1. Rapid Analysis of Perfluorinated Compounds Using Triple Quadrupole LC/MS/MS
Brahm Prakash Tairo Ogura and William Lipps
Shimadzu Scientific Instruments, Inc., Columbia MD
Promise not to call on anyone.
“How many can differentiate between chromatographic resolution and mass resolution?”
“How many here are very comfortable with their understanding of MRM, or an MRM transition on an LCMS TQ?”
“How many here understand why a TQ is more sensitive than a SQ even though ion intensity goes down?”
“How many would enjoy a debate on whether ESI is solution phase chemistry or gas phase chemistry?”
“How many have extensive TQ operation experience?”
“How many have run PFCs on a TQ?”

2. EPA Method 537 is an older method, uses SPE and does not take advantage of the power of modern LCMSMS

3. ASTM D7979 Extraction procedure eliminates SPE step
5 ml Sample
Surrogate
5 ml MeOH
EPA Method 537 is a solid phase extraction method. In this method you collect sample in a 250 milliliter bottle. Weigh the bottle. Add surrogate to the bottle then pass through the SPE cartridge at 10 milliliters per minute (This will take 25 minutes at minimum). Then rinse the bottle with reagent water and pass that through the SPE cartridge. Then add 4 milliliters of methanol to the sample bottle, rinse the sides, and pass the methanol dropwise through the SPE cartridge. This will take about one hour. Place the extract in a 60 C bath and evaporate (under nitrogen) the methanol extract to dryness. Add 1 milliliter of 96% methanol in reagent water. Add internal standard and analyze. Reweigh the bottle empty to calculate sample volume.

4. ASTM D7979 Extraction procedure
LCMS-8060
10 µL Acetic Acid
EPA Method 537 is a solid phase extraction method. In this method you collect sample in a 250 milliliter bottle. Weigh the bottle. Add surrogate to the bottle then pass through the SPE cartridge at 10 milliliters per minute (This will take 25 minutes at minimum). Then rinse the bottle with reagent water and pass that through the SPE cartridge. Then add 4 milliliters of methanol to the sample bottle, rinse the sides, and pass the methanol dropwise through the SPE cartridge. This will take about one hour. Place the extract in a 60 C bath and evaporate (under nitrogen) the methanol extract to dryness. Add 1 milliliter of 96% methanol in reagent water. Add internal standard and analyze. Reweigh the bottle empty to calculate sample volume.

5. In Water, Sludge, Influent, Effluent and Wastewater
Method Validation: Direct Inject Method ASTM D Standard Test Method for Determination of Perfluorinated Compounds by LCMS – TQ8060
In Water, Sludge, Influent, Effluent and Wastewater
500ng/L, high calibration point (XRODS-3x50x2.4_PhenylHexyl 2.1x100x3 u)
Chromatogram Obtained under optimized conditions – only half of the target compounds were spiked, awaiting other standards to arrive.
Chromatogram Obtained under optimized conditions

6. Glass Vial Poly Propylene
Standard Stability Study GLASS Vials Vs Poly Propylene Vials- 10% Methanol 90% water
Glass Vial
Poly Propylene

7. Poly Propylene Vials Glass Vials
Standard Stability Study GLASS Vials Vs Poly Propylene Vials- 30% Methanol 70% water
Poly Propylene Vials
Glass Vials

8. Glass Vials Poly Propylene Vials
Standard Stability Study GLASS Vials Vs Poly Propylene Vials- 50% Methanol 50% water
Glass Vials
Poly Propylene Vials

9. Glass Vials Poly Propylene Vials
Standard Stability Study GLASS Vials Vs Poly Propylene Vials- 70% Methanol 30% water
Glass Vials
Poly Propylene Vials

10. Glass Vials Poly Propylene Vials
Standard Stability Study GLASS Vials Vs Poly Propylene Vials % Methanol 10% water
Glass Vials
Poly Propylene Vials

11. Sample Vials - Vortex Effect
PFCs Could Float on the top of Sample Vials and be not injected… resulting in poor recoveries or could go as non detect.
Same Vial After Vortex
Before Vortex

12. Optimization of Interface Temperature, Desolvation line (DL) Temperature and heat Block Temperature
IF100_DL150_HB250 oC

13. Optimization of Interface Temperature, Desolvation line (DL) Temperature and heat Block Temperature
IF200_DL150_HB250 oC

14. Optimization of Interface Temperature, Desolvation line (DL) Temperature and heat Block Temperature
IF300_DL150_HB250

15. IF300_DL100_HB200 : Optimized Conditions
Optimization of Interface Temperature, Desolvation line (DL) Temperature and heat Block Temperature
IF300_DL100_HB200 : Optimized Conditions

16. Optimization of Interface Temperature, Desolvation line (DL) Temperature and heat Block Temperature
IF400_DL300_HB400

17. Target List Including Newly Added by Office of Superfund Remediation and Technology Innovation (OSRTI)

18. In Water, Sludge, Influent, Effluent and Wastewater
Direct Inject Method ASTM D Standard Test Method for Determination of Perfluorinated Compounds by LCMS – TQ8060 : Chromatogram at 100 ppt Using Optimized Conditions
In Water, Sludge, Influent, Effluent and Wastewater
100ng/L, high calibration point (XRODS-3x50x2.4_PhenylHexyl 2.1x100x3 u)

