environmental forensics - dioxin conference plenary 2011

Court D. Sandau, PhDChemistry Matters

Dioxin 2011 – Plenary – August 21, 2011

Environmental Forensics and POPs – Merging Fields of Expertise

Plenary Presentation at Dioxin 2011

• Honoured to be presenting a plenary talk at Dioxin 2011 in Belgium.

• Pictures from the talk below.

Dioxin 2011 Plenary - August 22nd, 2011, Belgium 2

Environmental Forensics

• What is it?

• Types of Environmental Forensics investigations

• Techniques used in EFIs

• POPs and EFIs

3www.chemistry-matters.com

Dioxin 2011 Plenary - August 22nd, 2011, Belgium

Environmental Forensic InvestigationThe systematic examination of environmental information, which may be used in litigation, to allocate responsibility for contamination

Success relies on an understanding of a variety of disciplines and knowing which tools are best suited for a particular case

“The application of scientific methods used to identify the origin and timing of a contaminant release”

Dioxin 2011 Plenary - August 22nd, 2011, Belgium 4www.chemistry-matters.com

What do POP researchers have to offer?

- Leading edge research- Finding compounds since

PCBs were discovered- Knowledge of contaminants

- biotransformation, air dispersion, multi-component mixtures, impurities in mixtures

- Breadth of knowledge of analytical and environmental chemistry (and related sciences)

After discovering PCBs because they were interfering with his DDT chromatograms….Methyl-sulphonyl metabolites of PCBs discovered by leaving GC on, chart recorder kept reading all night, caught late eluting chemicals. Soren Jensen. 1976.



Birthplace of Environmental Forensics

Exxon Valdez 1989Term “Environmental Forensics” coined in Dr. Robert Morrison’s first books since it was used in a peer reviewed publication.



Crime Scene Investigations



Environmental Forensic Investigation

Heisenberg is out for a drive when he's stopped by a police officer. The cop says: " Do you know how fast you were going? Heisenberg replies: "No, but I know where I am".



Two Categories of EFIs

Academic & Research

Liability Driven



Litigation and Liability Driven EFIs

• Adversarial process• Goal – make you look bad or incompetent while

making themselves look good/sympathetic• They are trying to tell if you are telling the truth or not• Good science does not necessarily prevail• Unfortunately, science is complicated (scientific

literacy 28% in 2007)• Need to prove it beyond a reasonable doubt



Differences Between Legal and Research

• Documentation, Documentation, Documentation

Project id.

Address

Project id.DateDate

Date

Notes and pictures/videos




• Documentation• Sample Handling• Chain of Custody




• Documentation• Sample Handling• Chain of Custody• Laboratory Competence• Standard Procedures• Communicating Results



Carpet Dust Sampling

• Must follow standardized procedures where available

• Difficult to change approaches

• Validation required if methodology changes

HVS-3 Vacuum Cleaner



Contaminants where Environmental Forensics has been used

• Petroleum Hydrocarbons (refined/unrefined)…• PAHs• Chlorinated solvents (PERC)• PCBs/PCDDs/PCDFs• Metals (selected)• VOCs• Methane (coal bed methane)• Pesticides (persistent/non-persistent)• Other persistent organic pollutants – soon…



Chemical Fingerprinting

Retention Time

Relative Response

S

S

Molecular fingerprint

=

Chemical Signature

DNA

Isotope Profiling=




Retention Time

Relative Response

S

S


=

Chemical Signature

Chemical Profiles

Marker compounds

- additives, indicator compounds, impurities



Marker Compounds in Petroleum Hydrocarbons

Note that the sample is severely

weathered.

Biomarkers have become the

predominant peaks

Isoprenoid / Hopane

Petroleum Biomarker

Standard

Site Sample

Soil samples from boggy areas have phytogenic

hydrocarbons



• Proprietary additives are compounds added to hydrocarbon products for specific purposes.

• Additives often have discrete time intervals during which they were introduced into a formulation.

