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EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017 117
Sampling, monitoring and source
tracking of dioxins in the environment
of an incinerator in the Netherlands
A Arkenbout1 and K H Esbensen2,3,4,5
ABSTRACT
In the north of the Netherlands, in the environment of Harlingen, an industrial waste incinerator
was installed in 2011. The population in the region is concerned about adverse health effects
related to possible emission of dioxins. This study compares the relative merit and reliability of
four sampling approaches aimed at quantifying the possible pollution patterns surrounding the
incinerator: short-term versus long-term isokinetic ue gas sampling, spatial sampling of grasses
and a novel approach developed by the Toxicowatch Foundation, based on local composite
sampling of ten eggs from backyard chickens (local area pollution load averaging by accumulating
bio-entities). Based on the latter approach results of a new analytical bioassay, DR CALUX®, from
15 locations near Harlingen harbour show distinct above-threshold dioxin levels in the environs
of the incinerator. Currently only short-term ue gas sampling is mandated by the authorities;
based hereupon, under normal operating conditions, the incinerator appears to be compliant
with emission standards. This short-term sampling scheme is seriously awed, however, in
that it only demands one continuous 12-hour sampling period per annum – an extreme grab
sampling transgression in the time domain. In starkest possible contrast, signicantly elevated
dioxins emissions were measured in ue gas during events of unstable combustion conditions
by continuous long-term measurements. The dioxin congener patterns from long-term ue gas
sampling show similar patterns as the congeners found in backyard chicken eggs and grass,
evidence that elevated dioxins in eggs is due to emissions from the incinerator. These results
make it mandatory to perform long-term, continuous measurements for all sources where
similar high-temperature combustion/emission processes take place. Backyard chicken eggs are
a prevalent regional bio-accumulating sample type in rural and agricultural regions, which is
relevant to air pollution emission monitoring. There are interesting relationships between this
type of sampling bio-sensors and the special bioassay DR CALUX® features, which, although
quite outside traditional contexts, never-the-less all can be well understood in the context of
the Theory of Sampling. We venture to unravel the complex interactions between correct and
incorrect sampling errors and the extraordinary complicated analytical issues involved in
monitoring and source tracking using accumulating bio-sensors. The present contribution is but
a rst foray indicating the general total sample error scope and total analytical error framework
needed for this approach.
INTRODUCTION
Harlingen is a small city of approximately 16 000 inhabitants,
situated near the Wadden Sea, inscribed in the world
heritage list of the United Nations Educational, Scientic
and Cultural Organisation (UNESCO). It is an agricultural
area with only smaller industries (relatively low-prole,
mainly concentrated near the harbour), comprised by an oil
company, shipyards, shing industry, a plastic company and
a waste incinerator, the latter built in 2011. In the area of this
research are several small villages Midlum, Wijnaldum and
Kimswerd, respectively 700, 100 and 600 inhabitants. In this
area it is common to keep backyard chicken for private use
or for small-scale marketing. The chicken are free-ranging
and stay out more than eight hours a day. Below we rst
describe the details and specics of the analytical procedures
and analytes used in the environmental-pollution survey,
many of which will be unknown or unconventional for
traditional TOS audiences, followed by the specic sampling
procedures developed.
1. Toxicologist, CEO at ToxicoWatch Foundation, Harlingen, The Netherlands. Email:arkenbout@toxicowatch.org
2. KHE Consulting, Copenhagen, Denmark. Email: khe.consult@gmail.com
3. Adjunct Professor: Aalborg University, Denmark; Geological Survey of Denmark and Greenland (GEUS), Denmark.
4. Associate Professor, Université de Québec à Chicutimi (UQAC), Quebec, Canada.
5. Guest Professor (2017): École de Technologie Supérieure (ÉTS), Montreal, Canada; Recinto Universitario de Mayaguez, Puerto Rico; University College of Southeast Norway (HSN), Porsgrunn, Norway.
EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017
A ARKENBOUT AND K H ESBENSEN
118
MATERIALS AND ANALYTICAL METHODS
Backyard chicken eggs were sampled at 15 different
geographical locations near Harlingen and analysed
for PolyChlorinated Dibenzo-p-Dioxins (PCDD), Poly-
Chlorinated DibenzoFurans (PCDF) and dioxin-like
PolyChlorinated Biphenyls (dl-PCB). Two analytical methods
are used, a novel bioanalytical screening and a conrmatory
chemical analytical reference method.
