1. Analysis of Per/Polyfluoroalkyl Substances (PFAS) in Drinking Water Using the Agilent Ultivo Triple Quadrupole LC/MS
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11/29/2018
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Terri Sosienski, Ph.D.
LC/MS Applications Scientist
Agilent Technologies, Santa Clara, CA
Tarun Anumol, Ph.D.
Agilent Technologies, Wilmington, DE
Dorothy Yang, Ph.D.

2. Per/Poly Fluoroalkyl Substances (PFAS) in Water
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PFASs widely used and can be sourced from:
Fire-fighting foams
Non-stick cookware
Food packaging
Manufacturing
USEPA drinking water health advisories for PFOA and PFOS 70 ng/L in drinking water
Individual US State guidelines 13 – 32 ng/L PFOS and PFOA in water.
US EPA Method 537 14 PFAS compounds
PFAS are very common compounds which aren’t limited to their use as surfactants fire retardants, and even in non-stick cookware.
They are consistently found in environmental waters, as they are very stable compounds which do not break down nor degrade easily.
There are some regulatory guidelines for PFOA and PFOS levels in drinking water set at 70 nanograms per liter
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3. Overview
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Two methods for detecting PFAS compounds in drinking water using the Ultivo LC/TQ
Method 1
17 PFAS Compounds
EPA 537 sample preparation
Method 2
51 Trace Organic Contaminants Including 10 PFAS compounds
Online SPE
PFAS are very common compounds which aren’t limited to their use as surfactants fire retardants, and even in non-stick cookware.
They are consistently found in environmental waters, as they are very stable compounds which do not break down nor degrade easily.
There are some regulatory guidelines for PFOA and PFOS levels in drinking water set at 70 nanograms per liter
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4. The New Ultivo Triple Quad LC/MS
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5. Ultivo: Space-saving, Sensitive, Robust & Powerful
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HED Detector
(accessible in stack)
VacShield
(no-vent maintenance)
Vortex Collision Cell
(twisted hexapole)
Cyclone Ion Optics
(dual twisted hexapoles)
Agilent Jet Stream Ion Source
Hyperbolic Quadrupoles
Virtual DC Filters
(ultra thin)
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6. Ultivo – Reduced Downtime
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Fast Autotune <10 minutes!
Vac Shield: Change capillary without venting
Accessible maintenance features
Some of the other innovations included in ULTIVO were made to reduce the instrument downtime.
We have a new swarm tune that can autotuned the instrument in less than 10 minutes!
Our new vac shield technology allows the user to change the capillary without venting the instrument. Here we have a rendering of the ion injector installed with the vac shield open, and the ion injector removed with the vac shield closed, maintaining the vacuum pressure in the instrument.
Since Ultivo is designed to be in the LC stack, we needed to make sure that the maintenance features were accessible from the sides of the instrument. So we have pictures our easy change detector assembly, which simply pulls out from the side panel of the instrument.
New Vac Shield Technology
Easy Change Detector Assembly
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7. Software Features for High Throughput
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Real time diagnostics
Capillary health
Spray stability
Source Cleanliness
Clearer callouts for entry errors
We didn’t stop with hardware, we also made some changes to our software because it’s the way that users actually interact with the instrument.
Our acquisition software has a fresh look and feel
We also incorporated clearer callouts so you know when there was an entry error, so errors can be fixed quickly.
We now have real time diagnostics which let the user know the health of the capillary, the stability of the spray and the cleanliness of the source.
We also made changes to our quantitative analysis software and we are introducing quant my way, which is a customizable version of our quant software. This version allows an administrator to create different versions or “flavors” of quant for a specific method or workflow, ultimately eliminating errors and streamlining the analysis process.
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8. Software Features for High Throughput
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11/29/2018
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Quant-My-Way: customize the user interface for data analysis.
We didn’t stop with hardware, we also made some changes to our software because it’s the way that users actually interact with the instrument.
