Analytical chemistry 
l‐sampling strategies
Dr. Pernilla Carlsson; carlsson@recetox.muni.cz, room 209, RECETOX
OutlineOutline
S li t t i• Sampling strategies
‐general limitations
‐bioaccumulation and sampling strategybioaccumulation and sampling strategy
• Your hypothesis and how to address it
• ”Chemical tools”• Chemical tools
‐chirality: environmental processes and different 
sourcessources
‐transport processes
‐fate of POPs
• Case study discussions
General issues
A t d t f d t• Amount and type of data
• Seasonal distribution
• Instrumental limitations and 
infrastructureinfrastructure
• Legislation
Sampling strategy ‐
biomagnificationbiomagnification
Cover the whole food web!
Biomagnification
PFOS in the eastern CanadianPFOS in the eastern Canadian
Arctic marine food web
• Who eats who?
• Benthic/pelagic couplings?
AMAP 2009
Your hypothesis and how to address itYour hypothesis and how to address it
• Clear research questions!
• Data neededData needed
• Financial and time frames
Your hypothesis and how to address itYour hypothesis and how to address it
• An example:
Does fish from the Baltic Sea contain higherDoes fish from the Baltic Sea contain higher
concentrations of PCB and dioxins compared to
fish from the Atlantic ocean?”fish from the Atlantic ocean?
Fish and POPsFish and POPs
• Study design:
• Fish –speciesFish species
• Sampling: fishing or buying?
• Transport
• AnalysesAnalyses
• Data interpretation
• Literature comparison
Chemical ”tools”Chemical tools
• Relative distribution 
• Degradation and metabolitesDegradation and metabolites
• Chirality
• Isotopes
Relative distributionRelative distribution
• PCBs: different mixtures had different ratio
between congenersg
• Low‐chlorinated PCBs: Easily undergo long‐
range transportrange transport.
PCBsPCBs
AMAP
http://www.amap.no/
Relative distribution –technical PCB 
mixtures
Arochlor: USA
Takasuga et al., 2006
PCB flux to the Arctic
ΣPCB13
(pg/L)
Altitude/
depth
Carrizo, D.; Gustafsson, O. Distribution and Inventories of Polychlorinated Biphenyls in the Polar Mixed Layer of Seven Pan‐
Arctic Shelf Seas and the Interior Basins. ES&T 2011.
Carrizo, D.; Gustafsson, O. Pan‐Arctic River Fluxes of Polychlorinated Biphenyls. ES&T 2011
PCB flux to the Arctic 
Altitude/
depth
 Tri‐PCB
 Tetra‐PCB
 Penta‐PCB
 H PCB Hexa‐PCB
 Hepta‐PCB
 Octa‐PCB
Carrizo, D.; Gustafsson, O. Distribution and Inventories of Polychlorinated Biphenyls in the Polar Mixed Layer of Seven Pan‐
Arctic Shelf Seas and the Interior Basins. ES&T 2011.
Carrizo, D.; Gustafsson, O. Pan‐Arctic River Fluxes of Polychlorinated Biphenyls. ES&T 2011
Relative distributionRelative distribution
• Long‐range transport: Will favour light, easy
volatile compoundsp
=>Changed ratio in Arctic compared to
technical mixturetechnical mixture
=>”Non‐changed ratio”: Local sources
Degradation and metabolitesDegradation and metabolites
d b li f• Hydroxy‐PCBs: Metabolism of PCBs to 
facilitate excreation of the compounds
• Oxychlordane: Common, stable metabolite 
from trans‐/cis‐nonachlor and –chlordane/
• DDE: Stable metabolite from DDT. Affect egg‐
shell thickness among birdsshell thickness among birds
Degradation of POPsg
• Cytochrome P450 (commonly known as CYP)y ( y )
• A superfamily of proteins (enzymes) found in the entire p y p ( y )
kingdom of life
• They can transform the structure of organic chemicals
CY 1A1 ibl f id i f O• CYP1A1 responsible for oxidation of many POPs
B ff ti h i l t t CYP k i• By affecting chemical structures, CYP may make a given 
compound nontoxic.