DR CALUX® is an abbreviation for a novel analytical
bioassay method (‘Dioxine Responsive Chemical Activated
LUciferace gene eXpression’), which is described in detail
(Besselink et al, 2004).
Briey, H4IIE cells stably transfected with an AhR-
controlled luciferase reporter gene construct were cultured
in α-MEM medium supplemented with ten per cent (v/v)
FCS under standard conditions (37°C, ve per cent CO2,
100 per cent humidity). Cells were exposed in triplicate on 96-
well microtiter plates containing the standard 2,3,7,8-TCDD
calibration range, a DMSO blank. Following an incubation
period of 24 hours, cells were lysed. A solution containing
luciferin (Glow Mix) was added and the luminescence was
measured using a luminometer (Berthold Centro XS3).
DR CALUX® analysis is carried out by the laboratories
of BioDetection Systems BV, Amsterdam and RIKILT,
Wageningen, both certied ISO 17025.
The GC/HRMS analysis on eggs were performed by MAS
(Münster, Germany), RIKILT Wageningen and Nofalab
Schiedam (both in the Netherlands).
Flue gas: AMESA® continuous sampling by the Environment
SA (Reinmann, Weber and Haag, 2010), analyses of the
samples by Eurons, Germany. AMESA® is a registered
trademark of an EU approved long-term sampling method.
These mandated analyses were done by SGS (Belgium). Grass
samples by the RIVM (National Institute for Public Health
and Environment in the Netherlands), analyses RIKILT,
Wageningen. All labs are ISO 17025 accredited according to
the European Commission (EC) 252/2012 guideline.
Methodological approach – eggs
Carefully selected biological entities used as accumulating
bio-sensors for specic purposes have been reported in the
literature, eg mussels or other bivalves for environmental
monitoring of sea water calibrated with regard to oil spillage
(Fitzpatrick et al, 1997; Dragsund et al, 2013), giving a relevant
backdrop for the recently invoked free-ranging chicken eggs
used in the present work (Arkenbout, 2014; Petrlík, 2015;
Polder et al, 2016). For each case of such bio-sensor application,
there are specic critical issues that must be honoured before
conventional quantitative analysis can be carried out without
ambiguity. In the present case egg yolk is used as the analytical
sample medium for lipophilic substances such as dioxins,
furans and dioxin-like polychlorobiphenyls (PCDD/F/dl-
PCBs). The processes of biotransformation, bioaccumulation
and active migration of these types of substances towards
the egg (a kind of detoxication of the chicken itself) makes
the egg, and especially the backyard/free range/biological
egg6, a simple but very relevant target for quantifying local
pollution levels a.o. However unusual, or complex-looking
from the point of view of traditional TOS audiences, in the
6. Clarication: backyard chicken, free-range chicken and biological chicken. In the
Netherlands ‘biological’ means chickens that are fed with biological (controlled) feed,
with the somewhat paradoxical result that a backyard chicken is not a biological chicken.
The expression ‘free-range’ is used commercially and here means chickens that have the
possibility to go outdoors (but most of the chickens stay inside). Backyard chickens have
to be out more than eight hours per day.
end the outcome of analysing egg yolk for highly specic
organic analytes will be like ‘conventional’ concentrations or
grades which can be used as in standard quantitative surveys,
for example delineating geographical patterns or localised
temporal patterns.
The novel egg sampling procedure is highly relevant for
ultra-low concentration dioxin quantication, but there are
also critical success factors that need to be covered before
conventional sampling parallels can be applied: the concept of
using a composite sample consisting of eggs from free-ranging
chickens is the logical approach for an effective accumulating
analytical medium (yolk); especially the stipulation to use (at
least) ten eggs (other approaches suggest 20). From a strict
composite sampling perspective (TOS), the higher the number
of eggs (each egg is an increment), the better – but tempered
by local logistical and practical sampling conditions. It is
clear that considerable biological knowledge is at a premium
regarding the details of how chickens ‘cover the ground’ and
what their detailed menus are. Backyard chickens eat worms
(Lumbricus terrestris), snails, insects (Tenebrio molitor), plants,
humus and soil particles. There’s a varying relation between
the dioxin contamination in worms, soil and eggs (Kijlstra,
Traag and Hoogenboom, 2008) so judicious averaging is
called for. Another factor contributing to the high sensibility
of backyard chicken is that egg yolks from these hens contain
a greater variety of fatty acids than do egg yolks from caged
hens (Jones, 1968).
