Featured Application: Decrease Analysis Time for Fluorochemicals with no Sacrifice in Chromatographic Performance.

Rapid Perfluorinated Alkyl Acid Analysis by LC-MS/MS Increases Sample Throughput


  • Raptor C18 SPP 5 µm core-shell silica particle columns offer excellent resolution for fluorochemicals with short total cycle times.
  • Meets EPA Method 537 requirements.
  • Unique, robust Raptor C18 column design increases instrument uptime.

Perfluorinated alkyl acids are man-made fluorochemicals used as surface-active agents in the manufacture of a variety of products, such as firefighting foams, coating additives, textiles, and cleaning products. They have been detected in the environment globally and are used in very large quantities around the world. These fluorochemicals are extremely persistent and resistant to typical environmental degradation processes. As a result, they are widely distributed across the higher trophic levels and are found in soil, air, groundwater, municipal refuse, and landfill leachates. The toxicity, mobility, and bioaccumulation potential of perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA), in particular, pose potential adverse effects for the environment and human health.

Perfluorinated alkyl acid analysis can be challenging because these compounds are chemically different from most other environmental contaminants. They are difficult to quantify because some are more volatile than others, and they also tend to be more hydrophilic and somewhat reactive. In addition, fluorochemicals are present in polytetrafluoroethylene (PTFE) materials, so excluding the use of any PTFE labware throughout the sampling and analytical processes (including HPLC solvent inlet tubing) is essential for accurate analysis. Typically, perfluorinated alkyl acids are analyzed by LC-MS/MS methods, such as EPA Method 537, but long analysis times can significantly limit sample throughput.

As written, the EPA 537 requires a 27-minute cycle per sample, but the method does allow flexibility in the column used as long as there is sufficient resolution for the MS dwell time for all compounds in a specific retention time window. As shown here, all target perfluorinated alkyl acids were analyzed on a Raptor C18 column with a total cycle time of 9 minutes—resulting in a three-fold faster analysis time than the EPA method. While this analysis is significantly faster, there is no sacrifice in peak resolution or selectivity, meaning all fluorochemicals are easily identified and they elute as highly symmetrical peaks that can be accurately integrated and quantified by MS/MS. In addition, this method resolves branched isomers of PFHxS and PFOS to satisfy EPA 537 method requirements. If PFOA and PFOS are the only target fluorochemicals, the chromatographic conditions can be further optimized. A fast, <2-minute separation with a total cycle time of just 4.5 minutes can be achieved with a 2.1 x 100 mm column as shown in the second chromatogram. Alternatively, a 2.1 x 50 mm column can be used for an even faster analysis.

Whether labs conducting perfluorinated alkyl acid analysis by LC use longer target analyte lists or focus just on PFOA and PFOS, the excellent peak shapes and separations achieved here result in consistent, accurate quantification with much shorter analysis times. By switching to a Raptor C18 column, labs can process more samples per hour while still meeting fluorochemical method requirements.


PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct Ion
1.Perfluorobutanesulfonic acid (PFBS)1.9810299.1079.99
2.Perfluoro-n-[1,2-13C2]hexanoic acid (13C-PFHxA)2.9210315.13270.13
3.Perfluorohexanoic acid (PFHxA)2.9310313.10269.12
4.Perfluoroheptanoic acid (PFHpA)4.005363.16319.09
5.Perfluorohexanesulfonic acid (PFHxS)4.1410399.1379.98
6.Perfluoro-[1,2-13C2]octanoic acid (13C-PFOA)4.835415.13370.10
7.Perfluorooctanoic acid (PFOA)4.835413.16369.10
8.Perfluorononanoic acid (PFNA)5.505463.16419.19
9.Perfluoro-1-[1,2,3,4-13C4]octanesulfonic acid (13C-PFOS)5.545503.1379.98
10.Perfluorooctanesulfonic acid (PFOS)5.5410499.1779.98
11.Perfluoro-n-[1,2-13C2]decanoic acid (13C-PFDA)6.065515.17470.17
12.Perfluorodecanoic acid (PFDA)6.075513.17469.16
13.N-deuteriomethylperfluoro-1-octanesulfonamidoacetic acid (d3-NMeFOSAA)6.285573.23419.15
14.N-methyl perfluorooctanesulfonamidoacetic acid (NMeFOSAA)6.285570.20419.17
15.N-deuterioethylperfluoro-1-octanesulfonamidoacetic acid (d5-NEtFOSAA)6.525589.27419.11
16.N-ethyl perfluorooctanesulfonamidoacetic acid (NEtFOSAA)6.525584.20419.18
17.Perfluoroundecanoic acid (PFUnA)6.565563.23519.24
18.Perfluorododecanoic acid (PFDoA)6.985613.23569.19
19.Perfluorotridecanoic acid (PFTrDA)7.355663.23619.21
20.Perfluorotetradecanoic acid (PFTA)7.675713.23669.23
Perfluorinated Alkyl Acids on Raptor C18 by LC-MS/MS for EPA Method 537
LC_EV0548
ColumnRaptor C18 (cat.# 9304552)
Dimensions:50 mm x 2.1 mm ID
Particle Size:5 µm
Pore Size:90 Å
Temp.:40 °C
Sample
Diluent:96:4 Methanol:water
Conc.:5–10 ng/mL
Inj. Vol.:2 µL
Mobile Phase
A:5 mM ammonium acetate in water
B:Methanol
Time (min)Flow (mL/min)%A%B
0.000.47525
8.000.41585
8.010.47525
9.000.47525
DetectorMS/MS
Ion Source:Electrospray
Ion Mode:ESI-
Mode:MRM
InstrumentWaters Acquity

PeakstR (min)Conc.
(ng/mL)
Precursor IonProduct Ion
1.Perfluoro-[1,2-13C2]octanoic acid (13C-PFOA)1.3810414.8369.8
2.Perfluorooctanoic acid (PFOA)1.385412.7368.8
3.Perfluoro-1-[1,2,3,4-13C4]octanesulfonic acid (13C-PFOS)1.6510502.779.9
4.Perfluorooctanesulfonic acid (PFOS)1.655498.779.9
Perfluorooctanoic Acid (PFOA) and Perfluorooctanesulfonic Acid (PFOS) on Raptor C18 by LC-MS/MS
LC_EV0536
ColumnRaptor C18 (cat.# 9304512)
Dimensions:100 mm x 2.1 mm ID
Particle Size:5 µm
Pore Size:90 Å
Temp.:40 °C
Sample
Diluent:Water:methanol (50:50)
Conc.:5-10 ng/mL
Inj. Vol.:5 µL
Mobile Phase
A:5 mM ammonium acetate in water
B:Methanol
Time (min)Flow (mL/min)%A%B
0.000.44060
2.500.4595
2.510.44060
4.500.44060
DetectorMS/MS
Ion Mode:ESI-
Mode:MRM
InstrumentUHPLC

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