in Table 1, covering a
concentration range of 0.2-20 ppt.
PFBA
PFHpS
H4PFOS
PFOA
PFOS
PFPeA
PFHxA
PFHpA
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 11
PFNA
PFDA
PFDS
PFUnA
PFDoA
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 12
Appendix D
MRL and DL Data
Table 6. MRL and DL Recoveries for EVOLUTE® PFAS 150 mg/6 mL columns.
Conc.
(ng/L)
1
(ng/L)
2
(ng/L)
3
(ng/L)
4
(ng/L)
5
(ng/L)
6
(ng/L)
7
(ng/L)
8
(ng/L)
x̅
(ng/L)
x̅
(%)
s
(ng/L)
CV
(%)
DL
(ng/L)
PFBA*
1.77
1.88
1.83
1.87
1.94
1.83
1.86
1.79
1.83
1.85
104
0.05
2.56
0.14
PFPeA
2.00
1.96
1.92
1.96
2.05
1.97
1.93
1.86
1.96
1.95
98
0.05
2.79
0.16
PFBS
2.00
1.87
1.76
1.84
1.73
1.77
1.81
1.73
1.75
1.78
89
0.05
2.83
0.15
PFHxA
2.00
2.09
2.00
2.04
2.10
2.03
2.09
1.98
2.08
2.05
103
0.05
2.20
0.14
PFHpA
2.00
2.08
1.96
1.98
1.91
2.04
1.95
2.01
2.01
1.99
100
0.06
2.76
0.17
PFHxS*
1.90
1.75
1.83
1.80
1.85
1.66
1.74
1.77
1.74
1.77
93
0.06
3.34
0.18
PFHpS*
1.91
1.77
1.78
1.85
1.79
1.64
1.66
1.69
1.64
1.73
91
0.08
4.69
0.24
H4PFOS*
1.90
1.91
1.84
1.97
1.92
2.05
2.06
2.01
2.11
1.98
104
0.09
4.58
0.27
PFOA
2.00
1.96
1.99
2.01
2.08
1.97
2.05
1.91
1.97
1.99
100
0.05
2.59
0.15
PFOS*
1.92
2.00
1.89
1.93
2.01
1.95
1.95
1.98
1.96
1.96
102
0.04
1.99
0.12
PFNA
2.00
2.08
1.94
2.02
2.01
2.04
2.08
2.11
2.02
2.03
102
0.05
2.59
0.16
PFDA
2.00
1.99
1.89
1.96
1.99
1.91
2.03
2.07
2.04
1.98
99
0.06
3.12
0.19
PFDS
2.00
1.92
2.08
1.81
2.04
2.01
1.93
2.04
1.90
1.97
98
0.09
4.61
0.27
PFUnA
2.00
2.09
1.97
1.93
1.95
1.89
1.97
1.95
1.98
1.97
98
0.06
2.94
0.17
PFDoA
2.00
2.08
1.91
2.02
2.05
2.02
2.03
2.04
2.04
2.02
101
0.05
2.52
0.15
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
Table 5. MRL and DL Recoveries for EVOLUTE® PFAS 500 mg/6 mL columns.
Conc.
(ng/L)
1
(ng/L)
2
(ng/L)
3
(ng/L)
4
(ng/L)
5
(ng/L)
6
(ng/L)
7
(ng/L)
x̅
(ng/L)
x̅
(%)
s
(ng/L)
CV
(%)
DL
(ng/L)
PFBA*
1.77
1.67
1.71
1.67
1.68
1.81
1.60
1.80
1.71
96
0.08
4.43
0.24
PFPeA
2.00
1.85
1.83
1.92
1.97
1.88
1.75
1.91
1.87
94
0.07
3.81
0.22
PFBS
2.00
1.73
1.90
1.82
1.70
1.73
1.65
1.95
1.78
89
0.11
6.12
0.34
PFHxA
2.00
1.80
1.92
1.89
1.80
1.98
1.75
1.92
1.87
93
0.08
4.50
0.26
PFHpA
2.00
1.96
2.06
2.04
1.94
2.01
1.80
2.08
1.99
99
0.09
4.75
0.30
PFHxS*
1.90
1.85
1.78
1.77
1.70
1.82
1.73
1.77
1.77
94
0.05
2.84
0.16
PFHpS*
1.91
1.90
1.90
1.86
1.65
1.92
1.51
1.73
1.78
93
0.16
8.79
0.49
H4PFOS*
1.90
1.80
1.87
1.88
1.86
1.87
1.68
1.86
1.83
96
0.07
4.02
0.23
PFOA
2.00
1.80
1.86
1.93
1.91
1.94
1.67
1.94
1.86
93
0.10
5.34
0.31
PFOS*
1.92
1.96
1.89
1.79
1.89
1.92
1.69
1.89
1.86
97
0.09
4.98
0.29
PFNA
2.00
1.88
1.92
1.95
1.90
1.94
1.79
2.07
1.92
96
0.08
4.30
0.26
PFDA
2.00
1.87
1.89
1.87
1.83
2.01
1.79
2.01
1.89
95
0.09
4.55
0.27
PFDS
2.00
2.10
2.06
1.95
2.14
1.70
1.64
1.93
1.93
97
0.19
9.98
0.61
PFUnA
2.00
1.89
1.91
1.90
1.81
1.88
1.75
1.94
1.87
93
0.07
3.51
0.21
PFDoA
2.00
1.85
1.96
1.96
1.81
2.00
1.75
1.96
1.90
95
0.10
5.04
0.30
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 13
Appendix E
PFAS Background Study
Table 7. Results of PFAS Background Study for EVOLUTE® PFAS 500 mg/6 mL columns (recoveries in ng/L).