It would appear that the traditional lot consists of the sum-
total of available free-ranging chicken eggs in a dened
geographical area, to be sampled in the 2D plane. But there
is more than what meets the eye. What would constitute the
‘heterogeneity’ of such a lot?
Organisms higher in the food chain are subject to
bioaccumulation and biomagnication. Chicken accumulates
dioxins because they are only very little metabolised in the
lipophilic state, but instead stored in the adipose tissue;
notably they are not secreted like hydrophilic material in urine
or faeces. So, dioxin bioaccumulation level grows with chicken
age. The process of egg-laying is a kind of detoxing process
for the chicken itself (Kijlstra, Traag and Hoogenboom, 2006).
The rst egg after an egg-breeding stop can therefore be high
in dioxins, a factor to be taken into account when sampling
for the composite sample for analysis. Another factor is the
different varieties of chickens, which can display different
behaviour or metabolism. The biomagnication of the other
important group of analytes persistent organic pollutants
(POPs) takes place because the chicken eats organisms like
worms which are contaminated with POPs.
So, an egg is not necessarily an egg (is not necessarily an
egg) – it is necessary to assess the parental chicken(s) actively
involved in laying eggs. A really unlucky (or distinctly ill-
informed) sampling would easily willy-nilly include such
over-the-top eggs; there are shades of IDE and IEE here, not
to mention a signicant risk of GSE, all inating the total
sampling variance. The eld sampler must be professionally
educated in these biological and physiological issues in order
to be able to carry out the imperative CSE and ISE reduction.
These issues weigh in heavily when carrying out the novel
sampling approach reported on here.
Free-range chickens have a soil uptake of ten to 30 per cent
of their feed, which translates to approximately 12 to 36 g
of soil/day. The correlation between pollution level in soil
and in chicken eggs was outlined in Arkenbout (2014),
Kijlstra, Traag and Hoogenboom (2006) and van Eijkeren
et al (2006). When chicken forage on soil with contaminant
levels above 2 pg TEQ/g ds, this result in dioxin levels above
EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017
SAMPLING, MONITORING AND SOURCE TRACKING OF DIOXINS IN THE ENVIRONMENT OF AN INCINERATOR IN THE NETHERLANDS
119
the threshold value of 2.5 pg TEQ PCDD/F (or >5 pg TEQ
PCDD/F/dl-PCBs) in eggs. Weber et al (2014) relates that
the problematic levels in soils are 1.4 to 4.2 ng TEQ/kg for
PCDD/F and 2.8 to 8.4 for sum PCDD/F+PCB. These levels
of dioxins are common in many areas of industrial emissions
and residential areas.
Such dioxin contributions represent a varying, or persistent
background level, no doubt involved in laying the basis for
ofcial background levels and critical thresholds. Should there
exists localised areas of enhanced dioxin loads the analytical
result of a composite sample would also be enhanced. This
effect could be viewed as a conventional GSE.
A minimum of ten eggs per location were collected, because
the small size of household ocks in practice limits a delivery of
more than ten simultaneous eggs. The ten-increment composite
sampling is also used in other researches, but different numbers
are in use; there is unfortunately no international sampling
harmonisation – yet (Overmeire et al , 2006; Petrlík and Behnisch,
2015). In standard analysis for commercial eggs a total number
of increment equalling 20 eggs is used. Special care was taken
to collect a composite sample of ten eggs from different hens,
because of the evidence of intra-sample variation for dioxins
in eggs. Kijlstra, Traag and Hoogenboom (2006) explain this
by different potential contact of hens with the environment.
Eggs were collected from a biological chicken farm 35 km from
Harlingen (well outside the incinerator area studied here) to be
used as analytical reference blanks (see Figure 1).
