Final day of the Dioxin 2016 conference in Florence, Italy, Dr. Court Sandau presented a summary of the analytical talks and posters presented at the Dioxin conference this year. Discussed the biggest highlights on PFAs, PBDEs, the history of Dioxin analysis and his impressions of where biomonitoring is going.

1. © 2016
Chemistry Ma2ers Inc. 1 Dioxin 2016 – Florence, Italy
Making chemistry data meaningful
New Methods and Perspec=ve of Analysis
Court Sandau, PhD
Donald G. Pa2erson Jr., PhD2
September 2, 2016
Dioxin 2016, Florence, Italy

2. © 2016
Chemistry Ma2ers Inc.
Looking for Inspira=on
2 Dioxin 2016 – Florence, Italy

3. © 2016
Chemistry Ma2ers Inc. 3
Quote for the day
A people without the knowledge of their past
history, origin and culture is like a tree without
roots. – Marcus Garvey
Dioxin 2016 – Florence, Italy

4. © 2016
Chemistry Ma2ers Inc.
History is Important
4
Clement R et al., Analytical Chemistry, 1991, 63, 1130A–1139A.
1971
1988
Dioxin 2016 – Florence, Italy

5. © 2016
Chemistry Ma2ers Inc.
Analy=cal Chemistry of the Time (1960s)
5
• GC-ECD (only seeing electronega@ve
compounds) and GC-FID very common
• Mass spectrometry s@ll quite specialized
(but needed for iden@fica@on)
• Most analy@cal standards not made yet,
need to be synthesized
• Packed GC columns provided very limited
resolving power
• No personal computers or google searches
• Literature search began in library, with
paper journals and took days/weeks
Dioxin 2016 – Florence, Italy

6. © 2016
Chemistry Ma2ers Inc.
PCB Metabolites Discovered
6
• A decade later (1976), Dr. Soren Jensen s@ll
analyzing PCBs in biota
• Le the laboratory in a hurry one evening
• Data was collec@ng on a Bal@c Seal sample
• A group of peaks elu@ng late from the GC-
ECD
• Chemically synthesized standards to confirm
iden@fica@on
• Iden@fied new compound group -
metabolites of PCBs
– methylsulfonyl-PCBs
– Opened new research into the field of POP
metabolites
• Comparing PCBs to metabolites, need as
many PCBs as possible measured in the
samples
Jensen , Ambio 1976 Vol. 5, No. 5/6, 257-260
Swedish Environmental Protection Agency, Persistent
Organic Pollutants, 1998, pp. 152.
Dioxin 2016 – Florence, Italy

7. © 2016
Chemistry Ma2ers Inc.
• Chromatogram of early dioxin
analysis of Agent Orange
• Inves@gators began by
iden@fying all congeners for
chlorinated dioxins and furans in
the chromatograms
– 210 congeners exist
• Using mass spectrometry and
glass packed columns
• Synthesized compounds for
posi@ve iden@fica@on
7
Rappe C et al., Chemosphere, 1978, 5, 431–438.
Dioxin 2016 – Florence, Italy
Rappe C, Gara A, Buser H.R, Chemosphere, 1978, 12, 981 – 991.
Early Inves=ga=ons in Dioxin Contamina=on
in Agent Orange and Chlorinated Phenols

8. © 2016
Chemistry Ma2ers Inc.
DIOXIN-2016 Analy=cal Techniques
GCxGC-TOFMS
“Simpler”
GCxGC-MS/MS
Convergence Chromatography
Timed-Cryogenic Zone
Compression HRMS
NCI-GC-HRMS
LC-MS/MS
APGC-MS/MS
Bio-Assays
GC-MS/MS
“FASTER”
GCxGC-QMS
GCxGC-HRTOFMS
No Targeted Analysis
Parallel Dual-Column
GCxGC-TOFMS
LC-APCI-MS/MS
Dioxin 2016 – Florence, Italy
Supercri=cal
Fluid DP-APCI/APPI-HRTOF-MS
HRGC-HRMS
8

9. © 2016
Chemistry Ma2ers Inc.
Analy=cal Content at Dioxin2016
9
• 118 talks and
posters covered
analytical
subject matter
• 21% of all
posters/talks
Dioxin 2016 – Florence, Italy