19. Direct Inject Method ASTM D Standard Test Method for Determination of Perfluorinated Compounds by LCMS – TQ8060 : Chromatogram at 1.0 ppt Using Optimized Conditions
1.0 ng/L, high calibration point (XRODS-3x50x2.4_PhenylHexyl 2.1x100x3 u)

20. ASTM D7979 by 10 uL Direct Injection at 40 ppt Using LCMS– TQ8060 MFPBA at 40 ppt
Level
Concentration 1
5 ppt 2
10 ppt 3
20 ppt 4
40 ppt 5
60 ppt 6
80 ppt 7
100 ppt 8
150 ppt 9
200 ppt
0.2
0.5
1.0
2.0
5.0
10.0
25.0
50.0
75.0
100.0
Calibration Curves
Calibration curves were produced in the range of 5 ng/L to 200 ng/L for the PFCs.
All curves had a regression coefficient higher than Curves for a selection of target compounds are plotted

21. ASTM D7979 by 10 uL Direct Injection at 40 ppt Using LCMS– TQ8060 PFPeS at 40 ppt
Level
Concentration 1
5 ppt 2
10 ppt 3
20 ppt 4
40 ppt 5
60 ppt 6
80 ppt 7
100 ppt 8
150 ppt 9
200 ppt
0.2
0.5
1.0
2.0
5.0
10.0
25.0
50.0
75.0
100.0
Calibration Curves
Calibration curves were produced in the range of 5 ng/L to 200 ng/L for the PFCs.
All curves had a regression coefficient higher than Curves for a selection of target compounds are plotted

22. ASTM D7979 by 10 uL Direct Injection at 40 ppt Using LCMS– TQ8060 PFNA at 40 ppt
Level
Concentration 1
5 ppt 2
10 ppt 3
20 ppt 4
40 ppt 5
60 ppt 6
80 ppt 7
100 ppt 8
150 ppt 9
200 ppt
0.2
0.5
1.0
2.0
5.0
10.0
25.0
50.0
75.0
100.0
Calibration Curves
Calibration curves were produced in the range of 5 ng/L to 200 ng/L for the PFCs.
All curves had a regression coefficient higher than Curves for a selection of target compounds are plotted

23. ASTM D7979 by 10 uL Direct Injection at 40 ppt Using LCMS– TQ8060 PFTRE at 40 ppt
Level
Concentration 1
5 ppt 2
10 ppt 3
20 ppt 4
40 ppt 5
60 ppt 6
80 ppt 7
100 ppt 8
150 ppt 9
200 ppt
0.2
0.5
1.0
2.0
5.0
10.0
25.0
50.0
75.0
100.0
Calibration Curves
Calibration curves were produced in the range of 5 ng/L to 200 ng/L for the PFCs.
All curves had a regression coefficient higher than Curves for a selection of target compounds are plotted

24. ASTM D7979 by 10 uL Direct Injection at 40 ppt Using LCMS– TQ8060 PFTreA at 40 ppt
Level
Concentration 1
5 ppt 2
10 ppt 3
20 ppt 4
40 ppt 5
60 ppt 6
80 ppt 7
100 ppt 8
150 ppt 9
200 ppt
0.2
0.5
1.0
2.0
5.0
10.0
25.0
50.0
75.0
100.0
Calibration Curves
Calibration curves were produced in the range of 5 ng/L to 200 ng/L for the PFCs.
All curves had a regression coefficient higher than Curves for a selection of target compounds are plotted

25. MRL’s and MDL’s for ASTM D7979- Shimadzu LCMS-TQ8060
Minimum Reporting Level (ng/L) n=8
Compound
Ret. Time
Fortified Conc. (ng/L)
MDL (ng/L)
Calibration Range (ng/L)
PFBA 5
4.1
MPFBA
5.0
PFPeA
0.9
M5PFPeA
0.6
4-2 FTS
1.7
M4-2 FTS
1.2
PFHxA
1.3
M4PFHxA
1.1
PFBS
1.5
M3PFBS
FHUEA
2.6
FHEA
100
32.5
PFHpA
1.4
M4PFHpA
0.7
PFPeS
6-2 FTS
2.5
M6-2 FTS
1.8
PFOA
5.1
M8PFOA

26. MRL’s and MDL’s for ASTM D7979- LCMS-TQ8060
Minimum Reporting Level (ng/L) n=8
Compound
Ret. Time
Fortified Conc. (ng/L)
MDL (ng/L)
Calibration Range (ng/L)
FhpPA 5
9.4
5 -200
FOEA
100
48.3
FOUEA
1.6
PFHxS
1.5
M3PFHxS
1.7
PFNA
M9PFNA
8-2 FTS
3.2
M8-2 FTS
1.8
PFHpS
N-MeFOSAA
3.6
MN-MeFOSAA
5.4
PFDA
2.3
M6PFDA
1.1
FDEA
35.5
N-EtFOSAA
5.3
MN-EtFOSAA
4.2
PFOS
3.0
M8PFOS