In this study eggs were collected exclusively from backyard
chicken so from relatively small ocks (<30 chickens). In the
Netherlands the enforcement of commercial eggs starts at a
ock size above 200 chickens (IKB, Integrale Ketenbeheersing).
In Kijlstra, Traag and Hoogenboom (2006) a negative
correlation between the level of dioxin pollution and ock
size in commercialised chicken farms was observed. Also
the Danish study by Sørensen et al (2013) shows more
examples of threshold-exceeding of dioxin levels when
the ock size is smaller. Kijlstra, Traag and Hoogenboom
(2006) explains the correlation between ock size and lower
dioxin contamination as follows: the larger the ock, the less
chickens go outside and will therefore have less contact with
the polluted environment.
Chickens (and their eggs) are therefore complex ‘active
sampling agents’ (sensors) but potentially excellent indicator
species for contaminated soils when all the above caveats
have been properly counteracted. But there are as yet only
very few systematic studies linking pollution sources and
loads, related soil levels and the resultant contaminant
levels in bio-accumulating bio entities used as food (Weber
et al, 2014). There is a clear need for a detailed mapping of
TOS’ standard error types onto this fascinating area in order
to reduce GSE, IDE, IEE and IPE where ever possible – and
to subject the optimised sampling_plus_analysis pathway
to a rigorous replication experiment (RE) characterisation
of the effective reproducibility of the (10,20) egg composite
sampling approach, DS 3077 (2013). These verication studies
are in the pipeline at present.
Methodological approach – grass
Sampling of grass was carried out by selecting two contiguous
sections of 25 m2 each. In each section approximately 15 evenly
spaced grass tufts were identied from which grass was cut
and aggregated to form a composite sample. The other section
yielded an archival sample which can be used for a double-
sampling RE, DS 3077 (2013).
RESULTS AND DISCUSSION
The start of the present research in the neighbourhood of
Harlingen was the advent of a new analysis approach, the DR
CALUX® bioassay, performed experimentally at 15 locations,
Figure 1 shows the results of this bioanalytical screening of
the composite samples of backyard chicken eggs. The results
of this test when expressed in bioanalytical equivalents (BEQ)
give an indication of the TEQ level (EU 709/2014). Extracts
from the yolk are split into fractions containing PCDD/
PCDFs and dioxin-like PCBs, allowing a separate indication
of PCDD/PCDFs and dioxin-like PCB TEQ levels (in BEQ).
The results show a highly systematic dioxin pollution
pattern in the environment of the incinerator. In the region
of Harlingen all samples within a radius of 2 km (n = 11)
were above the action limits set for the screening test of DR
CALUX® of 1.7 pg BEQ/g fat PCDD/F and above the limit
of 3.3 pg BEQ/g fat. The EU 709/2014 demands (in case of
commercial eggs) a conrmatory analysis with a chemical
method. On eight locations (n = 8) the DR CALUX® results
were veried with gas chromatography/mass spectrometry
analysis (GC/MS). The chemical analyses conrm the above-
threshold dioxin levels in backyard chicken eggs; the levels of
dioxins and dioxin-like PCBs increase towards the location of
the incinerator.
But also eggs of backyard chicken in other places in
the Netherlands tend to have elevated levels of dioxins,
underlining the general risk of consuming backyard chicken
eggs. Consumers of backyard chicken eggs must be aware
that ‘biological’ does not necessarily mean ‘less toxic’.
A subsequent comparative study in other parts of the
Netherlands by Hoogenboom et al (2016) shows 48 per cent
of the eggs of backyard chicken are compliant below the limit
value of dioxins in food, which means that one in two eggs
is below the regulatory threshold EU-value of dioxins in
typical regional areas. In Harlingen none of the egg samples
were below this limit value. A disadvantage in the study of
Hoogenboom et al (2016) is the lack of exact sample location
and corresponding analytical ngerprints. No conclusions can
be drawn whether or not a relationship is with the emission
of potential dioxin sources like incinerators. A request for
releasing more precise data of locations and ngerprints has
so far gone unanswered by the Ministry of Economic Affairs.
