1. Quantification
of short and medium chain chlorinated
paraffin residues in selected
environmental samples
Majlinda R. Lahaniatis
Ispra-Italy, 9 May 2003
TU MunichTU Munich
Weihenstephan Scientific Centre for Nutrition,Weihenstephan Scientific Centre for Nutrition,
Land Use and Environment,Land Use and Environment,
Chair of Ecological Chemistry and EnvironmentalChair of Ecological Chemistry and Environmental
Analysis, Weihenstephaner Steig 23Analysis, Weihenstephaner Steig 23
85350 Freising-Weihenstephan, Germany85350 Freising-Weihenstephan, Germany
(e-mail: lahaniatis@mesaep.org)(e-mail: lahaniatis@mesaep.org)

2. Short storyShort storyShort storyShort story
18401840
August-Wilhelm von Hofmann
(1818-†1892)
Founder and President of
DEUTSCHEN CHEMISCHEN
GESELLSCHAFT
(German Chemical Society)
TuilerienTuilerien

3. Jean Baptiste-Andre DUMAS
Chemist
(1800-†1884)
Alexandre BRONGNIART
director of the porcelain factory of Sèvres
(1770-†1847)
Short storyShort storyShort storyShort story

4. P. BOLLEY
Synthesis of long chain
chlorinated paraffins
"Chloraffin"
empirical formula CxHn-xClx
Short storyShort storyShort storyShort story
1. World War1. World War:: antiseptic solutions of Chlorcosans (first commer-first commer-
cial use of chlorinated paraffinscial use of chlorinated paraffins)
2. World War:2. World War: chlorinated paraffins are used as flame retardants
1993:1993: global consumption 300 kt
Nowadays:Nowadays: it is believed that the demand for PCAs will likely
continue to grow at about 1%/year

5. Industrial productionIndustrial productionIndustrial productionIndustrial production
CCnnHH2n+22n+2 +Cl+Cl22  CCnnHH2n+12n+1Cl + HClCl + HCl
nn-Alkane-Alkane
C10 C‑ 13, C14 C‑ 17, C>17
 , h
ClCl22 PCAPCA+
Chlorination:Chlorination:
radical substitutionradical substitution
reactionreaction
Propagation steps:Propagation steps:
CCnnHH2n+22n+2 + Cl+ Cl   CCnnHH2n+12n+1  + HCl+ HCl
CCnnHH2n+12n+1  + Cl+ Cl22  CCnnHH2n+12n+1Cl + ClCl + Cl 
CCnnHH2n+2-y2n+2-yClClyy + Cl+ Cl   CCnnHH2n+1-y2n+1-yClClyy  + HCl+ HCl
CCnnHH2n+1-y2n+1-yClClyy  + Cl+ Cl22  CCnnHH2n-y2n-yClCl y+1y+1 + Cl+ Cl 
Propagation steps:Propagation steps:
CCnnHH2n+22n+2 + Cl+ Cl   CCnnHH2n+12n+1  + HCl+ HCl
CCnnHH2n+12n+1  + Cl+ Cl22  CCnnHH2n+12n+1Cl + ClCl + Cl 
CCnnHH2n+2-y2n+2-yClClyy + Cl+ Cl   CCnnHH2n+1-y2n+1-yClClyy  + HCl+ HCl
CCnnHH2n+1-y2n+1-yClClyy  + Cl+ Cl22  CCnnHH2n-y2n-yClCl y+1y+1 + Cl+ Cl 
Chain termination steps:Chain termination steps:
ClCl  + Cl+ Cl   ClCl22
CCnnHH2n+1-x2n+1-xClClxx  + C+ CnnHH2n+1-y2n+1-yClClyy   CC2n2nHH4n4n+2-x-y+2-x-yClClx+yx+y
CCnnHH2n+1-y2n+1-yClClyy  + Cl+ Cl   CCnnHH2n+1-y2n+1-yClCly+1y+1
Chain termination steps:Chain termination steps:
ClCl  + Cl+ Cl   ClCl22
CCnnHH2n+1-x2n+1-xClClxx  + C+ CnnHH2n+1-y2n+1-yClClyy   CC2n2nHH4n4n+2-x-y+2-x-yClClx+yx+y
CCnnHH2n+1-y2n+1-yClClyy  + Cl+ Cl   CCnnHH2n+1-y2n+1-yClCly+1y+1
Initiation stepInitiation step::
ClCl22  2 Cl2 Cl 
Initiation stepInitiation step::
ClCl22  2 Cl2 Cl 

6. Long chainLong chain
(C(C>17>17) PCA) PCA
Medium chainMedium chain
(C(C1414 C‑C‑ 1717) PCA) PCA
Short chainShort chain
(C(C1010 C‑C‑ 1313) PCA) PCA
Chlorine content of theChlorine content of the
formulation (40-72 %, w/w)formulation (40-72 %, w/w)
and
Commercial PCA areCommercial PCA are
characterised by:characterised by:
Chain length ofChain length of
n-alkanen-alkane
PCAPCA consist ofconsist of mixtures of thousand componentsmixtures of thousand components
withwith different chain lengthdifferent chain length andand different chlorination degree.different chlorination degree.
Characterisation of technical productsCharacterisation of technical productsCharacterisation of technical productsCharacterisation of technical products

7. PCA
Carbon chain
C10
-C13
short
chain
C14
-C17
medium chain
C>17
long chain
liquid liquid liquid Solid
Chlorine
content (%)
48-71 40-59 26 59‑ 10 20 and 69-72‑
CAS Nr. 85535-84-8 85535-85-9 - 63449-39-8
EINECS Nr. 287-476-5 287-477-0 - 264-150-0
EINECS
Alkane C10
-C13
,
Chlor‑
Alkane
C14
-C17
, Chlor‑
Alkane C>17
,
Chlor‑
Paraffin wax and
hydrocarbon wax,‑
Chlor‑
IUPAC-Name Chloralkane
C10
-C13
Chloralkane
C14
-C17
Chloralkane
C>17
Chloroalkane C>17
(solid)
Other
Chloro-
paraffine
C10
C‑ 13
Chloro-
paraffine
C14
C‑ 17
Chloro-
paraffine
C>17
(liquid)
Chlorparaffine C>17
(solid)
Nomenclature of technical productsNomenclature of technical productsNomenclature of technical productsNomenclature of technical products
Overview of the numbers and designations used for PCA in different sources

8.  pH-neutral, chemically inertpH-neutral, chemically inert
 Low water solubility (WS).Low water solubility (WS).
 Chlorine substitution had significant effects on the waterChlorine substitution had significant effects on the water
solubility.solubility.
 Soluble in organic solvents, as well as in most of mineral,Soluble in organic solvents, as well as in most of mineral,
animal and vegetable oils.animal and vegetable oils.
 Among themselves miscible.Among themselves miscible.
 Prolonged heating at elevated temperatures results inProlonged heating at elevated temperatures results in
the release of HCl (>200 °C).the release of HCl (>200 °C).
Physical and Chemical Properties of PCAPhysical and Chemical Properties of PCAPhysical and Chemical Properties of PCAPhysical and Chemical Properties of PCA

9. Plasticiser for
PVC
Extreme
pressure
additive for
metal-working
fluids
Fire retardant
additive for
polymers
Plasticiser and
extender for
paint resins
Plasticiser for
sealants
and additives
Fat liquor for leather
processing
Use applicationsUse applicationsUse applicationsUse applications