Additives to Gasolines

Type of Additive Examples

Antiknock Alkyl Leads

Antioxidants p-phenylenediamine; alkyl-substituted phenols

Metal Deactivators Disalicylpropanediamine

Corrosion Inhibitors Carboxylic acids and diimides

Anti-icers Short-chained n-alcohols (freeze point depressants); amines and ethoxylated alcohols with long hydrocarbon chains.

Oxygenates Methanol, methyl-tertiary butyl ether (MTBE)

Lead Scavengers Ethylene dibromide, Ethylene dichloride

1980s62%

18%

2%



Dating with Additives

Historical trend in lead usage in gasoline

Phased out



Formulations of Aroclor 1254

- Dated Aroclor production changes

- Late production Aroclor - 1972

- Gives potential dating mechanism of PCB sources (unweathered)

G. Frame (1999) Improved Procedure for Single DB-XLB Column GC-MS-SIM Quantitation of PCB Congener Distributions and Characterization of Two Different Preparations Sold as “Aroclor 1254”Journal of High Resolution Chromatography 22:10, 533–540.

Johnson, G.W. et al (2007). Chapter 6: Principal components analysis and receptor models in environmental forensics. In: An Introduction to Environmental Forensics. 2nd Edition. (R. Morrison and B. Murphy, eds.). Elsevier. Amsterdam. pp. 207-272.

Dr. Glenn Johnson, Energy & Geoscience InstituteUniversity of Utah




Retention TimeRelative Response

S

S


=

Chemical Signature

Chemical Profiles



Limitations of Standard Practices

• Dioxin analyses focus on the dirty 17 congeners– Health driven, not forensically driven– PCBs went differently

• 193 other congeners to elucidate patterns of contamination

• More data is better for fingerprinting• Not limited to 2378 congeners• The other peaks are integrated for total

homologues

Dr. Walt Shields (Exponent) and Dr. Yves Tondeur (Analytical Perspectives) havepresented on patterns of dioxins (all congeners). Dioxin 2010.

- QA-QC of other congeners (standards!)



Dioxin Fingerprinting

Unstandardized Fingerprint



Normalizing Data

2378-Sum Relative Homologue

Relative TEQ Total Homologue



From Literature

Ba

rre

l B

urn

ing

Die

se

l F

ue

led

Tru

ck

Cre

ma

tori

um

2,3,7,8 SumRelative

Homologue Relative TEQTotal

Homologue



Case Studies

Inci

nera

tor

Unk

now

nC

reos

ote2,3,7,8 Sum

Relative Homologue Relative TEQ Total Homologue



Interpretation and Visualization

Stout et al. 2002, Intro to Enviro Forensics

Courtesy of ALS Laboratories, Edmonton



PCB Case Study

• PCBs measured in sediments from a man-made lake in USA

• Local authorities insisted that source was from my client

*13 sediments samples were analyzed for PCBs (congener specific analysis)



Measuring PCBs

• First measured on packed GC columns – Aroclor pattern analysis– Limited or no standards

• High resolution GC coupled with mass spectrometry (low / high resolution)– surrogates / isotope labels



Aroclor Differences

• Degree of chlorination shifts chromatogram to the right

• Most extensively used mixture was 1254– Two different formulations

• Weathering will also shift chromatogram to right

• Less time in environment = looks more like the source pattern and closer to the source

1242

1254

1260



Cosine Theta Analysis

• An analysis of the matrix of similarity coefficients between two different samples

• Compared each lake sediment sample congener pattern to each of the Aroclor patterns



Cosine Theta Analysis

Aroclor 12420.08, 85o

Sample X

Aroclor 1254

Aroclor 12480.20, 78o

Aroclor 12620.68, 47o

Aroclor 1260

0.91, 25o

0.75, 41o

Ort

hog

onal

Dis

sim

ilar

Co-linear, similar compositions Theoretical Vector1.00, 0o

Theoretical Vector0.00, 90o

• Normalize data into vectors

• Angles between the vectors are calculated

• Cosines of these angles represent the proportional similarity between the samples