Grass
Figure 2 shows two maps with grass sample locations and the
incinerator (see arrow) in 2014 and 2015 respectively. Grass
was deliberately sampled in March after the winter period
for optimal accumulation conditions, specically to avoid the
FIG 1 – DR CALUX® dioxin concentrations in eggs from backyard
chickens near Harlingen, the Netherlands. Concentration values
based on ten-increment composite samples, see text for details.
EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017
A ARKENBOUT AND K H ESBENSEN
120
inuence of grazing of cattle (or sheep) and mowing activities.
In 2014 an elevated dioxin deposition was measured near the
incinerator of 1.2 pg TEQ/g grass. In 2015 67 per cent of the
grass samples were above the 0.75 pg TEQ/g, the maximum
limit for dioxins in cattle feed. Near the incinerator the
dioxins are a factor 2–4 higher than the reference locations.
In December 2015 all the grass samples near the incinerator
were elevated with a factor 3 compared to the reference
locations. This was remarkable because these sample of grass
were only six weeks old (from 28 October no sheep and grass
was mowed and removed). There was a huge dioxin emission
event in October, but most was emitted to the Wadden Sea
(North East wind). Reference locations were 1.5–11 km from
the location of the incinerator. None of the reference locations
were above the 0.75 pg TEQ/g. It’s obvious the highest levels
are near the incinerator, pointing this source as the most likely
source of the found PCDD/Fs deposition.
Flue gas
The waste-to-energy incinerator in Harlingen was built
2011 and promoted as the most modern installation in
the Netherlands (state-of-the-art). It was issued a dioxin
emission permit threshold of 0.01 ng TEQ/Nm3, a factor 10
lower than the European standard. The air pollution control
devices (APCDs) are based on a dry scrubber system: an
electrostatic precipitator, fabric lter, additives-injection
with sodium bicarbonate and activated carbon and selective
catalytic reduction. The reason for selecting a dry scrubber
system (besides a lower cost) was to avoid a visible plume
at the shoreline of the Wadden Sea. However, the plume
of the Harlingen incinerator is very often visible and many
complaints have been raised by inhabitants of Harlingen
because of the ‘acid or irritating plastic smell’. Although the
incinerator is a modern installation, many failures, shutdown
and start-ups have taken place (more than 40 times after the
start in 2011). Besides the lack of efcient control of APCDs
(many fall-outs), other factors like a too short chimney
(44 m), windmills in the proximity and a (frequent) coastline
fumigation leads to insufcient spreading and dilution of ue
gas in the air.
Although today’s dioxins emissions from Municipal Waste
Incinerators (MWIs) are much lower than in the twentieth
century, there are documented reports of high dioxin
emissions during transient operations ie start-up and shutdown.
The regulation of 0.1 ng TEQ/Nm3 is meant for steady state
incineration. Tejima et al (2007) found high dioxin emission
during the start-up of the incinerator. These researchers didn’t
nd high level of dioxins during a regular shutdown, but in
the case of the REC Harlingen most often shutdowns occur
due to technical failures. In the case of October 2015 there
were several failures of the APCDs, resulting in high dioxin
emissions. Any short-time monitoring is highly likely only to
take place under the presumed steady state conditions, but this
will not be able to further reliable estimates of the much more
dangerous emission situation at transient conditions.
To better characterise the dioxin emissions of the Harlingen
incinerator, a long-term sampling program (AMESA) was
initiated and used to characterise the ue gas directly in the
incinerator chimney. The regulatory 12 hour measurement
period, which represents only 0.1 per cent of the yearly
operating time, is mandated by the government as a
means to determine the annual dioxin emissions (by linear
upscaling). Figure 3 shows results of short-term sampling
(eight measurements of six hours each in four years) as it is
to be carried out according to the laws of the Netherlands.
It is obvious that this short-term sampling approach does
not show a consistent pattern, also because most of the
critical ngerprint congeners cannot even be detected (below
detection limit). These results underline the need for profound
changes in the enforcement rules to eliminate unintentional
dioxin production. The mandated short-term measurements
are to be performed under normal operating conditions
(NOC), in connection with inspections which are announced
many months ahead – and thus specically not during
unstable combustion. Even under such optimal conditions
(high caloric waste, no wet digestate), the incinerator could
not produce consistent results of short-term sampling of
PCDD/F fractions.
