Dioxin/PCB crisis in feed/food A decade of DR CALUX testing
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From this document you will learn the answers to the following questions:
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1 6 th BioDetectors Meeting 2012 Dioxin/PCB crisis in feed/food A decade of DR CALUX testing Peter A. Behnisch BioDetection Systems Science Park, Amsterdam 1
2 Outline of Presentation Current situation in Europe German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Taiwan feed and food monitoirng BDS Client survey 2005 Other food monitoring programs using DR CALUX 2
3 Latest dioxin crisis 2010 Maize from Ukraine (Nederlands) 2011 eggs (Belgium/Netherlands) 2011 eggs and milk (Italy) 2011 Biodiesel used as feed led to 25% polluted market eggs (Germany) 2011 Sugar molasses (Germany) 2012 Red colorants (The Netherlands) Nearly weekly egg recalls in Germany by KAT 3
4 New EC guidelines 252/2012, 277/2012 and 278/2012 for screening for dioxins/pcbs by DR CALUX Cell based screening tests can now report samples for dioxins and dioxin-like PCBs as compliant or suspected to be non-compliant (suspected) New WHO-TEFs are more close to DR CALUX-REPs, lowering the amount of false negative for DR CALUX in case of high PCB contaminated samples False-compliant rate with respect to maximum levels is below 5% (before it was 1%) Repeatability RSD below 20% 20 confirmed results per matrix group demanded for evaluation of false negative rate Suitable cut-off level at 2/3rd of maxiumum level 4
5 New EC 277 and 278/2012 guidelines: Cut-off levels for DR CALUX Matrix Old Total- TEQ Old cutoff New Dioxin New Total- TEQ Poultry Bovine Fish Milk Eggs Plant oils New Cut- Off Babyfood no no
6 BDS DR CALUX vs HRGC/HRMS 170 comparisons: < 1% false negativ and < 8% false positive Matrix N False negative False positive egg fish Poultry meat Ruminant meat Pig meat fish meal Fish oil milk Feed, plant origin Feed, vegetable oil
7 Z-SCORE Proficiency test performance BDS ( ) Orebro univeristy, round 1 European Commission, Joint Research Centre BICS 2005 Goeyens, Belgium Norwegian Institute of Public Health in Oslo, Norway Orebro univeristy, round Dutch National Institute for Coastal and Marine Management Quasimeme round 48 7
8 Acceptance by International Quality systems MVO Code of dioxins: 100% of batches for coconut, thermic treated oils and oils direct used as animal feed have to be tested by dioxins/pcbs Private laboratories have also to report in case of higher dioxin/pcb levels Increase quality product labels on EU market such as IKB (eggs), QS (feed) German Meat Association VDF since Jan 2011, or GMP Plus 8
9 Modernisation by High-Through Put Robot (HTPS) 40 samples/hour dosing on 96 well plates 9
10 High-Through Put Robot (HTPS) 40 samples/hour dioxin and separate PCB-TEQ measurement 10
11 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project BDS Client survey 2005 Other food monitoring programs using DR CALUX 11
12 German Dioxin Crisis
13 Situation in January farms at first closed issue taken up by international media 70% of Germans doubted, if their breakfast eggs are dioxin-free Ca. 70% lower revenues for farmers/industry rough calculations talked from 20 Mio Euro/week damage Most local testing needed > 10 days = lack of local testing capacity Screening via DR CALUX offers time and cost saving More than 95% of the ca 1800 samples from Germany tested as express analysis in 2-3 days 13
14 Situation in January/February 2011 More than 15 countries banned German meat German Meat Association (VDF) in cooperation with German Authorities (BMELV) and the Russian Import Authorities accepted more than 40 labs for the German meat exports only 4 non-german laboratories BDS only accepted non-german lab using screening technology DR CALUX Regarding acceptance of ISO accredited labs please notice that according EC/764/2008 all reports of such accredited labs have to be accepted in all EC member states BDS ISO accredited since 2005 and new approved in mid 2011 until
