behnisch german crisis dr calux v2 - biodetection · pdf file1 behnisch, peter a.; besselink,...
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![Page 1: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/1.jpg)
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Behnisch, Peter A.; Besselink, Harrieand Brouwer, Bram
GERMAN DIOXIN CRISIS 2011 –
EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ
IN FOOD AND FEED MATRICES
BY THE HTPS DR CALUX®METHOD
DIOXIN 2011, Brussels
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![Page 3: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/3.jpg)
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Situation in January2011
•4700 farmsat firstclosed–issuetakenup by
international media
•70% of Germansdoubted, iftheirbreakfasteggsare
dioxin-free
•Ca. 70% lowerrevenuesforfarmers/industry–rough
calculationstalkedfrom20 MioEuro/weekdamage
•Most localtestingneeded> 10 days= lack of local
testingcapacity
•Screeningvia DR CALUX offerstime and costsaving
•Morethan95% of theca1800 samplesfrom
Germany testedas express analysisin 2-3 days
![Page 4: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/4.jpg)
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WHO-TEF/DR CALUX REP valuesof pollutedfoodfat
83
18
1800
H7D
197
121
SUM
Low riskof false
negative!
123
SUM All
18
27
270
123789-H6D
85
61
610
123678-H6D
3,6
1,2
12
123478-H6D
7,6
14
14
12378-P5D
DR CALUX-TEQ
WHO-TEQ
Conc.
![Page 5: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/5.jpg)
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Situation in January/February2011
•Morethan15 countriesbannedGerman meat
•German Meat Association (VDF) in cooperationwith
German Authorities(BMELV) and theRussianImport
Authoritiesacceptedmorethan40 labsforthe
German meatexports–only4 non-German
laboratories
•BDS onlyacceptednon-Germanlab usingscreening
technology DR CALUX
•Regardingacceptanceof ISO 17025 accreditedlabs
pleasenoticethataccordingEC/764/2008 all reports
of such accreditedlabshaveto beacceptedin all EC
memberstates
•BDS ISO 17025 accreditedsince2005 and new
approvedin mid2011 until2014
![Page 6: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/6.jpg)
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BDS ISO 17025 accreditation(Dutch RvA)
Quantitative screeninganalysisaccordingto current
EC guidelines:
•BDS newlyaccreditedin mid2011 byDutch RvA
•BDS twosuccessfullPT in 2010
•BDS usesseveralrelevant referencematerialswith
precisionbelow30%
•Linearity(R
2>0.98), sensitivity(1/5th EC level) and
precision(below30%)
•All in 2011 byHRGC/HRMS cross-checkednegative
samples(2%) didnotshowanyfalsenegative results
byDR CALUX
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BDS DR CALUX vsHRGC/HRMS –
170 comparisons: <1% falsenegativ and <8% falsepositive
Matrix
N False negative
False positive
egg
18
0 3
fish
14
0 0
Poultry
meat
9 0
1
Rumina
nt me
at 6
0 0
Pig m
eat
14
0 1
fish m
eal
8 0
0
Fish o
il 15
0
1
milk
33
0 3
Feed
, plan
t orig
in 18
0
4
Feed
, veg
etable
oil
7 0
1
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Examplesof DR CALUX usagein such
crisissituationsorforNational Monitoringprojects
•I) Examplesof such crisissituationsin whichtheDR CALUX
performedbyBDS has beeninvolved:
–Ireland(2009)
–Food/Feed
–Chile (2009)
–Food/Feed
–Italy(2008)
–Milk
–Netherlands(2005) –
Milk
–Belgium(1998)
-Eggs
•II) Examplesof governmentalmonitoringprojectsof feed/food
in whichDR CALUX byBDS has beeninvolved:
Chile (2008), Israel (2011), Morocco(2011), Mauretania(2010),
Netherlands(m
anyyears)
![Page 9: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/9.jpg)
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EarlierDutch reportsaboutdioxinsin eggs
analysedbyDR CALUX and HRGC/HRMS
•Controlling egg dioxin levels from laying hens with outdoor run by A. Kijlstra,
W. Traag and R. Hoogenboom (2008)
0,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
16,0
18,0
-4,0
-2,0
0,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
16,0
18,0
GC/MS (pg TEQ/g vet)
CALUX (pg TCDD eq/g vet)
•Bio-eggsin Dutch farmsbyJ. de Vries (2002)
At 2002 from 68 “biological”eggs 9 (13%) eggswere suspected in the DR-CALUX.
HRGC/MS showed, that total TEQ varied between 2.6 and 14.9 pg TEQ per gram of fat.
