An Online-SPE Approach to Analyzing Broad Classes of Emergent … · 2017. 8. 9. · An online-SPE...
Transcript of An Online-SPE Approach to Analyzing Broad Classes of Emergent … · 2017. 8. 9. · An online-SPE...
An online-SPE approach to analyzing broad classes of Emergent Contaminants
David Schiessel, Senior [email protected] Laboratories, Inc.6100 Quail Valley CourtRiverside, CA 92507951-653-3351 ext.268
Outline
Previous methodology and What’s new What is online-SPE Optimizing online-SPE Applications – PPCPs and Perfluorinated Compounds Improving methods for matrix interference Optimizing Mass Spectrometer parameters
Closing Comments
Previous Methodologyfor PPCP/CEC analysis
EPA 1694 (Dec. 2007) Four analyte classes – ESI(+) Formate buffer, MeOH:ACN ESI(+) Oxalic acid, MeOH:ACN ESI(-) Acetate buffer, MeOH:ACN ESI(+) HILIC, Basic extraction
Newer Developments
Eluent systems that improve ESI. More variety of SPE sorbents (polymeric, WAX) Online-SPE, decreasing analysis time Analytes of interest has changed Sample collection/preservation (Vanderford1) EPA method 543 (Online SPE specific) Improvements to Instrumentation Polarity switching (ESI+/-) High duty cycles with smaller FWHM
What is Online-SPE
Automated solid phase extraction at the instrument Uses 2 HPLC pumps and 2 injection valves; sample load
and LC run; can be multiplexed. Specialized LC column: 20-50mm, 12-40um, 2.1mm Loading a 2-10mL aqueous sample at 1-5mL/min Elution by backflushing the SPE sorbent with mobile phase *Turbulent flow: Specialized online SPE (Thermo Transcend
Turboflow)
Equan MAX System
Optimizing online SPE
Wash step – Allows the system to rinse the media to eliminate preservative and/or matrix salts. 2-3 sample volumes.
Elute focus – Step gradient organic to elute highly hydrophobic analytes from C18.
Mobile phase background elimination – Works well for PFC analysis. Mobile phase 90% organic while online SPE sample loading.
Use short analytical columns (50mm) unless increased retention is required.
Use fast gradients – online SPE adds retention
Compound Classes using online SPE
Pharmaceuticals (Acetaminophen) Hormones (17B-Estradiol) Organophosphate Flame Retardants (TCEP) Perfluorinated Compounds (PFOA) Artificial Sweeteners (Sucralose) Iodinated Contrast Media (Iopromide) Antimicrobials (Triclosan) Pesticides (Pyrethroids down to 1.0ng/L) Others (nonylphenols, Bisphenol-A, Phthalates, NP
Pesticides)*LIMITED by ESI more than SPE*
RT: 0.00 - 15.00
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100 NL: 1.93E5TIC F: + c ESI SRM ms2 152.100 [110.199-110.201] MS Test01
NL: 6.86E6TIC F: + c ESI SRM ms2 192.100 [118.999-119.001] MS Test01
NL: 1.97E5TIC F: + c ESI SRM ms2 195.000 [138.099-138.101] MS Test01
NL: 2.15E4TIC F: - c ESI SRM ms2 205.100 [160.999-161.001] MS Test01
NL: 1.05E4TIC F: - c ESI SRM ms2 227.100 [212.099-212.101] MS Test01
NL: 9.62E4TIC F: - c ESI SRM ms2 229.100 [169.099-169.101] MS Test01
NL: 6.95E5TIC F: + c ESI SRM ms2 233.000 [72.199-72.201] MS Test01
NL: 9.51E6TIC F: + c ESI SRM ms2 237.100 [194.099-194.101] MS Test01
Example Chromatogram at 50ng/L
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NL: 4.66E5TIC F: + c ESI SRM ms2 254.100 [92.099-92.101] MS Test01
NL: 4.59E3TIC F: - c ESI SRM ms2 271.100 [145.099-145.101] MS Test01
NL: 4.51E5TIC F: + c ESI SRM ms2 285.000 [222.999-223.001] MS Test01
NL: 8.00E2TIC F: - c ESI SRM ms2 287.000 [35.199-35.201] MS Test01
NL: 2.43E3TIC F: - c ESI SRM ms2 295.100 [145.099-145.101] MS Test01
NL: 5.14E5TIC F: + c ESI SRM ms2 296.000 [213.999-214.001] MS Test01
NL: 2.51E5TIC F: + c ESI SRM ms2 559.200 [440.299-440.301] MS Test01
Acetaminophen
Deet
Caffeine
Ibuprofen
BPA
Naproxen
Diuron
Carbamazepine
Gemfibrozil
Sulfamethoxazole
Estradiol
TCEP
Triclosan
Ethynylestradiol
Diclofenac
Lipitor
RT: 0.00 - 15.00 SM: 3B
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NL: 1.49E4TIC F: + c ESI SRM ms2 791.800 [299.699-299.701, 559.099-559.101] MS ICIS Test01
NL: 4.09E4TIC F: + c ESI SRM ms2 414.000 [198.999-199.001] MS ICIS Test01
Example Chromatogram at 50ng/L
Iopromide
Sucralose[M+NH3+H+]
Polymeric (Strata-X) online SPE retains highly polar analytes
PPCP analysis conditions
Sample: Prepreserved with sodium azide and ascorbic acid. Add 3 drops formic acid per 40mL prior to analysis
Mobile Phase: 300ul/min 5mM Ammonium Hydroxide with 10-80% Methanol.