15 15
16 German pig meat > 97,6% negative; less than 2.5 % false positive by DR CALUX PCB - TEQ PCDD/F -TEQ Total- TEQ Sample compliant PCDD/F Sample complian t TOTAL Sample compliant PCDD/F Sample compliant TOTAL - 25% - 25% - 50% - 50% Pig Meat; N Pig Meat; Mean ,8% 97,6 94% 89% 0,23 0,21 0,43 Pig Meat; Range 0,1-1,1 0,1-0,77 0,1-1,47 16
17 PCDD/D/dl-PCB-TEQ by DR CALUX DR CALUX analysis for PCB + PCDD/F-TEQ (# 502 samples): German pig meat Distribution 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 1,60 1,40 1,20 1,00 0,80 0,60 0,40 0,20 0,00 EC guided level 1.5 ng PCDD/F/PCB-TEQ/kg fat Numbers of samples 17
18 PCDD/F-TEQ by DR CALUX DR CALUX analysis for PCDD/Fs (# 496 samples): German pig meat Distribution Distribution PCDD/F-TEQ by DR CALUX for German pig meat in Jan/Feb ,80 EC guided level 1.0 ng PCDD/F-TEQ/kg fat 0,70 0,60 0,50 0,40 0,30 0,20 0,10 0, Number of tested samples 18
19 German pig liver > 97% negative; less than 3 % false positive by DR CALUX PCB- TEQ PCDD/F -TEQ Total- TEQ Sample compliant PCDD/F Sample compliant TOTAL Sample compliant PCDD/F Sample compliant TOTAL - 25% - 25% - 50% - 50% Pig Liver; N Pig Liver; Mean % 97% 93% 90% 0,80 0,78 1,57 Pig Liver; Range 0,1-2,1 0,1-3,4 0,1-5,2 19
20 German poultry meat > 98% negative; less than 2 % false positive by DR CALUX PCB -TEQ PCDD/F -TEQ Total- TEQ Sample compliant PCDD/F Sample compliant TOTAL Sample compliant PCDD/F Sample compliant TOTAL - 25% - 25% - 50% - 50% Chicken Meat; N Chicken Meat; Mean ,5% 98% 93% 88% 0,74 0,56 1,3 Chicken Meat; Range 0,3-3,3 0,3 1,7 0,6-3,8 20
21 PCDD/F/dl-PCB-TEQ by DR CALUX in ng TEQ/kg fat DR CALUX analysis for PCDD/F/PCB-TEQ (# 207 samples): German poultry meat Distribution PCDD/F/dl-PCB-TEQ by DR CALUX for German poultry meat in Jan/Feb 2011 EC guided level 4 ng PCDD/F/PCB-TEQ/kg fat Number of samples 21
22 PCDD/F-TEQ by DR CALUX in ng TEQ/kg fat DR CALUX analysis for PCDD/F-TEQ (# 201 samples): German poultry meat Distribution PCDD/F-TEQ by DR CALUX of German poultry in Jan/Feb 2011 (cut-off 1,4 pg PCDD/F-TEQ/gr fat) 2 EC guided level 2 ng PCDD/F-TEQ/kg fat 1,5 1 0, Number of samples 22
23 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Kuwait Monitoring of imported feed/food 2009 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project BDS Client survey
24 ng TEQ/kg product Total DR CALUX distribution for FEED samples, 2009 Asprocer Program, Chile % above max EU PCDD/F-PCB limit max EU limit PCDD/F-PCB = 1.5 ng TEQ/kg product % above max EU PCDD/F limit max EU limit PCDD/F = 0.75 ng TEQ/kg product 75% max EU PCDD/F limit 0.2 Santiago, Chile, 22 March # of analyses 24
25 ng TEQ/kg product Total DR CALUX distribution for MINERAL samples, 2009 Asprocer Program, Chile 3 38 ng TEQ/kg product m ineral clay 2.5 zinc oxide % above max EU PCDD/F-PCB limit max EU limit PCDD/F-PCB = 1.5 ng TEQ/kg product zinc oxide zinc oxide m ineral clay zinc oxide 1 7.6% above max EU PCDD/F limit max EU limit PCDD/F = 0.75 ng TEQ/kg product m ineral clay % max EU PCDD/F limit # of analyses Santiago, Chile, 22 March
26 ng TEQ/kg product Total DR CALUX distribution for FISH OIL samples, 2009 Asprocer Program, Chile % above max EU PCDD/F-PCB limit max EU limit PCDD/F-PCB = 24 ng TEQ/kg product % above max EU PCDD/F limit max EU limit PCDD/F = 6 ng TEQ/kg product 5 75% max EU PCDD/F limit # of analyses Santiago, Chile, 22 March
27 ng TEQ/kg product Total DR CALUX distribution for ANIMAL OIL samples, 2009 Asprocer Program, Chile % above max EU PCDD/F-PCB limit max EU limit PCDD/F-PCB = 3 ng TEQ/kg product % above max EU PCDD/F limit max EU limit PCDD/F = 2 ng TEQ/kg product % max EU PCDD/F limit # of analyses Santiago, Chile, 22 March
