Modernizing Persistent Organic Pollutants (POPs) Analysis · Modernizing Persistent Organic...
Transcript of Modernizing Persistent Organic Pollutants (POPs) Analysis · Modernizing Persistent Organic...
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Modernizing Persistent Organic Pollutants (POPs) Analysis
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• Introduction to POPs
• Dioxin analysis with the DFS
• Recent regulatory changes:• Dioxin analysis in food with the Thermo Scientific™ TSQ™
9000 triple quadrupole GC-MS/MS system
• Analysis of PBDEs with the Thermo Scientific™ TSQ™ 9000 triple quadrupole GC-MS/MS system
• Analysis of PBDEs with the Thermo Scientific™ Q Exactive™ GC Orbitrap™ GC-MS/MS
• Analysis of SCCPs wit the Thermo Scientific™ Q Exactive™ GC Orbitrap™ GC-MS/MS
• Conclusions
Agenda
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• POPs are toxic chemicals that adversely affect humans• Bioaccumulate and transferred via the environment and the
food chain• Long half lives and difficult to break down• POPs are regulated by the Stockholm convention
• 2,3,7,8-TCCD is know the most toxic compound in the world
What are Persistent Organic Pollutants (POPs)
Aldrin A
Chlordane A
DDT B
Dieldrin A
Endrin A
Heptachlor A
Hexachlorobenzene A and
CMirex A
Toxaphane A
Polychlorinated biphenyls (PCB)
A and
C Polychlorinated dibenzo-p-dioxins (PCDD)
C
Polychlorinated dibenzofurans (PCDF)
C
Alpha hexachlorocyclohexane A
Beta hexachlorocyclohexane A
Chlordecone A
Hexabromobiphenyl A
Hexabromocyclododecane A
Hexabromodiphenyl ether and heptabromodiphenyl ether (commercial “Octa”)
A
Hexachlorobutadiene A
Lindane A
Pentachlorobenzene A and C
Pentachlorophenol and its salts and esters
A
Perfluorooctane sulfonic acid, its salts and perfluorooctane sulfonyl fluoride
A
Polychlorinated naphthalenes A and C
Technical Endosulfan and related isomers
A
Tetrabromodiphenyl ether and pentabromodiphenyl ether (commercial “Penta”)
AViktor Yushchenko exposure to dioxins causing chloracne
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• Emerging POPs of interest:• Mixed halogenated dioxins and furans• Short-chain chlorinated paraffins (SCCPs)
• Challenges with the analysis of POPs:• Low regulatory limits• Varied matrices • Covering expanding lists of target analytes• Producing consistent results
• Need instrumentation suitable for the analysis and the regulation
Emerging POPs and Analysis Challenges
4000+ congenersof mixed halogenateddioxins and furans
Short chainedchlorinated paraffins
(SCCPs)
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Thermo Scientific DFS Magnetic Sector GC-HRMS – worldwide compliance
• Global compliance with any official Dioxin, PCB, or PBDE method (e.g. EPA 1613, 1668..)
• Proven leadership with robust sensitivity for routine applications thanks to large-volume ion source
• Exceptional productivity and flexibility with New DualData XL option, for up to doubled sample throughput
• Future committed for Dioxins and POPs regulations compliance today and tomorrow
thermoscientific.com/GoldStandard
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Worldwide Compliance - Official Methods Recognizing Magnetic Sector Technology
Do you have international customers? Are you working globally? Already now or in the future…
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15.2 MS Resolution - A static resolving power of at least 10,000 (10% valleydefinition) must be demonstrated at the appropriate m/z before any analysis isperformed. Static resolving power checks must be performed at the beginning andat the end of each 12 hour shift according to procedures in Section 10.1.2. Correctiveactions must be implemented whenever the resolving power does not meet therequirement.
DFS
Here: EPA 1613 compliance
10.2.1.2 The mass spectrometer shall be operated in a mass-drift correction mode,using perfluorokerosene (PFK) to provide lock m/z's. The lock-mass for each groupof m/z's is shown in Table 8. Each lock mass shall be monitored and shall not varyby more than ±20% throughout its respective retention time window. Variations of thelock mass by more than 20% indicate the presence of coeluting interferences that maysignificantly reduce the sensitivity of the mass spectrometer.