10. © 2016
Chemistry Ma2ers Inc.
Some of the Highlights
10 Dioxin 2016 – Florence, Italy

11. © 2016
Chemistry Ma2ers Inc.
Bioanaly=cal Developments
• CALUX Bioassay
becomes more
sensi@ve
• 4x increase in
response
11 Dioxin 2016 – Florence, Italy

12. © 2016
Chemistry Ma2ers Inc.
Finding the Best Method for PFAs
12 Dioxin 2016 – Florence, Italy

13. © 2016
Chemistry Ma2ers Inc.
Upda=ng Methods/Instrumenta=on
13 Dioxin 2016 – Florence, Italy
• Using nitrogen carrier
gas and atmospheric
pressure ioniza@on
– Measuring PBDEs

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Revised MDL Study, Manuscript in preparation
Dioxin 2016 – Florence, Italy 14
Comparing Instrumenta=on

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Chemistry Ma2ers Inc.
Firefighter helmet
wipes
• Model the direct
exposure of
firefighters to contact
with PXDD/F
compounds
Anal. Chem., (2015) 87 (20), 10368-77
Dioxin 2016 – Florence, Italy 15

16. © 2016
Chemistry Ma2ers Inc.
Interlaboratory Studies
16
Histogram
2,3,4,7,8-PeCDF
Sugar Beat Pulp Results - 2,3,4,7,8-PeCDF
Reported results: 87
Report results > LOQ: 86
Robust mean: 0.88 ng/kg (CV: 30%)
Median: 0.89 ng/kg
Modes: 0.81 ng/kg, 1.03 ng/kg
Contribution to WHO-PCDD/F-TEQ: ca. 50 %
Dioxin 2016 – Florence, Italy