In contrast the results of long-term sampling (Figure 4) eight
samples with long sampling periods (650–692 hours) are far
more consistent in PCDD/F/dl-PCBs patterns. Also shown
are the relative high contribution of the non-mandatory dl-
FIG 2 – Dioxin concentration levels in grass around the incinerator REC, Harlingen for the years 2014 and 2015 (arrow points to the incinerator).
FIG 3 – Short-term sampling of the incinerator.
Measurement period: March 2012– March 2015.
EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017
SAMPLING, MONITORING AND SOURCE TRACKING OF DIOXINS IN THE ENVIRONMENT OF AN INCINERATOR IN THE NETHERLANDS
121
PCBs in the total TEQ. Long-term sampling give a far more
realistic picture of emissions than short-term could ever give.
Long-term sampling gives more realistic information about
the combustion process, the functionality of APCDs during
imperfect combustion, start-ups and shutdowns. Long-term
sampling delivers the tools to actually meet the Stockholm
Convention to reduce or minimise the dioxin emission
towards zero.
Combustion failures with high risk emissions are not
included in the ofcial short-term measurements. One failure
event may, in just a few hours, exceed the annual permitted
dioxin emissions by a factor 2, expressed in TEQ, or in
concentrations PCDD/F up to 1000 times higher than normal
operation (Reinmann, Weber and Haag, 2010). The exclusion
of failure notications allows a wide-open loophole for non-
disclosure of exceeding and harmful emissions of dioxins.
Continuous measurements also make it possible to measure
POPs emissions during unstable combustion events of start-
ups, shutdowns or even failures in the APCD can be detected.
In October 2015, a conglomerate of events produced a
signicant emission 0.17 ng TEQ/Nm3, exceeding the general
European standard of 0.1 ng TEQ/Nm3, ie exceeding the
local licensed emissions of 0.01 ng TEQ/Nm3 by a factor 17.
This threshold transgression was measured in a cassette
representing 672 hours of continuous sampling (28 days).
Analyses of the emission patterns show several possible
sources/events of this type of high dioxin emission. The
governmental enforcement relates these emissions to a
40 hour cascade of failures of the APCD. This means that the
emission permit could have been exceeded by a factor of more
than 300 in this special failure event. The incinerator plant
management has promised improvement, but after several
occasions of similar events, results still show threshold
transgressions.
Biomarker ngerprinting
A scientic way of source tracking dioxins is by way of
‘ngerprinting’ a sensitive assemblage of 17 dioxin and furan
congeners of selected biomarkers in samples from the environs
and from a possible source. This set of congeners is the result
of international agreement. Figure 5 shows this approach
illustrating this ngerprint match-up of eggs collected from
within a radius of 2 km of the incinerator (n = 6), grass sampled
within in a radius of 1 km (n = 12) and ue gas sampled at the
REC itself (n = 15). The ngerprints are essentially identical.
Only minor details differ, and one can for example explain
the abundancy of low-chlorinated congeners by processes of
biotransformation (Fiedler, 2003). But the long-term sampling
also show that the incinerator emissions are not complying
with a constant ngerprint, especially not during failure
FIG 4 – Long-term sampling of the incinerator.
Measurement period: August 2015 – March 2016.
FIG 5 – Dioxin congener pattern match-up for backyard chicken eggs, grass and ue gas from the Harlingen incinerator REC, indicating a common source.
EIGHTH WORLD CONFERENCE ON SAMPLING AND BLENDING / PERTH, WA, 9–11 MAY 2017
A ARKENBOUT AND K H ESBENSEN
122
of a certain APCD, or due an incomplete combustion when
congeners can be released to the environment with an excess
factor of up to 1000 in the worst cases. These scenarios will
only be exacerbated by continuously changing proportions of
waste inputs and loads of industrial waste and sewage sludge.
For full verication the question arises if such closely
matching ngerprints can be the result of temporary
incinerator failure events – or is this an expression of a more
structural process, in fact of a direct causal link? In order to
answer this salient question, a further exposé of the analytical
intricacies is needed.
dl-PCBs
In the literature (Weber et al, 2014), the contributions of dl-
PCBs in the total TEQ of the ue gas is only three per cent.