DFS DFS DFS
DFS & EPA 1613:
• 10.000 resolution • Resolution checks• Use PFK• mass-drift correction• lock mass monitoring
Official Method Compliance – EPA 1613
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Ultimate Sensitivity Combined with Robustness
RT: 23.82 - 27.83
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24.0823.90 25.52 27.40 27.7527.2324.19 24.36 27.0726.2925.3724.54 25.63 26.6625.83 26.9226.4626.0324.74
RT: 25.22MA: 432
24.13 24.52 27.6424.95 27.2926.7726.22 27.0526.0325.54 25.85 26.55 27.5126.9026.3524.67
RT: 25.20AA: 172156
RT: 25.20AA: 218158
RT: 25.81MA: 165
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27.6724.17 27.1625.26 27.4024.60 26.1825.57 26.9926.6624.87 25.9624.39 26.46
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26.73 27.2527.7725.57 27.5424.61 27.1224.32 26.6026.2224.02 25.26 25.4624.96 27.0125.70 26.4626.05
RT: 25.22AA: 153519
RT: 25.79AA: 116608
RT: 25.22AA: 198314 RT: 25.79
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NL: 1.13E2m/z= 303.8868-303.9172 MS PCDDF_sample_219448-703_2ul_01
NL: 1.22E2m/z= 305.8837-305.9143 MS PCDDF_sample_219448-703_2ul_01
NL: 4.01E4m/z= 315.9262-315.9578 MS ICIS PCDDF_sample_219448-703_2ul_01
NL: 5.11E4m/z= 317.9231-317.9549 MS ICIS PCDDF_sample_219448-703_2ul_01
NL: 7.50E1m/z= 319.8810-319.9130 MS PCDDF_sample_219448-703_2ul_01
NL: 7.00E1m/z= 321.8779-321.9101 MS PCDDF_sample_219448-703_2ul_01
NL: 3.52E4m/z= 331.9204-331.9536 MS ICIS PCDDF_sample_219448-703_2ul_01
NL: 4.62E4m/z= 333.9173-333.9507 MS ICIS PCDDF_sample_219448-703_2ul_01
Real sample: serum extract (col.: 60m x 0.25 (0.25))
3 fg TCDF
2 fg TCDD
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10.91 11.0910.299.09 11.609.268.81 11.268.78 10.04 10.539.749.338.478.137.68 7.92 9.638.407.59
20 fg 2378-TCDD SN ≥ 200:1 (PtP, 4σ)
Best Dioxin specification on the market
Proof spec demonstrated to customers during DFS GC-HRMS installation
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Ultimate Sensitivity for Confidence
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Quantification mass trace
Qualifier (ratio) mass trace
100 fg
25 fg
50 fg
10 fg5 fg2 fg
1368-TCDD1379-TCDD
1378-TCDD
1478-TCDD
1234-TCDD
2378-TCDD
6 different native tcdd isomers at different concentrations (2 – 100 fg/ul)
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RT: 21.36 - 26.69
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RT: 25.22AA: 4505
RT: 24.69AA: 2042
RT: 24.30AA: 651
RT: 22.98MA: 356RT: 22.68
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RT: 25.81AA: 8332
RT: 25.25AA: 4356
RT: 24.72AA: 2250RT: 24.32
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RT: 23.01AA: 472RT: 22.68
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RT: 25.81AA: 8621
RT: 25.22AA: 4231
RT: 24.69AA: 2097RT: 24.30
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RT: 23.00AA: 440RT: 22.66
MA: 130
RT: 25.79AA: 8982
RT: 25.23AA: 4343
RT: 24.70AA: 2172RT: 24.29
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RT: 22.99AA: 419RT: 22.68
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RT: 25.80AA: 8955
RT: 25.24AA: 4324
RT: 24.71AA: 1797RT: 24.30
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RT: 22.98AA: 392RT: 22.71
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RT: 25.79AA: 8448
RT: 25.23AA: 4063
RT: 24.70AA: 2075RT: 24.29
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RT: 22.97AA: 515
RT: 22.66AA: 152
NL: 2.13E3m/z= 321.8876-321.9004 MS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_01
NL: 1.93E3m/z= 321.8876-321.9004 MS ICIS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_02
NL: 2.04E3m/z= 321.8876-321.9004 MS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_03
NL: 2.29E3m/z= 321.8876-321.9004 MS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_04
NL: 2.24E3m/z= 321.8876-321.9004 MS ICIS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_05
NL: 2.12E3m/z= 321.8876-321.9004 MS ICIS Dioxin_2-5-10-25-50-100fg_TCDD_1ul_06
x5
Inj. 1
Inj. 2
Inj. 3
Inj. 4
Inj. 5
Inj. 6
- zoomed - all data unsmoothed!