• Or drastically increase its toxicityOr, drastically increase its toxicity…
CYP1A
• Most important subfamily among the P450.
• 2,3,7,8‐TCDD, benzo(a)pyrene and dioxin‐like PCBs 
(non‐ and mono‐ortho‐PCBs) can induce the 
transcription of CYP1A genes via the aryl 
hydrocarbon (Ah) receptor.
2,3,7,8‐TCDD and PCB‐81.  www.chemspider.com
IsotopesIsotopes
Who eats who?
Trophic positions: “place in the food web”Trophic positions:  place in the food web
15N/14N=δ15N 
δ15N 
Illustration: Eldbjørg Heimstad
Transport processes
‐tracking sources
Grannas et al. (2013),  Jantunen et al .(2008),  
Kallenborn et al, (2012a), Noyes et al. (2009), 
Wong et al. (2011)
Chiralityy
–tracing environmental processes and sources
Chiral pesticides: a‐HCH, trans‐, cis‐, and oxychlordanep , , , y
Cl
Cl Cl
H
Cl
Cl
Cl
Cl
Cl
Cl
Cl
H
Enantiomer selective analyses
α‐HCH: ‐, + +Oxychlordane (OC), ‐OC, +trans‐chlordane (TC), +cis‐chlordane (CC), ‐CC, ‐TC
ance
e
Time (min)
Relativeabunda
Relativeabundance
Time (min)
Enantiomer fractions of chiral pesticides in plankton as a tool to 
differentiate between water masses
Chiral pesticides
non‐racemic EFs: indicatenon racemic EFs: indicate
biological transformation 
processes
Plankton
non‐selective metabolism, 
reflect EFs of pesticides in thereflect EFs of pesticides in the 
water mass
Peter Leopold © 
Carlsson et al, 2014
Sample
area
2007 2008 2009 2010 2011 Dominating
water mass
Winter ice
conditions
©Norwegian Polar Institute
area water mass conditions
Kongs-
fjorden
Hallanger et
al., 2011a
Hallanger et
al., 2011b
Carlsson et
al., 2014
Carlsson
et al., 2014
Atlantic None/partly
covered
Liefde- Hallanger et Carlsson et Atlantic/ March-JulyLiefde
fjorden
Hallanger et
al., 2011b
Carlsson et
al., 2014
Atlantic/
Arctic
March July
Rijp-
fjorden
Carlsson et
al., 2014
Carlsson
et al., 2014
Polar surface/
Arctic
Ice cover until
July
Pack ice Carlsson
et al.,
2014
Carlsson
et al., 2014
Meltwater/
Atlantic
Annual ice
cover
Median enantiomeric fractions in Calanus spp.
RF
2011
2011
KF ICE
RF
ICE
KF
KF= Kongsfjorden 
LF= Liefdefjorden
ICE = Ice stations (also represented by 
ag=Apherusa glacialis, gw=Gammarus
RF= Rijpfjorden 
ag Apherusa glacialis, gw Gammarus
wilkitzkii in 2010).
Carlsson et al, 2014
Chiral pesticides and water masses
•Ice cover and α‐HCH
(hindering of) volatilisation(hindering of) volatilisation
•Chlordanes and 2011
Large deviations from racemic trans‐chlordane
W d i i f h EF di ib i•Water masses and ice cover are important for the EF distribution.
•Impact from benthic‐pelagic processes not completely understoodImpact from benthic pelagic processes not completely understood
yet.
Case studies of environmental pollution
PCBs in a changing Arctic: Towards understanding their
input, transfer and uptake into Arctic biota and humans
under climate changeunder climate change
• Eva Brorström‐Lundén
• John Munthe, Hanna Andersson, Crispin Halsall, Roland Kallenborn, Pernilla Carlsson,  Henry Wöhrnschimmel, 
Matthew MacLeod, Ian Cousins, Deguo Kong, Gerhard Lammel, Arja Rautio, Khaled Abass 
Aim of PCB-case study:
•Synthesize ArcRisk results using PCBs as model substances
•Link sources and pathways to transfer processes and uptake
in biota and assess human exposure and effectsin biota and assess human exposure and effects
•Effect of future climate change on uptake processes
Eva Brorstrøm-Lundén, Swedish Environmental Research Institute, IVL
Describing the full sequence from emissions to exposure
R i f h• Review of the current status
• What will happen when the climate changes?