DR CALUX-TEQ
HRGC-HRMS TEQ
![Page 10: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/10.jpg)
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DIOXIN 2011 e-poster: ISZ Brescia, Italy
DR CALUX vsHRGC/HRMS:
100% of milk samplesin compliantwindow–
91% of egg samplesin non-compliantwindow
APPLICATION OF DR-CALUX TO MILK AND EGG SAMPLES:
COMPARISON BETWEEN HRGC-HRMS AND SCREENING DATA
Gasparini , Bertasi and Ferretti et al
![Page 11: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/11.jpg)
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HTPS DR CALUX®Methode
I) Rapid HTP Extraktion
30 samples/person/3 hrs
![Page 12: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/12.jpg)
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A) Clean-up:15 Proben/hrs
Low costs: ca. <15 Euro/sample
HTPS DR CALUX®Methode
II)Clean-Up and PCB/PCDD/PCDF Separation
B) DL-PCB and PCDD/F separation
(alumina oxide method according to
DIN 2464): ca. 15 Proben/hr
Low costs: <15 Euro/sample
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HTPS DR CALUX®Methode
III) Robot forSeeding cells
and Dosingstandards/samples
40 samples/hr
Hamilton MicrolabStarlet
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Evaluation seeding/dosingvia robot
vs. Manual analysis
Dose-response curve DR CALUX
LOG[TCDD] (M)
Signal
-14
-13
-12
-11
-10
-9
0510
MICROLA
B STARlet
Man
ual
Samples
![Page 15: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/15.jpg)
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HTPS DR CALUX®Methode
IV) MeasurementPCDD/F-and separate PCB-TEQ
DR CALUX®:
40 sample/hrforseparate
Dioxin and PCB-TEQ
(TristarMultimode Reader
Berthold Technologies
withstacker)
For comparison:
HRGC/HRMS:
0.5 sample/hrforseparate
Dioxin and PCB-TEQ
![Page 16: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/16.jpg)
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German DIOXIN Crisis2011 –
ResultsbyDR CALUX testing
![Page 17: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/17.jpg)
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German pigmeat> 97,6% negative;
lessthan2.5 % falsepositive byDR CALUX
0,1-
1,47
0,1-
0,77
0,1-
1,1
Pig
Meat;
Range
0,43
0,21
0,23
Pig
Meat;
Mean
89%
94%
97,6
99,8%
502
496
496
Pig
Meat; N
Sample
compliant
TOTAL
-50%
Sample
compliant
PCDD/F
-50%
Sample
complian
tTOTAL
-25%
Sample
compliant
PCDD/F
-25%
Total
-TEQ
PCDD/F
-TEQ
PCB
- TEQ
![Page 18: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/18.jpg)
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DR CALUX analysisforPCB + PCDD/F-TEQ(# 502 samples):
German pigmeatDistribution
Total-PCDD/F/dl-PCB-TEQ by DR CALUX for German pig
meat in Jan/Feb 2011 in pg PCDD/F/dl-PCB-TEQ/g fat
0,00
0,20
0,40
0,60
0,80
1,00
1,20
1,40
1,60
0100
200
300
400
500
600
Numbers of samples
PCDD/D/dl-PCB-TEQ by
DR CALUX
EC guidedlevel1.5 ng PCDD/F/PCB-TEQ/kg fat
![Page 19: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/19.jpg)
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DR CALUX analysisforPCDD/Fs(# 496 samples):
German pigmeatDistribution
Distribution PCDD/F-TEQ by DR CALUX for German pig meat in Jan/Feb 2011
0,00
0,10
0,20
0,30
0,40
0,50
0,60
0,70
0,80
0100
200
300
400
500
600
Number of tested samples
PCDD/F-TEQ by DR CALUX
EC guidedlevel1.0 ng PCDD/F-TEQ/kg fat
![Page 20: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/20.jpg)
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German pigliver> 97% negative;
lessthan3 % falsepositive byDR CALUX
0,1-
5,2
0,1-
3,4
0,1-
2,1
PigLiver;
Range
1,57
0,78
0,80
PigLiver;
Mean
90%
93%
97%
99%
34
37
32
PigLiver;
N
Sample
compliant
TOTAL
-50%
Sample
compliant
PCDD/F
-50%
Sample
compliant
TOTAL
-25%
Sample
compliant
PCDD/F
-25%
Total-
TEQ
PCDD/F
-TEQ
PCB-
TEQ
![Page 21: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/21.jpg)
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German poultrymeat> 98% negative;
lessthan2 % falsepositive byDR CALUX
0,6-
3,8
0,3 –
1,7
0,3-
3,3
Chicken
Meat;
Range
1,3
0,56
0,74
Chicken
Meat;
Mean
88%
93%
98%
99,5%
207
201
201
Chicken
Meat; N
Sample
compliant
TOTAL
-50%
Sample
compliant
PCDD/F
-50%
Sample
compliant
TOTAL
-25%
Sample
compliant
PCDD/F
-25%
Total-
TEQ
PCDD/F
-TEQ
PCB
-TEQ
![Page 22: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/22.jpg)