• Nearest to one is closest resemblance



Sediment Pattern Similarity to Aroclor 1254

• Sediments were most similar to Aroclor 1254

• Aroclor 1254 most likely source

• Very little difference between early and late production Aroclor 1254 for cosine theta

• Closest to one is more resemblance to Aroclor 1254

Dam

0.63

0.64

0.670.67

0.75

0.780.77

0.76

0.740.67

0.790.85

0.86

0.89

Outfall




DNA

Isotope Profiling=



Isotopes

What is an isotope? - has same number of protons and electrons,

but different number of neutrons.

Protium Deuterium Tritium

Proton

Electron

Neutron

1H (99.98%) 2H (0.015%) 3H T1/2 = 12.32 years

Stable UnstableRadioactive3He



Common Stable Isotopes Used in EFIs

Hydrogen 1H, 2H

Carbon 12C, 13C

Nitrogen 14N, 15N

Oxygen 16O, 17O, 18O

Sulfur 32S, 34S

Chlorine 35Cl, 37Cl



Bacterial MF

Clarke Diagram for 2D Fingerprinting

2H-CH4 (‰)

13C

-CH

4 (

‰)

Bacterial Carbonate Reduction

Bacterial

Mix

and

Tr

ansi

tion

Early Mature

Thermogenic

after Whiticar, M.J., 1999 Organic matter

Stable Isotope Analysis

38

O’Sullivan, G, et al. (2010) Methane forensic geo-gas investigation: building an urban reference library. Environmental Forensics 11:1, 1-9.

www.chemistry-matters.comDioxin 2011 Plenary - August 22nd, 2011, Belgium

Stable Isotopes

-40

-35

-30

-25

-20

-15

δ 1

3 C (

‰)

Compound

• Stable isotope data gives a “signature” to samples

• Samples may be linked or differentiated

O

AWMA – Vapour Intrusion 2007



Radiocarbon Isotope 14C

Naturally occurring isotope with a half life of 5730 yrs

Measure in Percent Modern Carbon (pMC)



Radiocarbon Dating

Relies on the radioactive nature and influence of events such as nuclear testing and fossil fuel burning



Carbon 14 and Natural Brominated Compounds

Compound Δ14C (pMC)

MeO-BDE47 +103

MeO-BDE68 +119

Bromkal 70 -998

Aroclor 1260 -999

Teuten et al (2005) Two Abundant Bioaccumulated Halogenated Compounds Are Natural Products, Science, 307 (5711) p.917-920.

Can use natural abundance 14C measurements to uncover the relative contribution of industrial and natural sources of halogenated organic compounds to a variety of environmental media



Pushing Environmental Forensics Forward

What are the limits?

• How low can you go?• Complex mixtures?



m/z 321.8936 [M+2]

12C-2378-TCDD

in Serum

Using cryo-focusing and HRMS, can achieve 540 ag 2378-TCDD, assuming 70 % recoveryS/N 474 (4 Sigma)

m/z 319.8965 [M]

How Low Can We Go?

Dr. Donald G. Patterson, Jr and Wayman Turner

Zoex - loopmodulator



Extreme Sensitivity(2,3,7,8-TCDD)

Weight Moles MoleculesMicrogram (10-6) 3 Nanomoles (10-9) 2,000,000,000,000,000

Nanogram (10-9) 3 Picomoles (10-12) 2,000,000,000,000

Picogram (10-12) 3 Femtomoles (10-15) 2,000,000,000

Femtogram (10-15) 3 Attomoles (10-18) 2,000,000

Attogram (10-18) 3 Zeptomoles (10-21) 2,000

Zeptogram (10-21) 3 Yaktomoles (10-24) 2

Yaktogram (10-24) 3 Fantomoles (10-27) 0.002

Currently at the attogram level with high resolution mass spectrometry

Dr. Donald G. Patterson, Jr.