PCB emissions and pathways to the Arctic
• Sources, releases,
• Air and water transport and deposition fluxes
• Key intercompartmental transfer
PCB occurrence in the ArcticPCB occurrence in the Arctic
• Levels in abiotic matrices and biotic matrices (key biota)
• Time-series of concentrations
Human exposure of PCB
Policy, directives
WHY PCBs?
 Banned, but still present in large quantities in the environment. Banned, but still present in large quantities in the environment.
 Studied extensively in the Arctic for more than three decades.
 W ll d t d h i l h i l ti i i th d Well-understood physical-chemical properties, emissions, pathways and
environmental concentrations
=> PCBs are perfect to evaluate models and as a benchmark
for assessment of how other substances are influenced by
climate change.
PCB emissions (tons/year)
Global primary emissions of PCB-28 and PCB-153 to air (high-end estimate),
• Primary emissions from constructions and waste dumps
p y ( g ),
according to Breivik et al. (2007).
Primary emissions from constructions and waste dumps.
• Secondary emissions of PCBs will become more important than
primary sources.
• Climate change will most likely increase these secondary emissions.
Identification of the most important pathways
of PCBs to the Arctic
 Atmospheric long range transport is the major route for global
of PCBs to the Arctic
 Atmospheric long range transport is the major route for global
distribution of PCBs to the Arctic.
 Long-range transport via oceanic currents is also of high Long range transport via oceanic currents is also of high
importance.
 Climate change is likely to affect these pathways and thereforeg y p y
the environmental fate of PCBs.
Atmospheric concentrations of PCBs
Long term monitoring of PCBs are carried out in Arctic and EuropeLong term monitoring of PCBs are carried out in Arctic and Europe
AMAP and EMEP databases hosted by NILU EBASE (www ebas nilu no)AMAP- and EMEP databases hosted by NILU EBASE (www.ebas.nilu.no).
Atmospheric concentrations of PCBs
- Long term monitoringLong term monitoring
Arctic inter-compartmental transfer of PCBs
-Processes of importance for climate changep g
 Precipitation: Important factor for PCB levels in snowpack surface
 Climate change related effects: Changes in precipitation patterns and snow
melt periods
Arctic inter-compartmental transfer of PCBs
-Processes of importance for climate changep g
 Accumulation of PCBs in sea ice =>”chemical storage”
 Ice melting: Release of PCBs to the water and the marine food web, especially
during early spring.
 Climate change will affect pathways and mobility of PCBs in Arctic sea ice Climate change will affect pathways and mobility of PCBs in Arctic sea ice.
Arctic inter-compartmental transfer of PCBs
-Processes of importance for climate changep g
PCB-52 declines when the
air temperatures increase
above 0oCabove 0 C.
The heavier PCB-153/-132
do not show this trend.
Concentrations of PCB-52 and PCB-153/132 in the ice-rafted snowpack
f h f S (C d ) d lof the Beaufort Sea (Canada) during late winter.
What will happen when the climate changes?
 Model results: relative increases in PCB concentrations in the Arctic
atmosphere and ocean. Mainly due to the temperature effect on volatilisation.
 The absolute concentrations by 2090 are forecasted to be several orders of
magnitude below present levels in all scenarios.
 Atmospheric PCB concentrations in the Arctic have shown a continuous
decreasing trend over the past decades.
f
What will happen when the climate changes?
Modeled relative change of PCB-153 and PCB-28
concentrations in the Arctic and European atmosphere.
  PCB153 PCB28 
Arctic AEurope 
 
PCB-153 in the Arctic: relative increase of ~1.5.
PCB-153 in Europe:relative increase of 2.
PCB 28 l iti t li t h th ff t i ithi th tPCB-28: less sensitive to climate change, the effect is within the parameter
uncertainties.
Concentrations of PCBs in biota (food items)
Median (ng/g lw). Samples from Nuuk, Greenland (2010).