10. Compared to the other
anthropogenic compounds, world wide
relatively little useful datarelatively little useful data are available
on PCA concentrations in
the ecosphere.
Analytic:Analytic: Determination of PCA in environmentalDetermination of PCA in environmental
samplessamples
Analytic:Analytic: Determination of PCA in environmentalDetermination of PCA in environmental
samplessamples

11. ProblemsProblems by the identification and quantification:
 no chromatographic methodno chromatographic method able to separate all PCA
components according to their chain length and chlorine content.
ProblemsProblems by the identification and quantification:
 no chromatographic methodno chromatographic method able to separate all PCA
components according to their chain length and chlorine content.
Analytic: Complex composition of PCA mixturesAnalytic: Complex composition of PCA mixturesAnalytic: Complex composition of PCA mixturesAnalytic: Complex composition of PCA mixtures
large number of homologueslarge number of homologues andand isomersisomers
high molecular weight,high molecular weight, low volatility,low volatility,
non polar characternon polar character of PCA componentsof PCA components
very difficult the measurementvery difficult the measurement
of PCA at low concentrationsof PCA at low concentrations
complex constitution of the components in the mixturescomplex constitution of the components in the mixtures

12. Analytic: Gas chromatographic separation of PCA mixturesAnalytic: Gas chromatographic separation of PCA mixturesAnalytic: Gas chromatographic separation of PCA mixturesAnalytic: Gas chromatographic separation of PCA mixtures
4035302520
Zeit (min)
45
Year 1972
Gas chromatogram of a commercialGas chromatogram of a commercial
CC1010 C‑C‑ 1616 PCA with 50 % Cl (w/w)PCA with 50 % Cl (w/w)
after separation with aafter separation with a
packedcolumnpackedcolumn
((glass column: length 280cm; 4 mm i.D.glass column: length 280cm; 4 mm i.D.
material: 1% Silicon OV 1in Chromosorb‑material: 1% Silicon OV 1in Chromosorb‑
6 AW, DMCS 70/80mesh6 AW, DMCS 70/80mesh))
5045403530252015
Zeit (min)
5045403530252015
Zeit (min)
Year 2000
ca. 30 yearsca. 30 years
latterlatter
? ?
Gas chromatogram (ECD) of aGas chromatogram (ECD) of a
commercial Ccommercial C1010 C‑C‑ 1313 PCA mix,‑PCA mix,‑
63 % Cl (w/w) after the sepa-63 % Cl (w/w) after the sepa-
ration with a 30 m long fusedration with a 30 m long fused
silicacapillarysilicacapillary
columncolumn
(DB 5).‑(DB 5).‑

13. 0
20
40
60
80
%age
300 350 400 550450 500 600
347
366
386
403
405
439
451
473
485
499
0
20
40
60
80
300 350 400 550450 500 600
368
380
403 405
421
439
453 473
487
C - C - PCA
52 % chlorine content (w/w)
14 17
327
341
359
397
431
445
459 477 483
361 375
0
20
40
60
80
%age
300 350 400 550450 500 600
411
C - C - PCA
60 % chlorine content (w/w)
10 13
333 360
368
389
403
405
439
453
473
487
507
0
20
40
60
80
%age
300 350 400 550450 500 600
Fish feed
Cod liver oil
%age
Cod liver oilCod liver oil
CC1010-C-C1313 PCA, 60% Cl (w/w)PCA, 60% Cl (w/w)
Different constitutionDifferent constitution ofof
PCA components inPCA components in
technical productstechnical products
as compared to theas compared to the
environmental samples.environmental samples.
Analytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samples
ProblemProblem
by theby the sample preparationsample preparation
recommended clean-up
procedures are generally
unspecificunspecific

14. The concentrations published up to
now are determined with differentdifferent
analytic proceduresanalytic procedures and because of their
missing comparability are hardly usableare hardly usable
for the estimation of the PCA
contamination in the environment.HRGC-ECD chromatograms of several
commercial PCA
identificationidentification and quantificationquantification
normally incorrectincorrect
Commercial PCA as external standardsCommercial PCA as external standards
co-elution of many homologues and
isomers
30,683
32,747
Zeit (min)
10 15 20 25 30 35 40 45 50
33,430
A
B
C
C10-C13 PCA,
63 % Cl (w/w)
C10-C13 PCA,
63 % Cl (w/w)
C10-C13 PCA,
70 % Cl (w/w)
C10-C13 PCA,
70 % Cl (w/w)
C14-C17 PCA,
52 % Cl (w/w)
C14-C17 PCA,
52 % Cl (w/w)
Time (min)
Analytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samplesAnalytic: Determination of PCA in environmental samples

15. QuantificationQuantification of PCA residues in cleaned-up sample
extracts with independent detection methodsindependent detection methodsand
different external standards.different external standards.
22
A newselectiveclean-up procedurenewselectiveclean-up procedurefor the separation
of C10-C13 PCA and C14-C17 PCA from lipids and interfering
chlorinated organic compounds, especially from toxaphene,
in selected fatty foods (altogether 56 samples).
11
Here we present:Here we present:Here we present:Here we present:
TheresultsTheresultsthus obtained with the different methods are
comparedcomparedwith each other and their environmental relevance
is assessedassessed.
33

16. Clean-up: 1. StepClean-up: 1. StepClean-up: 1. StepClean-up: 1. Step
Eluent: n-Hexane:DCM (1:1)
Eluent [ml] : Probe [g] = 10 : 1
Eluent: n-Hexane:DCM (1:1)
Eluent [ml] : Probe [g] = 10 : 1
Na2SO4 [g] =0.1 x Sample [g]Na2SO4 [g] =0.1 x Sample [g]
Sample [g] : Na2SO4 [g] : sea sand =1 : 4 : 2Sample [g] : Na2SO4 [g] : sea sand =1 : 4 : 2
Na2SO4 [g] =0.4 x Sample [g]Na2SO4 [g] =0.4 x Sample [g]
SiO2 [g] = 0.2 x Sample [g]SiO2 [g] = 0.2 x Sample [g]
SiO2 [g] = 0.3 x Sample [g]SiO2 [g] = 0.3 x Sample [g]
Sea sandSea sand
Glas woolGlas wool
SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w) and sea sand
SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w) [g] = 0.12  fish [g]  fat [%]
sea sand [g] = 0.08  fish [g]  fat [%]
SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w) and sea sand
SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w) [g] = 0.12  fish [g]  fat [%]
sea sand [g] = 0.08  fish [g]  fat [%]
lipidlipid
extractionextraction
isolation of theisolation of the
lipid-solublelipid-soluble
contaminantscontaminants
SimultaneousSimultaneous
SimultaneousSimultaneous
decompositiondecomposition
of lipidsof lipids

17. Clean-up: 1. StepClean-up: 1. StepClean-up: 1. StepClean-up: 1. Step
After the 1. Step:
lipid removal 99.9% and
the extract contains:
- rest of lipids: 0.1%
- all PCAs (recovery > 90%)
- other H2
SO4
resistant COS,
incl. toxaphene, PCBs etc.
 Reduction of necessary analysis time.
 Savings of substantial energy and water.
 Elimination of many manual operations and sources of
contamination.
 Avoidance of possible PCA decomposition and losses.
 Treatment of samples even with higher quantities of lipids.
ExtractionExtraction
ExtractionExtraction
LipidLipid
decompositiondecomposition
LipidLipid
decompositiondecomposition
PCAPCA
isolationisolation
PCAPCA
isolationisolation