28 ng TEQ/kg product Total DR CALUX distribution for VEGATABLE OIL samples, 2009 Asprocer Program, Chile % above max EU PCDD/F-PCB limit max EU limit PCDD/F-PCB = 1.5 ng TEQ/kg product 1 18% above max EU PCDD/F limit max EU limit PCDD/F = 0.75 ng TEQ/kg product % max EU PCDD/F limit # of analyses Santiago, Chile, 22 March
29 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project BDS Client survey 2005 Other food monitoring programs using DR CALUX 29
30 TEQ (pg/g fat) Dutch Dioxin crisis 2004: Clay effecting potato peeling used a animal feed. total DR CALUX TEQ distribution for milk samples, 2004 Dutch clay crisis % above 6 pg TEQ/g % above 3 pg TEQ/g % below 1.5 pg TEQ/g reported TEQ (sorted from lowest to highest) 30
31 Outline of Presentation International food/feed crisis Compliance with EU regulations Chemical TEQ vs. DR CALUX TEQ a few examples Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Other Monitoring Programs 31
32 Australia 2006 Soils Recycled Organics & Remediation Technologies Unit, Sydney, Australia Dioxins: A fishy business; the analysis of dioxins in seafood from Sydney Harbour 32
33 Sydney Harbour; preliminary results 33
34 Time line rapid implementation Week training CALUX chain of custody optimalisation lab QA/QC data set total TEQ testing PCB TEQ testing evaluation accreditation fish samples done analiysis done
35 Sydney harbour: Comparison of DR CALUX and HR-GCMS DR CALUX (log scale) HRGCMS (log scale) 35
36 Outline of Presentation International food/feed crisis Compliance with EU regulations Chemical TEQ vs. DR CALUX TEQ a few examples Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Other food monitoring programs using DR CALUX 36
37 total DR CALUX TEQ/max EC total TEQ level total DR CALUX TEQ/max EC total TEQ level National Monitoring Program from State Veterinary and Food Service of Slovak Republic Ratio: Total DR CALUX TEQ vs. accepted European Total-TEQ 1.25 Bovine Meat 1.25 Pork Meat Location Location CALUX Exchange 2006 Presentation 37
38 total DR CALUX TEQ/max EC total TEQ level total DR CALUX TEQ/max EC total TEQ level National Monitoring Program from State Veterinary and Food Service of Slovak Republic Ratio: Total DR CALUX TEQ vs. accepted European Total-TEQ 1.25 Egg 1.25 Milk Location Location CALUX Exchange 2006 Presentation 38
39 total DR CALUX TEQ/max EC total TEQ level total DR CALUX TEQ/max EC total TEQ level National Monitoring Program from State Veterinary and Food Service of Slovak Republic Ratio: Total DR CALUX TEQ vs. accepted European Total-TEQ 1.25 Fish 1.25 Poultry Location 10: Ratio 3.4 due to former PCB production Location Location CALUX Exchange 2006 Presentation 39
40 Outline of Presentation International food/feed crisis Compliance with EU regulations Chemical TEQ vs. DR CALUX TEQ a few examples Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Czech Food/Feed monitoring Norway Fish monitoring school project Taiwan Feed/Food Monitoring Other food monitoring programs using DR CALUX 40
41 School students working as environmental scientists- Global POP Dioxins in fish with BDS CALUX Eldbjørg Sofie Heimstad 1, Gaute Grønstøl 2, Karl Torstein Hetland 3, Javier Martinez Alarcon 1, Charlotta Rylander 1,4 and Espen Mariussen 1,5 1 Norwegian Institute for Air Research (NILU) 2 University of Bergen, Bergen. 3 Norwegian Centre for Science Education 4 Institute of Community Medicine, University of Tromsø 5 Norwegian Defence Research Establishment (FFI) 41
42 Global POP a school project Kids participate with field work and data publishing in a research project Investigate dioxins and dioxin-like compounds (CALUX) in fish common for consumption Schools from Arctic areas will be encouraged to participate. TEQ will be determined by NILU in fish that were caught by the students Investigate dioxins and dioxin-like compounds (CALUX) in fish common for consumption 42
43 Norwegian Fish Monitoring Project; Polar Bear 43