2 – 100 fg TCDD on 60 m Column
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Customer Lab in Japan (2010)
D:\Xcalibur\...\tcdd_001 3/9/2010 1:48:35 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.10 - 11.78
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10.7210.836.49 11.508.98 10.34 11.209.949.347.04 8.334.42 9.925.975.82 6.524.47 8.868.297.877.616.445.715.26
NL:7.34E2m/z= 321.48-322.48 MS tcdd_001
Base Peak Retention Time : 10.78 Scan : 1028 Intensity : 734
Calculated values Noise Range : 7.11 - 8.94 minutes (279 scans) Baseline : 19.3 Signal To Noise : 212.6 (4 sigma) Noise : 0.840 Algorithm: Statistical
212 : 1D:\Xcalibur\...\tcdd_007 3/9/2010 3:56:56 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.04 - 11.58
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NL:1.21E3m/z= 321.48-322.48 MS tcdd_007
Base Peak Retention Time : 10.84 Scan : 1036 Intensity : 1214
Calculated values Noise Range : 6.27 - 7.62 minutes (205 scans) Baseline : 19.6 Signal To Noise : 217.3 (4 sigma) Noise : 1.374 Algorithm: Statistical
217 : 1D:\Xcalibur\...\tcdd_008 3/9/2010 4:16:44 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
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14.0213.849.81 15.3910.92 11.466.13 8.34 14.886.47 7.39 9.775.955.064.44
NL:1.24E3m/z= 321.48-322.48 MS tcdd_008
Base Peak Retention Time : 10.84 Scan : 1036 Intensity : 1244
Calculated values Noise Range : 8.48 - 9.80 minutes (200 scans) Baseline : 19.5 Signal To Noise : 204.6 (4 sigma) Noise : 1.497 Algorithm: Statistical
205 : 1
# tcdd S/N (4σ)1 20 fg 212 : 12 20 fg 217 : 13 20 fg 205 : 1
D:\Xcalibur\...\tcdd_009 3/9/2010 5:19:31 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.06 - 12.02
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NL:1.04E3m/z= 321.48-322.48 MS tcdd_009
Base Peak Retention Time : 10.85 Scan : 1039 Intensity : 1040
Calculated values Noise Range : 7.56 - 8.56 minutes (152 scans) Baseline : 19.3 Signal To Noise : 237.3 (4 sigma) Noise : 1.075 Algorithm: Statistical
237 : 1
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D:\Xcalibur\...\tcdd_005 3/9/2010 3:15:13 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.04 - 11.96
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10.915.50 11.097.355.804.84 10.299.09 11.606.39 9.268.818.78 10.044.48 5.25 8.477.397.215.83 6.554.11
NL:1.44E3m/z= 321.48-322.48 MS tcdd_005
Base Peak Retention Time : 10.83 Scan : 1033 Intensity : 1444
Calculated values Noise Range : 9.37 - 10.04 minutes (101 scans) Baseline : 19.5 Signal To Noise : 241.6 (4 sigma) Noise : 1.474 Algorithm: Statistical
242 : 1
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D:\Xcalibur\...\tcdd_010 3/9/2010 5:39:08 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.10 - 11.84
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NL:1.06E3m/z= 321.48-322.48 MS tcdd_010
Base Peak Retention Time : 10.84 Scan : 1035 Intensity : 1062
Calculated values Noise Range : 6.59 - 7.97 minutes (209 scans) Baseline : 19.3 Signal To Noise : 235.8 (4 sigma) Noise : 1.106 Algorithm: Statistical
236 : 1
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D:\Xcalibur\...\tcdd_004 3/9/2010 2:54:44 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.07 - 11.80
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NL:1.20E3m/z= 321.48-322.48 MS tcdd_004
Base Peak Retention Time : 10.82 Scan : 1033 Intensity : 1200
Calculated values Noise Range : 10.06 - 10.46 minutes (61 scans) Baseline : 19.6 Signal To Noise : 212.2 (4 sigma) Noise : 1.391 Algorithm: Statistical
212 : 1
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D:\Xcalibur\...\tcdd_006 3/9/2010 3:36:00 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.04 - 11.77
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NL:1.14E3m/z= 321.48-322.48 MS tcdd_006
Base Peak Retention Time : 10.84 Scan : 1036 Intensity : 1136
Calculated values Noise Range : 8.30 - 9.17 minutes (132 scans) Baseline : 19.5 Signal To Noise : 234.9 (4 sigma) Noise : 1.188 Algorithm: Statistical
235 : 1D:\Xcalibur\...\tcdd_003 3/9/2010 2:35:42 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.04 - 11.69
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10.82 11.029.715.514.05 10.314.34 9.926.31 7.66 7.85 9.504.51 9.428.425.20 8.727.406.876.355.74
NL:1.27E3m/z= 321.48-322.48 MS tcdd_003
Base Peak Retention Time : 10.78 Scan : 1028 Intensity : 1274
Calculated values Noise Range : 8.87 - 9.45 minutes (87 scans) Baseline : 19.6 Signal To Noise : 222.3 (4 sigma) Noise : 1.411 Algorithm: Statistical
222 : 18 20 fg 235 : 19 20 fg 222 : 1
D:\Xcalibur\...\tcdd_002 3/9/2010 2:13:49 AM 20fg tcddinj. volume: 1ul column: Thermo TR-Dioxin 30m ID: 0.25mm film: 0.1um oven: 120(2)-15-240(0)-6-280(1)
RT: 4.04 - 11.81
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10.82 11.168.16 10.649.266.64 7.14 7.895.735.63 9.926.21 11.749.754.84 7.46 8.204.59 8.644.48
NL:1.06E3m/z= 321.48-322.48 MS tcdd_002
Base Peak Retention Time : 10.79 Scan : 1026 Intensity : 1055
Calculated values Noise Range : 8.17 - 9.02 minutes (129 scans) Baseline : 19.5 Signal To Noise : 227.2 (4 sigma) Noise : 1.139 Algorithm: Statistical
227 : 110 20 fg 227 : 1
10 times repeated TCDD spec
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Dioxins & POPs Productivity – Thermo Scientific DFS DualData XL Option
32 samples in 12 hour shift16 samples in 12 hour shift
Technology based on Magnetic Sector MS provides a solution for maximum productivity in terms of number of samples analyzed per time.
With DualData XL Option you can double your throughput, even with mixed Applications such as Dioxins, PCBs, PBDEs, OCPs and other.
• same hardware • same software • same lab space
= same cost of ownership as standard magnetic sector MS
...but with up to double productivity
DualDataXL
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Thermo Scientific TSQ 9000 Triple Quadrupole GC-MS/MS System
Feature BenefitHighly efficient
ionization A greater ion flux reaching
the detectorA more tightly focused
ion beam Less ion burn and a higher
degree of robustness
Thermo Scientific™ TSQ™ 9000 triple quadrupole GC-MS/MS system
Thermo Scientific™ Advanced Electron Ionization (AEI) source
IDLRSD = 120 ag
n = 8 injectionsOFN1 fg on-column
2.400 2.500 2.600 2.700 2.800 2.900 3.000500
1,000
1,500
2,000
2,500
3,000
3,500
4,000
4,500
min
counts
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Ion Source Quadrupole 1 (Q1)
Ion Source
Ion Transfer Optics
Quadrupole
Single Quadrupole GC-MS
Triple Quadrupole GC-MS/MS
MS
MS MS
Quadrupole 2 (Q2)
Collision CellQuadrupole 3 (Q3) EM Detector
EM Detector
Why Triple Quadrupole GC-MS/MS?
Ion Transfer Optics
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• Low detection limits• Optimized sample
preparation• Consolidated analytical
methods• Faster, automated data
processing
...it’s a high selectivitytechnique...
GC-MS/MS – What’s So Special?
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EU Compliance – Thermo Scientific TSQ 9000 triple quadrupole GC-MS/MS system
EU regulations amended in 2014 (589/2014 & 709/2014)Allowing the use of GC-MS/MS for confirmatory methods providing the following criteria are met:
Regulatory compliant sensitivity•10fg on-column TCDD with ion ratios (2 specific SRM transitions) within ±15% tolerance.
•Routine method (all 17 toxic PCDDFs + 4 non-ortho dl-PCBs).
Chromatographic and mass spectral resolution•Peak to peak separation of HxCDF isomers <25% (5.9%)•Q1 and Q3 both set to 0.7Da @FWHM – giving EU compliant separation between masses
LOQ compliance•For GC-MS/MS European guidance is to demonstrate LOQ throughout sequence (as opposed to using S:N ratios.•±15% ion ratios.•≤30% deviation from average RF.
•Demonstrated over multi-site validation study on three separate systems and ~2 weeks continuous runtime.
21 CFR, part 11 compliance•Chromeleon software has the tools to be fully compliant with FDA requirements, including audit trails.
•Real time results updates including SUM WHO-PCDD/F-TEQ values during data review.
•All data can be digitally reviewed and signed.
Resolution ≤1(separation of two peaks one mass
unit apart) for each quadrupole
Ion ratio ± 15% max deviation from theoretical
value for product ions of the two
transitions
2 specific SRM transitions
for all labelled and native congeners.
Chromatographic separation of Heptafurans
better than 25% peak to peak.
17
• … An online search engine for Thermo Fisher Scientific applications• … Provides comprehensive application information and ready-to-run analytical methods• … A central repository for Thermo Scientific chromatography and MS application information
Method Development Resource: The AppsLab Library of Analytical Applications
Visit: www.thermofisher.com/appslab
18
LOQ Consistency for Dioxin Analysis in Matrix
22
23
53
66
79
92
101
88.33 71.76 94.59 93.48 67.18 96.42 101.79 62.34 92.28 104.45 69.42 97.60 107.36 60.28 91.57 97.07 73.03 100.00 90.36 67.25 89.31
LOQ
/4LO
Q/4
LOQ
/4LO
Q/4
LOQ
/4LO
Q/4
LOQ
/4
2378-TCDD 123478-HxCDD OCDDInjection #
10 fg on-column 40 fg on-column 160 fg on-column
• Consistent results at the LOQ• Reproducibility within the regulatory
requirements
19.275 19.400 19.500 19.600 19.700 19.800 19.900 20.0006575100125150175200225250275300325350375400425450475500
2378-TCDD
n = 8 injections 2,3,7,8-TCDD5fg on-column
IDL0.58 fg
19
Reporting to Regulations Made Easy with Chromeleon
RT:20.86
ISTD
Con
f 1
RT:20.86
ISTD
Qua
n
RT:20.88
Con
f 1
RT:20.88
Qua
n
RT:25.96
ISTD
Con
f 1
RT:25.96
ISTD
Qua
n
RT:25.98
Con
f 1
RT:25.97
Qua
n
RT:38.39
ISTD
Con
f 1
RT:38.39
ISTD
Qua
n
RT:38.40
Con
f 1
RT:38.40
Qua
n
2,3,7,8-TCDD 12378-PeCDD OCDD
Automatic flagging of results outside tolerance
20
Agreement of Total TEQ with Reference Animal Fat Sample
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
WHO
-PCD
D/F-
TEQ
cont
ribut
ion
(pg/
g)
QK 1 - Reference value QK1 Avg (n=14)
21
Long Term Robustness for Dioxin Analysis
100
1000
10000
0 20 40 60 80 100 120 140 160
Peak
are
a(c
pm)
Injection number
Peak area repeatability
2378-TCDD - 6.4% RSD 0.17 pg 12378-PeCDF - 6.3% RSD 0.36 pg 23478-PeCDF - 4.2% RSD 0.92 pg
12378-PeCDD - 6.0% RSD 0.21 pg 234678-HxCDF - 4.4% RSD 0.31 pg 123789-HxCDD - 5.3% RSD 0.2 pg
1234678-HpCDD - 3.5% RSD 0.71 pg OCDD - 2.8% RSD 6.12 pg
A
22
LOQ Calculation Reproducibility
0
0.002
0.004
0 20 40 60 80 100 120 140 160Calc
ulat
ed a
mou
nt
(pg/
µL)
Injection number
2378-TCDD RF repeatability (n=39) 2378-TCDD
40
90
140
0 20 40 60 80 100 120 140 160
Qua
lifie
r Ion
Rat
io
(%)
Injection number
2378-TCDD IR stability (n=39) 2378-TCDD
B
23
4.10 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.47-5.0e70.0e05.0e71.0e81.5e82.0e82.5e83.0e83 5e8
BD
E-3
BB
DE
-15
BD
E-1
7B
DE
-28
BD
E-4
9B
DE
-71
BD
E-4
7B
DE
-66
BD
E-7
7B
DE
-100
BD
E-1
19B
DE
-99
BD
E-8
5B
DE
-126
BD
E-1
54B
DE
-153
BD
E-1
38B
DE
-156 B
DE
-184
BD
E-1
83B
DE
-191
BD
E-1
97B
DE
-196
BD
E-2
07B
DE
-206
BD
E-2
09
PBDE Analysis - Highest Calibration Standard
TetraTriDiMono Penta Hexa Hepta Octa Nona Deca
800 pg oc* 1600 pg OC 4000 pg OCRT (minutes)
Cou
nts
BD
E-7
1
BD
E-4
9
BD
E-2
09BDE-49/71 % Resolution = 0.4%<40% = PASS
BDE-209 USP Tf @10% height = 1.4<3 = PASS
BDE-99 USP Tf @10% height = 1.1<3 = PASS
BD
E-9
9
24
4.10 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.47-1.0e50.0e01.0e52.0e53.0e54.0e55.0e56.0e57.0e5
BD
E-3
BDE-
15
BDE-
17BD
E-28
BDE-
49BD
E-71 BD
E-47
BDE-
66BD
E-77 BD
E-10
0BD
E-11
9BD
E-99
BDE-
85BD
E-12
6 BDE-
154
BDE-
153
BDE-
138
BDE-
156
BDE-
184
BDE-
183
BDE-
191
BDE-
197
BDE-
196
BDE-
207
BDE-
206
BDE-
209
PBDE Analysis - Lowest Calibration Standard
• *oc = on column, solvent standard, overlay of quantification SRM transitions. (No signal normalization)
TetraTriDiMono Penta Hexa Hepta Octa Nona Deca
2 pg oc* 4 pg OC 10 pg OC
BD
E-2
09
RT (minutes)
Cou
nts
876
P
BD
E-4
9
BD
E-7
1
BDE-49/71 % Valley = 0.6 % <40% = PASS(EPA-1614 criteria)
BDE-209 USP Tf @10% height = 1.3<3 = PASS
BDE-99 USPTf @10% height = 0.8<3 = PASS
25
Sensitivity
• Solvent standard 0.25 pg/uL
• t-score = 2.624
• n=15 injections
• n=14 degrees of freedom
• 99% confidence level
• Peak area % RSD < 15%
Excellent sensitivity for PBDEs using the AEI source
9.6459.750-2.5e2
1.0e32.0e33.0e34.0e35.0e36.0e37.0e38.0e39.0e39.8e3counts
BDE-209, 500 fg OC, n=15 injections% RSD peak area = 7.7 %
Calculated IDL = 100 fg OC (0.1 ng/Kg in sample)
RT (minutes)
Cou
nts
26
Peak Area Repeatability (IDL concentration)
6%4%
7%9%
11% 11%9%
7%
11%10% 11%
6%
9%
6%
11% 10%9%
13% 13%
6% 6% 7%9% 10%
5% 6%8%
0%
5%
10%
15%
20%
25%
30%
35%
40%
45%
50%
Peak area % RSD
Peak
are
a %
RSD
Compound
Peak area % RSD, n=15 injectionsPB
DE-3
PBDE
-7
PBDE
-15
PBDE
-17
PBDE
-28
PBDE
-49
PBDE
-71
PBDE
-47
PBDE
-66
PBDE
-77
PBDE
-100
PBDE
-119
PBDE
-99
PBDE
-85
PBDE
-126
PBDE
-154
PBDE
-153
PBDE
-138
PBDE
-156
PBDE
-184
PBDE
-183
PBDE
-191
PBDE
-197
PBDE
-196
PBDE
-207
PBDE
-206
PBDE
-209
• Solvent standards 0.02-0.25 pg/uL, t-score = 2.624, n=15 injections, n=14 degrees of freedom, 99% confidence level and peak area % RSD < 15%.
27
Instrument Detection Limit (IDL)
8.5 % 5.2 %
6 2 4 5 6 6 5 3 6 8 6 3 5
15
2821 19
2720
9 10 1115 15
71
82
100
0
20
40
60
80
100
120
IDL (fg OC)
IDL
(fgO
C)
Compound
Instrument detection limit (fg OC)
BDE-
3
BDE-
7
BDE-
15
BDE-
17
BDE-
28
BDE-
49
BDE-
71
BDE-
47
BDE-
66
BDE-
77
BDE-
100
BDE-
119
BDE-
99
BDE-
85
BDE-
126
BDE-
154
BDE-
153
BDE-
138
BDE-
156
BDE-
184
BDE-
183
BDE-
191
BDE-
197
BDE-
196
BDE-
207
BDE-
206
BDE-
209
Mon
o
Di
Tri Te
tra
Pent
a
Hex
a
Hep
ta
Oct
a
Non
a Dec
a
28
Example Linearity of Response
Excellent linearity, R² > 0.98 and RRF % RSD < 10%
• Solvent standards
• 1.0-2000 pg/uL
• Internal standard adjusted with 13C BDE stds
• No weighting applied as RRF was used
• Triplicate injection per level
29
Linearity of Response
Excellent low end precision and accuracy for triplicate standard injections
30
Robustness
1.E+03
1.E+04
1.E+05
1.E+06
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70 73 76 79 82 85 88 91 94 97 100 103
Peak
are
a co
unts
.s-1
Injection number
BDE-7
BDE-49
BDE-119
BDE-153
BDE-183BDE-209
453 fg OC, % RSD= 7
255 fg OC, % RSD= 6
1.4 pg OC, % RSD= 5
2.0 pg OC, % RSD= 5
9.1 pg OC, % RSD= 3
11 fg OC, % RSD= 9
Peak area repeatability: n=105 unspiked extracted fish matrix injections
Peak area stable (no inlet, column, MS maintenance or tuning)
31
ASMS 2015:Thermo Scientific™ Q Exactive™ GC Orbitrap™ GC-MS/MS system
Unprecedented Depth in Analysis
RP 120,000 (FWHM @ m/z 200)
EI (VeV) / CI; Full-scan, Timed-SIM
MS/MS capability
Redefining Routine GC-MS
RP 60,000 (FWHM @ m/z 200)
EI (VeV) / CI; Full-scan; Timed-SIM
NACRW 2016:Thermo Scientific™ Exactive™ GC Orbitrap™ GC-MS system
Orbitrap GC-MS Family
Instrument.com 2015 China Outstanding New Instrument
32
Well Established and Proven Technologies
NeverVent™ technology
ExtractaBrite™ ion source
V-Lock source plug
Orbitrap mass analyzer
Incredible HR/AM performance
Highly regarded Q Exactive platform
TRACE 1310 GC
Unique modular injector and detector design
Rapid heat cycling
33
Thermo Scientific Q Exactive GC Orbitrap GC-MS/MS System : The Technology Inside
ExtractaBrite Ion Source
BentFlatapole
Curved Linear Trap(C-TRAP)
Orbitrap Mass Analyzer
HCD CellMS/MS fragmentation
AQT Quadrupole(Precursor ion isolation)
34
EI 70eVVeV
Orbitrap GC System Highlights
SensitivityMass AccuracyResolving Power Dynamic Range
Up to 120,000 at m/z 200
< 1ppm ppt >6 orders
Harder Softer
CI
Ionization Range
• Standard library searchable spectra • Molecular ion
confirmation (incl. adducts)
35
Achieve Maximum Resolution and Maintain Sensitivity
15K 30K 60K
High selectivity analysis all of the time
• TIC signal intensity vs. scan number
• Increase the resolving power during acquisition
• Negligible drop in sensitivity
36
Advantages for POPs analysis
Key BenefitsFast instrument and method set-up
Method consolidation
High efficiency data processing
Quantitative and qualitative
information in a single injection
Adjustable scope of analysis
Retrospective data analysis
37
Polybrominated Diphenyl Ethers
Persistent Organic
Pollutants
“GC-Orbitrap is a useful tool for control labs,
outstanding sensitivity…high selectivity when
analyzing complex matrices…high robustness
…helps to fulfill new regulations with
extremely low limits (e.g. PCBs, PBDEs)”
Practical Experiences of Implementing POPs Methods using Orbitrap™ GC-MS. Nuria Cortés-Francisco, BFR 2017
38
PBDE Analysis of Environmental Samples
4.06 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.25-4.5e10
-3.8e10
-2.5e10
-1.3e10
0.0e0
1.3e10
2.5e10
3.8e10
5.0e10
6.3e10
7.5e10
BDE -
7
BDE -
15
BDE -
17BD
E -28
BDE -
49BD
E -71
BDE -
47BD
E -66
BDE -
77
BDE -
100 BD
E -11
9
BDE -
99
BDE -
85 BDE -
154
BDE -
153
BDE -
138
BDE -
184
BDE -
183
BDE -
191
BDE -
197
BDE -
196
BDE -
207
BDE -
206
BDE -
209
BDE -
3
BDE -
156
-1.4e50.0e0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
3.5e6
3.9e6
di-BDEs
tetra-BDEs
tri-BDEs
penta-BDEs
hexa-BDEs
hepta-BDEs
octa-BDEs
nona-BDEs
deca-BDE
mono-BDE
BDE-
261
39
Chromatography – Extracted Ion Chromatogram
40
Sensitivity
41
Tuna Sample Spiked 0.01 ng/g PBDEs
x=1-5, y=0-5
42
Mass Accuracy and Isotope Pattern T:FTMS + p EI Full ms [75.0000-1050.0000]
949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966m/z
05
101520253035404550556065707580859095
100
Rela
tive
Abun
danc
e
959.16665C12OBr5
81Br5= 959.16751-0.89 ppm
957.16858C12OBr681Br4= 957.16956
-1.01 ppm 961.16437
C12OBr4 81Br6= 961.16546-1.13 ppm
955.17037C12OBr7 81Br3= 955.17160
-1.28 ppm 963.16210C12OBr3 81Br7= 963.16342
-1.36 ppm
953.17196C12OBr881Br2= 953.17365
-1.70 ppm965.16011
C12OBr281Br8= 965.16137-1.30 ppm
951.17438C12OBr981Br= 951.17570
-1.30 ppm
957.16960
949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966m/z
05
101520253035404550556065707580859095
100
Rela
tive
Abun
danc
e
959.16758C12OBr5
81Br5
12OBr681Br4 961.16556
C12OBr4 81Br6
955.17163C12OBr781Br3 963.16356
C12OBr381Br7
953.17367C12OBr881Br2 965.16159
C12OBr281Br8
951.17570C12OBr981Br
BDE-209acquired isotopic pattern
BDE-209theoretical
isotopic pattern
C
BDE-209
43
POPs in Salmon: A New Approach using High Resolution Full Scan
SCCPsMCCPsPCBs,Pesticides Toxaphenes
5.2 5.4 5.6 5.8 6.0 6.2 6.4 6.6 6.8 7.0 7.2Time (min)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
SCCP TIC
C10H16Cl6m/z 312.9671
C11H17Cl7m/z 360.9437
C12H19Cl7m/z 374.9589
C13H20Cl8m/z 422.9361
C12H18Cl8m/z 408.9204
C13H19Cl9m/z 458.8936
C11H18Cl6m/z 326.9822
MCCP TIC
C14H24Cl6m/z 368.0218
C15H26Cl6m/z 382.0375
C15H25Cl7m/z 417.0058
C16H27Cl7m/z 431.0220
44
HRAM Selectivity 63% C10-C13 Technical Mix, NCI, 60k Resolution
C10H15Cl6
C11H17Cl6
10.15
C11H16Cl7
10.73
C12H18Cl7
10.69
11.21
10.22
11.70
9.72
RT:6.48 - 14.43
6.5 7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time (min)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rel
ativ
e A
bund
ance
NL:2.00E8TIC MS
NL:2.00E7m/z= 346.92652
NL:3.00E7m/z= 360.94208
NL:3.00E7m/z= 394.90330
NL:2.00E7m/z= 408.91872
NL:2.00E7m/z= 444.87597
NL:2.00E7m/z= 458.89157
NL:2.00E7m/z= 492.85283
TIC
11.20
C12H17Cl8
11.73C13H19Cl8
C13H18Cl9
45
Table 2. SCCP Linearity
• Linearity was assessed using the following dilution series for each SCCP chlorinationdegree (63% and 55.5%):
• 25• 50• 100• 250• 500• 1,000• 5,000• 10,000
• Concentrations as pg/µL.• In each standard, ~400 pg/µL p-Terphenyl-d14 was added and used as an IS
10,000 ppb
5000 ppb
46
Negative Chemical Ionization: Linearity for SCCP 63% C10
R² = 0.9998
R² = 0.9998
R² = 0.9997
R² = 0.9996
0.00E+00
1.00E+07
2.00E+07
3.00E+07
4.00E+07
5.00E+07
6.00E+07
0 1000 2000 3000 4000 5000 6000
peak
are
a co
unts
Concentration [pg/µL]
m/z 312.9665
m/z 346.9275
m/z 380.8886
m/z 416.8466
Excellent compound linearity (25-5,000 ppb) was obtained for these C10 chlorinated homologues present in the SCCP 63% technical mixture.
47
Peak Area Repeatability
• Peak area repeatability using full-scan NCI across n=10 repeatinjections of 25 ppb solvent standard (63% C10-C13).
inj. no m/z 492.8546(C13H18Cl9)
m/z 458.8936(C13H19Cl8)
1 765881 1308232
2 822551 1428540
3 795041 1361253
4 781911 1363928
5 776597 1321808
6 731874 1250508
7 761201 1305483
8 749797 1284342
9 737987 1257718
10 757772 1286412
mean 768061 1316822
StDev 27217.2 54540.3
%RSD 3.5 4.1
48
Chlorinated Paraffins, PCBs, toxaphenes, pesticides
Persistent Organic
Pollutants
“using high resolution, accurate mass Orbitrap-MS enables much deeper insights into the pattern and content of CPs without having to fear mass interferences from other CPs or halogenated compounds such as PCBs. Preliminary results suggest that determination of both CPs and PCBs in the same sample in one run is possible, representing a potential for shorter sample preparation and quicker analyses of these types of POPs in food”
HIGH RESOLUTION ACCURATE MASS SCREENING FOR CHLORINATED PARAFFINS IN FOOD SAMPLES USING GC-ORBITRAP MASS SPECTROMETRY (Kerstin Krätschmer et al. Dioxin 2017)
49
Summary - POPs Portfolio
Thermo Scientific™ TSQ™ 9000 GC-MS/MS
Orbitrap GC-MS
Magnetic Sector GC-HRMS
Thermo Scientific™ DFS™ Magnetic Sector GC-HRMS
GC-MS Triple Quadrupole
• Target compound analysis in routine • World-Wide compliance (e.g. EPA 1613) • Sensitivity combined with robustness for routine analysis• Robust-by-size with large volume ion source
• Target compound analysis in routine• Ease-of-use • Compliant with EU regulations for Dioxin food & feed analysis • Great price/performance ratio
Thermo Scientific™ Q Exactive™ GC Orbitrap™ GC-MS/MS
• Untargeted compound analysis • Method consolidation• Highest resolution and mass accuracy • Unique unknown identification capabilities
• Analysis of POPs is a challenging and varied application
• Legislation changes are allowing methods to be run on different instrumentation
• Thermo offers instrumentation to meet all the requirements for current and future analysis
50
Thank You!
Q & A
51
Thank You
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