18. 2. Fraction (7 ml2. Fraction (7 ml nn-Hexane:CH-Hexane:CH22ClCl22 1:1)1:1)
90 % of C10-C13 and C‑ 14-C17 PCA mixtures
 ‑,  und‑ -HCH 10-20 % p,p´ DDD, p,p´ DDT‑ ‑
 rest of Toxaphen (ca. 10 %)
1. Fraction (8 ml n-Hexane)1. Fraction (8 ml n-Hexane)
 < 90 % of the Toxaphene components
 ca. 90 % p,p´ DDT‑
 ca. 80 % p,p´ DDD‑
 < 98 % PCT, HCB, -Chlorden, OCS,
-cis-Chlordan, p,p´-DDE, PCB 209
Clean-up: 2. StepClean-up: 2. StepClean-up: 2. StepClean-up: 2. Step
Eluent:
1. Fraction: 8 ml n-Hexane
2. Fraction: 7 ml n-Hexane:DCM (1:1)
Eluent:
1. Fraction: 8 ml n-Hexane
2. Fraction: 7 ml n-Hexane:DCM (1:1)
0.1 g SiO2
/H2
O(d)
(3 % H2
O(d)
, w/w)0.1 g SiO2
/H2
O(d)
(3 % H2
O(d)
, w/w)
1 g SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w)1 g SiO2
/H2
SO4
(44 % H2
SO4(c)
, w/w)
1 g SiO2
/H2
O(d)
(30 % H2
O(d)
, w/w)1 g SiO2
/H2
O(d)
(30 % H2
O(d)
, w/w)
0.3 g Na2
SO40.3 g Na2
SO4
sea sandsea sand
glass woolglass wool
Column: 22.0 cm length  0.7 cm i.d.
* particle size 0,063-0,200 mm
** particle size 0,2 0,5 mm‑
decomposition ofdecomposition of
the remaining lipidsthe remaining lipids
SimultaneousSimultaneous
SimultaneousSimultaneous
separation of the PCAsseparation of the PCAs
from the most part of thefrom the most part of the
interfering compoundsinterfering compounds

19. Clean-upClean-upClean-upClean-up
1
Zeit (min)
10 15 20 25 30 35 40 45 50
Zeit (min)
10 15 20 25 30 35 40 45 50
1
Zeit (min)
10 15 20 25 30 35 40 45 50
Zeit (min)
10 15 20 25 30 35 40 45 50
1
1
1
Zeit (min)
10 15 20 25 30 35 40 45 50
Figure: HRGC-ECD chromatograms of the separation
PCA/Toxaphene with adsorption chromatography. 1: -HCH
CB Standard C10-C13-PCA, 63 % Cl (w/w)
C10-C13-PCA, 63 % Cl (w/w)
and CB Standard
1. Fraction
1. Fraction
1. Fraction
2. Fraction
2. Fraction
2. Fraction
SiOSiO22/H/H22O, 10 % HO, 10 % H22OO
SiOSiO22/H/H22O, 30 % HO, 30 % H22OO
SiOSiO22/H/H22SOSO44,,
44 % H44 % H22SOSO4 conc.4 conc.
Vor theVor the
separationseparation
After theAfter the
separationseparation

20. Recovery:Recovery:
C10-C13 PCA with 45 % Cl (w/w): ca. 90 %
C10-C13 PCA with 56 % Cl (w/w): ca. 92 %
C10-C13 PCA with 63 % Cl (w/w): ca. 94 %
C14-C17 PCA with 52 % Cl (w/w): ca. 93 %
THF
Material: Phenogel 5 µ, 5Å
Clean-up: 3. StepClean-up: 3. StepClean-up: 3. StepClean-up: 3. Step

21. Figure: Clean-up procedure
and the corresponding
HRGC/ECD chromatograms.
1: HCB; 2: -HCH
2. Step2. Step2. Step2. Step
1. Step1. Step1. Step1. Step
3. Step3. Step3. Step3. Step
Commercial Toxaphene
(24 ng/µl)
C10-C13 PCA,
63 % Cl (w/w)
Extract after the
1. Step of the clean-up
2. Fraction1. Fraction
GPC Fr. 27-32 minGPC Fr. 23-27 min
2
1
Zeit (min)
15 20 25 30 35 40 45
Zeit (min)
15 20 25 30 35 40 45
1
2
Zeit (min)
15 20 25 30 35 40 45
Zeit (min)
15 20 25 30 35 40 45
2
2
Zeit (min)
15 20 25 30 35 40 45
1
Zeit (min)
15 20 25 30 35 40 45
Zeit (min)
15 20 25 30 35 40 45
Clean-upClean-upClean-upClean-up

22. SamplesSamplesSamplesSamples
Mussels
(1 sample)
Baltic Tell-mussels
(1 sample)
23 Fish samples
Deep sea shrimp
(1 sample)
16 Cod liver oil
samples
14 Fish oil samples
Fish feed
(1 sample)

23. Fish or fish feed sample homogenised
Sample [g] : Na2SO4 [g] : Sea sand [g] = 1 : 4 : 2
STEP I : Column extraction of POPs and the
fatty material, decomposition of lipids with
SiO2/H2SO4 (C), and isolation of H2SO4 resistant
POPs including PCAs, simultaneously
SiO2/H2SO4 (C) [g] = 0,12 x Sample [g] x Fat [%]
n-Hexane : DCM (1:1) [ml] : Sample [g] = 10 : 1
STEP II: Column decomposition of the remaining lipids (< 0.1 %) and
separation of PCAs from the most part of interfering POPs
1 g SiO2/H2O (30 % H2O, w/w) and 1 g SiO2/H2SO4 (C), (44 % H2SO4 (C), w/w)
Eluents: 8 ml n-Hexane (1. Fraction) and 7 ml n-Hexane/DCM=1:1 (2. Fraction)
Extract drying, solution in 300 µl n-Hexane
STEP III: GPC separation of PCAs from the rest of interfering POPs
HPLC-Column (300 x 7,8 mm): Phenogel 5 µ, 50 Å; 0,2 ml/min THF;
Injection 200µl; Temp. 25 °C; PCA-Fraction 23-27min
PCA extract (Fr. 2) drying with N2, solution in 200µl THF
Oil solved in n-Hexane / DCM (1:1)
n-Hexane : DCM [ml] : Oil [g] = 10 : 1
STEP I: Column decomposition of lipids with
SiO2/H2SO4 (C), and isolation of H2SO4 esistant
POPs including PCAs, simultaneously
SiO2/H2SO4 (C) [g] : Oil [g] = 10 : 1
n-Hexane : DCM (1:1) [ml] : Oil [g] = 15 : 1
Clean-up technique for the PCA determination inClean-up technique for the PCA determination in
fish, fish oils and fish feedfish, fish oils and fish feed
Clean-up technique for the PCA determination inClean-up technique for the PCA determination in
fish, fish oils and fish feedfish, fish oils and fish feed