The REC Harlingen has a much higher percentage of dl-
PCBs in the total concentration of PCDD/F/dl-PCBs in ue
gas (33 per cent) and the coplanar PCB 126 accounts for an
additional ten per cent of the total TEQ. The incinerator in
Harlingen produces ten to 15 per cent TEQ PCBs even under
normal conditions (n = 15). The relatively high concentrations
of PCBs can be due to an incomplete combustion condition
with too low combustion temperatures, – to a short residence
time in the combustion-zone, or to a heterogeneous
combustion due to waste overload on the re grid (‘cold
spots’). The ndings of relative high percentage of dl-PCBs
may also indicate that otherwise effective combustion
temperatures do in fact not achieve the regulatory demands
of 850°C/1100°C for two-second periods. Verication of
combustion temperature regimes is mandated by the EU
regulations with both full 100 per cent, as well as 80 per cent
loads measured under circumstances characteristic of the
worst combustion conditions.
The incinerator management makes public calculated
temperatures produced by an unveried polynomial data
tting approach, and only measured at 100 per cent load.
Although dioxin-like PCBs are not included in EU regulation
of dioxins, long-term sampling shows a contribution of
dl-PCBs which can be considerable. It’s a fundamental
enforcement error, not to involve the contribution of PCB’s
(or even other POPs, see below) in the total TEQ accounting.
This underlines the need for a revision of the regulation of
POPs. Elimination of POPs in modern waste incineration
monitoring turns out to be a critical issue.
Other persistent organic pollutants
A few remarks on the biotechnology of the DR CALUX®
analytical method. In some papers (Hoogenboom et al,
2016), some results of the DR CALUX® are much higher than
corresponding GC/MS analysis. This would indicate that
there are several inuencing factors behind DR CALUX®. An
explanation can be that DR CALUX® measures the biological
reaction of the cells on a whole group of substances with much
the same reaction mechanism. Thus, while chemical analysis
pinpoints and quanties a single dioxin compound, it is likely
that DR CALUX® also measure reactions of other TEQ-related
dioxin-like compounds, eg PCBs, brominated and uorinated
compounds (see below). Conceptually, CALUX® characterises
the overall biological activity or induction of gene expression
caused by all AhR ligands (agonists and antagonists) present
in the sample extract, while a chemical analysis focuses only
on a selected number of compounds. This makes DR CALUX®
a well suited bio-accumulating pollution monitor – but also
emphasises that the complete analytical reaction framework
needs to be fully understood. Thus, TAE needs very careful
specication in the present case where several tens of analytes
are in play simultaneously.
Polybrominated, or mixed halogenated dioxins and furans
(PBDD/F and PXDD/F), are not yet regulated in foodstuffs,
sewage sludge, soils or ue gas of an incinerator. Due to the
increasing stocks of PBDD/F precursors (PBDEs and other
brominated aromatic chemicals) and increasing thermal
treatment (incineration) of ame-retarded waste, these should
also be evaluated for possible consideration in regulatory
development, because of their contribution to the total dioxin-
like toxicity.
Harlingen sampling and analysis in the
contextof the Theory of Sampling
Backyard chickens were introduced as novel local area
pollution load averaging bio-sensors with a time-accumulating,
magnifying mechanism ending up with – eggs. Professional
(biological, physiological) competence is needed when
sampling. The above detailed exposé documents that the
many biological and physiological links in this process are
reasonably well known at present to be able meaningfully
to direct sampling of sets of eggs with this provenance –
composite samples (one egg from each individual chicken is an
increment). Informed competence regarding key biological
and physiological issues is critical in order to reduce or
avoid signicant ISE and CSE. While this ‘sampling process’
lie outside most traditional TOS experiences, it was shown
how its elements never-the-less can be identied within the
traditional TOS sampling error framework. TOS is helping to
focus the sampling process in this application area.
A novel twist is that analyte quantication of the specic
organic compounds used in dioxin, POP and DI-PCB
characterisations, are only arrived at as end-products of
quite complex bioassay approaches involving several tens
of analytes simultaneously – different from traditional
inorganic chemico-physical) ‘analysis’ of a singular (geo)
chemical analyte. There are quite specic, novel TAE issues
to be mastered here. In the end however, the end results are
identical to other types of quantitative analytical data which
can be used in spatial and/or time-dependent contexts for
environmental monitoring and source tracking purposes.