Online SPE media: Thermo Hypersil C18 and Strata-X (polymeric) for Sucralose/Iopromide (can use HyperSepRetain PEP7)
MS parameters: TSQ Quantum Ultra. Vap/Cap Temps at 250C, Sheath=35, Aux=15, CID=1.0mTorr Argon, Q1=0.4 FWHM
Quantitation method is strict isotope dilution
Example: 50ng/L C4-C14 PFCAs
RT: 5.00 - 10.00 SM: 5B
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RT: 7.03
RT: 7.38
RT: 7.54
RT: 8.79
RT: 7.66
RT: 7.77
RT: 7.88
RT: 8.00
NL: 4.16E4TIC F: - c ESI SRM ms2 213.000 [168.999-169.001] MS ICIS Test03
NL: 9.11E4TIC F: - c ESI SRM ms2 263.000 [218.999-219.001] MS ICIS Test03
NL: 3.19E5TIC F: - c ESI SRM ms2 313.000 [268.999-269.001] MS ICIS Test03
NL: 3.13E5TIC F: - c ESI SRM ms2 363.000 [318.999-319.001] MS ICIS Test03
NL: 3.71E5TIC F: - c ESI SRM ms2 413.000 [368.999-369.001] MS ICIS Test03
NL: 2.63E5TIC F: - c ESI SRM ms2 463.000 [418.999-419.001] MS ICIS Test03
NL: 2.20E5TIC F: - c ESI SRM ms2 513.000 [468.999-469.001] MS ICIS Test03
NL: 1.81E5TIC F: - c ESI SRM ms2 563.000 [518.999-519.001] MS ICIS Test03
RT: 5.00 - 10.00 SM: 5B
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RT: 8.13
RT: 8.41RT: 8.28
RT: 8.45
NL:1.40E5TIC F: - c ESI SRM ms2 613.000 [568.999-569.001] MS ICIS Test03
NL:1.06E5TIC F: - c ESI SRM ms2 663.000 [618.999-619.001] MS ICIS Test03
NL:1.07E5TIC F: - c ESI SRM ms2 713.000 [668.999-669.001] MS ICIS Test03
PFBA
PFPeA
PFHxA
PFHpA
PFOA
PFNA
PFDA
PFUnDA
PFDoDA
PFTrDA
PFTeDA
Example: 50ng/L C4-C10 PFSAs
RT: 5.00 - 10.00 SM: 3B
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0Time (min)
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RT: 7.42
RT: 7.55
RT: 7.67
RT: 7.76
RT: 7.87
RT: 7.99
NL: 3.67E4TIC F: - c ESI SRM ms2 299.000 [79.999-80.001] MS ICIS Test03
NL: 4.62E4TIC F: - c ESI SRM ms2 349.000 [79.999-80.001] MS ICIS Test03
NL: 4.28E4TIC F: - c ESI SRM ms2 399.000 [79.999-80.001] MS ICIS Test03
NL: 3.74E4TIC F: - c ESI SRM ms2 449.000 [79.999-80.001] MS ICIS Test03
NL: 2.28E4TIC F: - c ESI SRM ms2 499.000 [79.999-80.001] MS ICIS Test03
NL: 2.41E4TIC F: - c ESI SRM ms2 549.000 [79.999-80.001] MS ICIS Test03
NL: 2.08E4TIC F: - c ESI SRM ms2 599.000 [79.999-80.001] MS ICIS Test03
PFBS
PFPeS
PFHxS (branched)
PFHpS
PFOS (branched)
PFNS
PFDS
RT: 5.00 - 10.00 SM: 3B
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RT: 9.16RT: 7.65
RT: 7.88
RT: 7.94
RT: 8.55RT: 8.01
RT: 8.63RT: 7.78
RT: 8.20
NL: 5.74E4TIC F: - c ESI SRM ms2 327.000 [306.999-307.001] MS ICIS Test03
NL: 5.65E4TIC F: - c ESI SRM ms2 427.000 [406.999-407.001] MS ICIS Test03
NL: 4.00E4TIC F: - c ESI SRM ms2 527.000 [506.999-507.001] MS ICIS Test03
NL: 1.22E5TIC F: - c ESI SRM ms2 570.000 [418.999-419.001] MS ICIS Test03
NL: 9.67E4TIC F: - c ESI SRM ms2 584.000 [418.999-419.001] MS ICIS Test03
NL: 2.89E4TIC F: - c ESI SRM ms2 498.000 [77.999-78.001] MS ICIS Test03
Example: 50ng/L other PFCs
4:2 FTS
6:2 FTS
8:2 FTS
N-MeFOSAA
N-EtFOSAA
FOSA
PFC analysis conditions
Sample: Prepreserved with 5g/L Trizma Mobile Phase: 300ul/min 0.24-0.08% Ammonium Hydroxide
with 40-80% Methanol. Online SPE media: Strata-X-AW (mixed mode weak anion)
works especially well for PFBA/PFPeA LC Column: C18 EVO (stable at high pH) MS parameters: TSQ Quantum Ultra. Vap/Cap Temps at
250/150C, Sheath=40, Aux=40, CID=1.0mTorr Argon, Q1=0.4 FWHM