Our acquisition software has a fresh look and feel
We also incorporated clearer callouts so you know when there was an entry error, so errors can be fixed quickly.
We now have real time diagnostics which let the user know the health of the capillary, the stability of the spray and the cleanliness of the source.
We also made changes to our quantitative analysis software and we are introducing quant my way, which is a customizable version of our quant software. This version allows an administrator to create different versions or “flavors” of quant for a specific method or workflow, ultimately eliminating errors and streamlining the analysis process.
Faster, easier, error-free data analysis, review and reporting by novice operators.
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9. Quant-My-Way Presentation Title
11/29/2018
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We didn’t stop with hardware, we also made some changes to our software because it’s the way that users actually interact with the instrument.
Our acquisition software has a fresh look and feel
We also incorporated clearer callouts so you know when there was an entry error, so errors can be fixed quickly.
We now have real time diagnostics which let the user know the health of the capillary, the stability of the spray and the cleanliness of the source.
We also made changes to our quantitative analysis software and we are introducing quant my way, which is a customizable version of our quant software. This version allows an administrator to create different versions or “flavors” of quant for a specific method or workflow, ultimately eliminating errors and streamlining the analysis process.
11/29/2018
NEMC 2018

10. Presentation Title
11/29/2018
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Analysis of Per/Polyfluoroalkyl Substances (PFAS) in Drinking Water Using the Agilent Ultivo Triple Quadrupole LC/MS Application Note: EN
We wanted to show how great Ultivo will be for environmental applications, and how detection limits can be enhanced with agilents innovate sample prep and automated sample pre-concentration products.
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11. Analysis Method 17 PFAS compounds in drinking water.
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17 PFAS compounds in drinking water.
All 14 from USEPA 537 method.
Additional compounds PFDoA, PFPeA, and PFBA
Full list included in Application note EN
250mL water samples
Extracted with Agilent Weak Anion Exchange (WAX) cartridges.
Similar to US EPA 537 Method Conditions
Extracts concentrated to dryness with nitrogen
Reconstituted w/ 96:4 (v/v) Methanol/Water
IS added
For this application we analyzed 17 pfas in drinking water, which included all 14 from the USEPA 537 method.
250 mL of water was extracted using agilent extraction kits.
The total runtime for the method was 19 minutes using the agilent 1290 Infinity II HPLC, and the Ultivo Triple Quad MS.
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12. Instrumentation 1290 Infinity II LC
Analytical Column: Zorbax Eclipse plus
C18 3.0x50mm, 1.8µm
Mobile Phase A:H2O and
Mobile Phase B: 95:5 MeOH/H2O, 5mM Ammonium Acetate
Flow Rate: 0.4 ml/min
Injection Volume: 5µL
Ultivo Triple Quadrupole MS in negative ionization mode.
Dynamic MRM
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13. Overcoming Potential Contamination
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PFASs could potentially be used in LC and component manufacturing processes
Changes to the system:
Delay Column: Zorbax Eclipse plus C18 4.6x50mm, 3.5µm
Vials: 1 mL polypropylene vials
Caps: Snap top polypropylene caps
For this application we analyzed 17 pfas in drinking water, which included all 14 from the USEPA 537 method.
250 mL of water was extracted using agilent extraction kits.
The total runtime for the method was 19 minutes using the agilent 1290 Infinity II HPLC, and the Ultivo Triple Quad MS.
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14. PFAS Chromatography
1 ppb standard solution of PFAS in water.
Good chromatographic separation and S/N with 1 ppb standard solution.
EPA Method 537  35 to 37 min analytical run time
This method is less than 22 minutes.
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15. Recovery of PFAS Compounds in Water
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Recovery % for all compounds at 1 µg/L and 5 µg/L spiking levels
RSD%: % for all compounds at both spiking levels (5 replicates).
We achieved recoveries of 0-120% for all compounds at both 1ug/l and 5ug.l levels
Along with excellent sensitivity and linearity at a range of ug/L with R2 values greater than 0.99
Great precision, with rsd% between 0.3 and 10.8% for all compounds in the method.