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DR CALUX analysisforPCDD/F/PCB-TEQ(# 207samples):
German poultrymeatDistribution
PCDD/F/dl-PCB-TEQ by DR CALUX for German
poultry meat in Jan/Feb 2011
01234
050
100
150
200
250
Number of samples
PCDD/F/dl-PCB-TEQ by
DR CALUX in ng
TEQ/kg fat
Reeks1
EC guidedlevel4 ng PCDD/F/PCB-TEQ/kg fat
![Page 23: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/23.jpg)
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DR CALUX analysisforPCDD/F-TEQ(# 201 samples):
German poultrymeatDistribution
PCDD/F-TEQ by DR CALUX of German poultry in
Jan/Feb 2011 (cut-off 1,4 pg PCDD/F-TEQ/gr fat)
0
0,51
1,52
050
100
150
200
250
Number of samples
PCDD/F-TEQ by DR
CALUX in ng TEQ/kg
fat
EC guidedlevel2 ng PCDD/F-TEQ/kg fat
![Page 24: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/24.jpg)
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German eggsand feedingstuff> 98% negative;
lessthan2 % falsepositive byDR CALUX
100%
100%
16
Feed;
N
0,23
Feed;
Mean
0,1-
0,65
Feed;
Range
0,1-
2,6
0,1-
2,3
0,1-
1,0
Eggs;
Range
1,48
1,21
0,67
Eggs;
Mean
89%
94%
98%
99,8%
36
99
Eggs;
N
Sample
compliant
TOTAL
-50%
Sample
compliant
PCDD/F
-50%
Sample
compliant
TOTAL
-25%
Sample
compliant
PCDD/F
-25%
Total-
TEQ
PCDD/F-
TEQ
PCB-
TEQ
![Page 25: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/25.jpg)
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DR CALUX –higherfood/feedqualitythrougha highersafety
bycoveringalso PBDD/Fs, PBBs, PBDE, PCNs…….
0.0004
2,3,6,7-TCN
0.080
PBB-77
0.16
PBB-126
4.4E-4
Benzo[a]pyrene
0.00013
PBDE 126
0.0011
8-Iod-2,3,4-TriCDF
0.0002
8-Fluor-2,3,4-TriCDF
0.01
8-Methyl-2,3,7-Tri CDD
0.1
2,3,4,7,8-PBDF
0.60
2,3,7,8-TBDF
0.28
1-Br-2,3,7,8-TCDD
0.21
1,2,3,7,8-PBDD
0.67
2-Br-3,7,8-triCDD
0.86
2,3-diBr-7,8-diCDD
0.77
2,3,7,8-TetraBrDD
0.033
2,3,7-TriBrDD
0.0015
2,3,7-TriCDD
DR CALUX REPs
= chemical
analysis
below10%
false
negative
results
regarding
dioxin-like
toxicity
![Page 26: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/26.jpg)
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Take homemessage
•During the German DIOXIN crisis in February-March 2011 more than 1800
samplesfrom Germany have been tested by DR CALUX in just 8 weeks time
with an average TAT of 3 days –a significant time/cost saving!
•Here we report results of ca. 500 samples of pig meat, 200 of chicken meat, 40
of pig liver, several egg and feed samples
•For more than 90%of the samples analyzed by BDS, the PCB-TEQand
PCDD/F-TEQhave been reported separately by using a modified protocol of
the DIN 2464 method.
• •In case of feeding materials and premixes tested by DR CALUX, all samples
have been complaintwith levels below 50% of the in the EC/152/2009 regulated
levels.
•In case of the analyzed pig and poultry samples, more than 97%of the samples
have been complaint
•In case of the analyzed pig and poultry samples more than 88%of the samples
have been even lower than 50%of the EC regulation levels.
![Page 27: Behnisch German Crisis DR CALUX V2 - Biodetection · PDF file1 Behnisch, Peter A.; Besselink, Harrieand Brouwer, B ram GERMAN DIOXIN CRISIS 2011 – EXPRESS ANALYSIS OF PCB-AND PCDD/F-TEQ](https://reader031.vdocuments.net/reader031/viewer/2022021818/5aa46a807f8b9a185d8c00cf/html5/thumbnails/27.jpg)
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•In case of compliant DR CALUX results you are also able to exclude
relevant effects of PBDD/Fs, PBBsor PCNs= higher feed/food quality
and safety!!!
•Now please join us to break the 100 €/140 USD/11000 Yen cost barrier
of PCDD/F/PCB-TEQ analysis (under ISO 17025 mode) via Robotics!
•For the price of a typical HRGC/HRMS analysis you are now able to
analyze PXDD/PXDF/PXB as well as obesogens(PPAR CALUX),
hormones (CALUX panel) and all kinds of endocrine disrupting
chemicals (CALUX panel) –to obtain a higher quality of feed/food
safety!
•The results of this study shows again that the DR CALUX®bioassay
for screening of PCDD/Fs and dioxin-like PCBs in feed and food is an
important device to identify the few percentage of the EU limit
exceeding samples among the bulk of the compliant samples….!!!
Take homemessage
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•Thankyouverymuchforyourtime and interest…
•Herzlichen Dank für Ihre Zeit…
•Dank u wel…
•Mercibeaucouppour votretemps..
•Domo arrigato…
•Muchasgracias
………………..