17. © 2016
Chemistry Ma2ers Inc.
c:Xcalibur...cdx_240315cdx_240315-16 3/25/2015 1:34:24 AM Zuckerrübenschnitzel-sugar beet pulp_t
150141982-001
RT: 19.31 - 21.21
19.4 19.5 19.6 19.7 19.8 19.9 20.0 20.1 20.2 20.3 20.4 20.5 20.6 20.7 20.8 20.9 21.0 21.1 21.2
Time (min)
0
20
40
60
80
100
0
20
40
60
80
100
0
20
40
60
80
100
RelativeAbundance
0
20
40
60
80
100
RT: 19.46
AA: 727412
RT: 20.94
AA: 584377
RT: 19.52
AA: 580258 RT: 20.44
AA: 389185
RT: 19.94
AA: 256338 RT: 21.01
AA: 251896RT: 20.26
AA: 169059
RT: 20.03
AA: 182541 RT: 20.12
AA: 90417
RT: 19.82
AA: 83916
RT: 19.66
AA: 53010
RT: 20.59
AA: 31000
RT: 19.46
AA: 455035 RT: 20.93
AA: 360979
RT: 19.52
AA: 352698 RT: 20.44
AA: 238310
RT: 19.93
AA: 161784 RT: 21.01
AA: 155133RT: 20.26
AA: 106435
RT: 20.03
AA: 106185 RT: 20.11
AA: 57517
RT: 19.82
AA: 50238
RT: 19.66
AA: 31708
RT: 20.59
AA: 18870 RT: 21.15
AA: 755
RT: 21.01
AA: 181912
RT: 20.25
AA: 53012
RT: 20.44
AA: 1830
RT: 20.93
AA: 862
RT: 21.15
AA: 441
RT: 19.93
AA: 340
RT: 19.52
AA: 422
RT: 19.39
AA: 284
RT: 21.01
AA: 113616
RT: 20.24
AA: 30769
RT: 20.46
AA: 431
RT: 20.88
AA: 507
RT: 20.41
AA: 276
RT: 20.59
AA: 283
RT: 20.01
AA: 252
RT: 19.95
AA: 170
RT: 19.47
AA: 98.68
NL:
1.77E5
m/z=
339.8595-
339.8601
MS ICIS
cdx_24031
5-16
NL:
1.09E5
m/z=
341.8566-
341.8572
MS ICIS
cdx_24031
5-16
NL:
5.86E4
m/z=
351.8996-
351.9004
MS ICIS
cdx_24031
5-16
NL:
3.33E4
m/z=
353.8966-
353.8974
MS ICIS
cdx_24031
5-16
It S=ll Comes Down to Chromatography
2,3,4,7,8-PeCDF
DB-5 MS (Agilent)
c:Xcalibur...cdx_200215cdx_200215-31 3/4/2015 8:28:08 PM 1 Animal Feed 1501-AFB-74.1_t
AFB-74.1
RT: 26.77 - 34.51
40
60
80
100
0
20
40
60
80
100
0
20
40
60
80
100
RelativeAbundance
0
20
40
60
80
100
RT: 28.17
AA: 28645 RT: 33.31
AA: 28793RT: 27.44
AA: 33942 RT: 28.66
AA: 24124
RT: 30.64
AA: 16621
RT: 27.75
AA: 13713 RT: 30.59
AA: 6037
RT: 27.22
AA: 6541 RT: 30.00
AA: 4640
RT: 31.59
AA: 3998
RT: 33.74
AA: 3370
RT: 31.19
AA: 2599
RT: 33.53
AA: 441
RT: 28.17
AA: 18462 RT: 33.31
AA: 18170RT: 27.45
AA: 21557
RT: 28.66
AA: 15302
RT: 30.64
AA: 10406
RT: 27.75
AA: 8864
RT: 30.59
AA: 3999
RT: 27.23
AA: 4251
RT: 30.00
AA: 2758
RT: 31.59
AA: 2671
RT: 33.75
AA: 1359
RT: 31.19
AA: 1785
RT: 33.55
AA: 250
RT: 30.74
AA: 164
RT: 33.29
AA: 34396
RT: 27.41
AA: 7541
RT: 28.02
AA: 287
RT: 33.40
AA: 124
RT: 32.78
AA: 124
RT: 28.65
AA: 237
RT: 32.99
AA: 105
RT: 33.27
AA: 21044
RT: 27.42
NL:
5.58E3
m/z=
339.8595-
339.8601
MS ICIS
cdx_20021
5-31
NL:
3.69E3
m/z=
341.8566-
341.8572
MS ICIS
cdx_20021
5-31
NL:
6.28E3
m/z=
351.8996-
351.9004
MS ICIS
cdx_20021
5-31
NL:
3.57E3
m/z=
353.8966-
353.8974
MS ICIS
cdx_20021
5-31
Rtx-2330 (Restek)
• Values from ‘expert’ labs similar to other labs, therefore
even experienced labs were not getting the identification
correct.
• Analysts need to know their chromatograms.
• High TEQ, makes this compound important for studies.
2,3,4,7,8-PeCDF
Dioxin 2016 – Florence, Italy 17

18. © 2016
Chemistry Ma2ers Inc.
Comprehensive Analysis + Metabolomics
18
• 209 PCB Analysis (159
unique peaks w/ co-elutions)
• Surprising congeners in
samples
• Need to measure all potential
compounds
Dioxin 2016 – Florence, Italy

19. © 2016
Chemistry Ma2ers Inc.
Non-Targeted Analysis – Data Issues
19 Dioxin 2016 – Florence, Italy

20. © 2016
Chemistry Ma2ers Inc.
Where are We Going?
20
Multi-residue extraction method
Acidic, Basic and Neutral compounds
Non-targeted/Ultra Trace
Quantitation or
Semi-Quantitation
Multiple Methods?
Gene Array/Gene Expression
Quantitative Biomarker/Metabolites
Dioxin 2016 – Florence, Italy
Large Data Sets
Data Handling/Storage
Statistical Interpretation

21. © 2016
Chemistry Ma2ers Inc.
Big Data is Coming – Are you Ready?
21
http://itknowledgeexchange.techtarget.com/IT-watch-blog/files/2013/05/shutterstock_134968730.jpg
Dioxin 2016 – Florence, Italy

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Chemistry Ma2ers Inc.
Thank you for your attention
Contact Info:
Chemistry Matters Inc.
Court Sandau
Cell: 403.669.8566
Email: csandau@chemistry-matters.com
URL: chemistry-matters.com
Visit my website and sign up for email for automatic
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Twitter: @Chem_Matters
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