24. PCA determination in real environmental samplesPCA determination in real environmental samplesPCA determination in real environmental samplesPCA determination in real environmental samples Pag
Time (min)
0 5 10 15 20 25 30 35 40 45 50
A
B
Time (min)
0 5 10 15 20 25 30 35 40 45 50
A
B
Fish feedFish feed
GERMANYGERMANY
Cod liver oilCod liver oil
U.S.AU.S.A
AfterAfter
1. Step of1. Step of
clean-upclean-up
AfterAfter
3. Step of3. Step of
clean-upclean-up
Figure: HRGC/ECD chromatograms of real sample extracts after extraction and lipid
decomposition (1. Step of clean-up) and after complete elimination of interferences
(3. Step of clean-up) 1: HCB; 2: -HCH

25. Identification and quantification of PCAIdentification and quantification of PCAIdentification and quantification of PCAIdentification and quantification of PCA
SCGC/LRMS-ECNISCGC/LRMS-ECNI
in full scan modein full scan mode
HRGC/HRMS-ECNI-SIMHRGC/HRMS-ECNI-SIM
with time windowswith time windows
most recently developedmost recently developed
analytical methodsanalytical methods
for PCA analysisfor PCA analysis
HRCG/ECDHRCG/ECD

26. Analytic: HRGC/HRMS-ECNI-SIM with time windowsAnalytic: HRGC/HRMS-ECNI-SIM with time windowsAnalytic: HRGC/HRMS-ECNI-SIM with time windowsAnalytic: HRGC/HRMS-ECNI-SIM with time windows
CC1010
(Cl(Cl55 to Clto Cl1010))
CC1111
(Cl(Cl55 to Clto Cl1010))
CC1212
(Cl(Cl66 to Clto Cl1010))
CC1313
(Cl(Cl77 to Clto Cl99))
Ion - 279.0055
Ion - 293.0211
Ion - 312.9665
Ion - 326.9822
Ion - 346.9275
Ion - 360.9433
Ion - 394.9042
Ion - 416.8467
Ion - 430.8623
Ion - 450.8077
Ion - 464.8233
Ion - 380.8886
5:00 10:00 15:00 30:00
Zeit (min)
25:0020:00
5:00 10:00 15:00 30:00
Zeit (min)
25:0020:00
Ion - 340.9978
Ion - 374.9588
Ion - 408.9199
Ion - 444.8779
Ion - 478.8394
Ion - 388.9745
Ion - 422.9355
Ion - 458.8936
Figure: HRGC/LRMS elution range of selected
C12 and C13 -[M‑Cl]¯-ions of a commercial
C10-C13 PCA, 63 % Cl (w/w)
Figure: HRGC/LRMS elution range of selected
C10 and C11 [M‑Cl]¯-ions of a commercial
C10-C13 PCA, 63 % Cl (w/w)

27. Analytic:Analytic:
HRGC/HRMS-ECNIHRGC/HRMS-ECNI
SIM with timeSIM with time
windowswindows
Analytic:Analytic:
HRGC/HRMS-ECNIHRGC/HRMS-ECNI
SIM with timeSIM with time
windowswindows
SIM window
Nr. min:sek Quantification ion/ Confirmation ion (compound)
1 00:00-16:00 279,0055/277,0084 (C10H17Cl5); 293,0211/291,0241 (C11H19Cl5);
312,9665/314,9636 (C10H16Cl6); 326,9822/328,9792 (C11H18Cl6);
340,9978/342,9949 (C12H20Cl6); 346,9275/348,9246 (C10H15Cl7)
2 16:00-17:10 312,9665/314,9636 (C10H16Cl6); 326,9822/328,9792 (C11H18Cl6);
340,9978/342,9949 (C12H20Cl6); 346,9275/348,9246 (C10H15Cl7);
360,9432/362,9402 (C11H17Cl7); 380,8886/382,8856 (C10H14Cl8)
3 17:10-18:20 326,9822/328,9792 (C11H18Cl6); 340,9978/342,9949 (C12H20Cl6);
346,9275/348,9246 (C10H15Cl7); 360,9432/362,9402 (C11H17Cl7);
374,9588/376,9559 (C12H19Cl7); 380,8886/382,8856 (C10H14Cl8);
394,9042/396,9013 (C11H16Cl8)
4 18:20-20:20 326,9822/328,9792 (C11H18Cl6); 340,9978/342,9949 (C12H20Cl6);
360,9432/362,9402 (C11H17Cl7); 374,9588/376,9559 (C12H19Cl7);
380,8886/382,8856 (C10H14Cl8); 388,9745/390,9715 (C13H21Cl7);
394,9042/396,9013 (C11H16Cl8); 408,9199/410,9169 (C12H18Cl8);
416,8467/414,8496 (C10H13Cl9); 430,8623/428,8656 (C11H15Cl9)
5 20:20-21:00 374,9588/376,9559 (C12H19Cl7); 388,9745/390,9715 (C13H21Cl7);
394,9042/396,9013 (C11H16Cl8); 408,9199/410,9169 (C12H18Cl8);
416,8467/414,8496 (C10H13Cl9); 422,9355/424,9326 (C13H20Cl8);
430,8623/428,8656 (C11H15Cl9); 444,8779/442,8809 (C12H17Cl9)
6 21:00-22:40 374,9588/376,9559 (C12H19Cl7); 388,9745/390,9715 (C13H21Cl7);
394,9042/396,9013 (C11H16Cl8); 408,9199/410,9169 (C12H18Cl8);
416,8467/414,8496 (C10H13Cl9); 422,9355/424,9326 (C13H20Cl8);
430,8623/428,8656 (C11H15Cl9); 444,8779/442,8809 (C12H17Cl9);
450,8077/448,8106 (C10H12Cl10); 464,8233/462,8263 (C11H14Cl10)
7 22:40-25:00 388,9745/390,9715 (C13H21Cl7); 408,9199/410,9169 (C12H18Cl8);
416,8467/414,8496 (C10H13Cl9); 422,9355/424,9326 (C13H20Cl8);
430,8623/428,8656 (C11H15Cl9); 444,8779/442,8809 (C12H17Cl9);
450,8077/448,8106 (C10H12Cl10); 458,8936/456,8966 (C13H19Cl9);
464,8233/462,8263 (C11H14Cl10); 478,8390/476,8491 (C12H16Cl10)
8
25:00-35:00
422,9355/424,9326 (C13H20Cl8); 444,8779/442,8809 (C12H17Cl9);
450,8077/448,8106 (C10H12Cl10); 458,8936/456,8966 (C13H19Cl9);

28. Analytic:Analytic:
HRGC/HRMS-ECNI SIMHRGC/HRMS-ECNI SIM
with time windowswith time windows
Analytic:Analytic:
HRGC/HRMS-ECNI SIMHRGC/HRMS-ECNI SIM
with time windowswith time windows
Relativeconcentration(%)Relativeconcentration(%)Relativeconcentration(%)
Figure 1: Profile of components in selected
short chain commercial PCA mixtures
A
0
5
10
15
20
25
30
10 11 12 13 Z K
B
0
5
10
15
20
25
30
10 11 12 13 Z K
5 Cl
6 Cl
7 Cl
8 Cl
9 Cl
10 Cl
C
0
5
10
15
20
25
30
10 11 12 13 Z KC
C
C
CC1010-C-C1313 PCAPCA
60% Cl (w/w)60% Cl (w/w)
CC1010-C-C1313 PCAPCA
63% Cl63% Cl
(w/w)(w/w)
CC1010-C-C1313 PCAPCA
70% Cl70% Cl
(w/w)(w/w)Relativeconcentration(%)Relativeconcentration(%)
Figure 2: Profile of PCA components in a
environmental sample
0
5
10
15
20
10 11 12 13 Z KC
Relativeconcentration(%)
Cod liver oil
ENGLAND
Relativeconcentration(%)