TurboVap® LV
Laboratory Reagent Blanks
Replicate
1
2
3
4
5
6
7
8
1
2
3
4
PFBA*
0.00
0.00
0.00
0.00
0.00
0.10
0.05
0.04
0.09
0.06
0.06
0.06
PFPeA
0.02
0.03
0.03
0.03
0.03
0.06
0.02
0.03
0.04
0.04
0.02
0.04
PFBS
0.01
0.00
0.00
0.00
0.01
0.00
0.01
0.01
0.00
0.00
0.00
0.06
PFHxA
0.01
0.02
0.00
0.01
0.00
0.02
0.01
0.02
0.02
0.02
0.02
0.02
PFHpA
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
PFHxS*
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.01
0.00
0.01
0.01
0.01
PFHpS*
0.00
0.02
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
H4PFOS*
0.02
0.00
0.00
0.00
0.00
0.00
0.00
0.01
0.05
0.06
0.07
0.07
PFOA
0.02
0.00
0.00
0.00
0.03
0.00
0.01
0.00
0.00
0.00
0.00
0.00
PFOS*
0.00
0.00
0.00
0.00
0.03
0.00
0.00
0.02
0.01
0.02
0.01
0.01
PFNA
0.00
0.00
0.00
0.01
0.00
0.00
0.00
0.00
0.02
0.00
0.01
0.00
PFDA
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.02
0.00
0.00
0.00
0.00
PFDS
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
PFUnA
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
PFDoA
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
Table 8. Results of PFAS Background Study for EVOLUTE® PFAS 150 mg/6 mL columns (recoveries in ng/L).
TurboVap® LV
Laboratory Reagent Blanks
Replicate
1
2
3
4
1
2
3
4
PFBA*
0.06
0.02
0.01
0.00
0.11
0.09
0.09
0.10
PFPeA
0.03
0.03
0.03
0.02
0.05
0.06
0.04
0.04
PFBS
0.01
0.01
0.00
0.00
0.00
0.01
0.01
0.01
PFHxA
0.02
0.02
0.01
0.00
0.03
0.03
0.02
0.02
PFHpA
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
PFHxS*
0.01
0.01
0.00
0.00
0.01
0.01
0.01
0.01
PFHpS*
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
H4PFOS*
0.01
0.00
0.00
0.00
0.03
0.03
0.04
0.02
PFOA
0.00
0.02
0.03
0.00
0.00
0.00
0.05
0.00
PFOS*
0.00
0.00
0.01
0.01
0.00
0.00
0.00
0.00
PFNA
0.00
0.01
0.00
0.00
0.00
0.00
0.01
0.00
PFDA
0.00
0.02
0.00
0.00
0.00
0.00
0.00
0.02
PFDS
0.00
0.00
0.01
0.00
0.00
0.00
0.00
0.00
PFUnA
0.00
0.00
0.00
0.00
0.02
0.00
0.00
0.00
PFDoA
0.00
0.06
0.00
0.00
0.00
0.00
0.00
0.00
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 14
Appendix F
IDP and IDA Data
Table 9. Results of IDP and IDA for EVOLUTE® PFAS 500 mg/6 mL columns (15 ng/L, n=4).