It is obvious that the above-threshold transgressing dioxins
emissions in ue gas would never have been detected
without a program of continuous long-term measurements
(sampling). This makes it highly necessary to perform
continuous measurements for all industries where similar
combustion processes take place (mainly incinerator plants).
It is clear that short-term measurements cannot give a fair
rendering of the actual dioxin emissions. The incinerator
of Harlingen has had 35 start-ups since the installation in
2011, which makes it likely that emission thresholds of POPs
have in reality been exceeded many times. Mandating only
a period of 12 hours continuous measurement per annum
can only be characterised as a agrant grab sampling
misunderstanding in the time domain; 12 hours corresponds
to 1/(2 × 365) ~0.14 per cent coverage of a whole year.
The above descriptions and discussions, substantiated by
recent reports (Arkenbout, 2014; Petrlík, 2015; Reinmann,
Weber and Haag, 2010), underline the need for a careful,
much improved sampling and analyses regimen for this
type of complex, time-changing waste incineration emission
monitoring. On all accounts (logical, scientic, incinerator
plant performance) the current mandated protocols must be
repealed and replaced by a more comprehensive measurement
regimen, either a signicantly longer contiguous period and/
or a suitable time domain composite sampling scheme, the
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SAMPLING, MONITORING AND SOURCE TRACKING OF DIOXINS IN THE ENVIRONMENT OF AN INCINERATOR IN THE NETHERLANDS
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details of which can easily be set out based on the extensive
existing information. This is the only way in which to be able
to honour TOS’ fundamental sampling principle (FSP) in the
time domain.
CONCLUSIONS
The results of the present research show that composite
samples made up of eggs from individual backyard chickens
yield highly sensitive analytical biomarkers in the form of
dioxins and PCBs. This is also conrmed in other publications
of contamination in the vicinity of potential sources (harbour
activity, waste incinerator, landll). In the special case of
the incinerator REC in Harlingen, grass could also be used
as a biomarker sample type, a reection of the short distance
deposition of dioxins by this installation (short chimney,
windmills and shoreline fumigation). The detailed congener
patterns of ue gas match the patterns found in eggs and
grass closely, which is ‘a smoking gun’ as identication of
pollutants source go. The ndings of highly polluted eggs of
backyard chickens and matching congener patterns identify
the incinerator as the most likely source.
Long-term sampling is far more efcient than short-term
measurement of ue gas can ever be. Under NOC, the incinerator
complies with emission standards, but numerous inadequate
start-up procedures, shutdowns and failures in APCDs never-
the-less make incinerators a substantial source of PCDD/F/dl-
PCBs, brominated and uorinated compounds. Unwillingness
to release the actually measured critical temperature data also
seems contrary to the international agreements of the Aarhus
Convention (EC 2003/4/EC) and the Stockholm Convention
(EC 850/2004).
This study is an appeal to the scientic world community
to work together and nd solutions for a serious ongoing
pollution problem. Many different scientic elds are needed
in careful orchestration: biology, poultry science, analytical
chemistry in general, advanced analysis of dioxin in particular
– as well as the Theory of Sampling (TOS). It is challenging,
and satisfying, that one of the largest critical success factors
in this context is amenable to analysis and remediation by
invoking TOS’ principles to the decidedly novel ‘analytical
sample’ used here: composite increment sets, comprised by
10(20) eggs from individual free-ranging chickens (backyard
or otherwise). The present contribution is but a rst foray
indicating the general total sample error (TSE) scope and
complex TAE framework for this approach. Much interesting
work remains not least the intricate TAE issues interacting
with more traditional TOS and Measurement Uncertainty
(MU) quests (Esbensen and Wagner, 2014).
ACKNOWLEDGEMENTS
Citizens concerned about industrial pollution in their
environment fund this NGO Toxicowatch Foundation
project. The government of the Netherlands funds continuous
sampling of the incinerator and the study of grass. KHE
Consulting supports in part Toxicowatch’s inroads into the
application of TOS in the areas of environmental monitoring
and international scientic cooperation on conning the use
of environmental pollutants.
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