Bioavailability, food web transfer of PCBs
Bioavailability
Major climate change impact on bioavailability: distribution changes in
matrices, changed partitioning of PCBs in water due to increased temperatures.
Svalbard marine food web
Food web transfer acts as a link between the abiotic environment
and human exposureand human exposure
Health effects: Human exposure of PCBs
PCB-153 TREND DATA IN PLASMA LIPIDS (µg/kg), IN PREGNANT WOMEN LIVING IN
DISKO BAY (GREENLAND), NUUK (GREENLAND), AND NUNAVIK (QUEBEC,
CANADA) DURING YEARS 1992 – 2007
• In general, levels of PCB in humans
populations have declined over the
CANADA) DURING YEARS 1992 2007.
populations have declined over the
past 20-30 years.
• Declining trends of PCB-153 are
seen in all these regions in plasma
lipids in Inuit women.
Health effects
-Human exposure of PCBs in a warmer ArcticHuman exposure of PCBs in a warmer Arctic
 The impact of changes in contaminant concentrations in food will be small
l ti t th i t f h i di t b h irelative to the impact of changes in dietary behaviour.
 Consumption and type of fish and other seafood are the most important factors
for contaminant exposure, also in the future.p ,
Summary and conclusions
-PCBs in a changing Arctic
 ArcRisk results provides an overall picture of PCBs in the Arctic.
 Understanding of relationships between sources, transport, bioaccumulation, exposure
d h lth i t f PCB i l ti t li t hand health impacts of PCBs in relation to climate change.
 Long term monitoring of PCBs and other POPs in air, sea water and biota is important
both for evaluation of time trends and for detection of responses to environmental
hchange.
 Models are useful tools to predict future concentrations of contaminants in
environmental media and human exposure.
 Future needs: High quality datasets with simultaneously sampled biotic and abiotic
samples, which can be used to evaluate and improve model performance.
Contaminants in farmed versus wild fish
Salmon
Lowering risk of cardiovascular diseases.
Omega-3 fatty acids
S l f i=>Salmon farming
Farmed salmon (Hites et al 2004)Farmed salmon (Hites et al. 2004)
Higher concentrations of POPs compared to wild salmon
Cause: the feed used
Oceans and Human Health
Contaminants in farmed versus wild fish
Risk assessment
¤ Only cancer considered as effect
¤ Positive outcome of fish eating not included¤ Positive outcome of fish eating not included
¤ PCB and dieldrin concentrations ok according to US Food and Drug
Administration
¤ EPA guidlines: Combined concentrations of PCBs, toxaphenes andg , p
dieldrin were cause for concern.
¤ =>Eating farmed salmon 0.5-2 meals/month ok.g
Contaminants in farmed versus wild fish
But…
The combined effect of PCBs dieldrin and toxaphenes are not wellThe combined effect of PCBs, dieldrin and toxaphenes are not well
known.
Benefits of omega-3 fatty acids (reducing cardiovascular diseases) are
larger compared to the backdraws of POPslarger compared to the backdraws of POPs
Contaminants in farmed versus wild fish
Salmon feed today (Norway)
M t bili f d l POP d t fi h b d f dMore vegetabilic feed =>less POPs compared to fish-based feed.
However, also less fatty acids…
Norwegian farmed salmon in 2004:Norwegian farmed salmon in 2004:
ΣPBDE: 2.5 ng/g ww in salmon, BDE-47 and -99: 1.9 ng/g ww.
BDE-47 and -99 in (wild) mackerel and herring: 0.5-1.8 ng/g ww.
PCB-153
BDE-47
Norwegian Scientific Committee for Food Safety www.vkm.no
Study design of your projects
•Laboratory/field experiment
•Analytical methods•Analytical methods
•Interpretation of data –limitations?
•Time limitations?Time limitations?
•Suggestions for larger scale project
Summary ‐sampling strategies
• Plan your work: clear hypothesis, limitations of data, seasonality, time 
and financial frames
Ch i l “ l ” bl i hi li l i di ib i• Chemical “tools”: stable isotopes, chirality, relative distribution
• Important to link models with empirical data
• Identify limitiations in literaturey
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