Replicate
1 (%)
2 (%)
3 (%)
4 (%)
x̅ (%)
s (%)
CV (%)
PFBA*
93.97
93.52
97.66
94.56
94.93
1.87
1.97
PFPeA
91.10
99.84
99.34
92.16
95.61
4.62
4.83
PFBS
88.73
88.20
91.76
86.48
88.79
2.20
2.47
PFHxA
94.83
95.12
99.13
95.85
96.23
1.98
2.06
PFHpA
104.38
106.43
104.54
106.18
105.38
1.07
1.02
PFHxS*
95.20
99.75
101.57
95.96
98.12
3.04
3.10
PFHpS*
89.31
87.18
98.22
79.57
88.57
7.67
8.66
H4PFOS*
96.43
102.56
100.83
101.53
100.34
2.70
2.69
PFOA
94.28
94.77
96.59
92.79
94.61
1.56
1.65
PFOS*
91.58
93.17
98.48
89.66
93.22
3.79
4.06
PFNA
93.76
98.27
100.96
96.89
97.47
2.99
3.07
PFDA
96.19
91.59
101.20
100.51
97.38
4.45
4.57
PFDS
96.50
102.88
108.47
98.56
101.60
5.29
5.21
PFUnA
88.89
85.05
87.96
90.74
88.16
2.37
2.69
PFDoA
96.65
99.00
102.74
98.79
99.30
2.53
2.55
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
Table 10. Results of IDP and IDA for EVOLUTE® PFAS 150 mg/6 mL columns (15 ng/L, n=4).
Replicate
1 (%)
2 (%)
3 (%)
4 (%)
x̅ (%)
s (%)
CV (%)
PFBA*
102.37
101.69
101.80
103.26
102.28
0.72
0.70
PFPeA
101.89
102.16
101.32
100.30
101.42
0.82
0.81
PFBS
99.30
102.15
99.56
102.63
100.91
1.72
1.71
PFHxA
100.58
103.25
99.56
100.27
100.92
1.61
1.60
PFHpA
102.78
102.88
102.64
102.10
102.60
0.35
0.34
PFHxS*
97.07
98.52
98.87
97.40
97.96
0.86
0.88
PFHpS*
91.57
92.08
90.92
93.18
91.94
0.95
1.04
H4PFOS*
99.40
99.12
104.89
102.46
101.47
2.74
2.70
PFOA
104.83
105.70
104.59
106.65
105.44
0.94
0.89
PFOS*
101.18
103.31
101.24
102.55
102.07
1.04
1.02
PFNA
102.52
103.28
101.34
101.57
102.18
0.90
0.88
PFDA
102.85
104.63
101.29
103.65
103.10
1.41
1.37
PFDS
105.46
102.18
103.54
106.38
104.39
1.89
1.81
PFUnA
98.55
97.67
97.27
98.90
98.10
0.76
0.77
PFDoA
105.74
103.29
103.54
105.36
104.48
1.25
1.19
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
© Biotage 2022
Manual Extraction of PFAS in Drinking Water Following DIN 38407-42 | Page 15
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Appendix G
Carryover Data
Table 11. Results of carryover study following four, 50 ng/L LFB samples using EVOLUTE® PFAS 500 mg/6 mL columns.
Replicate
1
(ng/L)
2
(ng/L)
3
(ng/L)
4
(ng/L)
x̅
(ng/L)
PFBA*
0.11
0.09
0.09
0.09
0.09
PFPeA
0.04
0.04
0.04
0.04
0.04
PFBS
0.02
0.01
0.02
0.01
0.02
PFHxA
0.02
0.02
0.03
0.03
0.03
PFHpA
0.00
0.00
0.00
0.00
0.00
PFHxS*
0.02
0.00
0.02
0.01
0.01
PFHpS*
0.00
0.00
0.00
0.00
0.00
H4PFOS*
0.04
0.04
0.04
0.04
0.04
PFOA
0.09
0.04
0.07
0.00
0.05
PFOS*
0.04
0.03
0.04
0.04
0.04
PFNA
0.00
0.00
0.00
0.00
0.00
PFDA
0.02
0.02
0.02
0.02
0.02
PFDS
0.03
0.00
0.00
0.00
0.01
PFUnA
0.00
0.00
0.03
0.01
0.01
PFDoA
0.00
0.00
0.00
0.00
0.00
*Analytes were used in salt form and calculated concentrations were corrected to compensate where needed.