29. e. St. external Standard (commercial C10-C13-PCA, 60 % Cl (w/w)
i. St. internal Standard (Pentachlorotoluol)
inj. Menge injected amount (ng/µl)
SIM (Probe) the highest integrated Signal in the sample
SIM (e. St.) the highest integrated Signal in the external Standard
Ffg Ffg = 0,01  (% RC)
MG molecular weight
Quantification:
 Integration of [M Cl]¯ Signals of the selected quantification ions (‑ ‑ SIM-Values)
 Calculation of TRIS–Values: TRIS = SIM/(0,01  Fna) (TRIS: true relative integrated signal)
 Calculation of RAIS- Values by normalising the TRIS–Values (RAIS: relative adjusted ion signal)
 Calculation of RC- Values: RC = RAIS/Cl-Atoms in the molecule (RC: relative concentration)
Analytic: HRGC/HRMS-ECNI-SIM with time windowsAnalytic: HRGC/HRMS-ECNI-SIM with time windows
St.)(e.MengeInj.
(Probe)MengeInj.

St.)(e.SIM
(Probe)SIM

(Probe)Fna
St.)(e.Fna

(Probe)Ffg
St.)(e.Ffg

St.)(e.MG
(Probe)MG

St.)e.inSt.(i.SIM
Probe)St.(i.SIM

30. Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples
interinter
laboratorylaboratory
teststests
independentindependent
methods ofmethods of
analysisanalysis
CorrectnessCorrectness
andand
repeatabilityrepeatability
of aof a
quantification methodquantification method
certifiedcertified
referencereference
materialsmaterials
no certified referenceno certified reference
materials are presentmaterials are present

31. HRGC/HRMS-ECNI-SIM with time windowsHRGC/HRMS-ECNI-SIM with time windows
Inter laboratory studyInter laboratory study
HRGC/HRMS-ECNI-SIM with time windowsHRGC/HRMS-ECNI-SIM with time windows
Inter laboratory studyInter laboratory study
CommercialCommercial
CC1010-C-C1313 PCA,PCA,
60 % Cl (w/w)60 % Cl (w/w)
Mix of pureMix of pure
synthesised Csynthesised C1010 PCAPCA
componentscomponents (PCA-1)(PCA-1)
Biological sampleBiological sample
CommercialCommercial
CC1010-C-C1313 PCA,PCA,
70 % Cl (w/w)70 % Cl (w/w)
True value:True value: 7474ng/µlng/µl
Measured:Measured: 8181ng/µl (ng/µl (99±2099±20ng/µl)ng/µl)
Error: 10%Error: 10%
True value:True value: 118118ng/µlng/µl
Measured:Measured: 309309ng/µl (ng/µl (297297±132±132ng/µl)ng/µl)
Error: 160 %Error: 160 %
True value: ? ng/µlTrue value: ? ng/µl
Measured:Measured: 5858 ng/µlng/µl
Error: ?? %Error: ?? %

32. C10= 12.26%
C11= 47.74%
Cl8 (18.79%)
C12= 35.80
C13= 4.19
0,00
2,00
4,00
6,00
8,00
10,00
12,00
14,00
16,00
18,00
20,00
22,00
10.5
10.6
10.7
10.8
10.9
1010
11.5
11.6
11.7
11.8
11.9
1110
12.6
12.7
12.8
12.9
12,10
13.7
13.8
13.9
%relativeadjustedionsignal
C:Cl numbers
CommercialCommercial
CC1010-C-C1313 PCA,PCA,
70 % Cl (w/w)70 % Cl (w/w)
4,38 %
23,00 %23.07 %
49.55 %
0,00
10,00
20,00
30,00
40,00
50,00
10.5
10.6
10.7
10.8
10.9
1010
%relativeadjustedionsignal
C:Cl numbers
Mix of pureMix of pure
synthesised Csynthesised C1010 PCAPCA
componentscomponents
C13 = 3.48%
C12 = 35.95%
C11 = 46.77%
Cl7 (18.71%)
C10 = 13.79%
0,00
2,00
4,00
6,00
8,00
10,00
12,00
14,00
16,00
18,00
20,00
10.5
10.6
10.7
10.8
10.9
1010
11.5
11.6
11.7
11.8
11.9
1110
12.6
12.7
12.8
12.9
12,10
13.7
13.8
13.9
%relativeadjustedionsignal
C:Cl numbers
CommercialCommercial
CC1010-C-C1313 PCA,PCA,
60 % Cl (w/w)60 % Cl (w/w)HRGC/LRMS-ECNI-SIM:HRGC/LRMS-ECNI-SIM:
inter laboratory studyinter laboratory study
HRGC/LRMS-ECNI-SIM:HRGC/LRMS-ECNI-SIM:
inter laboratory studyinter laboratory study
 a method errora method error
 use of an unsuitableuse of an unsuitable
standardstandard
 both factorsboth factors
?

33. TheThe extent of this deviationextent of this deviation depends ondepends on
thethe difference in the chlorine contentdifference in the chlorine content
between sample and standardbetween sample and standard..
HRGC/HRMS-ECNI-SIM with time windowsHRGC/HRMS-ECNI-SIM with time windows
Inter laboratory studyInter laboratory study
HRGC/HRMS-ECNI-SIM with time windowsHRGC/HRMS-ECNI-SIM with time windows
Inter laboratory studyInter laboratory study
Different commercial formulationsDifferent commercial formulations used asused as
standards would providestandards would provide quite different estimatesquite different estimates
of PCA concentrations.of PCA concentrations.
If the quantification standard has aIf the quantification standard has a
lower chlorine contentlower chlorine content than thatthan that of the sample quantifiedof the sample quantified,,
increased quantification results must be countedincreased quantification results must be counted..

34. SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected fish samplesin selected fish samples
SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected fish samplesin selected fish samples
0
20
40
60
80
%age
300 350 400 550450 500 600
329
359
375
395
409
431
443
457
479
495 511
0
20
40
60
80
%age
300 350 400 550450 500 600
336
347
361
375
397
411
429 443 459
482
MonkfishMonkfish
Northern North SeaNorthern North Sea
MonkfishMonkfish
Northern North SeaNorthern North Sea
FishFish
CanadaCanada
FishFish
CanadaCanada

