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Download tech note (PDF)
Rapid, low-level analysis using the SCIEX 7500 system
Per- and polyfluoroalkyl substances (PFAS) have excellent stain repellent properties and are thus widely used in packaging materials, firefighting foams and many other uses. As PFAS have been found in virtually every environmental matrix including drinking water supplies, making monitoring critical to ensure safety. Here, a comprehensive quantitative method for a suite of PFAS compounds was developed for analysis of water samples on the SCIEX 7500 system. This high sensitivity system provided S/N gains of ~5.5x on average across the 54 compounds analyzed versus the SCIEX 6500+ system.
Per- and polyfluoroalkyl substances (PFAS) are synthetic chemical compounds with properties that have excellent resistance to heat, water, oil and grease. PFAS are widely used in food and other product packaging materials, as water and stain repellents, in firefighting foams and within many industrial processes.1 There is now ample evidence that exposure to high levels of PFAS can lead to adverse health outcomes in humans.2 Because PFAS bio-accumulate, the risk is even more profound.
Studies over the last two decades have shown that PFAS can be found in virtually every environmental matrix. Many municipal drinking water supplies within the US and around the world contain PFAS contamination. As a result, the EU has set regulations for acceptable levels of PFAS.3 While the US Environmental Protection Agency (EPA) is still establishing maximum contaminant levels (MCLs) for PFAS, it has issued health advisories. To better evaluate the risks of PFAS to human health, it is important to monitor their levels within humans, water supplies and the environment.
In this technical note, the SCIEX 7500 system was evaluated for the quantification of a comprehensive suite of PFAS compounds in a sample of surface water. This system was designed to enable new levels of sensitivity for the quantification of previously undetectable compounds. Results from the SCIEX 7500 system were compared with results from the SCIEX Triple Quad 6500+ system, an established platform that is routinely used for low-level quantification of PFAS compounds. Figure 1 shows the comparison of a representative PFAS compound, perfluorohexanesulfonic acid (PFHxS).
Figure 1. MRM peaks for perfluorohexanesulfonic acid (PFHxS). The SCIEX 7500 system data (right) shows a 7.1x increase in S/N compared with the SCIEX Triple Quad 6500+ system data (left). Note: gray shaded areas represent the noise region used for the S/N calculation.
Sample preparation: A surface water sample and a low-level standard (50 ppt) were analyzed in this study. The standard consisted of a comprehensive suite of PFAS compounds, including perfluorinated carboxylic acids (PFCAs), fluorotelomer sulfonates (FTSs), sulfonamides and fluoroether acids (Wellington Laboratories, Guelph, ON). Table 1 contains a list of all compounds studied. The surface water sample was prepared by extraction of 500 mL of water through weak anion exchange (WAX) SPE cartridges. The eluent was reduced in volume under N2 gas and reconstituted in an 80:20 methanol and water mixture. Five replicates of the standard and 3 replicates of the sample were analyzed.
Chromatography: An ExionLC system was plumbed with a Phenomenex Gemini C18 (5 μm, 3 x 50 mm) column between the LC pumps and the autosampler. This column temporarily retains contaminants from the mobile phases and LC system and elutes them later in the gradient.5 A Phenomenex Gemini C18 (3 µm, 3 x 100 mm) column was used as the analytical column. An LC flow rate of 500 µL/min was used along with a 1 µL injection volume. Mobile phases were water and methanol, both amended with 10 mM ammonium acetate. The LC gradient is shown in Figure 2.
Figure 2. Flow program for analysis of PFAS compounds.
Mass spectrometry: A SCIEX 7500 system was used with an OptiFlow Pro ion source and an electrospray ionization (ESI) probe in negative ion mode. The source and MS parameters were as follows: TEM = 400°C, GS1 = 40, GS2 = 70, ISV = -2000, CUR = 40, CAD = 14. For comparison purposes, a SCIEX Triple Quad 6500+ system was used with the IonDrive Turbo V ion source. Primary (and, in most cases, secondary) multiple reaction monitoring (MRM) transitions were monitored for each PFAS compound based upon previous studies and were identical for both instruments. Each system was optimized for maximum sensitivity.
Data processing: Data acquisition and automatic data processing were performed using SCIEX OS software and analytics in SCIEX OS software. Flagging rules were specified within the analytics module to quickly flag good and bad data.
The SCIEX 7500 system was designed to push sensitivity boundaries beyond those previously established with the high-end SCIEX Triple Quad 6500+ system. These sensitivity improvements are realized through several areas.4 For example, the E Lens probe of the OptiFlow Pro ion source more efficiently breaks up droplets, releases ions and actively drives ions toward the orifice. The 4.3x increase in the area of the entrance orifice allows more ions to enter the instrument, improving ion sampling. The D Jet ion guide more efficiently captures and focuses ions from the ESI spray plume, allowing more ions to traverse the instrument to be analyzed and detected. All of these enhancements improve the total number of ions available for analysis in the system. Since the noise does not increase proportionally, the S/N ratio increases, quite dramatically for some compounds, for SCIEX 7500 system data.