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16. Linearity of PFAS Compounds
Excellent Linearity for 0.2 – 100 µg/L calibration curves.
R2 > 0.99 for all compounds, based on average of 3 calibration curves run over 1 month period.
Linearity over 1 month highly stable.
Compound R2
CCV RSD (%)
PFBA
0.9965
2.1
PFPeA
0.9996
2.7
PFBS
0.9964
3.1
PFHxA
0.9988
3.0
PFHpA
0.9985
3.4
PFHxS
0.9995
1.7
PFOA
0.9971
1.2
PFOS
0.9991
1.9
PFNA
0.9811
3.7
PFDA
0.9941
3.6
N-MeFOSAA
0.9999
PFDS
2.3
PFUdA
0.9940
N-EtFOSAA
5.2
PFDoA
0.9974
PFTrDA
0.9984
3.5
PFTeDA
0.9986
3.2
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17. Continuous Calibration Verification
Calibration sample run after every ten samples - required by US EPA 537.
CCV samples at 1 ppb level were calculated for a 26 hour batch.
94% of Analytes have less than 5% RSD at 1ppb.
Very stable over 26 hour experiment.  
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18. Analysis of Real Water Samples
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Regulatory statement (if applicable))
PFBS, PFOS, PFOA, PFNA were detected at 10, 4, 0.6, and 8, ng/L
Drinking water sample sourced from California
4 PFAS compounds detected
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19. Conclusions
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Regulatory statement (if applicable))
Ultivo can exceed the sensitivity requirements for PFAS detection in drinking water.
This method is robust both in standard curve and check sample evaluations.
Shorter runtime and less sample volume than traditional methods.
PFBS, PFOS, PFOA, PFNA were detected at 10, 4, 0.6, and 8, ng/L
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20. Presentation Title
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Sensitive Detection of Trace Organic Contaminants in Water Using Agilent 1290 Infinity FlexCube Online SPE and Agilent Ultivo Triple Quadrupole LC/MS Application Note: EN
We wanted to show how great Ultivo will be for environmental applications, and how detection limits can be enhanced with agilents innovate sample prep and automated sample pre-concentration products.
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21. Ultivo: Even more powerful with Flex Cube Online SPE
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11/29/2018
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Automated sample pre-concentration
Fits neatly into LC Stack
Operated using Agilent MassHunter Software
Many phases compatible with Online SPE format
With this method that we are about to present, we are using the Agilent flex cube online SPE, which is a tool that we use to automatically preconcentrate our sample.
The Flex cube fits right into the LC stack and is operated using our agilent masshunter software so it fits seamlessly into the LC/MS process.
We also have many phases of preconcentration columns which are compatible with the online SPE workflow.
Using the flexcube for sample preconcentration is not new, and we have lots of experience applying this process to various methods. Check out agilent’s application portfolio for more methods using our flex cube.
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22. Comparing Methods: Online SPE Wins: Sample volume Number of steps
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Conventional SPE:
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Water sample
Evaporation w/ Nitrogen
Benchtop SPE
Re-constitution
Analysis w/
LC/TQ
Online SPE can make your sample preparation more efficient
Here is the typical process for conventional SPE:
Now for the online SPE process:
Online SPE wins with sample volume,
Number of steps
And when you increase your number of steps, then you introduce more places for error in your process.
Online SPE can reduce the cost of materials
And ultimately it saves you time. There are many active steps with the traditional SPE, but with the Flex cube online SPE you can sit back, relax (or do other tasks) , and let your instrument do the work for you.