35. SCGC/LRMS-ECNI in full scan modeSCGC/LRMS-ECNI in full scan modeSCGC/LRMS-ECNI in full scan modeSCGC/LRMS-ECNI in full scan mode
313
349
383
419
349
385
421
313
313
347
381
417
313
347
383 417
347 381
417
453
345
381 417
451
483
C - PCA, 45 % Chlorgehalt (Gew.-%)
10
C - PCA, 50 % Chlorgehalt (Gew.-%)
10
C - PCA, 55 % Chlorgehalt (Gew.-%)
10
C - PCA, 60 % Chlorgehalt (Gew.-%)
10
C - PCA, 65 % Chlorgehalt (Gew.-%)
10
C - PCA, 70 % Chlorgehalt (Gew.-%)
10
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
341
373
377
411
445
341
373
377
409
443
341
373
375
411
445
479
341
375 411
445
479
375
407
443
479
513
547
375
409
445
479
513
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
0
20
40
60
80
%age
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
300 350 400 550450 500 600
C - PCA, 45 % Chlorgehalt (Gew.-%)
12
C - PCA, 50 % Chlorgehalt (Gew.-%)
12
C - PCA, 55 % Chlorgehalt (Gew.-%)
12
C - PCA, 60 % Chlorgehalt (Gew.-%)
12
C - PCA
65 % Chlorgehalt (Gew.-%)
12
C - PCA
70 % Chlorgehalt (Gew.-%)
12
CC1010 PCA, 45 % Cl (w/w)PCA, 45 % Cl (w/w)CC1010 PCA, 45 % Cl (w/w)PCA, 45 % Cl (w/w)
CC1010 PCA, 50 % Cl (w/w)PCA, 50 % Cl (w/w)CC1010 PCA, 50 % Cl (w/w)PCA, 50 % Cl (w/w)
CC1010 PCA, 55 % Cl (w/w)PCA, 55 % Cl (w/w)CC1010 PCA, 55 % Cl (w/w)PCA, 55 % Cl (w/w)
CC1010 PCA, 60 % Cl (w/w)PCA, 60 % Cl (w/w)CC1010 PCA, 60 % Cl (w/w)PCA, 60 % Cl (w/w)
CC1010 PCA, 65 % Cl (w/w)PCA, 65 % Cl (w/w)CC1010 PCA, 65 % Cl (w/w)PCA, 65 % Cl (w/w)
CC1010 PCA, 70 % Cl (w/w)PCA, 70 % Cl (w/w)CC1010 PCA, 70 % Cl (w/w)PCA, 70 % Cl (w/w)
CC1212 PCA, 45 % Cl (w/w)PCA, 45 % Cl (w/w)CC1212 PCA, 45 % Cl (w/w)PCA, 45 % Cl (w/w)
CC1212 PCA, 50 % Cl (w/w)PCA, 50 % Cl (w/w)CC1212 PCA, 50 % Cl (w/w)PCA, 50 % Cl (w/w)
CC1212 PCA, 55 % Cl (w/w)PCA, 55 % Cl (w/w)CC1212 PCA, 55 % Cl (w/w)PCA, 55 % Cl (w/w)
CC1212 PCA, 60 % Cl (w/w)PCA, 60 % Cl (w/w)CC1212 PCA, 60 % Cl (w/w)PCA, 60 % Cl (w/w)
CC1212 PCA,PCA,
65 % Cl (w/w)65 % Cl (w/w)
CC1212 PCA,PCA,
65 % Cl (w/w)65 % Cl (w/w)
CC1212 PCA,PCA,
70% Cl (w/w)70% Cl (w/w)
CC1212 PCA,PCA,
70% Cl (w/w)70% Cl (w/w)

36. 0
20
40
60
80
%age
300 350 400 550450 500 600
347
366
386
403
405
439
451
473
485
499
0
20
40
60
80
300 350 400 550450 500 600
368
380
403 405
421
439
453 473
487
C - C - PCA
52 % chlorine content (w/w)
14 17
327
341
359
397
431
445
459 477 483
361 375
0
20
40
60
80
%age
300 350 400 550450 500 600
411
C - C - PCA
60 % chlorine content (w/w)
10 13
333 360
368
389
403
405
439
453
473
487
507
0
20
40
60
80
%age
300 350 400 550450 500 600
Fish feed
Cod liver oil
%age
Fish feedFish feedFish feedFish feed
Cod liver oilCod liver oilCod liver oilCod liver oil
CC1414-C-C1717 PCAPCA
52 % Cl (w/w)52 % Cl (w/w)
CC1414-C-C1717 PCAPCA
52 % Cl (w/w)52 % Cl (w/w)
CC1010-C-C1313 PCAPCA
60 % Cl (w/w)60 % Cl (w/w)
CC1010-C-C1313 PCAPCA
60 % Cl (w/w)60 % Cl (w/w)
SCGC/LRMSSCGC/LRMS
ECNIECNI
mass spectramass spectra
SCGC/LRMSSCGC/LRMS
ECNIECNI
mass spectramass spectra

37. Identification and quantification by SCGC/LRMS-ECNIIdentification and quantification by SCGC/LRMS-ECNIIdentification and quantification by SCGC/LRMS-ECNIIdentification and quantification by SCGC/LRMS-ECNI
PCA
chain
length
Quantification ion
(Substance)
C10 313 (C10H16Cl6); 349 (C10H15Cl7); 379 (C10H14Cl8);
413 (C10H13Cl9); 447 (C10H12Cl10);
C11 327 (C11H18Cl6); 361 (C11H17Cl7); 397 (C11H16Cl8);
431 (C11H15Cl9); 463 (C11H14Cl10)
C12 341 (C12H20Cl6); 375 (C12H19Cl7); 411 (C12H18Cl8);
445 (C12H17Cl9); 481 (C12H16Cl10)
C13 353 (C13H22Cl6); 389 (C13H21Cl7); 423 (C13H20Cl8);
459 (C13H19Cl9); 493 (C13H18Cl10)
C14 369 (C14H24Cl6); 402 (C14H23Cl7); 438 (C14H22Cl8);
473 (C14H21Cl9)
C15 416 (C15H25Cl7); 453 (C15H24Cl8); 487 (C15H23Cl9)
Selected mass fragment ions for the
quantification of C10-C17 PCA by
SCGC/LRMS-ECNI
For each carbon chain lengthFor each carbon chain length
(i.e. C(i.e. C1010-C-C1313 PCA) only suchPCA) only such
m/z-fragment ions werem/z-fragment ions were
chosen, which were exactlychosen, which were exactly
in the gap between ECNIin the gap between ECNI
clusters of the other PCAclusters of the other PCA
carbon chain lengthcarbon chain length
(i.e. C(i.e. C1414-C-C1717 PCA).PCA).
The whole range CThe whole range C1010-C-C1313
and Cand C1414-C-C1717 PCA should bePCA should be
covered by SCGC/LRMS-ECNI.covered by SCGC/LRMS-ECNI.

38. Identification andIdentification and
quantificationquantification
byby
SCGC/LRMS-ECNISCGC/LRMS-ECNI
Identification andIdentification and
quantificationquantification
byby
SCGC/LRMS-ECNISCGC/LRMS-ECNI
327
341
359
397
431
445
459 477 483
361 375
0
20
40
60
80
%age
0
20
40
60
80
%age
300 350 400 550450 500 600
300 350 400 550450 500 600
0
20
40
60
80
%age
300 350 400 550450 500 600
0
20
40
60
80
%age
300 350 400 550450 500 600
359
361
375
395
409
431
445
463
479
481
495 513
409
324
341
359
361
375
395
425
443
457
477
493
495
411
368
380
403 405
421
439
453 473
487
C - C - PCA
60 % Chlorgehalt (Gew.-%)
10 13
C - C - PCA
63 % Chlorgehalt (Gew.-%)
10 13
C - C - PCA
70 % Chlorgehalt (Gew.-%)
10 13
C - C - PCA
52 % Chlorgehalt (Gew.-%)
14 17
CC1414HH2424ClCl66 m/z 369m/z 369
CC1414HH2121ClCl99 m/z 473m/z 473
CC1414HH2323ClCl77 m/z 402m/z 402 CC1414HH2323ClCl88 m/z 438m/z 438
CC1515HH2525ClCl77 m/z 416m/z 416
CC1515HH2323ClCl99 m/z 487m/z 487
CC1010-C-C1313 PCA withPCA with
60 % chlorine content (w/w)60 % chlorine content (w/w)
C10-C13 PCA withwith
63 % chlorine content (w/w)chlorine content (w/w)
CC1515HH2424ClCl88 m/z 453m/z 453
C10-C13 PCA withwith
70 % chlorine content (w/w)chlorine content (w/w)
C14-C17 PCA withwith
52 % chlorine content (w/w)chlorine content (w/w)
Figure: SCGC/LRMS
ECNI mass spectra of
commercial C10-C13 and
C14-C17 PCA mixtures with
different chlorine content
CC1616 and Cand C1717 PCAPCA
could not becould not be
identified.identified.
DisadvantagesDisadvantages

39. Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples
1. comparison N t-Test P-Value
A-B 26 6,9 <0,001
A-C 26 6,2 <0,001
B-C 25 -0,8 0,459
N Number of samples in the series
Hypothesis (HHypothesis (H00))Hypothesis (HHypothesis (H00))
Differences between
two series of data
amount to zero.
1.1. Different detection methodsDifferent detection methods
the same external standard.the same external standard.CC1010-C-C1313 PCAPCA
HRGC/ECDHRGC/ECD
(Methode A)(Methode A)
HRGC/ECDHRGC/ECD
(Methode A)(Methode A)
HRGC/LRMS-ECNIHRGC/LRMS-ECNI
in SIM modein SIM mode
(Methode B)(Methode B)
HRGC/LRMS-ECNIHRGC/LRMS-ECNI
in SIM modein SIM mode
(Methode B)(Methode B)
SCGC/LRMS-ECNISCGC/LRMS-ECNI
in full scan modein full scan mode
(Methode C)(Methode C)
SCGC/LRMS-ECNISCGC/LRMS-ECNI
in full scan modein full scan mode
(Methode C)(Methode C)
commercial
CC1010-C-C1313 PCAPCA
PCA residues:
Standard:

40. Quantification ofQuantification of
PCA residues inPCA residues in
cleaned-upcleaned-up
samplessamples
Quantification ofQuantification of
PCA residues inPCA residues in
cleaned-upcleaned-up
samplessamples
C10-C13 PCA residues (µg/Kg Fat) in
real sample extracts quantified by:
(A) HRGC/ECD
(B) HRGC/LRMS-ECNI-SIM‘
(C) SCGC/LRMS-ECNI
with a commercial C10-C13 PCA with
63 % Cl (w/w).
NN number of samples in the data
series
0
100
200
300
400
500
1 6 11 16 21 26
HRGC/LRMS-ECNI-SIM
HRGC/ECD
0
200
400
600
800
1 6 11 16 21 26 31
SCGC/LRMS-ECNI
HRGC/ECD
0
30
60
90
120
150
1 6 11 16 21
HRGC/LRMS-ECNI-SIM
SCGC/LRMS-ECNI
N
A - B
A - C
Concentration(µg/KgFat)Concentration(µg/KgFat)Concentration(µg/KgFat)
N
B - C
N
HRGC/LRMS-ECNI-SIMHRGC/LRMS-ECNI-SIM
HRGC/ECDHRGC/ECD
SCGC/LRMS-ECNISCGC/LRMS-ECNI
HRGC/ECDHRGC/ECD
HRGC/LRMS-ECNI-SIMHRGC/LRMS-ECNI-SIM
SCGC/LRMS-ECNISCGC/LRMS-ECNI

41. Quantification ofQuantification of
PCA residues inPCA residues in
cleaned-upcleaned-up
samplessamples
Quantification ofQuantification of
PCA residues inPCA residues in
cleaned-upcleaned-up
samplessamples
C10-C13 PCA residues (µg/Kg Fat) in
real sample extracts quantified by:
(A) HRGC/ECD
(B) HRGC/LRMS-ECNI-SIM‘
(C) SCGC/LRMS-ECNI
with a commercial C10-C13 PCA with
63 % Cl (w/w).
NN number of samples in the data
series
0
100
200
300
400
500
1 6 11 16 21 26
HRGC/LRMS-ECNI-SIM
HRGC/ECD
0
200
400
600
800
1 6 11 16 21 26 31
SCGC/LRMS-ECNI
HRGC/ECD
0
30
60
90
120
150
1 6 11 16 21
HRGC/LRMS-ECNI-SIM
SCGC/LRMS-ECNI
N
A - B
A - C
Concentration(µg/KgFat)Concentration(µg/KgFat)Concentration(µg/KgFat)
N
B - C
N
HRGC/LRMS-ECNI-SIMHRGC/LRMS-ECNI-SIM
HRGC/ECDHRGC/ECD
SCGC/LRMS-ECNISCGC/LRMS-ECNI
HRGC/ECDHRGC/ECD
HRGC/LRMS-ECNI-SIMHRGC/LRMS-ECNI-SIM
SCGC/LRMS-ECNISCGC/LRMS-ECNI

42. Quantification of PCA: HRGC/ECDQuantification of PCA: HRGC/ECDQuantification of PCA: HRGC/ECDQuantification of PCA: HRGC/ECD
CC1010-C-C1313 PCA concentrationsPCA concentrations
measured by HRGC/ECDHRGC/ECD
(Method A)
CC1010-C-C1313-PCA concentrations-PCA concentrations
measured by GC/MS-ECNIGC/MS-ECNI
(Method B und C)
>
ExplanationExplanation::  maybe in the extracts besides C10-C13 exist also C14-C17 or C>17 PCA
 the new developed clean-up procedure
 the gas chromatography
provide no separation ofprovide no separation of
CC1010-C-C1313 from the other PCAsfrom the other PCAs
accordingly by EC-detection Caccordingly by EC-detection C1414-C-C1717 or Cor C>17>17 PCA are quantified with.PCA are quantified with.
SinceSince
bothboth

43. HRGC/ECD is suitable only forHRGC/ECD is suitable only for
screening investigationsscreening investigations and/orand/or
for afor a raw estimation of the totalraw estimation of the total
amount PCA residuesamount PCA residues inin
environmental samples.environmental samples.
ConclusionConclusion
Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples

44. Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples
To whatextentis affectedtheTo whatextentis affectedthe
GC/MS-ECNI quantificationGC/MS-ECNI quantification
fromtheuseof commercialfromtheuseof commercial
PCA standardsPCA standards ??
pure synthesised
C10
, C11
, C12
, C13
PCA
with 45 to 70% Cl (w/w)
Commercial
C10-C13 PCA
63 % Cl (w/w)
SCGC/LRMS-ECNISCGC/LRMS-ECNI
in full scan modein full scan mode

45. Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples
Hypothesis (HHypothesis (H00))Hypothesis (HHypothesis (H00))
Differences between
two series of data
amount to zero.
2. comparison N t-Test P-Value
C-F 35 -4,8 <0,001
N Number of samples in the series
2.2. The same detection method butThe same detection method but
different external standards.different external standards.CC1010-C-C1313-PCA-PCA
CC1010-C-C1313 PCA,PCA,
63% Cl (w/w)63% Cl (w/w)
(Method C)(Method C)
CC1010-C-C1313 PCA,PCA,
63% Cl (w/w)63% Cl (w/w)
(Method C)(Method C)
pure laboratory synthesised Cpure laboratory synthesised C1010-, C-, C1111-,-,
CC1212-, C-, C1313-PCAs, 45 to 70% Cl (w/w)-PCAs, 45 to 70% Cl (w/w)
(Methode F)(Methode F)
pure laboratory synthesised Cpure laboratory synthesised C1010-, C-, C1111-,-,
CC1212-, C-, C1313-PCAs, 45 to 70% Cl (w/w)-PCAs, 45 to 70% Cl (w/w)
(Methode F)(Methode F)
SCGC/LRMS-ECNISCGC/LRMS-ECNI
PCA residues:
Detection method:

46. The quantification with pureThe quantification with pure
laboratory synthesised PCAlaboratory synthesised PCA
components (mixtures) leadscomponents (mixtures) leads
probably to more accurateprobably to more accurate
concentrations of residues inconcentrations of residues in
environmental samples.environmental samples.
ConclusionConclusion
Quantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samplesQuantification of PCA residues in cleaned-up samples

47. Quantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samples
Several samplesSeveral samples CC1414 PCAPCA
(i.e. Fish feed,(i.e. Fish feed,  are the mainare the main
cod liver oil etc).cod liver oil etc). Components.Components.
In 40 % of fish oilsIn 40 % of fish oils CC1111 PCAPCA
in 45 % of the water organismsin 45 % of the water organisms  are the mainare the main
in 50 % of cod liver oilsin 50 % of cod liver oils components.components.
Identified: both CIdentified: both C1010-C-C1313 and Cand C1414-C-C1717 PCA.PCA.
PCA residuesPCA residues  in ca. 63 % of the samples (116,0-1719,3 µg/kg fat ).in ca. 63 % of the samples (116,0-1719,3 µg/kg fat ).

48. Real samples
Depending on the sample art and the origin the relation :Depending on the sample art and the origin the relation :
CC1010-C-C1313 PCA / CPCA / C1010-C-C1717 PCAPCA  0.11 to 5.89.0.11 to 5.89.
0,00
20,00
40,00
60,00
80,00
100,00
0 5 10 15 20 25 30 35
Number of samples
C10-C13PCAcontent(%)
Figure: Content of C10-C13 PCA in % compared to the total amount of PCA
residues quantified in real sample extracts
Quantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samplesQuantification of PCA in selected environmental samples

49. Quantification of PCA in selected fish samplesQuantification of PCA in selected fish samplesQuantification of PCA in selected fish samplesQuantification of PCA in selected fish samples
0
100
200
300
400
500
600
700
Sprat
Sprat
Hake
Redfish
Redfish
Redfish
Redfish
Salmon
Salmon
Monkfish
Monkfish
Herring
Herring
Mackerel
Halibut
BalticTellmuschel
Mussel
Sardine
Sardine
Mackerel(grilled)
Breedtrout
PCAconcentration[µg/kgfat]
Short chain PCA
Medium chain PCA
Figure: Content of C10-C17 PCA quantified in fish, mussels and deep sea shrimps sample extracts

50. Quantification of PCAQuantification of PCA
in selected oil samplesin selected oil samples
Quantification of PCAQuantification of PCA
in selected oil samplesin selected oil samples
0
50
100
150
200
250
300
350
Germany
Germany
England
Island
Island
Short chain PCAs
Medium chain PCAs
Fish oilsFish oilsFish oilsFish oils
Short chain PCAs:Short chain PCAs:
44.9 to 321.4 µg/kg44.9 to 321.4 µg/kg
Medium chain PCAs:Medium chain PCAs:
63.1 and 190.9 µg/kg63.1 and 190.9 µg/kg
ExceptionsExceptions: fish oils: fish oils
originated from Germanyoriginated from Germany
0
100
200
300
400
500
600
700
France
England
England
England
England
Island
USA
USA
Short chain PCAs
Medium chain PCAs
Cod liver oilsCod liver oilsCod liver oilsCod liver oils
Short chain PCAs:Short chain PCAs:
57.9 and 644.457.9 and 644.4 µg/kgµg/kg
Medium chain PCAs:Medium chain PCAs:
94.6 and 538.594.6 and 538.5 µg/kgµg/kg
ExceptionsExceptions: cod liver oils: cod liver oils
from France and U.S.A.from France and U.S.A.

51. SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected oil samplesin selected oil samples
SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected oil samplesin selected oil samples
0
20
40
60
80
%age
300 350 400 550450 500 600
333
397
403
441
467
0
20
40
60
80
%age
300 350 400 550450 500 600
319
347 368
403
437
473
509
Fish oilFish oil
Salmon (capsules)Salmon (capsules)
GERMANYGERMANY
Cod liver oilCod liver oil
(capsules)(capsules)
FRANCEFRANCE

52. SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected environmental samplesin selected environmental samples
SCGC/LRMS-ECNI mass spectra of CSCGC/LRMS-ECNI mass spectra of C1010 C‑C‑ 1717 PCAPCA
in selected environmental samplesin selected environmental samples
0
20
40
60
80
%age
300 350 400 550450 500 600
347
366
386
403
405
439
451
473
485
499
360
368
389
403
405
439
453
473
487
507
0
20
40
60
80
%age
300 350 400 550450 500 600
Cod liver oilCod liver oil
(capsules)(capsules)
U.S.A.U.S.A.
Fish feedFish feed
GERMANYGERMANY

53. PerspectivePerspectivePerspectivePerspective
 certified reference materialscertified reference materials
 well defined Standardswell defined Standards
 validated clean-up andvalidated clean-up and
quantification methodsquantification methods
 certified reference materialscertified reference materials
 well defined Standardswell defined Standards
 validated clean-up andvalidated clean-up and
quantification methodsquantification methods
1. Problem:1. Problem:
Non-Non-
existenceexistence
Important and necessary:Important and necessary:
Well determined
study in internatio-
nal level.
What?What? How?How?
To establish well validated
analytical procedures for
this group of substances.
To establish well validated
analytical procedures for
this group of substances.
To produce well defined C10
to C17 PCA components and
certified reference materials
for different kind of environ-
mental matrices.
To produce well defined C10
to C17 PCA components and
certified reference materials
for different kind of environ-
mental matrices.
Why?Why?
In order to control
the accuracy,
reproducibility and
applicability of any
analytical procedure.
What?What?

54. The ecotoxikologic relevance of this group of chlorinated
substances should be clarified by toxicological studies
with single components or at least with pure synthesised
and well defined mixtures.
No possible to identify and quantify PCA with aNo possible to identify and quantify PCA with a
chain length >16 in environmental samples (lowchain length >16 in environmental samples (low
response factors and high molecular weight).response factors and high molecular weight).
No possible to identify and quantify PCA with aNo possible to identify and quantify PCA with a
chain length >16 in environmental samples (lowchain length >16 in environmental samples (low
response factors and high molecular weight).response factors and high molecular weight).
GC/ECD and/or GC/MS-ECNIGC/ECD and/or GC/MS-ECNI
detection very difficult.detection very difficult.
GC/ECD and/or GC/MS-ECNIGC/ECD and/or GC/MS-ECNI
detection very difficult.detection very difficult.
Alternative methodsAlternative methods
must be developed.must be developed.
Alternative methodsAlternative methods
must be developed.must be developed.
2. Problem:2. Problem:
PerspectivePerspectivePerspectivePerspective