27. MRL’s and MDL’s for ASTM D7979- LCMS-TQ8060
Minimum Reporting Level (ng/L) n=8
Compound
Ret. Time
Fortified Conc. (ng/L)
MDL (ng/L)
Calibration Range (ng/L)
PFUnA 5
2.9
M7PFUnA
1.5
PFNS
1.3
PFDoA
2.2
MPFDoA
0.8
FOSA
0.6
M8FOSA
1.6
PFDS
2.1
PFTriA
1.1
PFTreA
M2PFTreA
0.7

28. Method Detection Limit and Reporting Ranges ASTM Vs Shimadzu
Shimadzu LCMSMS-TQ8060
Analyte MDL(ng/L) Ranges
MDL(ng/L) Ranges (ng/L
PFTreA – –200
PFTriA –
PFDoA –
PFUnA
PFDA
PFDS –
PFOS –
PFNA –
PFNS –
PFOA –
PFHpS –

29. Method Detection Limit and Reporting Ranges ASTM Vs Shimadzu
Shimadzu LCMSMS-TQ8060
Analyte MDL(ng/L) Ranges (ng/L) MDL (ng/L) Ranges (ng/L)
PFHxS –
PFHpA –
PFHxA
PFBS –
PFPeS –
PFPeA –
PFBA
FOSA –
4:2 FTS –
6:2 FTS –
8:2 FTS –

30. Method Detection Limit and Reporting Ranges ASTM Vs Shimadzu
Shimadzu LCMSMS-TQ8060
Analyte MDL(ng/L) Ranges (ng/L) MDL (ng/L) Ranges (ng/L)
FHEA –
FOEA –
FDEA –
FOUEA –
FPpPA –
FHUEA –

31. Comparison MDL Data EPA M537 Vs Shimadzu Results Obtained using LCMS-TQ8060
Compound
EPA 537
Spike (ng/L)
Sample Extract vol250 mL
EPA M537
MDL (ng/L)
LCMRL(ng/L)
Shimadzu
LCMS-8060 Spike (ng/L)
LCMS-8060
DL (ng/L)
PFBA
9.1 5
4.1
PFPeA
0.9
PFHxA
5.0
0.5
2.9
1.5
PFBS
1.6
3.7
1.4
PFHpA
3.1
3.8
1.3
PFOA
4.6
1.7
5.1
PFNA
4.8
0.7
5.5
PFDA
2.3
PFOS
9.6
6.5
3.0
PFUnA
5.4
2.8
6.9
PFDoA
1.1
3.5
2.1
PFTriA
2.2
PFTreA
4.7
Qwing to the high sensitivity of the LCMS-TQ8060 system these low ng/L levels were easily obtained for all compounds

32. Precision & Accuracy for ASTM D7979 Using LCMS-TQ8060
Precision & Accuracy at 10ng/L (n=8)
Compound
Mean Concentration (ng/L)
% Recovery
% RSD
PFBA 16
164
10.5
MPFBA 11
111
9.0
PFPeA 10
104
4.7
M5PFPeA
101
3.1
4-2 FTS
103
3.4
M4-2 FTS 9 93
3.5
PFHxA
105
6.9
M4PFHxA
102
2.1
PFBS
9.6
M3PFBS
106
4.8
FHUEA
112
5.5
FHEA
211
PFHpA
7.5
M4PFHpA
2.7

33. Precision & Accuracy for ASTM D7979 Using Shimadzu LCMS-TQ8060
Precision & Accuracy at 20ng/L (n=8)
Compound
Mean Concentration (ng/L)
% Recovery
% RSD
PFBA
22.4
112
6.6
MPFBA
17.3 86
10.2
PFPeA
20.1
101
2.9
M5PFPeA
20.0
100
1.4
4-2 FTS
20.3
102
3.2
M4-2 FTS
18.4 92
3.0
PFHxA
20.2
3.9
M4PFHxA
2.3
PFBS
10.4
M3PFBS
19.6 98
4.1
FHUEA
21.6
108
5.6
FHEA
397.5 99
5.3
PFHpA
20.5
M4PFHpA
19.7

34. Summary and Conclusions
Under Optimized conditions-The Shimadzu LCMS-TQ8060 will exceed and meet performance criteria specified for EPA Method 537
Results demonstrated that high-sensitivity analysis with high repeatability is possible with Shimadzu’s triple quadrupole instruments
The method presented here used an GL Science InertSustain Phenylhexyl 3um 2.1x100mm C/N analytical column and a gradient that was designed to increase method throughput , while still providing sufficient chromatographic resolution.
Lower MRLS for most PFCs is possible using advanced state-of-the art technologies
Recommendation
Due to high-sensitivity performance of the LCMS-TQ8060, it is recommended for use for methods allowing direct injections of PFCs, ASTM D7979

35. Study Continues …on Method Development for PFCs
Thank You for attending…
For questions and to know more details about the Method conditions please contact
Brahm Prakash Phone: