analysis of dioxins and pops using atmospheric pressure gc/ms
TRANSCRIPT
Analysis of Dioxins and POPs using
Atmospheric Pressure GC/MS
Ingrid Ericson Jogsten
MTM research centre, Örebro University
2014-12-16 1
2014-12-16 2
Outline
POPs and related environmental pollutants
POPs analysis
APGC
Dioxins
Other POPs
APGC vs high res for PCBs and OCPs
BFRs
PBDEs
Other environmental pollutants
Progress of scientific knowledge
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Scientific publications
Knowledge on effects
"PCBs and effects"
"PFOS and effects"
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400 Scientific publications
Analytical development
"Analytical Chemistry"
Slide courtesy of Samira Salihovic
Environment and Health Research focussed on POPs
• Health and exposure
• Environmental Levels
• Chemical and Bioassay Analysis
• UNEP reference laboratory
Stockholm Convention
Parties (179), Signatories (152)
Chemical Parent POPs Transformation products
Aldrin aldrin
Dieldrin dieldrin
Endrin endrin
Chlordane cis- and trans-chlordane cis- and trans nonachlor, oxychlordane
Heptachlor heptachlor -heptachlorepoxide
DDT 4,4’-DDT, 2,4’-DDT 4,4’-DDE, 2,4’ DDE, 4,4’-DDD, 2,4’-DDD
HCB hexachlorobenzene
Mirex mirex
Toxaphene congeners P26, P50, P62
PCB Σ7 ‘marker’ congeners: #28,#52,#101,#118,#138,#153,#180
12 congeners with TEFs: #77, #81, #105, #114, #118, #123, #126, #156, #157,
#167, #169, #189
PCDD/PCDF 2,3,7,8-PCDD/PCDF (17 congeners)
New POPs on SC 2009
HCHs hexachlorocyclohexane
PCBz pentachlorobenzene
BFRs PBDEs (not DeBDE #209), PBB
PFAS PFOS and its salts, POSFs
Substances to Be Monitored
Substances to be monitored (GMP guidance)
ESI
APCI
APPI
GC/MS
MALDI PFOS (LC-MS/MS)
Dioxins (HRGC/HRMS)
PBDEs (NCI m/z 79/81 non-specific)
Toxaphene, Drins (large fragmentation EI -> NCI)
Different Ionization Techniques?
Universal MS platform for POPs
analysis
2014-12-16 8
Atmospheric Pressure Ionisation
APGC
Plasma
Corona Pin
Analyte Molecules
Sample Cone
Make-up gas (N2)
Mechanism of Ionization (I)
N2+●
N2 e-
2e-
2N2
N4+● M●+
M Corona Pin
M●+
M
Charge Transfer
“Dry” source conditions
Favored by relatively non-polar compounds
Horning et al. 1973 (Anal. Chem, 1973, 45, 936-943)
Mechanism of Ionization (II)
N2+●
N4+●
H2O
H2O+●
H2O
H3O+●
+OH●
[M+H]+
M
Protonation
Modified source conditions eg. with water or methanol present
Favored by relatively polar compounds
Corona Pin
Horning et al. 1973 (Anal. Chem, 1973, 45, 936-943)
Analytical difficulties using high
resolution magnetic sectors
instruments
Dioxin analysis on HRGC-HRMS
• GC separation
– Capillary column
• Ionisation
– EI
• Mass detection
– SIR
• Resolution
– > 10 000
– 0.01 amu
Sensitivity 100 fg and 10 fg TCDD on
column
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Sensitivity mix; TCDDs
100 fg, 50 fg, 25 fg, 10 fg, 5 fg, 2 fg
Dioxins on APGC
Congener Concentration (pg/µL)
Name 1/10 CSL* CSL CS0.5 CS1 CS2 CS3 CS4
TCDD 0.01 0.1 0.25 0.5 2 10 40
TCDF 0.01 0.1 0.25 0.5 2 10 40
PCDD 0.05 0.5 1.25 2.5 10 50 200
PCDF 0.05 0.5 1.25 2.5 10 50 200
HxCDD 0.05 0.5 1.25 2.5 10 50 200
HxCDF 0.05 0.5 1.25 2.5 10 50 200
HpCDD 0.05 0.5 1.25 2.5 10 50 200
HpCDF 0.05 0.5 1.25 2.5 10 50 200
OCDD 0.1 1 2.5 5 20 100 400
OCDF 0.1 1 2.5 5 20 100 400
APGC Dioxins
Dilution 1/10 of CSL (10-100 fg/ul), 1 ul injection
Comparing APGC vs HRMS
APGC HRMS
EURL 0.85 0.83 2%
(pg/g lipids) 0.69 0.72 -3%
1.24 1.31 -6%
1.07 1.14 -7%
1.86 1.89 -2%
3.46 3.39 2%
MTM 6.1 5.8 5%
(pg/PUF) 13.9 14.0 -1%
45.6 47.7 -5%
63.8 62.0 3%
172 168 3%
17.3 16.2 7%
CSIC/IUPA 2.19 2.12 3%
(pg/g) 0.40 0.41 -2%
0.62 0.59 4%
238 228 4%
3640 3470 5%
96.2 89.4 7%
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Favorable ionization for OCDD
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CS4_H2O
m/z310 320 330 340 350 360 370 380 390 400 410 420 430 440 450
%
0
100
m/z310 320 330 340 350 360 370 380 390 400 410 420 430 440 450
%
0
100
DIOXINS0065 6062 (30.654) MS2 AP+ 3.00e7
443.70
423.74407.74
405.74372.04355.01329.08315.08
301.23342.08 388.90
439.71445.69
446.71
DIOXINS0006 6054 (30.614) MS2 AP+ 3.00e7
443.68
441.68
439.69
423.73
422.72407.73
404.95385.78373.85371.12329.91314.95 333.02 348.96
436.71
445.67
446.69
448.67
H2O
N2
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Additional POPs
PCBs and organochlorine pesticides
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Instrumental comparison
APGC vs high resolution GC/MS
N=8
03-Nov-200910 ng/ul pestmix 13 coneflow at 30
Time9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50 13.00 13.50 14.00 14.50
%
0
100
XTQ_021109_010 1: MS2 AP+ TIC
2.62e8
11.96
11.66
9.26
9.19
10.91
10.47
9.66
9.53 10.17
10.03
10.67
11.16
11.54
11.70
12.48
12.14
12.38
13.39
13.0812.88
12.75
14.2314.06
13.95
14.75
Pest mix 13, 10 ng/ul, no modifier, fullscan APGC
Stockholm Convention POP mix (APCI) Pesticide-Mix 13
PCB No. 28
PCB No. 52
PCB No. 101
PCB No. 138
PCB No. 153
PCB No. 180
Aldrin
cis-Chlordane
trans-Chlordane
oxy-Chlordane
2,4’-DDD
4,4’-DDD
2,4’-DDE
4,4’-DDE
2,4’-DDT
4,4’-DDT
Dieldrin
alpha-Endosulfan
beta-Endosulfan
Endrin
alpha-HCH
beta-HCH
gamma-HCH
delta-HCH
epsilon-HCH
Heptachlor
cis-Heptachlor
Several halogenated Non-BDE Replacement Flame Retardants were
analysed using APGC by Reiner et al. (2010)
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Brominated flame retardants (BFRs)
- 2,2',4,4',5,5'-Hexabromobiphenyl (BB-153)
- Hexabromocylcododecane (HBCD) (α, β, γ)
- Tetrabromobisphenyl-A (TBBPA)
- Allyl 2,4,6-tribromophenyl ether (ATE)
- 2-Bromoallyl-2,4,6-tribromophenyl ether (BATE)
- 2,3-Dibromopropyl-2,4,6-tribromophenyl ether (DPTE)
- Octabromotrimethylphenylindane (OBIND)
- Pentabromoethylbenzene (PBEB)
- Hexabromobenzene (HBB)
- 1,2-Bis(2,4,6-tribromophenoxy) ethane (BTBPE)
- Decabromodiphenylethane (DBDPE)
- Dechlorane Plus (DP) (anti, syn)
- Hexachlorocyclopentadienyl-dibromocyclooctane (HCDBCO)
- 2-Ethylhexyl-2,3,4,5-tetrabromobenzoate (EHTeBB)
- Bis(2-ethly-1-hexyl)tetrabromophthalate (BEHTBP)
- 2,2',3,3',4,5,5',6,6'-Nonabromo-4'-chlorodiphenyl ether (4PC-BDE208)
- Dechloranes – 602, 603, 604
Polybrominated diphenyl ethers (PBDEs)
Optimization of collision energy
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CE 20 V CE 45 V
Total ion chromatogram by
APGC-MS/MS
RRF and r2 of the calibration curve
0.04pg/uL 0.6pg/uL 4pg/uL 12pg/uL 60pg/uL average STD RSD(%) r r2
BDE#28 8.247 7.233 7.507 7.536 8.415 7.788 0.51 6.6 0.9990 0.9980
BDE#47 5.57 4.89 4.586 5.096 4.559 4.940 0.42 8.4 0.9991 0.9982
BDE#66 4.526 4.049 4.167 4.761 4.612 4.423 0.30 6.8 0.9996 0.9992
BDE#100 2.026 2.177 2.205 2.26 2.34 2.202 0.12 5.3 0.9998 0.9997
BDE#99 2.477 2.094 2.211 2.13 2.506 2.284 0.19 8.5 0.9981 0.9963
BDE#85 5.44 5.372 5.447 5.591 6.693 5.709 0.56 9.7 0.9973 0.9945
BDE#154 2.175 2.432 2.42 2.478 3.312 2.563 0.43 17 0.9938 0.9876
BDE#153 2.694 2.184 2.233 2.522 3.001 2.527 0.34 13 0.9965 0.9930
BDE#138 1.692 1.731 1.663 1.877 2.074 1.807 0.17 9.4 0.9984 0.9969
2014-12-16
2014-12-16
Comparison on PBDEs concentrations
analyzed by APGC-MS/MS and HRGC/HRMS
Important to include BDE# 209
2014-12-16 J. Muñoz-Arnanz et al. / Environment International 37 (2011) 572–576
2014-12-16 33
PBDE analysis on APGC
• Problems in sample transfer – GC-MS
interface – probably activation of GC column
section in the interface region (atmospheric
conditions, cold spots), uneven heat
distribution.
• Sensitive compounds (DBDPE, octa-
decaBDEs) exhibit significant peak
broadening and delays
APGC problem solving
APGC problem solving
APGC problem solving
• Siltek deactivation showed promising
• Siltek deactivated capillary (0.25 mm id, same as column) and
Siltek pressfits (a bit tricky to connect)
• BDE#209 peaks ~15s wide, broadening after 40+ injection
APGC problem solving
APGC problem solving
Siltek capillary 0.25mm id, Siltek pressfit, after 35 injections,
He flow 3ml/min, interface 360C
Siltek capillary 0.25mm id, Siltek pressfit, after 76 injections,
He flow 3ml/min, interface 360C
APGC Final Settings
• Use of Siltek treated capillaries and unions (thermally resistant up to
min. 380°C)
• High column carrier gas flows and high temperatures (to overcome
uneven heat distribution and minimize “dwell time” of analytes in
interface region)
Optimized APGC Conditions
GC
Column Rtx®-1614, 15 m ×
0.25 mm, 0.10 μm
Carrier gas Helium 3 mL/min
Injector mode Pulsed Splitless,
450 kPa (1 min)
Column pneumatics Constant flow
Injection volume(μL) 1
Injector temp(℃) 280
2014-12-16
MS
Ionization APGC with Dry N2
Corona current(μA) 2.5
Source offset(V) 70
Cone voltage(V) 30
Source temp(℃) 150
Cone gas flow(L/hr) 160
Collision gas Argon at 3.5·10-3 mbar
Acquisition Multiple Reaction Monitoring
(MRM)
Transfer line
Column Siltek®-Deactivated Guard, 0.25mm(ID), 0.37±0.04 mm(OD)
Temp (℃) 360 Make up gas flow(L/hr) 350
RRF and r2 of the calibration curve
0.04pg/µL 0.6pg/µL 4pg/µL 12pg/µL 30pg/µL average STD RSD(%) r r2
BDE#28 8.612 8.886 9.425 10.217 9.832 9.394 0.66 7.0 0.9997 0.9994
BDE#47 8.72 7.867 7.677 9.536 8.378 8.436 0.74 8.8 0.9979 0.9954
BDE#66 6.401 6.605 6.775 7.377 7.32 6.896 0.43 6.3 0.9997 0.9994
BDE#100 5.22 5.182 5.791 5.747 6.13 5.614 0.41 7.2 0.9994 0.9989
BDE#99 4.981 4.923 5.765 5.437 5.635 5.348 0.38 7.1 0.9998 0.9995
BDE#85 2.623 2.835 2.937 3.562 3.363 3.064 0.39 13 0.9988 0.9975
BDE#154 2.139 2.245 2.308 2.69 2.541 2.385 0.23 9.5 0.9992 0.9983
BDE#153 1.76 1.802 1.861 2.294 2.077 1.959 0.22 11 0.9982 0.9965
BDE#138 1.375 1.435 1.578 1.869 1.649 1.581 0.19 12 0.9981 0.9961
BDE#183 1.056 1.098 1.193 1.376 1.337 1.212 0.14 12 0.9992 0.9985
BDE#209 1.018 1.089 1.06 1.099 1.041 1.061 0.03 3.2 0.9997 0.9994
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BDE#209 in osprey eggs
2014-12-16
PBDE results summarized
1. The APGC Xevo TQ-S is a higly sensitive instrument for
the analysis of PBDEs. The results of the APGC-MS/MS
on osprey eggs samples were in very good comparison
with the high res results from tri to hexa-BDEs.