Quantitation method is internal standard by compound class
Addressing Matrix issues
For samples containing high salts Cl- or SO42-, use
Dionex OnGuard II Ba/Ag/H. Check for losses. Have a wash step after sample loading (1-2 sample
volumes) Filter samples with appropriate media. Glass fiber
works well for PFCs, even for PFTeDA Use smaller FWHM; 0.2 or 0.4 instead of 0.7Da
After filtering >2000ppm Cl-
Ba
Ag
H+
RT: 0.08 - 11.72
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RT: 9.23AA: 4741SN: 74
NL: 2.00E4TIC F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test08b
NL: 2.00E4Base Peak m/z= 433.60-434.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test08b
NL: 2.00E4Base Peak m/z= 190.60-191.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test08b
Effect of FWHM
Cyfluthrin 451>191 + 454>434
Cyfluthrin 451>191 only
Cyfluthrin 451>434 only
Q1 FWHM = 0.7Da
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RT: 9.23AA: 5284SN: 52
RT: 7.90AA: 81926SN: 859
RT: 9.23AA: 4741SN: 74
RT: 7.94AA: 86433SN: 5614
Effect of FWHM
RT: 0.08 - 11.72
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RT: 1.50AA: 8478SN: 123
NL: 2.00E4TIC F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09b
NL: 2.00E4Base Peak m/z= 433.60-434.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09b
NL: 2.00E4Base Peak m/z= 190.60-191.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09b
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RT: 8.21
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RT: 7.98AA: 83268SN: 1107
RT: 8.02AA: 93917SN: 4608
Cyfluthrin 451>191 + 454>434
Cyfluthrin 451>191 only
Cyfluthrin 451>434 only
Q1 FWHM = 0.5Da
Effect of FWHM
Cyfluthrin 451>191 + 454>434
Cyfluthrin 451>191 only
Cyfluthrin 451>434 only
Q1 FWHM = 0.2Da
RT: 0.08 - 11.72
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RT: 6.68AA: 5276SN: 310
RT: 1.12AA: 7079SN: 567
RT: 6.57AA: 2191SN: 115
NL: 2.00E4TIC F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09a
NL: 2.00E4Base Peak m/z= 433.60-434.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09a
NL: 2.00E4Base Peak m/z= 190.60-191.60 F: + c ESI SRM ms2 451.100 [191.099-191.101, 434.099-434.101] MS ICIS Test09a
7 8 9
RT: 8.00AA: 147122SN: 4003
6.68 5276 310
RT: 8.00AA: 76383SN: 3291
57
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RT: 8.02AA: 70963SN: 8471
RT: 0.00 - 13.01
0 1 2 3 4 5 6 7 8 9 10 11 12 13Time (min)
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RT: 7.84AA: 20302SN: 2838
NL:2.04E3TIC F: + c ESI SRM ms2 451.100 [434.099-434.101] MS ICIS Test11b
NL:3.58E3TIC F: + c ESI SRM ms2 451.100 [191.099-191.101] MS ICIS Test11b
Effect of FWHM
Cyfluthrin 451>191 only
Cyfluthrin 451>434 only
Q1 FWHM = 0.04Da
Diminishing return on S/NDue to mass stability?
Sensitivity vs. FWHM
0%
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100%
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8
Intensity vs FWHM
295 Da
393 Da
491 Da
Mean
Power (Mean)
Mass Calibration
Lowering FWHM requires special attention to mass calibration frequency.
Mass stability for TSQ Series around ±0.05 Da There is a limit to lowering FWHM (0.2-0.3 Da) Using Phosphoric acid clusters for mass calibration
can improve mass calibration stability (9 mass cal points instead of 3.