44 Salvelinus alpinus Anguilla anguilla Gadus morhua Pleuronectes platessa Sebastes marinus Pollachius virens Salmo trutta Pollachius pollachius Perca fluviatilis Cyprinus caprio Coregonus lavaretus Platichtys flesus Scomber scombus Lota lota Melanogramus aeglefinus Molva molva Leuciscus cephalus Esox lucius Rutilus rutilus Blicca bjoerkna Limanda limanda Mugil cephalus Sparus aurata Acanthopagrus australis Acanthopagrus butcheri Without datasheet Global POP Number Number Gadus morhua, cod. Perca fluviatilis, European perch. Salmo trutta, trout. 44
45 EU wide Fish Monitoring from 54 schools 45
46 DL-PCBs+PCDD/Fs PCDD/Fs May 2009: Countries: 11 Schools: 51 Fish samples:
47 07' ' ' ' ' '333 08'337 08'347 08'339 08'343 08'361 08'365 08'369 08'373 08'388 08'392 08'396 08'400 08'484 08'507 08'546 08'621 08'645 08'356 08'659 08'748 08'752 08'756 08'760 08'805 08'809 08'819 08'877 08' ' ' ' ' ' '1596 TEQ(pg/g) Low average level of all samples: 0,42 (pg TEQ/g) 2,00 1,50 1,00 Average TEQ 0,50 0,00 Sample number 47
48 River fish higher than coastal fish N Norway: n=27, S Norway n=43 48
49 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Taiwan Monitoring of feed and food BDS Client survey 2005 Other food monitoring programs using DR CALUX 49
50 TACTRI/COA in Taiwan: Egg and Fish monitoring Sample Nr of samples Fat % Total-TEQ Eggs ( ) Eel-Feed 6 9 0,4 ( ) Seabass-Feed ( ) Tilapia-Feed ( ) Duck eggs ( ) Salted duck eggs ,2 ( ) 50
51 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Taiwan Monitoring of feed and food BDS Client survey 2005 Other food monitoring programs using DR CALUX 51
52 BDS clients survey in 2006: Dioxin/PCB-TEQ by DR CALUX TEQ in food samples (n=490) 52
53 BDS clients survey in 2006: Dioxin/PCB-TEQ by DR CALUX TEQ in feed samples (n=100) 53
54 Outline of Presentation Compliance with EU regulations German Dioxin Crisis 2011 Chile Food/Feed monitoring 2008 Dutch Clay/Potato peeling crisis 2004 Sydney harbor fish screening 2005 Slovak Republic Food/Feed monitoring Norway Fish monitoring school project Taiwan Feed/Food Monitoring BDS Client survey 2005 Other monitoring programs using DR CALUX 54
55 Other Monitoring Programs using DR CALUX I. Ireland 2008 used PCB electricity transformator oil in food recycling contaminated more than 80% of pig meat export Intensive monitoring program incl. DR CALUX. II. Italian Mozarella Crisis in 2008 Intensive monitoring program of local farmer association incl. DR CALUX III. E.g. Israel, Mauretania, Chile, Thailand, China, Korea, Spain. are currently running monitoring programs via DR CALUX 55
56 Mother milk in Hong Kong and China Comparison DR CALUX-Total-TEQ (BDS) and HRGC/HRMS WHO-Total-TEQ (WHO Reference lab) for pooled breast milk samples (pg/g fat) from Hui et al. Chemosphere 69, 1287 (2007) DR CALUX WHO Lab (TEF1997) WHO Lab (TEF2005) Hong Kong (eversmokers, n= 25; age 26.5) 2. Hong Kong (high dairy product intake,16; 31) 3. Hong Kong (high seafood intake, 17; 30.5) 4. Hong Kong (low dairy AND low seafood intake, 32; 31.4) 5. Hong Kong (high dairy OR high seafood intake, 28; 32.6) 6. Mainland China (low dairy AND low seafood intake, 21; 26.6) 7. Mainland China (high dairy OR high seafood intake, 34; 28.1) 8. China Immigrant (2 6 years stay in Hong Kong, 22; 27.6) 9. China Immigrant (7 years stay in Hong Kong or above, 23; 30.1) 10. Overseas (1 10 years of overseas stay, 18; 32.1) 11. Overseas (11 years of overseas stay or above, 10; 25.7) Overall (246; 30) 56
57 Take home message The DR CALUX cell-based screening method is used for feed/food testing since more than 15 years.and evaluated in many laboratory in many countries The new EC guidelines open now via the qualitative screening approach an easier and faster approach for the reporting of compliant or suspected samples BDS is further focusing to decrease the costs for the PCDD/F/PCB- TEQ analysis (under ISO mode) via easier clean-up methods and Robotics! The results of these studies 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.!!! 57
58 58
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