1. Derisking Offshore Oil Exploration with Lipidomics
G. Todd Ventura

2. Derisking Offshore Oil Exploration with Lipidomics
Petroleum Geochemistry
Bioassay 1
Aerobic HC Oxidizers
Bioassay 2
Anaerobic HC Oxidizers
Bioassay 3
Endospores
Lipidomics

3. How it works – Lipidomics and IPLs
Lipidomics – The large-scale study of the structure, pathways and networks of cellular lipids in biological and geochemical systems.

4. How it works – Lipidomics and IPLs
Extracellular fluid
Peripheral protein
Glycoprotein
Phospholipid
fatty acids
Glycolipids
Bilayer and
hydrophobic core
Cytoplasm

5. Intact Polar Lipids Bacterial IPLs – Archaeal IPLs – Head group
Glycerol
Aliphatic tail
Head group
ether
Isoprenoid tail

6. Diversity of IPLs
Aktas et al., 2017
Fredricks et al., 2004

7. Other Lipids of Consideration
Fossil Lipids:
Core lipids
Hydrocarbon biomarkers

8. Research Strategy
Proof of concept
Validation

9. Research Strategy – Proof of Concept
Carl has collected frozen piston core samples being stored at Bedford Institute of Oceanography (BIO).
Dr. Carl Peters
Mitacs Postdoctoral Fellow

10. Research Strategy – Proof of Concept
Matrix matched reference material – Bay of Fundy estuary mud
Total Lipid Extract (TLE) recovery:
Number of Repeats = 6

11. Bay of Fundy Mud – Matrix Matched Reference Material
500
750
1000
1250
1750
m/z 10 1 2 3 4 5
Int.
Bay of Fundy Mud – Matrix Matched Reference Material
(HILIC-uHPLC-1% TLE injected_qToF file 8159)
1G-Cren
HPH-Cren
1500
1G-GDGT-0
1G-OH-GDGT-0
2G-OH-GDGT
HPH-GDGT-0 BL
DPG
Lyso-DPG
2G-DAG
PC-DAG
PE-DAG OL
PG-DAG
PDME ?
&
Dimers
1G-DAG PI
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
Time [min]
22.5
BL – characteristic fragment in MS2 of m/z
1G-DAG – characteristic head group loss of 197 Da (sugar+NH4+)
PDME - characteristic head group loss of Da
PG - characteristic head group loss of Da
PC – characteristic fragment in MS2 of m/z
PE - characteristic head group loss of Da
PME - characteristic head group loss of Da
OL – characteristic fragment in MS2 of m/z
PI - characteristic head group loss of 277 Da

12. Research Strategy – Proof of Concept
Sample Sites
Modified from Fowler and Webb (2017)

13. Research Strategy – Proof of Concept
HC negative site
HC positive site
HC positive site

14. Research Strategy – Proof of Concept
A‘A‘‘
(HILIC-uHPLC - 10% TLE injected_qToF file 8166)
1G-GDGT-0
1G-Cren
1G-OH-GDGT-0
2G-OH-GDGT
1G-2OH-GDGT-0
2G-2OH-GDGT
2G-GDGT-0
500
750
1000
1250
1500
1750
m/z 10 1 2 3 4 5
Int.
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
Time [min]
22.5

15. Research Strategy – Proof of Conceptm
(HILIC-uHPLC-10% TLE injected_qToF file 8165)
500
750
1000
1250
1500
1750
m/z 10 1 2 3 4 5
Int.
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
Time [min]
1G-GDGT-0
1G-Cren
1G-OH-GDGT-0
2G-OH-GDGT
1G-2OH-GDGT-0
2G-2OH-GDGT
2G-GDGT-0
unknown
bacterial (?)
series
22.5

16. Research Strategy – Proof of Concept
Reproducible extraction method that is yielding lipids.
Extracts from HC positive Scotian shelf piston cores appear to be consistently greater than for a HC negative site.
The Scotian Shelf sediments have IPLs that are much less diverse than what we see for our reference standard.
Both HC positive and negative sites have Archaeal IPLs.
So far, the HC positive site also contains what are likely to be Bacterial IPLs.

17. Research Strategy – Proof of Concept
Reproducible extraction method that is yielding lipids.
Extracts from HC positive Scotian shelf piston cores appear to be consistently greater than for a HC negative site.
The Scotian Shelf sediments have IPLs that are much less diverse than what we see for our reference standard.
Both HC positive and negative sites have Archaeal IPLs.
So far, the HC positive site also contains what are likely to be Bacterial IPLs.
Research Strategy – Validation

18. Research Strategy - Validation
Dr. Casey Hubert and his group have conducted three sampling seasons.
2015 – Where, CCGS Hudson
2016 – Where, CCGS Hudson
2017 – Sydney Basin, RV Coriolis

19. Core Sampling Strategy
Research Strategy - Validation
Core Sampling Strategy
73 samples available
Representing:
8 HC positive sites:
Thermogenic
Biogenic
3 HC negative sites:

20. Research Strategy - Validation
Develop Scotian Shelf lipid library
Determine microbial substrate utilization via CSIA
Compare results to Calgary’s genomic data
SRB
ANME-2
ANME-1
1G-GDGT-0
1G-Cren
1G-OH-GDGT-0
2G-OH-GDGT
1G-2OH-GDGT-0
2G-2OH-GDGT
500
750
1000
1250
1500
1750
m/z 10 1 2 3 4 5
Int.
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
Time [min]
22.5

21. Conclusions
We believe we have the right strategy and support network to evaluate the use of IPLs as IHIs for offshore exploration in Nova Scotia.
Stay tuned for more!

22. Acknowledgements Collaborators:
Casey Hubert, Jayne Rattray, Julius Lipp, Florence Schubotz, and Kai Uwe-Hinrichs.
Funding Sources:
- Canadian Research Chair
- Canadian Foundation for Innovation – JELF
- Offshore Exploration Research Association
(Halifax, Nova Scotia, Canada)
- Department of Energy Nova Scotia
- Mitacs
- SMU FGSR

23. Thank you

24. How it works – Lipidomics and IPLs

25. Derisking Offshore Oil Exploration with Lipidomics
We are working to generate lipid profiles extracted from piston core sediments collected at known seep sites to target subsurface living microbial communities that are actively degrading petroleum-based hydrocarbons.
This project aims to compare the sensitivity of lipidomics with that of the University of Calgary’s bioassay technology.
The lipid assignments for hydrocarbon oxidation are equally to be compared with the petroleum geochemistry data produced by APT Ltd. Canada.

26. Microbial Hydrocarbon Oxidation
Contingent is the hypothesis working is that microbes are proliferating in seep sediments by anaerobically oxidizing petroleum forming hydrocarbons
How does this look:

27. Microbial Hydrocarbon Oxidation