Instrument Mass stability is the limiting factor to lowering FWHM.
Mass Stability and FWHM
Test04 #1 RT: 0.01 AV: 1 NL: 3.30E6T: + p ESI Q1MS [294.071-296.000, 392.071-394.000, 490.071-492.000]
294.5 295.0 295.5 296.0m/z
392.5 393.0 393.5 394.0m/z
490.5 491.0 491.5 492.0m/z
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294.7857
294.2143
392.7857
490.8572
491.7857
1 AV: 1 NL: 2.97E6 94.000-296.000, 392.000-394.000, 490.000-492.000]
FWHM = 0.70 DaMass when analyte was infused ± 0.05Da
Test08c #26 RT: 0.26 AV: 1 NL: 9.90E5T: + c ESI Q1MS [294.750-295.250, 392.750-393.250, 490.750-491.250]
294.8 295.0 295.2m/z
392.8 393.0 393.2m/z
490.8 491.0 491.2m/z
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Mass Stability and FWHM
FWHM = 0.06 Da
, , ]
295.0051
295.23980
392.9949
393.1684392.8928
490.9745
491.1684490.8418
Mass when analyte was infused ± 0.05Da
Mass Error vs FWHMBefore/After Mass Calibration
-600
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-400
-300
-200
-100
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Abs
Err
or (
ppm
)
FWHM (Da)
295 Abs Error
393 Abs Error
491 Abs Error
295 Abs Error
393 Abs Error
491 Abs Error
Red: old mass cal (2mo)Green: new mass cal
Data Applicability
Do results from online-SPE produce results that reflect bioavailable3 analyte only? Particles trapped in media Particles pass through (turbulent flow at high flows)
Analytes with high Koc might be trapped in suspended solids
Examples – Pyrethroids and some PFCs. Generally analytes that are VERY hydrophobic
Final Thoughts
Most analytes can be done by online-SPE which makes analysis much faster
There may be limitations on sensitivity and peak shape.
Coupling online-SPE with MS/MS is a sensitive analytical tool.
Online SPE may be adapted to soil analysis if aqueous extraction is used
References
1) Vanderford, Brett J., Douglas B. Mawhinney, Rebecca A. Trenholm, Janie C. Zeigler-Holady, and Shane A. Snyder. “Assessment of Sample Preservation Techniques for Pharmaceuticals, Personal Care Products, and Steroids in Surface and Drinking Water.” Analytical and Bioanalytical Chemistry 399, no. 6 (February 2011): 2227–34. doi:10.1007/s00216-010-4608-5.
2) Peake, Barrie M., Rhiannon Braund, Alfred Y.C. Tong, and Louis A. Tremblay. “Detection and Presence of Pharmaceuticals in the Environment.” In The Life-Cycle of Pharmaceuticals in the Environment, 77–107. Elsevier, 2016. doi:10.1016/B978-1-907568-25-1.00004-9.
3) Maund, Steve J., Mick J. Hamer, Mike C. G. Lane, Eamonn Farrelly, Jean H. Rapley, Una M. Goggin, and Wendy E. Gentle. “Partitioning, Bioavailability, and Toxicity of the Pyrethroid Insecticide Cypermethrin in Sediments.” Environmental Toxicology and Chemistry / SETAC 21, no. 1 (January 2002): 9–15.
4) EPA Method 537 v1.1, Determination Of Selected Perfluorinated Alkyl Acids In Drinking Water By Solid Phase Extraction And Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS), September 2009
5) EPA Method 543 v1.0, Determination of Selected Organic Chemicals in Drinking Water by On-line Solid Phase Extraction - Liquid Chromatography/Tandem Mass Spectrometry (On-line SPE - LC/MS/MS), March 2015
6) Yamazaki, Eriko, SachiTaniyasu, KodaiShimamura, Shunya Sasaki, and Nobuyoshi Yamashita. “Development of a Solid-Phase Extraction Method for the Trace Analysis of Perfluoroalkyl Substances in Open-Ocean Water.” Bunseki Kagaku 64, no. 10 (2015): 759–68. doi:10.2116/bunsekikagaku.64.759
7) Batchu, Sudha Rani, Natalia Quinete, Venkata R. Panditi, and Piero R. Gardinali. “Online Solid Phase Extraction Liquid Chromatography Tandem Mass Spectrometry (SPE-LC-MS/MS) Method for the Determination of Sucralose in Reclaimed and Drinking Waters and Its Photo Degradation in Natural Waters from South Florida” 7 (2013): 141.
QUESTIONS ?COMMENTS?
David Schiessel, Senior [email protected] Laboratories, Inc.6100 Quail Valley CourtRiverside, CA 92507951-653-3351 ext.268