Because of these changes in source and entrance optics, PFAS compounds were first evaluated using a range of source and entrance parameters to find optimum conditions. Although there were some dependencies on source temperature and ionization voltage as a function of PFAS class and chain length, conditions were found that produced good signal for all compounds. Interestingly, lower voltages were generally required using the OptiFlow Pro ion source than previous source designs due to the function of the E Lens probe.
The sensitivities of the SCIEX Triple Quad 6500+ system and the SCIEX 7500 system were evaluated by injecting the low-level standard onto each system. Table 1 lists all of the PFAS compounds contained within the low-level standard. The S/N was automatically calculated within SCIEX OS software using the “peak-to-peak” algorithm and is listed for each MRM transition. The table also shows the improvement factor in S/N for the SCIEX 7500 system versus the SCIEX Triple Quad 6500+ system. Improvements in S/N for the SCIEX 7500 system were on average around 5.5x but ranged anywhere from ~1x to more than 50x.
Figure 1, Figure 3 and Figure 4 show MRM peaks obtained for 3 representative PFAS compounds on the SCIEX 7500 system compared with the SCIEX Triple Quad 6500+ system. Integrated peak areas are shaded in purple while the gray highlighted areas represent the noise used by the S/N algorithm.
For the SCIEX 7500 system data, peak areas and total peak intensities are greater while the noise is maintained at relatively low levels, resulting in increased S/N in all cases. This ultimately results in improved sensitivity and lower limits of quantification (LOQ). For example, as shown in Figure 4, the increased S/N using the SCIEX 7500 system enables quantification of important, lower-level isomers for N-ethyl perfluorooctane sulfonamide (EtFOSA).
Table 1. Signal-to-noise for each PFAS MRM transition. Each PFAS was measured (using both the SCIEX Triple Quad 6500+ system and the SCIEX 7500 system) and the S/N was calculated. The “7500/6500+” column shows the improvement factor for the SCIEX 7500 system, ranging from ~1 to >50.
Figure 3. MRM peaks for perfluorooctanoic acid (PFOA). The SCIEX 7500 system data (right) shows a 13.5x increased S/N compared with the SCIEX Triple Quad 6500+ system data (left). Note: gray shaded areas represent the noise region used for the S/N algorithm.
Figure 4. MRM peaks for N-ethyl perfluorooctane sulfonamide (EtFOSA). The SCIEX 7500 system data (right) shows a 3.8x increased S/N compared with the SCIEX Triple Quad 6500+ system data (left). The increased S/N enables quantification of important lower-level EtFOSA isomers (red circle) that are missed using the SCIEX Triple Quad 6500+ system. Note: gray shaded areas represent the noise region used for the S/N algorithm.
Five replicate injections were made for the low-level standard and %CVs were calculated. As shown in Figure 5, precision was excellent with CVs of less than 10% for the SCIEX 7500 system data.
For the SCIEX Triple Quad 6500+ system data, CVs were generally less than 20%, with some outliers ranging as high as 30%. The lower %CV of the SCIEX 7500 system represents improved precision and therefore improved data quality near the lower LOQ.
Figure 5. Precision for 5 replicate injections of the low-level standard. The SCIEX 7500 system had CVs of <10% for all PFAS transitions (orange bars) while the CVs for the SCIEX Triple Quad 6500+ system were generally under 20%, but with some outliers ranging up to 30% (blue bars).
The surface water sample was analyzed in triplicate on both the SCIEX Triple Quad 6500+ system and the SCIEX 7500 system. As shown in Figure 6, Figure 7 and Figure 8, the higher sensitivity of the SCIEX 7500 system resulted in higher signals and greater S/N for all compounds, transitions and replicates.
For example, this increased sensitivity enabled the detection of low-level isomers of N-methylperfluorooctane sulfonamidoacetic acid, or MeFOSAA (Figure 7), and it increased confidence in the detection of perfluorotridecanoic acid, or PFTrA (Figure 8), a hydrophobic compound that normally would not be expected to be present in surface water.
Figure 6. Quantification of perfluoropentane sulfonic acid (PFPeS) in a surface water sample. Triplicate analysis comparing the SCIEX Triple Quad 6500+ system (top) and the SCIEX 7500 system (bottom). The higher S/N observed with the SCIEX 7500 system leads to higher precision and confidence in the analysis.
Figure 7. Quantification of N-methylperfluorooctane sulfonamidoacetic acid (MeFOSAA) in a surface water sample. Triplicate analysis comparing the SCIEX Triple Quad 6500+ system (top) and the SCIEX 7500 system (bottom). The higher S/N observed with the SCIEX 7500 system leads to the detection of low-level isomers in the analysis.
Figure 8. Quantification of perfluorotridecanoic acid (PFTrA) in a surface water sample. Triplicate analysis comparing the SCIEX Triple Quad 6500+ system (top) and the SCIEX 7500 system (bottom). The higher S/N observed with the SCIEX 7500 system leads to the detection of unexpected low-level analytes.
The increased sensitivity of the SCIEX 7500 system allows a new level of performance for the analysis of PFAS compounds.
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