One part-per-quadrillion(ppq or ag/µL)

is like measuring 1 second in 1 x 1015 seconds

… or 1 second in 32,000,000 years

One part-per-quadrillion(ppq or ag/µL)

is like measuring 1 second in 1 x 1015 seconds

… or 1 second in 32,000,000 years

Wayman Turner 46www.chemistry-matters.com


Complexity?Conventional GC Analysis

Unresolved Complex Matter

Nelson et al. Environmental Forensics, 7:33–44, 2006



Two-Dimensional Gas Chromatography

Nelson et al. Environmental Forensics, 7:33–44, 2006



β-HCCH

γ-HCCH

HCBPCB-18

PCB-28

PCB-52

PCB-49

PCB-44

PCB-74+

HeptachlorEpoxide

Oxychlordane

PCB-101

PCB-66

DDE

PCB-99

t-Nonachlor

PCB-87

Dieldrin

PCB-110

PCB-151

PCB-149

PCB-146

PCB-153

o,pDDT

p,pDDT

PCB-118

BDE-28

PCB-105

PCB-138

PCB-158

PCB-178

PCB-183

PCB-187

PCB-177

PCB-172

PCB-180

PCB-128

PCB-167

PCB-156

PCB-157

BDE-47

BDE-66

Mirex

PCB-201

PCB-196+

PCB-203

PCB-170

PCB-195

PCB-194

PCB-206

PCB-209

BDE-99

BDE-100

PCB-189

BDE-153

BDE-154

BDE-85 BB-153

β-HCCH

γ-HCCH

HCBPCB-18

PCB-28

PCB-52

PCB-49

PCB-44

PCB-74+

HeptachlorEpoxide

Oxychlordane

PCB-101

PCB-66

DDE

PCB-99

t-Nonachlor

PCB-87

Dieldrin

PCB-110

PCB-151

PCB-149

PCB-146

PCB-153

o,pDDT

p,pDDT

PCB-118

BDE-28

PCB-105

PCB-138

PCB-158

PCB-178

PCB-183

PCB-187

PCB-177

PCB-172

PCB-180

PCB-128

PCB-167

PCB-156

PCB-157

BDE-47

BDE-66

Mirex

PCB-201

PCB-196+

PCB-203

PCB-170

PCB-195

PCB-194

PCB-206

PCB-209

BDE-99

BDE-100

PCB-189

BDE-153

BDE-154

BDE-85 BB-153

59 Analytes

Dr. Jef Focant

Separation Power of GCxGC – POPs



Dust Extract D/F Fraction by GCxGC-TOFMS

TIC

Dr. Donald G. Patterson, Jr and Dr. Jef Focant



Local Supply of Diesel Fuel

Dr. Ralph Ruffolo



Comparison of Gasolines

Gas Station 1

Gas Station 2

Dr. Ralph Ruffolo



Concluding an EFI

*Communicating the results becomes the focus of most EFIs

*Complicated methodologies, analytical techniques and results of the data interpretation must be easy to understand

53

3D-GIS Movie

Removed for smaller file size

www.chemistry-matters.comDioxin 2011 Plenary - August 22nd, 2011, Belgium

Environmental Forensics Resources



Societies and Organizations

• International Network of Environmental Forensics (INEF)

• Association for Environmental Health & Sciences (AEHS) Foundation

• International Society of Environmental Forensics (ISEF)



University Degrees and Training

• Masters degree – University of Strathclyde• Bachelor Degree – Bangor University• University of Florida – Certificate program

• Many courses on Environmental Forensics offered at Universities

• Environmental Forensics certification coming soon (AEHS Foundation)



Acknowledgements

Contact Info:

Chemistry Matters Inc.

Court Sandau

[email protected]

www.chemistry-matters.com

Twitter – Chem_Matters



mailto:[email protected]

http://www.chemistry-matters.com/