Online SPE:
Online SPE Wins:
Sample volume
Number of steps
Potential error
Cost
Your time
900 µL
Water sample
Online SPE
Coupled w/ LC/TQ Analysis
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23. Valve System Presentation Title 11/29/2018 NEMC 2018 11/29/2018
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24. Excellent Reproducibility of Dual Cartridge System
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Chromatographic overlay of 6 injections at 20 ng/L with alternating enrichment cartridges
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25. Trace Organic Compounds in Drinking Water (51 Analytes)
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Pesticides/Metabolites
3-Hydroxy Carbofuran
Alachlor
DEET
Disulfoton-Sulfoxide
Ethion
Fenamiphos
Indoxacarb
Metalaxyl
Mevinphos (Cis)
Methomyl
Oxadixyl
Propoxur
Vamidothion sulfoxide
Blue=EPA 540
Pharmaceuticals
Carbamazepine
Diclofenac
Dilitiazem
Gemfibrozil
Hydrocortisone
Ibuprofen
Norgestrel
Primidone
Propanolol
Simazine
Sulfamethoxazole
Testosterone
Triclocarban
Trimethoprim
Other Chemicals
Caffeine
Benzotriazole
Diphenylhydramine
Propylparaben
TCEP
PFASs
(Neg ESI)
PFBS
PFDA
PFDoA
PFHpA
PFHxA
PFHxS
PFNA
PFOA
PFPeA
PFUdA
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26. Trace Organic Compounds in Drinking Water
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51 representative compounds
1260 Infinity II HPLC
Column: Poroshell 120 EC C18, 3.0 x 50 mm, 2.7 μm (p/n )
Mobile Phases: H2O (1mM NH4OAc + 0.1mM NH4F % HOAc) and ACN (0.01% HOAc)
Flow Rate: 0.4 ml/min
Flex Cube Online SPE
900uL injection volume.
No bench-top sample pretreatment.
SPE Cartridge: PLRP, 2.1x12.5mm
Load: H2O, 0.1% Formic Acid
Clean: 1:1 ACN/IPA
Fast 21 min total (including post time)!
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AJS Source and MS Parameters
Drying Gas Temp
250oC
Drying gas Flow
11L/min
Sheath Gas Temp
375oC
Sheath Gas Flow
12L/min
Nebulizer Pressure
45 psi
Nozzle Voltage (V)
500 (+); 500 (-)
Capillary Voltage (V)
4000 (+); 3500 (-)
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27. Excellent Signal Response
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Signal response at 10 ppt with 900 uL injection
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28. Excellent Dynamic Range (1.25 ng/L to 500 ng/L)
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3-Hydroxy Carbofuran
100
200
300
400
500 4
x10 2 6
R^2 = 0.999
Disulfoton-Sulfoxide 5 1 3
Methomyl
PFOA
0.5
Concentration (pg/ml)
Ibuprofen
R^2 = 0.997
Testosterone
0.2
0.6
R^2 = 0.995
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29. Detection Sensitivity
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Distribution of the limits of quantitation for 51 contaminants (accuracy %)
Analyte
LOQ (ng/L)
PFBS 5
PFDA 20
PFDoA 10
PFHpA
1.25
PFHxA
2.5
PFHxS
PFNA
PFOA
PFPeA
PFUdA 50
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30. %RSD at the lowest concentration for accurate quantitation (n=6)
Detection Precision
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%RSD at the lowest concentration for accurate quantitation (n=6)
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31. Conclusions PFAS compounds included in TOrC method.
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Regulatory statement (if applicable))
PFAS compounds included in TOrC method.
Online SPE is fast, and can streamline your laboratory workflows.
Only 900ul of sample required.
Sensitive and reproducible results.
Quantitation limits below 50 ppt – many in single digit ppt range
PFBS, PFOS, PFOA, PFNA were detected at 10, 4, 0.6, and 8, ng/L
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32. UNBELIEVABLY POWERFUL REMARKABLY SMALL Ultivo Triple Quad LC/MS
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UNBELIEVABLY POWERFUL
REMARKABLY SMALL
Ultivo Triple Quad LC/MS
Fit for Purpose!

33. Thank you Tarun Anumol and Dorothy Yang
References
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11/29/2018
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Thank you Tarun Anumol and Dorothy Yang
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