2. All target compounds were successfully detected in the
low level standards in a single run.
3. Excellent linearity was obtained for all compounds over
the range 40fg/µL to 30 pg/µL.
4. The detection of higher brmomated compounds in osprey
eggs looks promising.
2014-12-16
What is new?
Dave Stalling et al. 1982
Polybrominated dibenzo dioxins and
furans (PBDD/F)
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PBDD/F
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Overlaid chromatograms of
three consecutive injections of
500 fg TBDD on column
Precursor ion Product
ion
Area Time
499.7 392.8 792 8.25
499.7 392.8 739 8.25
499.7 392.8 760 8.25
Std dev (area) 26.7
RSD (area) 3.5%
PBDD/Fs 0.5-50 pg on column
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Late eluters
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2 µl – 1 050 000
1 µl – 570 000
3 µl – 1 520 000
4 µl – 2 000 000
APGC – Injection volume vs area
Fluorotelomer alcohols (FTOH) and
fluorinated sulfonamide/ethanols
(FOSA/FOSE)
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FTOH – APCI+
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70 V
30 V
[M-HF]+
[M-H]+
[M+H]+
100 fg FTOH on column
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FOSA/FOSE – APCI-
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Universal MS platform
UPLC analysis of PFASs
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Perfluoroalkylcarboxylic acids (PFCAs):
PFBA (C4)
PFPePA (C5)
PFHxA (C6)
PFHpA (C7)
PFOA (C8)
PFNA (C9)
PFDA (C10)
PFUnDA (C11)
PFDoDA (C12)
PFTrDA (C13)
PFTDA (C14)
Perfluoroalkylsufonic acids
(PFSAs):
PFBuS (C4)
PFHxS (C6)
PFOS (C8)
PFDS (C10)
Structural PFOS isomers:
L-PFOS (linear PFOS)
1-PFOS
6/2-PFOS
3/4/5-PFOS
4.4/4.5/5.5-PFOS
Fluorotelomer carboxylic acids (FTCA)
3:3 FTCA
5:3 FTCA
7:3 FTCA
Fluorotelomer unsaturated acids (FTUCA)
6:2 FTUCA
8:2 FTUCA
10:2 FTUCA
Polyfluoroalkyl phosphate surfactants (PAPS)
6:2 monoPAP, 8:2 monoPAP, 10:2 monoPAP 6:2 diPAP (including isomers 4:2/8:2, 2:2/10:2), 6:2/8:2 diPAP (including isomers 4:2/10:2) 8:2 diPAP (including isomers 6:2/10:2, 4:2/12:2), 8:2/10:2 diPAP (including isomer 6:2/12:2) 10:2 diPAP (including isomers 8:2/12:2, 6:2/14:2),
• APCI
– Similar to electrospray
– Soft ionisation
– Efficient ionisation
– Two mechanisms
• Charge transfer
• Protonation
– Interface
• TOF (High Res)
• MS/MS
• Universal instrumentation
for POPs analysis and
related compounds
– APGC • Dioxins (PCDD/Fs,
PBDD/Fs)
• PCBs
• Organochlorine pesticides
• Brominated flame retardants (PBDEs)
– UPLC-ESI • PFCAs/PFSAs
• FTCA/FTUCA
• PAPS
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Conclusions
Acknowledgements
2014-12-16 57
MTM research centre, Örebro University
Bert van Bavel, Dawei Geng, Filip Bjurlid
RECETOX; Petr Kukucka
Waters Corporation: Jody Dunstan, Keith Worrall, Rhys Jones
Laura Cherta, Jaime Nácher-Mestre, Tania Portolés, Manuela Ábalos, Jordi Sauló, Esteban
Abad, Jody Dunstan, Rhys Jones, Alexander Kotz, Helmut Winterhalter, Rainer Malisch, Wim
Traag, Joaquim Beltran, Félix Hernández
Thank you for your time!
Ingrid Ericson Jogsten
MTM research centre, Örebro University
2014-12-16 58